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So YH, Mishra D, Gite S, Sonawane R, Waite D, Shaikh R, Vora LK, Thakur RRS. Emerging trends in long-acting sustained drug delivery for glaucoma management. Drug Deliv Transl Res 2025; 15:1907-1934. [PMID: 39786666 PMCID: PMC12037438 DOI: 10.1007/s13346-024-01779-4] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 12/17/2024] [Indexed: 01/12/2025]
Abstract
Glaucoma is an optic neuropathy in which progressive degeneration of retinal ganglion cells and the optic nerve leads to irreversible visual loss. Glaucoma is one of the leading causes of blindness. The pathogenesis of glaucoma is determined by different pathogenetic mechanisms, including increased intraocular pressure, mechanical stress, excitotoxicity, resistance to aqueous drainage and oxidative stress. Topical formulations are often used in glaucoma treatment, whereas surgical measures are used in acute glaucoma cases. For most patients, long-term glaucoma treatments are given. Poor patient compliance and low bioavailability are often associated with topical therapy, which suggests that sustained-release, long-acting drug delivery systems could be beneficial in managing glaucoma. This review summarizes the eye's physiology, the pathogenesis of glaucoma, current treatments, including both pharmacological and nonpharmacological interventions, and recent advances in long-acting drug delivery systems for the treatment of glaucoma.
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Affiliation(s)
- Yin Ho So
- School of Pharmacy, Medical Biology Centre, Queen's University Belfast, Belfast, UK
| | - Deepakkumar Mishra
- School of Pharmacy, Medical Biology Centre, Queen's University Belfast, Belfast, UK
| | - Sandip Gite
- School of Pharmacy, Medical Biology Centre, Queen's University Belfast, Belfast, UK
| | - Rahul Sonawane
- School of Pharmacy, Medical Biology Centre, Queen's University Belfast, Belfast, UK
| | - David Waite
- School of Pharmacy, Medical Biology Centre, Queen's University Belfast, Belfast, UK
| | - Rahamatullah Shaikh
- Centre for Pharmaceutical Engineering Science, School of Pharmacy and Medical Sciences, University of Bradford, Bradford, BD7 1DP, UK
| | - Lalitkumar K Vora
- School of Pharmacy, Medical Biology Centre, Queen's University Belfast, Belfast, UK.
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Zhu Y, Liu W, Liang Y. Effects of small incision cataract surgery and routine ultrasonic phacoemulsification on vision recovery, anterior segment parameters and corneal regularity. Technol Health Care 2025; 33:1304-1309. [PMID: 40331547 DOI: 10.1177/09287329241293894] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/08/2025]
Abstract
ObjectiveTo compare the effects of minor incision cataract surgery (SICS) and routine ultrasonic phacoemulsification (PHACO) on vision recovery, anterior segment parameters and corneal regularity.MethodsNinety-two age-related cataract patients (100 eyes) were prospectively selected from January 2017 to December 2019. Fifty cases (50 eyes) were treated with SICS, while the remaining 42 cases (50 eyes) underwent PHACO. Their post-operative visual acuity, central corneal thickness (CCT), endothelial cell density, coefficient of variation and proportion of hexagonal cells were compared.ResultsCCT was more extensive in the two groups one week after the operation than before, while it returned to the pre-operative level 6 weeks after. Compared with before the operation, CCT had a significant difference in the two groups one week after the operation (P < 0.05). In contrast, there was no significant difference six weeks after the operation (P > 0.05). CCT had a substantial difference between the two groups 1-week after operation (P < 0.05). The endothelial cell density had a significant difference in PHACO group 1 and 6 weeks after operation compared to before (P < 0.01). In contrast, the two groups had no significant difference at each time point before and after operation (P > 0.05). There were no significant differences in the coefficient of corneal endothelial cell area variation or proportion of hexagonal cells between the two groups at each time point (P > 0.05).ConclusionsSICS and PHACO have comparable clinical effects. For patients with senile hard-nucleus cataracts, SICS is characterised by fewer surgical complications.
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Affiliation(s)
- Yueqing Zhu
- Department of Ophthalmology, The First People's Hospital of Jiashan County, Jiaxing, Zhejiang, China
| | - Weijie Liu
- Department of Ophthalmology, Lishui Huaxia Eye Hospital Co., Ltd, Lishui, Zhejiang, China
| | - Yong Liang
- Department of Ophthalmology, Pengzhou People's Hospital, Pengzhou, Sichuan, China
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Yuan Y, Yasuda S, Funk KL, Kao W, Saika S, Kaufman A, Liu CY. Smad4 deficiency ameliorates the progressive corneal stroma thinning caused by the loss of Tbr1. Ocul Surf 2025; 36:181-189. [PMID: 39894408 DOI: 10.1016/j.jtos.2025.01.013] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/15/2024] [Revised: 01/20/2025] [Accepted: 01/28/2025] [Indexed: 02/04/2025]
Abstract
PURPOSE To understand how Tbr1 and Smad4 play a pivotal role in controlling ECM synthesis versus degradation for maintaining corneal stromal homeostasis and otherwise leading to corneal ectasia. METHODS Keratocyte-specific and inducible knockout (iKO) of Tbr1, Smad4, or Tbr1/Smad4 double KO (iDKO) mice were generated. OCT was used to assess corneal thickness in vivo. Masson's trichrome and collagen hybridizing peptide stainings were performed to examine collagen expression. Immunostaining with an anti-cathepsin B antibody was used to assess ECM degradation. Cathepsin B inhibitor, CA-074Me, eyedrop was conducted to test its effect on treating stromal thinning in Tbr1 iKO mice. RESULTS Tbr1 iKO and Smad4 iKO displayed corneal thinning, but Tbr1 iKO revealed a progressive and more severe pathology than Smad4 iKO. Tbr1 iKO cornea lost most of its stroma and thus a dome shape. Collagen ECM is evenly distributed in Smad4 iKO as well as control littermates but was lost mainly in the anterior stroma of the Tbr1 iKO. Interestingly, Tbr1/Smad4 iDKO ameliorated Tbr1 iKO phenotype. The basal level of Cathepsin b (Ctsb) could be detected in the control stroma but was significantly increased in the Tbr1 iKO stromal cells and this effect was canceled in Tbr1/Smad4 iDKO. CA-074Me eyedrops administration significantly inhibited progressive corneal thinning caused by the Tbr1 iKO. CONCLUSION Our data from Tbr1/Smad4 iDKO argued that Smad4 played a pivotal role in controlling Tbr1-dependent ECM synthesis and Tbr1-independent ECM degradation to maintain corneal stromal integrity and homeostasis.
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Affiliation(s)
- Yong Yuan
- Edith Crawley Vision Research Center, Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH, USA
| | - Shingo Yasuda
- Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan
| | - Kaitlyn L Funk
- Edith Crawley Vision Research Center, Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH, USA
| | - Winston Kao
- Edith Crawley Vision Research Center, Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH, USA
| | - Shizuya Saika
- Department of Ophthalmology, Wakayama Medical University, Wakayama, Japan
| | - Adam Kaufman
- Edith Crawley Vision Research Center, Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH, USA
| | - Chia-Yang Liu
- Edith Crawley Vision Research Center, Department of Ophthalmology, College of Medicine, University of Cincinnati, Cincinnati, OH, USA.
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Peshkar-Kulkarni S, Chung DD, Aldave AJ. Antioxidant MitoQ increases viability of human corneal endothelial cells with congenital hereditary endothelial dystrophy-associated SLC4A11 mutations. Ophthalmic Genet 2025; 46:166-173. [PMID: 39834031 PMCID: PMC12003074 DOI: 10.1080/13816810.2025.2450455] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/07/2024] [Revised: 12/27/2024] [Accepted: 01/02/2025] [Indexed: 01/22/2025]
Abstract
PURPOSE To assess the impact of MitoQ, a mitochondria-targeted antioxidant, on viability of human corneal endothelial cell (hCEnC) lines expressing SLC4A11 mutations associated with congenital hereditary endothelial dystrophy (CHED) and Fuchs endothelial corneal dystrophy type 4 (FECD4). METHODS SLC4A11 wildtype (SLC4A11WT) and mutant (SLC4A11MU) hCEnC lines were created to express either SLC4A11 variant 2 (V2) or variant 3 (V3) by stable transduction of SLC4A11-/- hCEnC-21T with lentiviruses containing either SLC4A11WT or one of the following mutations: V2 (V3) mutants c.374 G>A (c.326 G>A) (CHED), c.1813C>T (c.1765C>T) (CHED), c.2263C>T (c.2215C>T) (CHED), or c.2224 G>A (c.2176 G>A) (FECD4). A SLC4A11-/- empty hCEnC line was created by stable transduction of SLC4A11-/- hCEnC-21T with an empty lentiviral plasmid. Cell viability was measured by exposing MitoQ treated and untreated cells to oxidative stress agent tert-butyl hydroperoxide (tBH) followed by performing XTT assays and spectrophotometry. RESULTS SLC4A11-/- empty, SLC4A11 V2WT, and SLC4A11 V3WT hCEnC exposed to ≤0.01 μM MitoQ retained over 90% of the viability of untreated SLC4A11-/- empty hCEnC. When treated with MitoQ, SLC4A11-/- empty was able to demonstrate partial restoration of cell viability. All CHED-associated mutant hCEnC lines treated with 0.01 μM MitoQ demonstrated increased viability compared to untreated following exposure to tBH. The FECD4-associated mutant hCEnC line treated with 0.01 μM MitoQ showed no significant increase in cell viability compared to untreated following exposure to tBH. CONCLUSIONS Media supplementation with antioxidant MitoQ has beneficial effects on cell viability in hCEnC harboring CHED-associated SLC4A11 mutations following exposure to tBH-induced oxidative stress.
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Affiliation(s)
| | - Doug D Chung
- Department of Ophthalmology, Stein Eye Institute at UCLA, Los Angeles, California, USA
| | - Anthony J Aldave
- Department of Ophthalmology, Stein Eye Institute at UCLA, Los Angeles, California, USA
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De Miguel MP, Cadenas-Martin M, Stokking M, Martin-Gonzalez AI. Biomedical Application of MSCs in Corneal Regeneration and Repair. Int J Mol Sci 2025; 26:695. [PMID: 39859409 PMCID: PMC11766311 DOI: 10.3390/ijms26020695] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/29/2024] [Revised: 01/10/2025] [Accepted: 01/11/2025] [Indexed: 01/27/2025] Open
Abstract
The World Health Organization estimates that approximately 285 million people suffer from visual impairments, around 5% of which are caused by corneal pathologies. Currently, the most common clinical treatment consists of a corneal transplant (keratoplasty) from a human donor. However, worldwide demand for donor corneas amply exceeds the available supply. Lamellar keratoplasty (transplantation replacement of only one of the three layers of the cornea) is partially solving the problem of cornea undersupply. Obviously, cell therapy applied to every one of these layers will expand current therapeutic options, reducing the cost of ophthalmological interventions and increasing the effectiveness of surgery. Mesenchymal stem cells (MSCs) are adult stem cells with the capacity for self-renewal and differentiation into different cell lineages. They can be obtained from many human tissues, such as bone marrow, umbilical cord, adipose tissue, dental pulp, skin, and cornea. Their ease of collection and advantages over embryonic stem cells or induced pluripotent stem cells make them a very practical source for experimental and potential clinical applications. In this review, we focus on recent advances using MSCs from different sources to replace the damaged cells of the three corneal layers, at both the preclinical and clinical levels for specific corneal diseases.
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Affiliation(s)
- Maria P. De Miguel
- Cell Engineering Laboratory, La Paz University Hospital Health Research Institute, IdiPAZ, 28046 Madrid, Spain; (M.C.-M.); (M.S.); (A.I.M.-G.)
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Xiao Y, McGhee CNJ, Zhang J. Posterior Limbal Mesenchymal Stromal Cells Promote Proliferation and Stemness of Transition Zone Cells: A Novel Insight Into Corneal Endothelial Rejuvenation. Invest Ophthalmol Vis Sci 2025; 66:44. [PMID: 39820276 PMCID: PMC11753478 DOI: 10.1167/iovs.66.1.44] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/01/2024] [Accepted: 12/21/2024] [Indexed: 01/19/2025] Open
Abstract
Purpose Progenitors for the corneal endothelium have been identified in the transition zone (TZ), but their cellular interactions remain undefined. Posterior limbal mesenchymal stromal cells (P-LMSCs) may support TZ cells in the posterior limbus. This study aims to characterize P-LMSCs and investigate their effects on TZ cells. Methods Human P-LMSCs and TZ cells were isolated by explant culture. P-LMSCs were characterized by comparing with anterior limbal mesenchymal stromal cells (A-LMSCs) using immunocytochemistry. TZ cells were cocultured with P-LMSCs in a Transwell, with TZ cell and A-LMSC coculture and TZ cells only as the control groups. The proliferation and wound healing capacity of TZ cells were assessed by EdU assay and scratch wound assay. Colony forming assay, droplet digital PCR, Western blotting, and immunocytochemistry were used to compare the stemness of TZ cells. The effect of P-LMSC conditioned medium on endothelial wound healing was evaluated in organ-cultured mouse corneas. Endothelial regeneration was measured by trypan blue staining. Results P-LMSCs expressed similar proteins (vimentin, Nestin, TRA-1-60, Oct3/4) as A-LMSCs. TZ cells cocultured with P-LMSCs had significantly higher proliferation, wound healing speed, and colony-forming efficiency than TZ cells only. TZ cells supported by P-LMSCs expressed higher levels of stem/progenitor markers (Nestin, Sox9, AP-2α, Pitx2) than the control groups. P-LMSC conditioned medium stimulated regeneration of mouse corneal endothelium from the TZ region. Conclusions The proliferation and stemness of TZ cells were enhanced by P-LMSCs in both cell and organ culture models. Our study provides an innovative strategy for corneal endothelial rejuvenation.
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Affiliation(s)
- Yuting Xiao
- Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand
| | - Charles N. J. McGhee
- Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand
| | - Jie Zhang
- Department of Ophthalmology, New Zealand National Eye Centre, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand
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Özer Ö, Doğan L, Baysal Z. The effect of adrenaline and trypan blue used during cataract surgery on anatomical and functional outcomes in pseudoexfoliation syndrome patients. Cutan Ocul Toxicol 2024; 43:287-292. [PMID: 39250681 DOI: 10.1080/15569527.2024.2402405] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/26/2024] [Revised: 08/06/2024] [Accepted: 09/04/2024] [Indexed: 09/11/2024]
Abstract
PURPOSE To evaluate the effect of the intracameral adrenaline and trypan blue used during cataract surgery on corneal endothelial parameters in pseudoexfoliation syndrome (PEX) patients. METHODS The patients were divided into four groups according to intraoperative use of agents during cataract surgery: intracameral adrenaline (1/10,000, 0.1 ml) (group 1), trypan blue (0.6 mg/ml, 0.1 ml) (group 2), combination of adrenaline and trypan blue (group 3) and none (group 4). RESULTS Preoperative ECD, CV, HEX and CCT parameters were similar between the groups. A mean loss of 12.7% in ECD was observed at the postoperative third months compared to the preoperative. In group 3, ECD was found to be lower in the postoperative third months compared to the preoperative (p = 0.014). In the other groups, no statistically significant difference was found in preoperative and postoperative comparisons. CONCLUSION In conclusion, the utilization of intracameral adrenaline or trypan blue did not cause a significant difference in corneal endothelium in PEX patients. However, their combined use may have a negative effect on endothelial cell density. In a cataract surgery performed in the presence of PEX, the increased likelihood of endothelial damage should be taken into consideration, and appropriate precautions should be taken preoperatively and intraoperatively.
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Affiliation(s)
- Ömer Özer
- Department of Ophthalmology, Niğde Ömer Halisdemir University, Niğde, Turkey
| | - Levent Doğan
- Department of Ophthalmology, Niğde Ömer Halisdemir University, Niğde, Turkey
| | - Zeki Baysal
- Department of Ophthalmology, Niğde Ömer Halisdemir University, Niğde, Turkey
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8
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Ge LY, Tian X, Guo HM, Yin X. Secondary childhood glaucoma in neurofibromatosis type 1: an unusual corneal leukoma case report. Front Oncol 2024; 14:1469969. [PMID: 39669368 PMCID: PMC11634849 DOI: 10.3389/fonc.2024.1469969] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/09/2024] [Accepted: 11/12/2024] [Indexed: 12/14/2024] Open
Abstract
Neurofibromatosis type 1 (NF1) is a rare autosomal dominant disorder that affects the skin, eyes and peripheral nervous system. It is rarely associated with glaucoma, especially in pediatric patients. We herein report an unusual case of corneal degeneration in a child with NF1, characterized by peripheral corneal leukoma and a membrane under Descemet's membrane. This finding offers new insights for the ophthalmic diagnosis of NF1.
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Affiliation(s)
| | | | | | - Xue Yin
- Department of Ophthalmology, the First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China
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9
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Yang GN, Sun YBY, Roberts PK, Moka H, Sung MK, Gardner-Russell J, El Wazan L, Toussaint B, Kumar S, Machin H, Dusting GJ, Parfitt GJ, Davidson K, Chong EW, Brown KD, Polo JM, Daniell M. Exploring single-cell RNA sequencing as a decision-making tool in the clinical management of Fuchs' endothelial corneal dystrophy. Prog Retin Eye Res 2024; 102:101286. [PMID: 38969166 DOI: 10.1016/j.preteyeres.2024.101286] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/17/2024] [Revised: 06/14/2024] [Accepted: 07/02/2024] [Indexed: 07/07/2024]
Abstract
Single-cell RNA sequencing (scRNA-seq) has enabled the identification of novel gene signatures and cell heterogeneity in numerous tissues and diseases. Here we review the use of this technology for Fuchs' Endothelial Corneal Dystrophy (FECD). FECD is the most common indication for corneal endothelial transplantation worldwide. FECD is challenging to manage because it is genetically heterogenous, can be autosomal dominant or sporadic, and progress at different rates. Single-cell RNA sequencing has enabled the discovery of several FECD subtypes, each with associated gene signatures, and cell heterogeneity. Current FECD treatments are mainly surgical, with various Rho kinase (ROCK) inhibitors used to promote endothelial cell metabolism and proliferation following surgery. A range of emerging therapies for FECD including cell therapies, gene therapies, tissue engineered scaffolds, and pharmaceuticals are in preclinical and clinical trials. Unlike conventional disease management methods based on clinical presentations and family history, targeting FECD using scRNA-seq based precision-medicine has the potential to pinpoint the disease subtypes, mechanisms, stages, severities, and help clinicians in making the best decision for surgeries and the applications of therapeutics. In this review, we first discuss the feasibility and potential of using scRNA-seq in clinical diagnostics for FECD, highlight advances from the latest clinical treatments and emerging therapies for FECD, integrate scRNA-seq results and clinical notes from our FECD patients and discuss the potential of applying alternative therapies to manage these cases clinically.
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Affiliation(s)
- Gink N Yang
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
| | - Yu B Y Sun
- Department of Anatomy and Development Biology, Monash University, Clayton, Australia
| | - Philip Ke Roberts
- Department of Ophthalmology, Medical University Vienna, 18-20 Währinger Gürtel, Vienna, Austria
| | - Hothri Moka
- Mogrify Limited, 25 Cambridge Science Park Milton Road, Milton, Cambridge, UK
| | - Min K Sung
- Mogrify Limited, 25 Cambridge Science Park Milton Road, Milton, Cambridge, UK
| | - Jesse Gardner-Russell
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
| | - Layal El Wazan
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
| | - Bridget Toussaint
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
| | - Satheesh Kumar
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
| | - Heather Machin
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia; Lions Eye Donation Service, Level 7, Smorgon Family Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia
| | - Gregory J Dusting
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
| | - Geraint J Parfitt
- Mogrify Limited, 25 Cambridge Science Park Milton Road, Milton, Cambridge, UK
| | - Kathryn Davidson
- Department of Anatomy and Development Biology, Monash University, Clayton, Australia
| | - Elaine W Chong
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia; Department of Ophthalmology, Royal Melbourne Hospital, Melbourne, Victoria, Australia
| | - Karl D Brown
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia
| | - Jose M Polo
- Department of Anatomy and Development Biology, Monash University, Clayton, Australia
| | - Mark Daniell
- Centre for Eye Research Australia, Level 7, Peter Howson Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia; Ophthalmology, Department of Surgery, University of Melbourne and Royal Victorian Eye and Ear Hospital, East Melbourne, Victoria, Australia; Lions Eye Donation Service, Level 7, Smorgon Family Wing, 32 Gisborne Street, East Melbourne, Victoria, Australia.
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Ma Z, Wang Y, He H, Liu T, Jiang Q, Hou X. Advancing ophthalmic delivery of flurbiprofen via synergistic chiral resolution and ion-pairing strategies. Asian J Pharm Sci 2024; 19:100928. [PMID: 38867804 PMCID: PMC11165342 DOI: 10.1016/j.ajps.2024.100928] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/08/2023] [Revised: 12/24/2023] [Accepted: 01/16/2024] [Indexed: 06/14/2024] Open
Abstract
Flurbiprofen (FB), a nonsteroidal anti-inflammatory drug, is widely employed in treating ocular inflammation owing to its remarkable anti-inflammatory effects. However, the racemic nature of its commercially available formulation (Ocufen®) limits the full potential of its therapeutic activity, as the (S)-enantiomer is responsible for the desired anti-inflammatory effects. Additionally, the limited corneal permeability of FB significantly restricts its bioavailability. In this study, we successfully separated the chiral isomers of FB to obtain the highly active (S)-FB. Subsequently, utilizing ion-pairing technology, we coupled (S)-FB with various counter-ions, such as sodium, diethylamine, trimethamine (TMA), and l-arginine, to enhance its ocular bioavailability. A comprehensive evaluation encompassed balanced solubility, octanol-water partition coefficient, corneal permeability, ocular pharmacokinetics, tissue distribution, and in vivo ocular anti-inflammatory activity of each chiral isomer salt. Among the various formulations, S-FBTMA exhibited superior water solubility (about 1-12 mg/ml), lipid solubility (1< lg Pow < 3) and corneal permeability. In comparison to Ocufen®, S-FBTMA demonstrated significantly higher in vivo anti-inflammatory activity and lower ocular irritability (such as conjunctival congestion and tingling). The findings from this research highlight the potential of chiral separation and ion-pair enhanced permeation techniques in providing pharmaceutical enterprises focused on drug development with a valuable avenue for improving therapeutic outcomes.
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Affiliation(s)
- Zhining Ma
- School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China
| | - Yuequan Wang
- Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, China
| | - Huiyang He
- Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, China
| | - Tong Liu
- Liaoning Provincial Institute of Drug Inspection and Testing, Shenyang 110036, China
| | - Qikun Jiang
- Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang 110016, China
| | - Xiaohong Hou
- School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China
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11
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Jin L, Zhang L, Yan C, Liu M, Dean DC, Liu Y. Corneal injury repair and the potential involvement of ZEB1. EYE AND VISION (LONDON, ENGLAND) 2024; 11:20. [PMID: 38822380 PMCID: PMC11143703 DOI: 10.1186/s40662-024-00387-0] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/27/2024] [Accepted: 05/07/2024] [Indexed: 06/03/2024]
Abstract
The cornea, consisting of three cellular and two non-cellular layers, is the outermost part of the eyeball and frequently injured by external physical, chemical, and microbial insults. The epithelial-to-mesenchymal transition (EMT) plays a crucial role in the repair of corneal injuries. Zinc finger E-box binding homeobox 1 (ZEB1), an important transcription factor involved in EMT, is expressed in the corneal tissues. It regulates cell activities like migration, transformation, and proliferation, and thereby affects tissue inflammation, fibrosis, tumor metastasis, and necrosis by mediating various major signaling pathways, including transforming growth factor (TGF)-β. Dysfunction of ZEB1 would impair corneal tissue repair leading to epithelial healing delay, interstitial fibrosis, neovascularization, and squamous cell metaplasia. Understanding the mechanism underlying ZEB1 regulation of corneal injury repair will help us to formulate a therapeutic approach to enhance corneal injury repair.
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Affiliation(s)
- Lin Jin
- Department of Ophthalmology, The Third People's Hospital of Dalian, Dalian Medical University, Dalian, 116033, China
| | - Lijun Zhang
- Department of Ophthalmology, The Third People's Hospital of Dalian, Dalian Medical University, Dalian, 116033, China
| | - Chunxiao Yan
- Department of Ophthalmology, The Third People's Hospital of Dalian, Dalian Medical University, Dalian, 116033, China
| | - Mengxin Liu
- Department of Ophthalmology, The Third People's Hospital of Dalian, Dalian Medical University, Dalian, 116033, China
| | - Douglas C Dean
- James Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY, 40202, USA.
- Department of Medicine, University of Louisville School of Medicine, Louisville, KY, 40202, USA.
| | - Yongqing Liu
- James Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY, 40202, USA.
- Department of Medicine, University of Louisville School of Medicine, Louisville, KY, 40202, USA.
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Mallareddy V, Daigavane S. Nanoparticle-Mediated Cell Delivery: Advancements in Corneal Endothelial Regeneration. Cureus 2024; 16:e56958. [PMID: 38665717 PMCID: PMC11044897 DOI: 10.7759/cureus.56958] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/15/2024] [Accepted: 03/26/2024] [Indexed: 04/28/2024] Open
Abstract
Corneal endothelial dysfunction poses significant challenges in ophthalmology, leading to corneal edema and vision loss. Traditional treatments, including corneal transplantation, are limited by donor scarcity and potential complications. Nanoparticle-mediated cell delivery emerges as a promising approach for corneal endothelial regeneration, offering targeted and minimally invasive solutions. This comprehensive review provides insights into the role of nanoparticles in enhancing cell survival, integration, and therapeutic efficacy. We discuss the current understanding of corneal endothelial dysfunction, emphasizing the importance of regeneration. Furthermore, we explore the potential implications of nanoparticle-mediated approaches in clinical practice, highlighting opportunities for personalized treatment strategies. Future directions are also discussed, including optimization of nanoparticle design and exploration of combination therapies. Overall, this review elucidates the promising advancements in nanoparticle-mediated cell delivery for corneal endothelial regeneration and underscores the importance of continued research efforts in this evolving field.
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Affiliation(s)
- Vijaya Mallareddy
- Ophthalmology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, IND
| | - Sachin Daigavane
- Ophthalmology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Higher Education and Research, Wardha, IND
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Boix-Lemonche G, Hildebrand T, Haugen HJ, Petrovski G, Nogueira LP. Contrast-enhanced Micro-CT 3D visualization of cell distribution in hydrated human cornea. Heliyon 2024; 10:e25828. [PMID: 38356495 PMCID: PMC10865036 DOI: 10.1016/j.heliyon.2024.e25828] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/09/2023] [Revised: 01/27/2024] [Accepted: 02/02/2024] [Indexed: 02/16/2024] Open
Abstract
Background The cornea, a vital component of the human eye, plays a crucial role in maintaining visual clarity. Understanding its ultrastructural organization and cell distribution is fundamental for elucidating corneal physiology and pathology. This study comprehensively examines the microarchitecture of the hydrated human cornea using contrast-enhanced micro-computed tomography (micro-CT). Method Fresh human corneal specimens were carefully prepared and hydrated to mimic their in vivo state. Contrast enhancement with Lugol's iodine-enabled high-resolution Micro-CT imaging. The cells' three-dimensional (3D) distribution within the cornea was reconstructed and analyzed. Results The micro-CT imaging revealed exquisite details of the corneal ultrastructure, including the spatial arrangement of cells throughout its depth. This novel approach allowed for the visualization of cells' density and distribution in different corneal layers. Notably, our findings highlighted variations in cell distribution between non-hydrated and hydrated corneas. Conclusions This study demonstrates the potential of contrast-enhanced micro-CT as a valuable tool for non-destructive, 3D visualization and quantitative analysis of cell distribution in hydrated human corneas. These insights contribute to a better understanding of corneal physiology and may have implications for research in corneal diseases and tissue engineering.
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Affiliation(s)
- Gerard Boix-Lemonche
- Center for Eye Research and Innovative Diagnostics, Department of Ophthalmology, Institute for Clinical Medicine, Faculty of Medicine, University of Oslo, Norway
| | | | | | - Goran Petrovski
- Center for Eye Research and Innovative Diagnostics, Department of Ophthalmology, Institute for Clinical Medicine, Faculty of Medicine, University of Oslo, Norway
- Department of Ophthalmology, and Norwegian Center for Stem Cell Research, Oslo University Hospital, Oslo, Norway
- Department of Ophthalmology, University of Split School of Medicine and University Hospital Centre, Split, Croatia
- UKLO Network, University St. Kliment Ohridski – Bitola, Bitola, Macedonia
| | - Liebert Parreiras Nogueira
- Oral Research Laboratory, Institute of Clinical Dentistry, Faculty of Dentistry, University of Oslo, Oslo, Norway
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Altamirano F, Ortiz-Morales G, O'Connor-Cordova MA, Sancén-Herrera JP, Zavala J, Valdez-Garcia JE. Fuchs endothelial corneal dystrophy: an updated review. Int Ophthalmol 2024; 44:61. [PMID: 38345780 DOI: 10.1007/s10792-024-02994-1] [Citation(s) in RCA: 5] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/12/2023] [Accepted: 10/19/2023] [Indexed: 02/15/2024]
Abstract
PURPOSE The present review will summarize FECD-associated genes and pathophysiology, diagnosis, current therapeutic approaches, and future treatment perspectives. METHODS Literature review. RESULTS Fuchs' endothelial corneal dystrophy (FECD) is the most common bilateral corneal dystrophy and accounts for one-third of all corneal transplants performed in the US. FECD is caused by a combination of genetic and non-heritable factors, and there are two types: early-onset FECD, which affects individuals from an early age and is usually more severe, and late-onset FECD, which is more common and typically manifests around the age of 40. The hallmark findings of FECD include progressive loss of corneal endothelial cells and the formation of focal excrescences (guttae) on the Descemet membrane. These pathophysiological changes result in progressive endothelial dysfunction, leading to a decrease in visual acuity and blindness in later stages. The present review will summarize FECD-associated genes and pathophysiology, diagnosis, current therapeutic approaches, and future treatment perspectives. CONCLUSION With the characterization and understanding of FECD-related genes and ongoing research into regenerative therapies for corneal endothelium, we can hope to see more significant improvements in the future in the management and care of the disease.
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Affiliation(s)
- Francisco Altamirano
- Tecnologico de Monterrey, School of Medicine and Health Sciences, Monterrey, Mexico
| | | | | | | | - Judith Zavala
- Tecnologico de Monterrey, School of Medicine and Health Sciences, Monterrey, Mexico
| | - Jorge E Valdez-Garcia
- Tecnologico de Monterrey, School of Medicine and Health Sciences, Monterrey, Mexico.
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Azevedo MG, Méndez NP, Cargnin LS, Rocha RS, Seibel MP, da Silva AF, Pigatto JAT. Specular microscopy of the corneal endothelial cells of bovines: an ex vivo study. Open Vet J 2023; 13:1554-1561. [PMID: 38292711 PMCID: PMC10824098 DOI: 10.5455/ovj.2023.v13.i12.5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/12/2023] [Accepted: 11/16/2023] [Indexed: 02/01/2024] Open
Abstract
Background The endothelium is the most posterior layer of the cornea and is essential for maintaining corneal transparency. Due to variations in corneal endothelial parameters among different species, knowledge of the normal parameters for each species is crucial. Aim To evaluate the corneal endothelium of bovines using contact specular microscopy. Methods Twenty eyeballs from 10 male Brangus (Bos taurus) aged 24 months were evaluated. Contact specular microscopy was performed on the central corneal area. The analyzed parameters were endothelial cell density (ECD) and endothelial cell morphology. Results The ECD in the central area was 1,277 cells/mm2. Regarding the morphology, mainly cells with six (74.3%), five (14.7%) and seven sides (10%) were found. There were no significant differences in ECD and morphology between left and right eyes. Conclusion Contact specular microscopy facilitated the analysis and measurement of corneal endothelial parameters in bovines. The data obtained will serve as a reference for the analysis of bovine corneal endothelium.
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Affiliation(s)
| | - Natália Pons Méndez
- College of Veterinary, Federal University of Rio Grande do Sul, UFRGS, Porto Alegre, Brazil
| | - Luísa Soares Cargnin
- College of Veterinary, Federal University of Rio Grande do Sul, UFRGS, Porto Alegre, Brazil
| | - Rafaella Silva Rocha
- College of Veterinary, Federal University of Rio Grande do Sul, UFRGS, Porto Alegre, Brazil
| | - Maiara Poersch Seibel
- College of Veterinary, Federal University of Rio Grande do Sul, UFRGS, Porto Alegre, Brazil
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Huang X, Li L, Chen Z, Yu H, You X, Kong N, Tao W, Zhou X, Huang J. Nanomedicine for the Detection and Treatment of Ocular Bacterial Infections. ADVANCED MATERIALS (DEERFIELD BEACH, FLA.) 2023; 35:e2302431. [PMID: 37231939 DOI: 10.1002/adma.202302431] [Citation(s) in RCA: 17] [Impact Index Per Article: 8.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/15/2023] [Revised: 05/15/2023] [Indexed: 05/27/2023]
Abstract
Ocular bacterial infection is a prevalent cause of blindness worldwide, with substantial consequences for normal human life. Traditional treatments for ocular bacterial infections areless effective, necessitating the development of novel techniques to enable accurate diagnosis, precise drug delivery, and effective treatment alternatives. With the rapid advancement of nanoscience and biomedicine, increasing emphasis has been placed on multifunctional nanosystems to overcome the challenges posed by ocular bacterial infections. Given the advantages of nanotechnology in the biomedical industry, it can be utilized to diagnose ocular bacterial infections, administer medications, and treat them. In this review, the recent advancements in nanosystems for the detection and treatment of ocular bacterial infections are discussed; this includes the latest application scenarios of nanomaterials for ocular bacterial infections, in addition to the impact of their essential characteristics on bioavailability, tissue permeability, and inflammatory microenvironment. Through an in-depth investigation into the effect of sophisticated ocular barriers, antibacterial drug formulations, and ocular metabolism on drug delivery systems, this review highlights the challenges faced by ophthalmic medicine and encourages basic research and future clinical transformation based on ophthalmic antibacterial nanomedicine.
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Affiliation(s)
- Xiaomin Huang
- Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University; Key Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, 200030, China
- Shanghai Research Center of Ophthalmology and Optometry, Shanghai, 200030, China
- Wenzhou Medical University, Wenzhou, Zhejiang, 325027, China
| | - Luoyuan Li
- Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University; Key Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, 200030, China
- Shanghai Research Center of Ophthalmology and Optometry, Shanghai, 200030, China
- The Eighth Affiliated Hospital Sun Yat-sen University, Shenzhen, Guangdong, 518033, P. R. China
| | - Zhongxing Chen
- Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University; Key Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, 200030, China
- Shanghai Research Center of Ophthalmology and Optometry, Shanghai, 200030, China
| | - Haoyu Yu
- The Eighth Affiliated Hospital Sun Yat-sen University, Shenzhen, Guangdong, 518033, P. R. China
| | - Xinru You
- Center for Nanomedicine and Department of Anesthesiology Brigham and Women's Hospital Harvard Medical School, Boston, MA, 02115, USA
| | - Na Kong
- Center for Nanomedicine and Department of Anesthesiology Brigham and Women's Hospital Harvard Medical School, Boston, MA, 02115, USA
| | - Wei Tao
- Center for Nanomedicine and Department of Anesthesiology Brigham and Women's Hospital Harvard Medical School, Boston, MA, 02115, USA
| | - Xingtao Zhou
- Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University; Key Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, 200030, China
- Shanghai Research Center of Ophthalmology and Optometry, Shanghai, 200030, China
| | - Jinhai Huang
- Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University; Key Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, 200030, China
- Shanghai Research Center of Ophthalmology and Optometry, Shanghai, 200030, China
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Aloy-Reverté C, Bandeira F, Otero N, Rebollo-Morell A, Nieto-Nicolau N, Álvaro P. Gomes J, Güell JL, Casaroli-Marano RP. Corneal Endothelial Cell Cultures from Organotypic Preservation of Older Donor Corneas Are Suitable for Advanced Cell Therapy. Ophthalmic Res 2023; 66:1254-1265. [PMID: 37722372 PMCID: PMC10614447 DOI: 10.1159/000533701] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2022] [Accepted: 08/09/2023] [Indexed: 09/20/2023]
Abstract
INTRODUCTION The purpose of this work was to evaluate the in vitro growth capacity and functionality of human corneal endothelial cells (hCEC) expanded from corneas of elderly (>60 years) donors that were preserved using an organotypic culture method (>15 days, 31°C) and did not meet the clinical criteria for keratoplasty. METHODS Cell cultures were obtained from prior descemetorhexis (≥10 mm) and a controlled incubation with collagenase type I followed by recombinant trypsin. Cells were seeded on coated plates (fibronectin-albumin-collagen I) and cultures were expanded using the dual supplemented medium approach (maintenance medium and growth medium), in the presence of a 10 μm Rho-associated protein kinase inhibitor (Y-27632). Cell passages were obtained at culture confluency (∼2 weeks). A quantitative colorimetric WST-1 cell growth assay was performed at different time points of the culture. Morphometric analysis (area assessment and circularity), immunocytochemistry (ZO-1, Na+/K+-ATPase α, Ki67), and transendothelial electrical resistance (TEER) were performed on confluent monolayers. RESULTS There was no difference between the cell growth profiles of hCEC cultures obtained from corneas older than 60 years, whether preserved cold or cultivated organotypic corneas. Primary cultures were able to maintain a certain cell circularity index (around 0.8) and morphology (hexagonal) similar to corneal endothelial mosaic. The ZO-1 and Na+/K+-ATPase pump markers were highly positive in confluent cell monolayers at 21 days after isolation (passage 0; P0), but significantly decreased in confluent monolayers after the first passage (P1). A weak expression of Ki67 was observed in both P0 and P1 monolayers. The P0 monolayers showed a progressive increase in TEER values between days 6 and 11 and remained stable until day 18 of culture, indicating a state of controlled permeability in monolayers. The P1 monolayers also showed some functional ability but with decreased TEER values compared to monolayers at P0. CONCLUSIONS Our results indicate that it is possible to obtain functional hCEC cultures in eye banks, using simplified and standardized protocols, from older donor corneas (>60 years of age), previously preserved under organotypic culture conditions. This tissue is more readily available in our setting, due to the profile of the donor population or due to the low endothelial count (<2,000 cells/mm2) of the donated cornea.
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Affiliation(s)
| | - Francisco Bandeira
- Department of Ophthalmology and Visual Sciences, Escola Paulista de Medicina (EPM), Universidade Federal de Sao Paulo (UNIFESP), Sao Paulo, Brazil
| | - Nausica Otero
- Barcelona Tissue Bank (BTB), Banc de Sang i Teixits (BST), Barcelona, Spain
| | | | | | - José Álvaro P. Gomes
- Department of Ophthalmology and Visual Sciences, Escola Paulista de Medicina (EPM), Universidade Federal de Sao Paulo (UNIFESP), Sao Paulo, Brazil
| | - José L. Güell
- Instituto de Microcirugía Ocular (IMO), IMO Foundation, Barcelona, Spain
| | - Ricardo P. Casaroli-Marano
- Barcelona Tissue Bank (BTB), Banc de Sang i Teixits (BST), Barcelona, Spain
- Department of Ophthalmology and Visual Sciences, Escola Paulista de Medicina (EPM), Universidade Federal de Sao Paulo (UNIFESP), Sao Paulo, Brazil
- Department of Surgery, School of Medicine and Health Sciences and Hospital Clinic de Barcelona, Universitat de Barcelona, Barcelona, Spain
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Zembala J, Forma A, Zembala R, Januszewski J, Zembala P, Adamowicz D, Teresiński G, Buszewicz G, Flieger J, Baj J. Technological Advances in a Therapy of Primary Open-Angle Glaucoma: Insights into Current Nanotechnologies. J Clin Med 2023; 12:5798. [PMID: 37762739 PMCID: PMC10531576 DOI: 10.3390/jcm12185798] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/05/2023] [Revised: 08/29/2023] [Accepted: 09/04/2023] [Indexed: 09/29/2023] Open
Abstract
Glaucoma is a leading cause of irreversible blindness and is characterized by increased intraocular pressure (IOP) and progressive optic nerve damage. The current therapeutic approaches for glaucoma management, such as eye drops and oral medications, face challenges including poor bioavailability, low patient compliance, and limited efficacy. In recent years, nanotechnology has emerged as a promising approach to overcome these limitations and revolutionize glaucoma treatment. In this narrative review, we present an overview of the novel nanotechnologies employed in the treatment of primary open-angle glaucoma. Various nanosystems, including liposomes, niosomes, nanoparticles, and other nanostructured carriers, have been developed to enhance the delivery and bioavailability of antiglaucoma drugs. They offer advantages such as a high drug loading capacity, sustained release, improved corneal permeability, and targeted drug delivery to the ocular tissues. The application of nanotechnologies in glaucoma treatment represents a transformative approach that addresses the limitations of conventional therapies. However, further research is needed to optimize the formulations, evaluate long-term safety, and implement these nanotechnologies into clinical practice. With continued advancements in nanotechnology, the future holds great potential for improving the management and outcomes of glaucoma, ultimately preserving vision and improving the lives of millions affected by this debilitating disease.
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Affiliation(s)
- Julita Zembala
- University Clinical Center, Medical University of Warsaw, Lindleya 4, 02-005 Warsaw, Poland
| | - Alicja Forma
- Department of Forensic Medicine, Medical University of Lublin, Jaczewskiego 8b, 20-090 Lublin, Poland; (G.T.); (G.B.)
| | - Roksana Zembala
- Faculty of Medicine, Cardinal Stefan Wyszynski University in Warsaw, Wóycickiego 1/3, 01-938 Warsaw, Poland;
| | - Jacek Januszewski
- Department of Human Anatomy, Medical University of Lublin, Jaczewskiego 4, 20-090 Lublin, Poland; (J.J.); (J.B.)
| | - Patryk Zembala
- Faculty of Medicine, Medical University of Warsaw, Banacha 1A, 02-097 Warsaw, Poland;
| | - Dominik Adamowicz
- University Clinical Center, Medical University of Warsaw, Banacha 1A, 02-097 Warsaw, Poland;
| | - Grzegorz Teresiński
- Department of Forensic Medicine, Medical University of Lublin, Jaczewskiego 8b, 20-090 Lublin, Poland; (G.T.); (G.B.)
| | - Grzegorz Buszewicz
- Department of Forensic Medicine, Medical University of Lublin, Jaczewskiego 8b, 20-090 Lublin, Poland; (G.T.); (G.B.)
| | - Jolanta Flieger
- Department of Analytical Chemistry, Medical University of Lublin, Chodźki 4A, 20-093 Lublin, Poland;
| | - Jacek Baj
- Department of Human Anatomy, Medical University of Lublin, Jaczewskiego 4, 20-090 Lublin, Poland; (J.J.); (J.B.)
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Ng XY, Peh GSL, Yam GHF, Tay HG, Mehta JS. Corneal Endothelial-like Cells Derived from Induced Pluripotent Stem Cells for Cell Therapy. Int J Mol Sci 2023; 24:12433. [PMID: 37569804 PMCID: PMC10418878 DOI: 10.3390/ijms241512433] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/30/2023] [Revised: 07/28/2023] [Accepted: 07/31/2023] [Indexed: 08/13/2023] Open
Abstract
Corneal endothelial dysfunction is one of the leading causes of corneal blindness, and the current conventional treatment option is corneal transplantation using a cadaveric donor cornea. However, there is a global shortage of suitable donor graft material, necessitating the exploration of novel therapeutic approaches. A stem cell-based regenerative medicine approach using induced pluripotent stem cells (iPSCs) offers a promising solution, as they possess self-renewal capabilities, can be derived from adult somatic cells, and can be differentiated into all cell types including corneal endothelial cells (CECs). This review discusses the progress and challenges in developing protocols to induce iPSCs into CECs, focusing on the different media formulations used to differentiate iPSCs to neural crest cells (NCCs) and subsequently to CECs, as well as the characterization methods and markers that define iPSC-derived CECs. The hurdles and solutions for the clinical application of iPSC-derived cell therapy are also addressed, including the establishment of protocols that adhere to good manufacturing practice (GMP) guidelines. The potential risks of genetic mutations in iPSC-derived CECs associated with long-term in vitro culture and the danger of potential tumorigenicity following transplantation are evaluated. In all, this review provides insights into the advancement and obstacles of using iPSC in the treatment of corneal endothelial dysfunction.
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Affiliation(s)
- Xiao Yu Ng
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (X.Y.N.); (G.S.L.P.); (G.H.-F.Y.)
| | - Gary S. L. Peh
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (X.Y.N.); (G.S.L.P.); (G.H.-F.Y.)
- Ophthalmology and Visual Sciences Academic Clinical Program, SingHealth and Duke-NUS Medical School, Singapore 169857, Singapore;
| | - Gary Hin-Fai Yam
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (X.Y.N.); (G.S.L.P.); (G.H.-F.Y.)
- Corneal Regeneration Laboratory, Department of Ophthalmology, University of Pittsburgh, 6614, Pittsburgh, PA 15260, USA
| | - Hwee Goon Tay
- Ophthalmology and Visual Sciences Academic Clinical Program, SingHealth and Duke-NUS Medical School, Singapore 169857, Singapore;
- Centre for Vision Research, DUKE-NUS Medical School, Singapore 169857, Singapore
| | - Jodhbir S. Mehta
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore 169856, Singapore; (X.Y.N.); (G.S.L.P.); (G.H.-F.Y.)
- Ophthalmology and Visual Sciences Academic Clinical Program, SingHealth and Duke-NUS Medical School, Singapore 169857, Singapore;
- Centre for Vision Research, DUKE-NUS Medical School, Singapore 169857, Singapore
- Department of Cornea and External Eye Disease, Singapore National Eye Centre, Singapore 168751, Singapore
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20
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Luo X, He X, Zhao H, Ma J, Tao J, Zhao S, Yan Y, Li Y, Zhu S. Research Progress of Polymer Biomaterials as Scaffolds for Corneal Endothelium Tissue Engineering. NANOMATERIALS (BASEL, SWITZERLAND) 2023; 13:1976. [PMID: 37446492 DOI: 10.3390/nano13131976] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/12/2023] [Revised: 06/11/2023] [Accepted: 06/27/2023] [Indexed: 07/15/2023]
Abstract
Nowadays, treating corneal diseases arising from injury to the corneal endothelium necessitates donor tissue, but these corneas are extremely scarce. As a result, researchers are dedicating significant efforts to exploring alternative approaches that do not rely on donor tissues. Among these, creating a tissue-engineered scaffold on which corneal endothelial cells can be transplanted holds particular fascination. Numerous functional materials, encompassing natural, semi-synthetic, and synthetic polymers, have already been studied in this regard. In this review, we present a comprehensive overview of recent advancements in using polymer biomaterials as scaffolds for corneal endothelium tissue engineering. Initially, we analyze and present the key properties necessary for an effective corneal endothelial implant utilizing polymer biomaterials. Subsequently, we focus on various emerging biomaterials as scaffolds for corneal endothelium tissue engineering. We discuss their modifications (including natural and synthetic composites) and analyze the effect of micro- and nano-topological morphology on corneal endothelial scaffolds. Lastly, we highlight the challenges and prospects of these materials in corneal endothelium tissue engineering.
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Affiliation(s)
- Xiaoying Luo
- State Key Laboratory of Metal Matrix Composite, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240, China
| | - Xin He
- State Key Laboratory of Metal Matrix Composite, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240, China
| | - Hui Zhao
- National Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai 200080, China
| | - Jun Ma
- UniSA STEM and Future Industries Institute, University of South Australia, Mawson Lakes, SA 5095, Australia
| | - Jie Tao
- National Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai 200080, China
| | - Songjiao Zhao
- National Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai 200080, China
| | - Yan Yan
- National Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai 200080, China
| | - Yao Li
- State Key Laboratory of Metal Matrix Composite, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240, China
| | - Shenmin Zhu
- State Key Laboratory of Metal Matrix Composite, School of Materials Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240, China
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Jiang GJ, You XG, Fan TJ. Carteolol triggers senescence via activation of β-arrestin-ERK-NOX4-ROS pathway in human corneal endothelial cells in vitro. Chem Biol Interact 2023; 380:110511. [PMID: 37120125 DOI: 10.1016/j.cbi.2023.110511] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/25/2023] [Revised: 04/16/2023] [Accepted: 04/25/2023] [Indexed: 05/01/2023]
Abstract
Carteolol is a commonly-used topical medication for primary open-angle glaucoma. However, long-term and frequent ocular application of carteolol entails its residuals at low concentration in the aqueous humor for a long duration and may exert latent toxicity in the human corneal endothelial cells (HCEnCs). Here, we treated the HCEnCs in vitro with 0.0117% carteolol for 10 days. Thereafter, we removed the cartelolol and normally cultured the cells for 25 days to investigate the chronical toxicity of carteolol and the underlying mechanism. The results exhibited that 0.0117% carteolol induces senescent features in the HCEnCs, such as increased senescence-associated β-galactosidase positive rates, enlarged relative cell area and upregulated p16INK4A and senescence-associated secretory phenotypes, including IL-1α, TGF-β1, IL-10, TNF-α, CCL-27, IL-6 and IL-8, as well as decreased Lamin B1 expression and cell viability and proliferation. Thereby, further exploration demonstrated that the carteolol activates β-arrestin-ERK-NOX4 pathway to increase reactive oxygen species (ROS) production that imposes oxidative stress on energetic metabolism causing a vicious cycle between declining ATP and increasing ROS production and downregulation of NAD+ resulting in metabolic disturbance-mediated senescence of the HCEnCs. The excess ROS also impair DNA to activate the DNA damage response (DDR) pathway of ATM-p53-p21WAF1/CIP1 with diminished poly(ADP-Ribose) polymerase (PARP) 1, a NAD+-dependent enzyme for DNA damage repair, resulting in cell cycle arrest and subsequent DDR-mediated senescence. Taken together, carteolol induces excess ROS to trigger HCEnC senescence via metabolic disturbance and DDR pathway.
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Affiliation(s)
- Guo-Jian Jiang
- College of Marine Life Sciences, Ocean University of China, Qingdao, Shandong province, 266003, China
| | - Xin-Guo You
- School of Bioscience and Technology, Weifang Medical University, Weifang, Shandong province, 261053, China
| | - Ting-Jun Fan
- College of Marine Life Sciences, Ocean University of China, Qingdao, Shandong province, 266003, China.
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Abstract
Corneal endothelium is the innermost layer of the cornea which has both barrier and pump function and very important to maintain cornea clarity. Unlike epithelium, endothelium does not have regenerative potential; hence, endothelial damage or dysfunction could lead to corneal edema and visual impairment. Advanced corneal transplantation which involves selective replacement of dysfunctional endothelium has led to improved and faster visual rehabilitation. But in recent times, alternative therapies in the management of corneal edema and endothelial diseases have been reported. In this review, we aim to give a comprehensive review of various strategies for the management of corneal endothelial dysfunction in order to give treatment which is precisely tailored for each individual patient. A review of all peer-reviewed publications on novel strategies for the management of endothelial dysfunction was performed. The various approaches to the management of endothelial dysfunction are compared and discussed. Shortage of human donor corneas globally is fuelling the search for keratoplasty alternatives. Corneal endothelial dysfunction can be caused following surgery, laser or corneal endothelial dystrophies which could be amenable to treatment with pharmacological, biological intervention and reverse the endothelial dysfunction in the early stages of endothelial failure. Pharmacological and surgical intervention are helpful in cases of good peripheral endothelial cell reserve, and advanced cases of endothelial cell dysfunction can be targeted with cell culture therapies, gene therapy and artificial implant. Treatment strategies which target endothelial dysfunction, especially FECD in its early stages, and gene therapy are rapidly evolving. Therapies which delay endothelial keratoplasty also are evolving like DSO and need more studies of long-term follow-up and patient selection criteria.
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Affiliation(s)
- Shalini Singh
- Academy of Eye Care Education, L V Prasad Eye Institute, Hyderabad, India.,The Cornea Institute, L V Prasad Eye Institute, Hyderabad, India
| | - Sunita Chaurasia
- The Cornea Institute, L V Prasad Eye Institute, Hyderabad, India
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23
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Jia S, Bu Y, Lau DSA, Lin Z, Sun T, Lu WW, Lu S, Ruan C, Chan CHJ. Advances in 3D bioprinting technology for functional corneal reconstruction and regeneration. Front Bioeng Biotechnol 2023; 10:1065460. [PMID: 36686254 PMCID: PMC9852906 DOI: 10.3389/fbioe.2022.1065460] [Citation(s) in RCA: 16] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/09/2022] [Accepted: 12/19/2022] [Indexed: 01/09/2023] Open
Abstract
Corneal transplantation constitutes one of the major treatments in severe cases of corneal diseases. The lack of cornea donors as well as other limitations of corneal transplantation necessitate the development of artificial corneal substitutes. Biosynthetic cornea model using 3D printing technique is promising to generate artificial corneal structure that can resemble the structure of the native human cornea and is applicable for regenerative medicine. Research on bioprinting artificial cornea has raised interest into the wide range of materials and cells that can be utilized as bioinks for optimal clarity, biocompatibility, and tectonic strength. With continued advances in biomaterials science and printing technology, it is believed that bioprinted cornea will eventually achieve a level of clinical functionality and practicality as to replace donated corneal tissues, with their associated limitations such as limited or unsteady supply, and possible infectious disease transmission. Here, we review the literature on bioprinting strategies, 3D corneal modelling, material options, and cellularization strategies in relation to keratoprosthesis design. The progress, limitations and expectations of recent cases of 3D bioprinting of artifial cornea are discussed. An outlook on the rise of 3D bioprinting in corneal reconstruction and regeneration is provided.
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Affiliation(s)
- Shuo Jia
- Department of Ophthalmology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, Hong Kong SAR, China
| | - Yashan Bu
- Department of Ophthalmology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, Hong Kong SAR, China
| | - Dzi-Shing Aaron Lau
- Department of Orthopedic and Traumatology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, Hong Kong SAR, China
| | - Zhizhen Lin
- Department of Ophthalmology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, Hong Kong SAR, China
| | - Tianhao Sun
- Department of Orthopedic and Traumatology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, Hong Kong SAR, China
- Shenzhen Gangqing Biomedical Technology Co. Ltd, Shenzhen, China
| | - Weijia William Lu
- Department of Orthopedic and Traumatology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, Hong Kong SAR, China
- Research Center for Human Tissues and Organs Degeneration, Institute of Biomedicine and Biotechnology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China
| | - Sheng Lu
- Department of Orthopedic Surgery, The First People’s Hospital of Yunnan Province, Kunming, China
| | - Changshun Ruan
- Research Center for Human Tissues and Organs Degeneration, Institute of Biomedicine and Biotechnology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China
| | - Cheuk-Hung Jonathan Chan
- Department of Ophthalmology, LKS Faculty of Medicine, University of Hong Kong, Hong Kong, Hong Kong SAR, China
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24
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Tavakkoli F, Eleiwa TK, Elhusseiny AM, Damala M, Rai AK, Cheraqpour K, Ansari MH, Doroudian M, H Keshel S, Soleimani M, Djalilian AR, Sangwan VS, Singh V. Corneal stem cells niche and homeostasis impacts in regenerative medicine; concise review. Eur J Ophthalmol 2023:11206721221150065. [PMID: 36604831 DOI: 10.1177/11206721221150065] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/07/2023]
Abstract
The limbal stem cells niche (LSCN) is an optimal microenvironment that provides the limbal epithelial stem cells (LESCs) and strictly regulates their proliferation and differentiation. Disturbing the LSCN homeostasis can lead to limbal stem cell dysfunction (LSCD) and subsequent ocular surface aberrations, such as corneal stromal inflammation, persistent epithelial defects, corneal neovascularisation, lymphangiogenesis, corneal opacification, and conjunctivalization. As ocular surface disorders are considered the second main cause of blindness, it becomes crucial to explore different therapeutic strategies for restoring the functions of the LSCN. A major limitation of corneal transplantation is the current shortage of donor tissue to meet the requirements worldwide. In this context, it becomes mandatory to find an alternative regenerative medicine, such as using cultured limbal epithelial/stromal stem cells, inducing the production of corneal like cells by using other sources of stem cells, and using tissue engineering methods aiming to produce the three-dimensional (3D) printed cornea. Limbal epithelial stem cells have been considered the magic potion for eye treatment. Epithelial and stromal stem cells in the limbal niche hold the responsibility of replenishing the corneal epithelium. These stem cells are being used for transplantation to maintain corneal epithelial integrity and ultimately sustain optimal vision. In this review, we summarised the characteristics of the LSCN and their current and future roles in restoring corneal homeostasis in eyes with LSCD.
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Affiliation(s)
- Fatemeh Tavakkoli
- Department of Community Health, College of Health Technology, Cihan University, Erbil, Iraq.,SSR Stem Cell Biology Laboratory, Brien Holden Eye Research Centre, Centre for Ocular Regeneration, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, Hyderabad, India.,Centre for Genetic Disorders, Banaras Hindu University, Varanasi, India
| | - Taher K Eleiwa
- Department of Ophthalmology, Benha University, Benha, Egypt
| | - Abdelrahman M Elhusseiny
- Department of Ophthalmology, Harvey and Bernice Jones Eye Institute, University of Arkansas for Medical Sciences, Little Rock, AR, USA
| | - Mukesh Damala
- SSR Stem Cell Biology Laboratory, Brien Holden Eye Research Centre, Centre for Ocular Regeneration, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, Hyderabad, India.,School of Life Sciences, University of Hyderabad, Hyderabad, India
| | - Amit K Rai
- Centre for Genetic Disorders, Banaras Hindu University, Varanasi, India
| | - Kasra Cheraqpour
- Translational Eye Research Center, Farabi Eye Hospital, 48439Tehran University of Medical Sciences, Tehran, Iran
| | - Mohammad H Ansari
- Ophthalmic Research Center, Department of Ophthalmology, Labbafinejad Medical Center, 556492Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Mohammad Doroudian
- Department of Cell and Molecular Biology, Faculty of Biological Sciences, 145440Kharazmi University, Tehran, Iran
| | - Saeed H Keshel
- Department of Tissue Engineering and Applied Cell Sciences, 556492Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Mohammad Soleimani
- Department of Ophthalmology, 159636Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA
| | - Ali R Djalilian
- Department of Ophthalmology, 159636Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, IL, USA
| | | | - Vivek Singh
- SSR Stem Cell Biology Laboratory, Brien Holden Eye Research Centre, Centre for Ocular Regeneration, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, Hyderabad, India
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25
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Vercammen H, Miron A, Oellerich S, Melles GRJ, Ní Dhubhghaill S, Koppen C, Van Den Bogerd B. Corneal endothelial wound healing: understanding the regenerative capacity of the innermost layer of the cornea. Transl Res 2022; 248:111-127. [PMID: 35609782 DOI: 10.1016/j.trsl.2022.05.003] [Citation(s) in RCA: 14] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 02/09/2022] [Revised: 04/14/2022] [Accepted: 05/18/2022] [Indexed: 12/13/2022]
Abstract
Currently, there are very few well-established treatments to stimulate corneal endothelial cell regeneration in vivo as a cure for corneal endothelial dysfunctions. The most frequently performed intervention for a damaged or dysfunctional corneal endothelium nowadays is corneal endothelial keratoplasty, also known as lamellar corneal transplantation surgery. Newer medical therapies are emerging and are targeting the regeneration of the corneal endothelium, helping the patients regain their vision without the need for donor tissue. Alternatives to donor tissues are needed as the aging population requiring transplants, has further exacerbated the pressure on the corneal eye banking system. Significant ongoing research efforts in the field of corneal regenerative medicine have been made to elucidate the underlying pathways and effector proteins involved in corneal endothelial regeneration. However, the literature offers little guidance and selective attention to the question of how to fully exploit these pathways. The purpose of this paper is to provide an overview of wound healing characteristics from a biochemical level in the lab to the regenerative features seen in the clinic. Studying the pathways involved in corneal wound healing together with their key effector proteins, can help explain the effect on the proliferation and migration capacity of the corneal endothelial cells.
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Affiliation(s)
- Hendrik Vercammen
- Antwerp Research Group for Ocular Science (ARGOS), Translational Neurosciences, Faculty of Medicine and Health Sciences, University of Antwerp, Wilrijk, Belgium
| | - Alina Miron
- Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, The Netherlands
| | - Silke Oellerich
- Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, The Netherlands
| | - Gerrit R J Melles
- Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, The Netherlands; Melles Cornea Clinic Rotterdam, The Netherlands
| | - Sorcha Ní Dhubhghaill
- Antwerp Research Group for Ocular Science (ARGOS), Translational Neurosciences, Faculty of Medicine and Health Sciences, University of Antwerp, Wilrijk, Belgium; Department of Ophthalmology, Antwerp University Hospital, Edegem, Belgium; Netherlands Institute for Innovative Ocular Surgery (NIIOS), Rotterdam, The Netherlands
| | - Carina Koppen
- Antwerp Research Group for Ocular Science (ARGOS), Translational Neurosciences, Faculty of Medicine and Health Sciences, University of Antwerp, Wilrijk, Belgium; Department of Ophthalmology, Antwerp University Hospital, Edegem, Belgium
| | - Bert Van Den Bogerd
- Antwerp Research Group for Ocular Science (ARGOS), Translational Neurosciences, Faculty of Medicine and Health Sciences, University of Antwerp, Wilrijk, Belgium.
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26
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Jiang GJ, You XG, Fan TJ. Ultraviolet B irradiation induces senescence of human corneal endothelial cells in vitro by DNA damage response and oxidative stress. JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY. B, BIOLOGY 2022; 235:112568. [PMID: 36137302 DOI: 10.1016/j.jphotobiol.2022.112568] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/03/2022] [Revised: 09/04/2022] [Accepted: 09/12/2022] [Indexed: 06/16/2023]
Abstract
The human corneal endothelial cells (HCEnCs) play a vital role in the maintenance of corneal transparency and visual acuity. In our daily life, HCEnCs are inevitably exposed to ultraviolet B (UVB) radiation leading to decreases of visual acuity and corneal transparency resulting in visual loss eventually. Therefore, understanding the UVB-induced cytotoxicity in HCEnCs is of importance for making efficient strategies to protect our vision from UVB-damage. However, in-depth knowledge about UVB-induced cytotoxicity in HCEnCs is missing. Herein, we pulse-irradiated the HCEnCs in vitro with 150 mJ/cm2 UVB (the environmental dose) at each subculture for 4 passages to explore the insights into UVB-induced phototoxicity. The results showed that the UVB-treated HCEnCs exhibit typical senescent characteristics, including significantly enlarged relative cell area, increased senescence-associated β-galactosidase positive staining, and upregulated p16INK4A and senescence associated secretory phenotypes (SASPs) such as CCL-27, IL-1α/6/8/10, TGF-β1 and TNF-α, as well as decreased cell proliferation and Lamin B1 expression, and translocation of Lamin B1. Furthermore, we explored the causative mechanisms of senescence and found that 150 mJ/cm2 UVB pulse-irradiation impairs DNA to activate DNA damage response (DDR) pathway of ATM-p53-p21WAF1/CIP1 with downregulated DNA repair enzyme PARP1, leading to cell cycle arrest resulting in DDR-mediated senescence. Meanwhile, UVB pulse-irradiation also elicits a consistent increase of ROS production to aggravate DNA damage and impose oxidative stress on energy metabolism leading to metabolic disturbance resulting in metabolic disturbance-mediated senescence. Altogether, the repeated pulse-irradiation of 150 mJ/cm2 UVB induces HCEnC senescence via both DDR pathway and energy metabolism disturbance.
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Affiliation(s)
- Guo-Jian Jiang
- College of marine life sciences, Ocean university of China, Qingdao, Shandong province 266003, China
| | - Xin-Guo You
- School of bioscience and technology, Weifang medical university, Weifang, Shandong province 261053, China
| | - Ting-Jun Fan
- College of marine life sciences, Ocean university of China, Qingdao, Shandong province 266003, China.
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27
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Montalvo-Parra MD, Ortega-Lara W, Loya-García D, Bustamante-Arias A, Guerrero-Ramírez GI, Calzada-Rodríguez CE, Torres-Guerrero GF, Hernández-Sedas B, Cárdenas-Rodríguez IT, Guevara-Quintanilla SE, Salán-Gomez M, Hernández-Delgado MÁ, Garza-González S, Gamboa-Quintanilla MG, Villagómez-Valdez LG, Zavala J, Valdez-García JE. Customizable Collagen Vitrigel Membranes and Preliminary Results in Corneal Engineering. Polymers (Basel) 2022; 14:polym14173556. [PMID: 36080636 PMCID: PMC9460691 DOI: 10.3390/polym14173556] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2022] [Revised: 07/28/2022] [Accepted: 08/24/2022] [Indexed: 11/16/2022] Open
Abstract
Corneal opacities are a leading cause of visual impairment that affect 4.2 million people annually. The current treatment is corneal transplantation, which is limited by tissue donor shortages. Corneal engineering aims to develop membranes that function as scaffolds in corneal cell transplantation. Here, we describe a method for producing transplantable corneal constructs based on a collagen vitrigel (CVM) membrane and corneal endothelial cells (CECs). The CVMs were produced using increasing volumes of collagen type I: 1X (2.8 μL/mm2), 2X, and 3X. The vitrification process was performed at 40% relative humidity (RH) and 40 °C using a matryoshka-like system consisting of a shaking-oven harboring a desiccator with a saturated K2CO3 solution. The CVMs were characterized via SEM microscopy, cell adherence, FTIR, and manipulation in an ex vivo model. A pilot transplantation of the CECs/CVM construct in rabbits was also carried out. The thickness of the CVMs was 3.65–7.2 µm. The transparency was superior to a human cornea (92.6% = 1X; 94% = 2X; 89.21% = 3X). SEM microscopy showed a homogenous surface and laminar organization. The cell concentration seeded over the CVM increased threefold with no significant difference between 1X, 2X, and 3X (p = 0.323). The 2X-CVM was suitable for surgical manipulation in the ex vivo model. Constructs using the CECs/2X-CVM promoted corneal transparency restoration.
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Affiliation(s)
- María Dolores Montalvo-Parra
- Tecnologico de Monterrey, Escuela de Ingenieria, 2501 Garza Sada Ave., Colonia Tecnologico. C.P., 64849 Monterrey, NL, Mexico
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Wendy Ortega-Lara
- Tecnologico de Monterrey, Escuela de Ingenieria, 2501 Garza Sada Ave., Colonia Tecnologico. C.P., 64849 Monterrey, NL, Mexico
| | - Denise Loya-García
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Andrés Bustamante-Arias
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Guillermo-Isaac Guerrero-Ramírez
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Cesar E. Calzada-Rodríguez
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Guiomar Farid Torres-Guerrero
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Betsabé Hernández-Sedas
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Italia Tatnaí Cárdenas-Rodríguez
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Sergio E. Guevara-Quintanilla
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Marcelo Salán-Gomez
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Miguel Ángel Hernández-Delgado
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Salvador Garza-González
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Mayra G. Gamboa-Quintanilla
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Luis Guillermo Villagómez-Valdez
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
| | - Judith Zavala
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
- Correspondence:
| | - Jorge E. Valdez-García
- Tecnologico de Monterrey, Escuela de Medicina, 3000 Morones Prieto Ave., Colonia Los Doctores. C.P., 64710 Monterrey, NL, Mexico
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28
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Corneal Regeneration Using Adipose-Derived Mesenchymal Stem Cells. Cells 2022; 11:cells11162549. [PMID: 36010626 PMCID: PMC9406486 DOI: 10.3390/cells11162549] [Citation(s) in RCA: 23] [Impact Index Per Article: 7.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/15/2022] [Revised: 08/12/2022] [Accepted: 08/12/2022] [Indexed: 11/28/2022] Open
Abstract
Adipose-derived stem cells are a subtype of mesenchymal stem cell that offers the important advantage of being easily obtained (in an autologous manner) from low invasive procedures, rendering a high number of multipotent stem cells with the potential to differentiate into several cellular lineages, to show immunomodulatory properties, and to promote tissue regeneration by a paracrine action through the secretion of extracellular vesicles containing trophic factors. This secretome is currently being investigated as a potential source for a cell-free based regenerative therapy for human tissues, which would significantly reduce the involved costs, risks and law regulations, allowing for a broader application in real clinical practice. In the current article, we will review the existing preclinical and human clinical evidence regarding the use of such adipose-derived mesenchymal stem cells for the regeneration of the three main layers of the human cornea: the epithelium (derived from the surface ectoderm), the stroma (derived from the neural crest mesenchyme), and the endothelium (derived from the neural crest cells).
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29
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Hyaluronic acid hydrogels crosslinked via blue light-induced thiol-ene reaction for the treatment of rat corneal alkali burn. Regen Ther 2022; 20:51-60. [PMID: 35402662 PMCID: PMC8971597 DOI: 10.1016/j.reth.2022.03.005] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/16/2021] [Revised: 02/23/2022] [Accepted: 03/08/2022] [Indexed: 02/05/2023] Open
Abstract
To assess corneal inflammation from alkali chemical burns, we examined the therapeutic effects of in situ-forming hyaluronic acid (HA) hydrogels crosslinked via blue light-induced thiol-ene reaction on a rat corneal alkali burn model. Animals were divided into three groups (n = 7 rats per group): untreated, treated with 0.1% HA eye drops, and treated with crosslinked HA hydrogels. Crosslinking of HA hydrogel followed by the administration of HA eye drops and crosslinked HA hydrogels were carried out once a day from days 0–4. Corneal re-epithelialization, opacity, neovascularization, thickness, and histology were evaluated to compare the therapeutic effects of the three groups. Further investigation was conducted on the transparency of HA hydrogels to acquire the practical capabilities of hydrogel as a reservoir for drug delivery. Compared to untreated animals, animals treated with crosslinked HA hydrogels exhibited greater corneal re-epithelialization on days 1, 2, 4, and 7 post-injury (p = 0.004, p = 0.007, p = 0.008, and p = 0.034, respectively) and the least corneal neovascularization (p = 0.008). Histological analysis revealed lower infiltration of stromal inflammatory cells and compact collagen structure in crosslinked HA hydrogel-treated animals than in untreated animals. These findings corresponded with immunohistochemical analyses indicating that the expression of inflammatory markers such as α-SMA, MMP9, and IL1-β was lower in animals treated with crosslinked HA hydrogels than untreated animals and animals treated only with 0.1% HA eye drops. With beneficial pharmacological effects such as re-epithelization and anti-inflammation, in situ-forming hyaluronic acid (HA) hydrogels may be a promising approach to effective drug delivery in cases of corneal burn injuries.
Corneal chemical injuries can induce corneal opacification, limbal ischemia, and loss of vision. Limitations for using topical eye drops includes maintaining the optimal concentration of the drug on the ocular surface. Crosslinked HA hydrogels achieved rapid corneal re-epithelialization and low-grade neovascularization after chemical injury.
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30
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Corneal Endothelial Cell Loss in Glaucoma and Glaucoma Surgery and the Utility of Management with Descemet Membrane Endothelial Keratoplasty (DMEK). J Ophthalmol 2022; 2022:1315299. [PMID: 35637682 PMCID: PMC9148223 DOI: 10.1155/2022/1315299] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/24/2021] [Accepted: 01/10/2022] [Indexed: 01/15/2023] Open
Abstract
The corneal endothelium has a crucial role in maintaining a clear and healthy cornea. Corneal endothelial cell loss occurs naturally with age; however, a diagnosis of glaucoma and surgical intervention for glaucoma can exacerbate a decline in cell number and impairment in morphology. In glaucoma, the mechanisms for this are not well understood and this accelerated cell loss can result in corneal decompensation. Given the high prevalence of glaucoma worldwide, this review aims to explore the abnormalities observed in the corneal endothelium in differing glaucoma phenotypes and glaucoma therapies (medical or surgical including with new generation microinvasive glaucoma surgeries). Descemet membrane endothelial keratoplasty (DMEK) is increasingly being used to manage corneal endothelial failure for glaucoma patients and we aim to review the recent literature evaluating the use of this technique in this clinical scenario.
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31
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Chen J, Ou Q, Wang Z, Liu Y, Hu S, Liu Y, Tian H, Xu J, Gao F, Lu L, Jin C, Xu GT, Cui HP. Small-Molecule Induction Promotes Corneal Endothelial Cell Differentiation From Human iPS Cells. Front Bioeng Biotechnol 2021; 9:788987. [PMID: 34976977 PMCID: PMC8714889 DOI: 10.3389/fbioe.2021.788987] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/04/2021] [Accepted: 11/25/2021] [Indexed: 12/13/2022] Open
Abstract
Purpose: Corneal endothelial cells (CECs) serve as a barrier and foothold for the corneal stroma to maintain the function and transparency of the cornea. Loss of CECs during aging or disease states leads to blindness, and cell replacement therapy using either donated or artificially differentiated CECs remains the only curative approach. Methods: Human induced pluripotent stem cells (hiPSCs) that were cultured in chemically defined medium were induced with dual-SMAD inhibition to differentiate into neural crest cells (NCCs). A small-molecule library was screened to differentiate the NCCs into corneal endothelial-like cells. The characteristics of these cells were identified with real-time PCR and immunofluorescence. Western blotting was applied to detect the signaling pathways and key factors regulated by the small molecules. Results: We developed an effective protocol to differentiate hiPSCs into CECs with defined small molecules. The hiPSC-CECs were characterized by ZO-1, AQP1, Vimentin and Na+/K+-ATPase. Based on our small-molecule screen, we identified a small-molecule combination, A769662 and AT13148, that enabled the most efficient production of CECs. The combination of A769662 and AT13148 upregulated the PKA/AKT signaling pathway, FOXO1 and PITX2 to promote the conversion of NCCs to CECs. Conclusion: We established an efficient small molecule-based method to differentiate hiPSCs into corneal endothelial-like cells, which might facilitate drug discovery and the development of cell-based therapies for corneal diseases.
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Affiliation(s)
- Jie Chen
- Department of Ophthalmology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China
| | - Qingjian Ou
- Department of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
| | - Zhe Wang
- Department of Ophthalmology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China
| | - Yifan Liu
- Department of Ophthalmology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China
| | - Shuqin Hu
- Department of Ophthalmology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China
| | - Yumeilan Liu
- Department of Ophthalmology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China
| | - Haibin Tian
- Department of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
| | - Jingying Xu
- Department of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
| | - Furong Gao
- Department of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
| | - Lixia Lu
- Department of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
- Department of Biochemistry and Molecular Biology, School of Medicine, Tongji University, Shanghai, China
| | - Caixia Jin
- Department of Ophthalmology, Shanghai Tenth People’s Hospital, Tongji University School of Medicine, Shanghai, China
- Department of Biochemistry and Molecular Biology, School of Medicine, Tongji University, Shanghai, China
| | - Guo-Tong Xu
- Department of Biochemistry and Molecular Biology, School of Medicine, Tongji University, Shanghai, China
| | - Hong-Ping Cui
- Department of Ophthalmology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai, China
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Improvement of an Effective Protocol for Directed Differentiation of Human Adipose Tissue-Derived Adult Mesenchymal Stem Cells to Corneal Endothelial Cells. Int J Mol Sci 2021; 22:ijms222111982. [PMID: 34769411 PMCID: PMC8585097 DOI: 10.3390/ijms222111982] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/08/2021] [Revised: 10/28/2021] [Accepted: 11/01/2021] [Indexed: 12/13/2022] Open
Abstract
Corneal disease affects 12.5 million individuals worldwide, with 2 million new cases each year. The standard treatment consists of a corneal transplantation from a human donor; however, the worldwide demand significantly exceeds the available supply. Lamellar endothelial keratoplasty, the replacement of only the endothelial layer of the cornea, can partially solve the problem. Progressive efforts have succeeded in expanding hCECs; however, the ability to expand hCECs is still limited, and new sources of CECs are being sought. Crucial advances have been achieved by the directed differentiation of embryonic or induced pluripotent stem cells, but these cells have disadvantages, such as the use of oncogenes, and are still difficult to establish. We aimed to transfer such knowledge to obtain hCECs from adipose tissue-derived adult mesenchymal stem cells (ADSC) by modifying four previously published procedures. We present several protocols capable of the directed differentiation of human ADSCs to hCECs. In our hands, the protocol by Ali et al. was the best adapted to such differentiation in terms of efficiency, time, and financial cost; however, the protocol by Wagoner et al. was the best for CEC marker expression. Our results broaden the type of cells of autologous extraocular origin that could be employed in the clinical setting for corneal endothelial deficiency.
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Spinozzi D, Miron A, Bruinsma M, Dapena I, Kocaba V, Jager MJ, Melles GRJ, Ni Dhubhghaill S, Oellerich S. New developments in corneal endothelial cell replacement. Acta Ophthalmol 2021; 99:712-729. [PMID: 33369235 DOI: 10.1111/aos.14722] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2020] [Accepted: 11/20/2020] [Indexed: 12/16/2022]
Abstract
Corneal transplantation is currently the most effective treatment to restore corneal clarity in patients with endothelial disorders. Endothelial transplantation, either by Descemet membrane endothelial keratoplasty (DMEK) or by Descemet stripping (automated) endothelial keratoplasty (DS(A)EK), is a surgical approach that replaces diseased Descemet membrane and endothelium with tissue from a healthy donor eye. Its application, however, is limited by the availability of healthy donor tissue. To increase the pool of endothelial grafts, research has focused on developing new treatment options as alternatives to conventional corneal transplantation. These treatment options can be considered as either 'surgery-based', that is tissue-efficient modifications of the current techniques (e.g. Descemet stripping only (DSO)/Descemetorhexis without endothelial keratoplasty (DWEK) and Quarter-DMEK), or 'cell-based' approaches, which rely on in vitro expansion of human corneal endothelial cells (hCEC) (i.e. cultured corneal endothelial cell sheet transplantation and cell injection). In this review, we will focus on the most recent developments in the field of the 'cell-based' approaches. Starting with the description of aspects involved in the isolation of hCEC from donor tissue, we then describe the different natural and bioengineered carriers currently used in endothelial cell sheet transplantation, and finally, we discuss the current 'state of the art' in novel therapeutic approaches such as endothelial cell injection.
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Affiliation(s)
- Daniele Spinozzi
- Netherlands Institute for Innovative Ocular Surgery Rotterdam The Netherlands
| | - Alina Miron
- Netherlands Institute for Innovative Ocular Surgery Rotterdam The Netherlands
| | - Marieke Bruinsma
- Netherlands Institute for Innovative Ocular Surgery Rotterdam The Netherlands
| | - Isabel Dapena
- Netherlands Institute for Innovative Ocular Surgery Rotterdam The Netherlands
- Melles Cornea Clinic Rotterdam The Netherlands
| | - Viridiana Kocaba
- Netherlands Institute for Innovative Ocular Surgery Rotterdam The Netherlands
- Melles Cornea Clinic Rotterdam The Netherlands
- Tissue Engineering and Stem Cell Group Singapore Eye Research Institute Singapore Singapore
| | - Martine J. Jager
- Department of Ophthalmology Leiden University Medical Center Leiden The Netherlands
| | - Gerrit R. J. Melles
- Netherlands Institute for Innovative Ocular Surgery Rotterdam The Netherlands
- Melles Cornea Clinic Rotterdam The Netherlands
- Amnitrans EyeBank Rotterdam The Netherlands
| | - Sorcha Ni Dhubhghaill
- Netherlands Institute for Innovative Ocular Surgery Rotterdam The Netherlands
- Melles Cornea Clinic Rotterdam The Netherlands
- Antwerp University Hospital (UZA) Edegem Belgium
| | - Silke Oellerich
- Netherlands Institute for Innovative Ocular Surgery Rotterdam The Netherlands
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Smeringaiova I, Paaske Utheim T, Jirsova K. Ex vivo expansion and characterization of human corneal endothelium for transplantation: a review. Stem Cell Res Ther 2021; 12:554. [PMID: 34717745 PMCID: PMC8556978 DOI: 10.1186/s13287-021-02611-3] [Citation(s) in RCA: 18] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2021] [Accepted: 07/26/2021] [Indexed: 12/13/2022] Open
Abstract
The corneal endothelium plays a key role in maintaining corneal transparency. Its dysfunction is currently treated with penetrating or lamellar keratoplasty. Advanced cell therapy methods seek to address the persistent global deficiency of donor corneas by enabling the renewal of the endothelial monolayer with tissue-engineered grafts. This review provides an overview of recently published literature on the preparation of endothelial grafts for transplantation derived from cadaveric corneas that have developed over the last decade (2010–2021). Factors such as the most suitable donor parameters, culture substrates and media, endothelial graft storage conditions, and transplantation methods are discussed. Despite efforts to utilize alternative cellular sources, such as induced pluripotent cells, cadaveric corneas appear to be the best source of cells for graft preparation to date. However, native endothelial cells have a limited natural proliferative capacity, and they often undergo rapid phenotype changes in ex vivo culture. This is the main reason why no culture protocol for a clinical-grade endothelial graft prepared from cadaveric corneas has been standardized so far. Currently, the most established ex vivo culture protocol involves the peel-and-digest method of cell isolation and cell culture by the dual media method, including the repeated alternation of high and low mitogenic conditions. Culture media are enriched by additional substances, such as signaling pathway (Rho-associated protein kinase, TGF-β, etc.) inhibitors, to stimulate proliferation and inhibit unwanted morphological changes, particularly the endothelial-to-mesenchymal transition. To date, this promising approach has led to the development of endothelial grafts for the first in-human clinical trial in Japan. In addition to the lack of a standard culture protocol, endothelial-specific markers are still missing to confirm the endothelial phenotype in a graft ready for clinical use. Because the corneal endothelium appears to comprise phenotypically heterogeneous populations of cells, the genomic and proteomic expression of recently proposed endothelial-specific markers, such as Cadherin-2, CD166, or SLC4A11, must be confirmed by additional studies. The preparation of endothelial grafts is still challenging today, but advances in tissue engineering and surgery over the past decade hold promise for the successful treatment of endothelial dysfunctions in more patients worldwide.
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Affiliation(s)
- Ingrida Smeringaiova
- Laboratory of the Biology and Pathology of the Eye, Institute of Biology and Medical Genetics, First Faculty of Medicine, Charles University and General University Hospital in Prague, Albertov 4, 128 00, Prague, Czech Republic
| | - Tor Paaske Utheim
- Department of Medical Biochemistry, Oslo University Hospital, Oslo, Norway.,Department of Plastic and Reconstructive Surgery, Oslo University Hospital, Oslo, Norway
| | - Katerina Jirsova
- Laboratory of the Biology and Pathology of the Eye, Institute of Biology and Medical Genetics, First Faculty of Medicine, Charles University and General University Hospital in Prague, Albertov 4, 128 00, Prague, Czech Republic.
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Jameson JF, Pacheco MO, Nguyen HH, Phelps EA, Stoppel WL. Recent Advances in Natural Materials for Corneal Tissue Engineering. Bioengineering (Basel) 2021; 8:161. [PMID: 34821727 PMCID: PMC8615221 DOI: 10.3390/bioengineering8110161] [Citation(s) in RCA: 12] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/26/2021] [Revised: 10/12/2021] [Accepted: 10/21/2021] [Indexed: 12/13/2022] Open
Abstract
Given the incidence of corneal dysfunctions and diseases worldwide and the limited availability of healthy, human donors, investigators are working to generate engineered cellular and acellular therapeutic approaches as alternatives to corneal transplants from human cadavers. These engineered strategies aim to address existing complications with human corneal transplants, including graft rejection, infection, and complications resulting from surgical methodologies. The main goals of these research endeavors are to (1) determine ideal mechanical properties, (2) devise methodologies to improve the efficacy of engineered corneal grafts and cell-based therapies, and (3) optimize transplantation of engineered tissue structures in the eye. Thus, recent innovations have sought to address these challenges through both in vitro and in vivo studies. This review covers recent work aimed at evaluating engineered materials, potential therapeutic cells, and the resulting cell-material interactions that lead to optimal corneal graft properties. Furthermore, we discuss promising strategies in corneal tissue engineering techniques and in vivo studies in animal models.
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Affiliation(s)
- Julie F. Jameson
- Department of Chemical Engineering, University of Florida, Gainesville, FL 32611, USA; (J.F.J.); (M.O.P.)
| | - Marisa O. Pacheco
- Department of Chemical Engineering, University of Florida, Gainesville, FL 32611, USA; (J.F.J.); (M.O.P.)
| | - Henry H. Nguyen
- Department of Materials Science and Engineering, University of Florida, Gainesville, FL 32611, USA;
| | - Edward A. Phelps
- J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL 32611, USA;
| | - Whitney L. Stoppel
- Department of Chemical Engineering, University of Florida, Gainesville, FL 32611, USA; (J.F.J.); (M.O.P.)
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36
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Mofidfar M, Abdi B, Ahadian S, Mostafavi E, Desai TA, Abbasi F, Sun Y, Manche EE, Ta CN, Flowers CW. Drug delivery to the anterior segment of the eye: A review of current and future treatment strategies. Int J Pharm 2021; 607:120924. [PMID: 34324989 PMCID: PMC8579814 DOI: 10.1016/j.ijpharm.2021.120924] [Citation(s) in RCA: 74] [Impact Index Per Article: 18.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/21/2021] [Revised: 07/02/2021] [Accepted: 07/05/2021] [Indexed: 01/03/2023]
Abstract
Research in the development of ophthalmic drug formulations and innovative technologies over the past few decades has been directed at improving the penetration of medications delivered to the eye. Currently, approximately 90% of all ophthalmic drug formulations (e.g. liposomes, micelles) are applied as eye drops. The major challenge of topical eye drops is low bioavailability, need for frequent instillation due to the short half-life, poor drug solubility, and potential side effects. Recent research has been focused on improving topical drug delivery devices by increasing ocular residence time, overcoming physiological and anatomical barriers, and developing medical devices and drug formulations to increase the duration of action of the active drugs. Researchers have developed innovative technologies and formulations ranging from sub-micron to macroscopic size such as prodrugs, enhancers, mucus-penetrating particles (MPPs), therapeutic contact lenses, and collagen corneal shields. Another approach towards the development of effective topical drug delivery is embedding therapeutic formulations in microdevices designed for sustained release of the active drugs. The goal is to optimize the delivery of ophthalmic medications by achieving high drug concentration with prolonged duration of action that is convenient for patients to administer.
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Affiliation(s)
| | - Behnam Abdi
- Institute of Polymeric Materials (IPM), Sahand University of Technology, New Town of Sahand, Tabriz, Iran; Faculty of Polymer Engineering, Sahand University of Technology, New Town of Sahand, Tabriz, Iran
| | - Samad Ahadian
- Terasaki Institute for Biomedical Innovation, Los Angeles, CA, USA
| | - Ebrahim Mostafavi
- Department of Medicine, Stanford University School of Medicine, Stanford, CA, USA; Stanford Cardiovascular Institute, Stanford University, CA, USA
| | - Tejal A Desai
- Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA, USA
| | - Farhang Abbasi
- Institute of Polymeric Materials (IPM), Sahand University of Technology, New Town of Sahand, Tabriz, Iran; Faculty of Polymer Engineering, Sahand University of Technology, New Town of Sahand, Tabriz, Iran
| | - Yang Sun
- Byers Eye Institute, Stanford University School of Medicine, Palo Alto, CA, USA.
| | - Edward E Manche
- Byers Eye Institute, Stanford University School of Medicine, Palo Alto, CA, USA.
| | - Christopher N Ta
- Byers Eye Institute, Stanford University School of Medicine, Palo Alto, CA, USA.
| | - Charles W Flowers
- USC Roski Eye Institute, University of Southern California, Los Angeles, CA, USA.
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Bertolin M, Lamon M, Franco E, Barbaro V, Ferrari S, Bovone C, Yu AC, Parekh M, Ponzin D, Busin M. Culture of corneal endothelial cells obtained by descemetorhexis of corneas with Fuchs endothelial corneal dystrophy. Exp Eye Res 2021; 211:108748. [PMID: 34461137 DOI: 10.1016/j.exer.2021.108748] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/23/2021] [Revised: 07/27/2021] [Accepted: 08/23/2021] [Indexed: 12/31/2022]
Abstract
Currently, endothelial keratoplasty is the gold standard for the surgical treatment of Fuchs endothelial corneal dystrophy (FECD). Despite the remarkable success in terms of surgical outcomes, a shortage of corneal donor tissue poses a limitation to performing endothelial keratoplasty in many parts of the world. Cell therapy is a potential alternative strategy to keratoplasty and is currently under investigation. Considering that corneas with FECD may contain relatively healthy endothelial cells, samples obtained by descemetorhexis of eyes undergoing EK for FECD can be used for ex vivo expansion of endothelial cells as an autologous cell culture. In this study, we established corneal endothelial cell cultures derived from 40 patients that underwent endothelial keratoplasty for advanced FECD. Several parameters were evaluated including patient characteristics such as age, gender, and endothelial cell density as well as various processing and cell culture protocols based on different combinations of shipping temperatures, stabilization periods and treatment methods for corneal endothelial cell dissociation. FECD cultures were classified into three groups as: (i) no cells, (ii) cell cultures with endothelial-like morphology or (iii) cell cultures with fibroblast-like features. Our data seem to suggest that some factors can influence FECD cell culture characteristics including young age, high paracentral endothelial cell density, low shipping temperature and short stabilization period prior to cell isolation. Treatment with type 1 collagenase for cell isolation can delay endothelial-to-mesenchymal transition, but does not increase proliferative capacity. Although heterologous corneal endothelial cultures from healthy donors have shown encouraging outcomes, the feasibility of autologous cell therapy as a potential treatment for FECD remains challenging. Low initial cell concentration as well as endothelial to mesenchymal transition are the main obstacles to the application of FECD cultures in the clinical setting.
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Affiliation(s)
| | - Mattia Lamon
- Fondazione Banca degli Occhi del Veneto, Venice, Italy
| | - Elena Franco
- University of Ferrara, Department of Translational Medicine, Ferrara, Italy; Department of Ophthalmology, Ospedali Privati Forlì "Villa Igea", Forlì, Italy; Istituto Internazionale per la Ricerca e Formazione in Oftalmologia (IRFO), Forlì, Italy
| | | | | | - Cristina Bovone
- University of Ferrara, Department of Translational Medicine, Ferrara, Italy; Department of Ophthalmology, Ospedali Privati Forlì "Villa Igea", Forlì, Italy; Istituto Internazionale per la Ricerca e Formazione in Oftalmologia (IRFO), Forlì, Italy
| | - Angeli Christy Yu
- University of Ferrara, Department of Translational Medicine, Ferrara, Italy; Department of Ophthalmology, Ospedali Privati Forlì "Villa Igea", Forlì, Italy; Istituto Internazionale per la Ricerca e Formazione in Oftalmologia (IRFO), Forlì, Italy
| | | | - Diego Ponzin
- Fondazione Banca degli Occhi del Veneto, Venice, Italy
| | - Massimo Busin
- University of Ferrara, Department of Translational Medicine, Ferrara, Italy; Department of Ophthalmology, Ospedali Privati Forlì "Villa Igea", Forlì, Italy; Istituto Internazionale per la Ricerca e Formazione in Oftalmologia (IRFO), Forlì, Italy
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Hatou S, Sayano T, Higa K, Inagaki E, Okano Y, Sato Y, Okano H, Tsubota K, Shimmura S. Transplantation of iPSC-derived corneal endothelial substitutes in a monkey corneal edema model. Stem Cell Res 2021; 55:102497. [PMID: 34411973 DOI: 10.1016/j.scr.2021.102497] [Citation(s) in RCA: 22] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/07/2021] [Revised: 07/02/2021] [Accepted: 08/05/2021] [Indexed: 12/13/2022] Open
Abstract
OBJECTIVE In order to provide regenerative therapy for millions of patients suffering from corneal blindness globally, we derived corneal endothelial cell substitute (CECSi) cells from induced pluripotent stem cells (iPSCs) to treat corneal edema due to endothelial dysfunction (bullous keratopathy). METHODS AND RESULTS We developed an efficient xeno-free protocol to produce CECSi cells from both research grade (Ff-MH09s01 and Ff-I01s04) and clinical grade (QHJI01s04) iPSCs. CECSi cells formed a hexagonal confluent monolayer with Na, K-ATPase alpha 1 subunit expression (ATP1A1), tight junctions, N-cadherin adherence junction formation, and nuclear PITX2 expression, which are all characteristics of corneal endothelial cells. CECSi cells can be cryopreserved, and thawed CECSi cell suspensions also expressed N-cadherin and ATP1A1. Residual undifferentiated iPSCs in QHJI01s04-derived CECSi cells was below 0.01%. Frozen stocks of Ff-I01s04- and QHJI01s04-derived CECSi cells were transported, thawed and transplanted into a monkey corneal edema model. CECSi-transplanted eyes significantly reduced corneal edema compared to control group. CONCLUSION Our results show a promising approach to provide bullous keratopathy patients with an iPS-cell-based cell therapy to recover useful vision.
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Affiliation(s)
- Shin Hatou
- Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan; Cellusion Inc, Tokyo, Japan
| | - Tomoko Sayano
- Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan; Cellusion Inc, Tokyo, Japan
| | - Kazunari Higa
- Department of Ophthalmology, Tokyo Dental College Ichikawa General Hospital, Ichikawa, Japan
| | - Emi Inagaki
- Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan
| | - Yuji Okano
- Department of Physiology, Keio University School of Medicine, Tokyo, Japan
| | - Yasunori Sato
- Department of Preventive Medicine and Public Health, Keio University School of Medicine, Tokyo, Japan
| | - Hideyuki Okano
- Department of Physiology, Keio University School of Medicine, Tokyo, Japan
| | - Kazuo Tsubota
- Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan
| | - Shigeto Shimmura
- Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.
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Incorporating Differential Gene Expression Analysis with Predictive Biomarkers to Identify Novel Therapeutic Drugs for Fuchs Endothelial Corneal Dystrophy. J Ophthalmol 2021; 2021:5580595. [PMID: 34258047 PMCID: PMC8260298 DOI: 10.1155/2021/5580595] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/28/2021] [Revised: 05/15/2021] [Accepted: 06/17/2021] [Indexed: 11/24/2022] Open
Abstract
Purpose Based on the differential gene expression analysis for predictive biomarkers with RNA-Sequencing data from Fuchs endothelial corneal dystrophy (FECD) patients, we are aiming to evaluate the efficacy of Library of Integrated Network-based Cellular Signatures (LINCS) perturbagen prediction software to identify novel pharmacotherapeutic targets that can revert the pathogenic gene expression signatures and reverse disease phenotype in FECD. Methods A publicly available RNA-seq dataset was used to compare corneal endothelial specimens from controls and patients with FECD. Based on the differential gene expression analysis for predictive biomarkers, we evaluated the efficacy of LINCS perturbagen prediction software to identify novel therapeutic targets that can revert the pathogenic gene expression signatures and reverse disease phenotypes in FECD. Results The RNA-seq dataset of the corneal endothelial cells from FECD patients revealed the differential gene expression signatures of FECD. Many of the differential expressed genes are related to canonical pathways of the FECD pathogenesis, such as extracellular matrix reorganization and immunological response. The expression levels of genes VSIG2, IL18, and ITGB8 were significantly increased in FECD compared with control. Meanwhile, the expression levels of CNGA3, SMOX, and CERS1 were significantly lower in the FECD than in control. We employed LINCS L1000 Characteristic Direction Signature Search Engine (L1000-CDS2) to investigate pathway-based molecular treatment. L1000-CDS2 predicted that small molecule drugs such as histone deacetylase (HDAC) inhibitors might be a potential candidate to reverse the pathological gene expression signature in FECD. Conclusions Based on differential gene expression signatures, several candidate drugs have been identified to reverse the disease phenotypes in FECD. Gene expression signature with LINCS small molecule prediction software can discover novel preclinical drug candidates for FECD.
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Pan SH, Zhao N, Feng X, Jie Y, Jin ZB. Conversion of mouse embryonic fibroblasts into neural crest cells and functional corneal endothelia by defined small molecules. SCIENCE ADVANCES 2021; 7:7/23/eabg5749. [PMID: 34088673 PMCID: PMC8177713 DOI: 10.1126/sciadv.abg5749] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 01/18/2021] [Accepted: 04/20/2021] [Indexed: 05/06/2023]
Abstract
Reprogramming of somatic cells into desired functional cell types by small molecules has vast potential for developing cell replacement therapy. Here, we developed a stepwise strategy to generate chemically induced neural crest cells (ciNCCs) and chemically induced corneal endothelial cells (ciCECs) from mouse fibroblasts using defined small molecules. The ciNCCs exhibited typical NCC features and could differentiate into ciCECs using another chemical combination in vitro. The resulting ciCECs showed consistent gene expression profiles and self-renewal capacity to those of primary CECs. Notably, these ciCECs could be cultured for as long as 30 passages and still retain the CEC features in defined medium. Transplantation of these ciCECs into an animal model reversed corneal opacity. Our chemical approach for direct reprogramming of mouse fibroblasts into ciNCCs and ciCECs provides an alternative cell source for regeneration of corneal endothelia and other tissues derived from neural crest.
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Affiliation(s)
- Shao-Hui Pan
- Institute of Stem Cell Research, The Eye Hospital, Wenzhou Medical University, Wenzhou 325027, China
| | - Ning Zhao
- Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology and Visual Science Key Laboratory, Beijing 100730, China
- Beijing Advanced Innovation Center for Big Data-Based Precision Medicine, Beihang University and Capital Medical University, Beijing Tongren Hospital, Beijing 100730, China
| | - Xiang Feng
- Institute of Stem Cell Research, The Eye Hospital, Wenzhou Medical University, Wenzhou 325027, China
| | - Ying Jie
- Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology and Visual Science Key Laboratory, Beijing 100730, China
| | - Zi-Bing Jin
- Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology and Visual Science Key Laboratory, Beijing 100730, China.
- Beijing Advanced Innovation Center for Big Data-Based Precision Medicine, Beihang University and Capital Medical University, Beijing Tongren Hospital, Beijing 100730, China
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Florou C, Aissopou E, Chalkiadaki E, Andreanos K, Koutsandrea C, Papaconstantinou D, Georgalas I. Corneal endothelial cells and central corneal thickness in patients with neurofibromatosis type 1. Indian J Ophthalmol 2021; 69:1522-1526. [PMID: 34011734 PMCID: PMC8302278 DOI: 10.4103/ijo.ijo_1967_20] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022] Open
Abstract
Purpose: The aim of this study was to evaluate the morphological properties of corneal endothelial cells and central corneal thickness (CCT) in patients with neurofibromatosis type 1 (NF1) and to compare them with age-matched healthy controls. Methods: Nineteen NF1 patients and 38 healthy individuals were recruited. All participants underwent complete ophthalmological examination as well as noncontact specular microscopy to measure endothelial cell density (ECD), average cell area (AVG), coefficient of variation of cell area (CV), the percentage of hexagonal cells, and CCT. Eyes with previous ocular trauma, inflammation or surgery, and preexisting corneal and ocular surface diseases were excluded. Results: NF1 patients had higher ECD compared to healthy controls of the same age (2764.2 ± 270.4 versus 2570.4 ± 449.2 cells/mm, respectively), although at a borderline level (P = 0.051). Patients with NF1 presented significantly lower CV and AVG when compared to controls (32.9 ± 4.6 versus 37.8 ± 9.5%, P = 0.011 and 364.9 ± 34.4 versus 406.0 ± 107.4 µm, P = 0.038, respectively). The NF1 group had significantly higher hexagonality in comparison with controls (55.7 ± 6.5 versus 50.5 ± 9.9%, P = 0.025). CCT was similar between the two groups (P = 0.955). Conclusion: Our results show that corneal endothelium has more favorable morphological characteristics in NF1 patients compared to healthy individuals of the same age.
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Affiliation(s)
- Chrysoula Florou
- First Department of Ophthalmology, National and Kapodistrian University of Athens, General Hospital "G. Gennimatas", Athens, Greece
| | - Evaggelia Aissopou
- Ophthalmologist in Private Office, Papadiamantopoulou 186, Athens, Greece
| | - Evangelia Chalkiadaki
- First Department of Ophthalmology, National and Kapodistrian University of Athens, General Hospital "G. Gennimatas", Athens, Greece
| | | | - Chrysanthi Koutsandrea
- First Department of Ophthalmology, National and Kapodistrian University of Athens, General Hospital "G. Gennimatas", Athens, Greece
| | - Dimitrios Papaconstantinou
- First Department of Ophthalmology, National and Kapodistrian University of Athens, General Hospital "G. Gennimatas", Athens, Greece
| | - Ilias Georgalas
- First Department of Ophthalmology, National and Kapodistrian University of Athens, General Hospital "G. Gennimatas", Athens, Greece
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Cetin EN, Bozkurt K, Akbulut S, Pekel G, Taşcı M, Çobankara V. Corneal and anterior chamber morphology in patients with nonınfectious ıntraocular ınflammation. Arq Bras Oftalmol 2021; 84:220-224. [PMID: 33567019 PMCID: PMC11826773 DOI: 10.5935/0004-2749.20210030] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/22/2019] [Accepted: 03/27/2020] [Indexed: 11/20/2022] Open
Abstract
PURPOSE To evaluate the corneal and anterior chamber morphology in phakic eyes with noninfectious intraocular inflammation. METHODS This study included 59 eyes with active uveitis, 62 with inactive uveitis, and 95 healthy eyes. Corneal endothelial cell density, hexagonal cell ratio, coefficient of variation (CV), corneal thickness and volume, maximum keratometry, and anterior chamber volume and depth (ACD) measurements were performed using a specular microscope and Pentacam HR. RESULTS The mean duration of uveitis was 24.6 ± 40.5 (0-180) months. The mean number of uveitis attacks was 2.8 ± 3.0 (1-20). Coefficient of variation was significantly higher in the active uveitis group compared with inactive uveitis group (p=0.017, Post Hoc Tukey). Anterior segment parameters other than coefficient of variation were not significantly different between active/inactive uveitis and control groups (p>0.05). Multiple linear regression analysis showed that coefficient of variation was greater in active uveitis compared with inactive uveitis after adjusting for the duration of uveitis, type of uveitis, having a rheumatologic disease, and having immunosuppressive treatment (p=0.003). The duration of uveitis and number of attacks were not significantly correlated with ocular parameters (p>0.05, Spearman's correlation). The difference in parameters was not significant based on uveitis type (p>0.05). CONCLUSIONS Coefficient of variation was higher in eyes with active uveitis than that in eyes with inactive uveitis, whereas corneal endothelial cell density and anterior chamber morphology did not significantly differ between active/inactive uveitis and control groups.
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Affiliation(s)
- Ebru Nevin Cetin
- Department of Ophthalmology, Pamukkale University, Denizli, Turkey
| | - Kerem Bozkurt
- Department of Ophthalmology, Denizli State Hospital, Denizli,
Turkey
| | - Selen Akbulut
- Department of Ophthalmology, Pamukkale University, Denizli, Turkey
| | - Gökhan Pekel
- Department of Ophthalmology, Pamukkale University, Denizli, Turkey
| | - Murat Taşcı
- Department of Rheumatology, Pamukkale University, Denizli, Turkey
| | - Veli Çobankara
- Department of Rheumatology, Pamukkale University, Denizli, Turkey
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43
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Öztürk-Öncel MÖ, Erkoc-Biradli FZ, Rasier R, Marcali M, Elbuken C, Garipcan B. Rose petal topography mimicked poly(dimethylsiloxane) substrates for enhanced corneal endothelial cell behavior. MATERIALS SCIENCE & ENGINEERING. C, MATERIALS FOR BIOLOGICAL APPLICATIONS 2021; 126:112147. [PMID: 34082958 DOI: 10.1016/j.msec.2021.112147] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/03/2020] [Revised: 04/05/2021] [Accepted: 04/26/2021] [Indexed: 12/29/2022]
Abstract
Low proliferation capacity of corneal endothelial cells (CECs) and worldwide limitations in transplantable donor tissues reveal the critical need of a robust approach for in vitro CEC growth. However, preservation of CEC-specific phenotype with increased proliferation has been a great challenge. Here we offer a biomimetic cell substrate design, by optimizing mechanical, topographical and biochemical characteristics of materials with CEC microenvironment. We showed the surprising similarity between topographical features of white rose petals and corneal endothelium due to hexagonal cell shapes and physiologically relevant cell density (≈ 2000 cells/mm2). Polydimethylsiloxane (PDMS) substrates with replica of white rose petal topography and cornea-friendly Young's modulus (211.85 ± 74.9 kPa) were functionalized with two of the important corneal extracellular matrix (ECM) components, collagen IV (COL 4) and hyaluronic acid (HA). White rose petal patterned and COL 4 modified PDMS with optimized stiffness provided enhanced bovine CEC response with higher density monolayers and increased phenotypic marker expression. This biomimetic approach demonstrates a successful platform to improve in vitro cell substrate properties of PDMS for corneal applications, suggesting an alternative environment for CEC-based therapies, drug toxicity investigations, microfluidics and organ-on-chip applications.
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Affiliation(s)
| | | | - Rıfat Rasier
- Department of Ophthalmology, Demiroglu Bilim University, Istanbul, Turkey
| | - Merve Marcali
- UNAM-National Nanotechnology Research Center, Institute of Materials Science and Nanotechnology, Bilkent University, Ankara, Turkey
| | - Caglar Elbuken
- UNAM-National Nanotechnology Research Center, Institute of Materials Science and Nanotechnology, Bilkent University, Ankara, Turkey; Faculty of Biochemistry and Molecular Medicine, Faculty of Medicine, University of Oulu, 90014 Oulu, Finland
| | - Bora Garipcan
- Institute of Biomedical Engineering, Boğaziçi University, Istanbul, Turkey.
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44
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Expression and Function of ZEB1 in the Cornea. Cells 2021; 10:cells10040925. [PMID: 33923743 PMCID: PMC8074155 DOI: 10.3390/cells10040925] [Citation(s) in RCA: 16] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/28/2021] [Revised: 04/12/2021] [Accepted: 04/14/2021] [Indexed: 12/13/2022] Open
Abstract
ZEB1 is an important transcription factor for epithelial to mesenchymal transition (EMT) and in the regulation of cell differentiation and transformation. In the cornea, ZEB1 presents in all three layers: the epithelium, the stroma and the endothelium. Mutations of ZEB1 have been linked to multiple corneal genetic defects, particularly to the corneal dystrophies including keratoconus (KD), Fuchs endothelial corneal dystrophy (FECD), and posterior polymorphous corneal dystrophy (PPCD). Accumulating evidence indicates that dysfunction of ZEB1 may affect corneal stem cell homeostasis, and cause corneal cell apoptosis, stromal fibrosis, angiogenesis, squamous metaplasia. Understanding how ZEB1 regulates the initiation and progression of these disorders will help us in targeting ZEB1 for potential avenues to generate therapeutics to treat various ZEB1-related disorders.
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45
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Kara Ö, Dereli Can G. Topographic and specular microscopic evaluation of cornea and meibomian gland morphology in children with isolated growth hormone deficiency. Int Ophthalmol 2021; 41:2827-2835. [PMID: 33818674 DOI: 10.1007/s10792-021-01839-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/05/2020] [Accepted: 03/29/2021] [Indexed: 10/21/2022]
Abstract
PURPOSE To evaluate whether the anterior segment topographic measurements, meibomian gland (MG), and non-invasive tear film break-up time (NITFBUT) differ between healthy children and children with isolated growth hormone deficiency (GHD). METHODS A total of 74 eyes of 37 children with GHD and 84 eyes of 42 age- and sex-matched healthy children were included in the study. The spherical equivalence (SE), mean keratometry (Km), corneal thickness, corneal volume (CV), anterior chamber depth (ACD), topographic NITFBUT, qualitative and quantitative MG measurements, corneal endothelial cell density (CD), and proportion of hexagonal cells (HG) were analysed. RESULTS The mean SE level of GHD group was similar between groups (p = 0.017). Back Km values were insignificantly steep in children with GHD (p = 0.004, with Bonferroni correction). Specular microscopy analysis was not different between groups. MG loss of GHD group were higher than control group (p < 0.001). The MG morphology analysis and distortion grade were not different between groups (p > 0.05). CONCLUSIONS Our results showed that the growth hormone (GH) may have an important role on the anterior segment parameters; however, it is not clear that this misregulation leads to a clinical scenario in childhood. Future studies investigating GHD and/or GH therapy on the ocular surface system are required to clearly demonstrate basic mechanism of GH action.
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Affiliation(s)
- Özlem Kara
- Department of Pediatric Endocrinology, Bursa Yuksek Ihtisas Training and Research Hospital, Bursa, Turkey.
| | - Gamze Dereli Can
- Department of Ophthalmology, Bursa City Training and Research Hospital, Bursa, Turkey
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46
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Ong HS, Ang M, Mehta J. Evolution of therapies for the corneal endothelium: past, present and future approaches. Br J Ophthalmol 2021; 105:454-467. [PMID: 32709756 PMCID: PMC8005807 DOI: 10.1136/bjophthalmol-2020-316149] [Citation(s) in RCA: 50] [Impact Index Per Article: 12.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/23/2020] [Revised: 05/16/2020] [Indexed: 12/13/2022]
Abstract
Corneal endothelial diseases are leading indications for corneal transplantations. With significant advancement in medical science and surgical techniques, corneal transplant surgeries are now increasingly effective at restoring vision in patients with corneal diseases. In the last 15 years, the introduction of endothelial keratoplasty (EK) procedures, where diseased corneal endothelium (CE) are selectively replaced, has significantly transformed the field of corneal transplantation. Compared to traditional penetrating keratoplasty, EK procedures, namely Descemet's stripping automated endothelial keratoplasty (DSAEK) and Descemet membrane endothelial keratoplasty (DMEK), offer faster visual recovery, lower immunological rejection rates, and improved graft survival. Although these modern techniques can achieve high success, there are fundamental impediments to conventional transplantations. A lack of suitable donor corneas worldwide restricts the number of transplants that can be performed. Other barriers include the need for specialized expertise, high cost, and risks of graft rejection or failure. Research is underway to develop alternative treatments for corneal endothelial diseases, which are less dependent on the availability of allogeneic tissues - regenerative medicine and cell-based therapies. In this review, an overview of past and present transplantation procedures used to treat corneal endothelial diseases are described. Potential novel therapies that may be translated into clinical practice will also be presented.
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Affiliation(s)
- Hon Shing Ong
- Corneal and External Diseases Department, Singapore National Eye Centre, Singapore, Singapore
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore, Singapore
- Department of Ophthalmology and Visual Science, Duke-National University of Singapore (NUS) Graduate Medical School, Singapore, Singapore
| | - Marcus Ang
- Corneal and External Diseases Department, Singapore National Eye Centre, Singapore, Singapore
- Department of Ophthalmology and Visual Science, Duke-National University of Singapore (NUS) Graduate Medical School, Singapore, Singapore
| | - Jodhbir Mehta
- Corneal and External Diseases Department, Singapore National Eye Centre, Singapore, Singapore
- Tissue Engineering and Cell Therapy Group, Singapore Eye Research Institute, Singapore, Singapore
- Department of Ophthalmology and Visual Science, Duke-National University of Singapore (NUS) Graduate Medical School, Singapore, Singapore
- School of Material Science & Engineering and School of Mechanical and Aerospace Engineering, Nanyang Technological University, Singapore, Singapore
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47
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Wang X, Zhou Q, Zhao C, Duan H, Li W, Dong C, Gong Y, Li Z, Shi W. Multiple roles of FGF10 in the regulation of corneal endothelial wound healing. Exp Eye Res 2021; 205:108517. [PMID: 33617851 DOI: 10.1016/j.exer.2021.108517] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/13/2020] [Revised: 02/11/2021] [Accepted: 02/15/2021] [Indexed: 12/29/2022]
Abstract
Corneal endothelial dysfunction usually induces corneal haze and oedema, which seriously affect visual function. The main therapeutic strategy for this condition is corneal transplantation, but the use of this strategy is limited by the shortage of healthy donor corneas. Compared with corneal transplantation, drug intervention is less invasive and more accessible; thus, finding an effective pharmaceutical alternative for cornea transplantation is critical for the treatment of corneal endothelial dysfunction. In this study, we established a rabbit scratch model to investigate the effect of fibroblast growth factor 10 (FGF10) on corneal endothelial wound healing. Results showed that FGF10 injection accelerated the recovery of corneal transparency and increased the protein expression levels of ZO1, Na+/K+-ATPase and AQP-1. Moreover, FGF10 significantly inhibited the expression levels of endothelial-to-mesenchymal transition proteins and reduced the expression levels of the proinflammatory factors IL-1β and TNF-α in the anterior chamber aqueous humour. FGF10 also enhanced the Na+/K+-ATPase activity by enhancing mitochondrial function as a result of its direct interaction with its conjugate receptor. Thus, FGF10 could be a new pharmaceutical preparation as treatment for corneal endothelial dysfunction.
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Affiliation(s)
- Xin Wang
- Department of Medicine, Qingdao University, Qingdao, China; State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China
| | - Qingjun Zhou
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China
| | - Can Zhao
- Shandong Eye Hospital, Shandong Eye Institute, Shandong First Medical University &Shandong Academy of Medical Sciences, China
| | - Haoyun Duan
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China
| | - Wenjing Li
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China
| | - Chunxiao Dong
- Department of Medicine, Qingdao University, Qingdao, China; State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China
| | - Yajie Gong
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China
| | - Zongyi Li
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China.
| | - Weiyun Shi
- State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao, China; Shandong Eye Hospital, Shandong Eye Institute, Shandong First Medical University &Shandong Academy of Medical Sciences, China.
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48
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Parekh M, Romano V, Hassanin K, Testa V, Wongvisavavit R, Ferrari S, Haneef A, Willoughby C, Ponzin D, Jhanji V, Sharma N, Daniels J, Kaye SB, Ahmad S, Levis HJ. Biomaterials for corneal endothelial cell culture and tissue engineering. J Tissue Eng 2021; 12:2041731421990536. [PMID: 33643603 PMCID: PMC7894589 DOI: 10.1177/2041731421990536] [Citation(s) in RCA: 27] [Impact Index Per Article: 6.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/04/2020] [Accepted: 01/08/2021] [Indexed: 12/20/2022] Open
Abstract
The corneal endothelium is the posterior monolayer of cells that are responsible for maintaining overall transparency of the avascular corneal tissue via pump function. These cells are non-regenerative in vivo and therefore, approximately 40% of corneal transplants undertaken worldwide are a result of damage or dysfunction of endothelial cells. The number of available corneal donor tissues is limited worldwide, hence, cultivation of human corneal endothelial cells (hCECs) in vitro has been attempted in order to produce tissue engineered corneal endothelial grafts. Researchers have attempted to recreate the current gold standard treatment of replacing the endothelial layer with accompanying Descemet's membrane or a small portion of stroma as support with tissue engineering strategies using various substrates of both biologically derived and synthetic origin. Here we review the potential biomaterials that are currently in development to support the transplantation of a cultured monolayer of hCECs.
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Affiliation(s)
- Mohit Parekh
- Faculty of Brain Sciences, Institute of Ophthalmology, University College London, London, UK.,International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, Italy
| | - Vito Romano
- St. Paul's Eye Unit, Royal Liverpool Broadgreen University Hospital, Liverpool, UK.,Instituto Universitario Fernandez-Vega, Universidad de Oviedo and Fundacion de Investigacion on Oftalmologica, Oviedo, Spain.,Department of Eye and Vision Science, Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, UK
| | - Kareem Hassanin
- St. Paul's Eye Unit, Royal Liverpool Broadgreen University Hospital, Liverpool, UK
| | - Valeria Testa
- Eye Clinic, Department of Neuroscience, Rehabilitation, Ophthalmology, Genetics, Maternal and Child Health, University of Genoa, Genoa, Italy.,Ospedale Policlinico San Martino IRCCS, Genoa, Italy
| | - Rintra Wongvisavavit
- Faculty of Brain Sciences, Institute of Ophthalmology, University College London, London, UK.,HRH Princess Chulabhorn College of Medical Sciences, Chulabhorn Royal Academy, Bangkok, Thailand
| | - Stefano Ferrari
- International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, Italy
| | - Atikah Haneef
- Department of Eye and Vision Science, Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, UK
| | - Colin Willoughby
- School of biomedical sciences, University of Ulster, Belfast, UK
| | - Diego Ponzin
- International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, Italy
| | - Vishal Jhanji
- Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA, USA
| | - Namrata Sharma
- Dr. Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India
| | - Julie Daniels
- Faculty of Brain Sciences, Institute of Ophthalmology, University College London, London, UK
| | - Stephen B Kaye
- St. Paul's Eye Unit, Royal Liverpool Broadgreen University Hospital, Liverpool, UK
| | - Sajjad Ahmad
- Faculty of Brain Sciences, Institute of Ophthalmology, University College London, London, UK.,Moorfields Eye Hospital NHS Trust Foundation, London, UK
| | - Hannah J Levis
- Department of Eye and Vision Science, Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, UK
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49
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Erkoc-Biradli FZ, Ozgun A, Öztürk-Öncel MÖ, Marcali M, Elbuken C, Bulut O, Rasier R, Garipcan B. Bioinspired hydrogel surfaces to augment corneal endothelial cell monolayer formation. J Tissue Eng Regen Med 2021; 15:244-255. [PMID: 33448665 DOI: 10.1002/term.3173] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/05/2020] [Revised: 12/08/2020] [Accepted: 12/26/2020] [Indexed: 11/06/2022]
Abstract
Corneal endothelial cells (CECs) have limited proliferation ability leading to corneal endothelium (CE) dysfunction and eventually vision loss when cell number decreases below a critical level. Although transplantation is the main treatment method, donor shortage problem is a major bottleneck. The transplantation of in vitro developed endothelial cells with desirable density is a promising idea. Designing cell substrates that mimic the native CE microenvironment is a substantial step to achieve this goal. In the presented study, we prepared polyacrylamide (PA) cell substrates that have a microfabricated topography inspired by the dimensions of CECs. Hydrogel surfaces were prepared via two different designs with small and large patterns. Small patterned hydrogels have physiologically relevant hexagon densities (∼2000 hexagons/mm2 ), whereas large patterned hydrogels have sparsely populated hexagons (∼400 hexagons/mm2 ). These substrates have similar elastic modulus of native Descemet's membrane (DM; ∼50 kPa) and were modified with Collagen IV (Col IV) to have biochemical content similar to native DM. The behavior of bovine corneal endothelial cells on these substrates was investigated and results show that cell proliferation on small patterned substrates was significantly (p = 0.0004) higher than the large patterned substrates. Small patterned substrates enabled a more densely populated cell monolayer compared to other groups (p = 0.001 vs. flat and p < 0.0001 vs. large patterned substrates). These results suggest that generating bioinspired surface topographies augments the formation of CE monolayers with the desired cell density, addressing the in vitro development of CE layers.
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Affiliation(s)
- Fatma Zehra Erkoc-Biradli
- (Bio)3 Research laboratory, Institute of Biomedical Engineering, Bogazici University, Istanbul, Turkey
| | - Alp Ozgun
- (Bio)3 Research laboratory, Institute of Biomedical Engineering, Bogazici University, Istanbul, Turkey
| | | | - Merve Marcali
- Institute of Materials Science and Nanotechnology, National Nanotechnology Research Center (UNAM), Bilkent University, Ankara, Turkey
| | - Caglar Elbuken
- Institute of Materials Science and Nanotechnology, National Nanotechnology Research Center (UNAM), Bilkent University, Ankara, Turkey.,Faculty of Biochemistry and Molecular Medicine, Faculty of Medicine, University of Oulu, Oulu, Finland
| | - Osman Bulut
- Faculty of Civil Engineering, Istanbul Technical University, Istanbul, Turkey
| | - Rıfat Rasier
- Department of Ophthalmology, Demiroglu Bilim University, Istanbul, Turkey
| | - Bora Garipcan
- (Bio)3 Research laboratory, Institute of Biomedical Engineering, Bogazici University, Istanbul, Turkey
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50
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Li PC, Chen SC, Hsueh YJ, Shen YC, Tsai MY, Hsu LW, Yeh CK, Chen HC, Huang CC. Gelatin scaffold with multifunctional curcumin-loaded lipid-PLGA hybrid microparticles for regenerating corneal endothelium. MATERIALS SCIENCE & ENGINEERING. C, MATERIALS FOR BIOLOGICAL APPLICATIONS 2021; 120:111753. [DOI: 10.1016/j.msec.2020.111753] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/15/2020] [Revised: 11/08/2020] [Accepted: 11/21/2020] [Indexed: 01/21/2023]
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