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Carstens KE, Gronskaya E, Jäckel D, Bertoli J, Cuevas KR, Dorier J, Wang S, Lopez-Rodriguez D, Shafer TJ, Zurich MG, Pamies D. Application of a high-density microelectrode array assay using a 3D human iPSC-derived brain microphysiological system model for in vitro neurotoxicity screening of environmental compounds. Arch Toxicol 2025:10.1007/s00204-025-04043-x. [PMID: 40293475 DOI: 10.1007/s00204-025-04043-x] [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: 11/19/2024] [Accepted: 03/27/2025] [Indexed: 04/30/2025]
Abstract
Unraveling the associations between human exposure to environmental chemicals and potential neurotoxicity presents significant challenges. Evaluation of neurotoxicity potential using animal testing is resource-intensive (financial, labor, and animal use) and faces uncertainties regarding biological relevance to human health outcomes. Therefore, there is a need to develop efficient and human-relevant in vitro new approach methodologies (NAMs) to screen and evaluate chemicals for neurotoxicity potential. Recording of neural network activity using microelectrode array (MEA) technology has been identified as a reliable and reproducible method for evaluating neurotoxicity. Much of this research has been performed in 2D rodent-derived cell models. The 'BrainSpheres MEA assay' described in this study offers a promising functional human induced pluripotent stem cell (iPSC)-derived 3D brain model comprising neurons, astrocytes, and oligodendrocytes. We demonstrate consistent spontaneous neuronal firing and network bursting parameters from 7-week-old BrainSpheres using a high-density MEA technology. The performance of this model as a human-relevant NAM was evaluated by conducting a multi-concentration, 13 day exposure study with a set of ten chemicals. Neural activity metrics were assessed and compared to results from a 2D-MEA assay using rodent cells. Loperamide and domoic acid (two assay positive controls) demonstrated similar bioactivity profiles in the BrainSphere MEA assay to the 2D-MEA assay, while acetaminophen (assay negative control) was inactive in both assays. The 2D-MEA model demonstrated more potent bioactivity for 4/7 chemicals that were active in both assays. In the future, reducing replicate variability and testing a larger set of chemicals will likely improve the accuracy and reliability of the assay. These preliminary findings suggest that the BrainSphere assay could be used alongside the rat network formation assay (rNFA) as part of a tiered strategy, where hits in the rNFA are confirmed and further characterized in the BrainSphere model, helping move toward animal-free toxicological testing.
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Affiliation(s)
- Kelly E Carstens
- Center for Computational Toxicology and Exposure, USA Environmental Protection Agency, Research Triangle Park, North Carolina, NC, 27707, USA
| | | | | | - Jessica Bertoli
- Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland
| | | | - Julien Dorier
- Bioinformatics Competence Center, University of Lausanne, CH- 1015, Lausanne, Switzerland
- Bioinformatics Competence Center, Ecole Polytechnique Fédérale de Lausanne, CH- 1015, Lausanne, Switzerland
| | - Shan Wang
- Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland
| | - David Lopez-Rodriguez
- Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland
- Swiss Centre for Applied Human Toxicology (SCAHT), Basel, Switzerland
- Institute of Earth Surface Dynamics, University of Lausanne, Canton de Vaud, Lausanne, Switzerland
| | - Timothy J Shafer
- Center for Computational Toxicology and Exposure, USA Environmental Protection Agency, Research Triangle Park, North Carolina, NC, 27707, USA
| | - Marie-Gabrielle Zurich
- Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland
- Swiss Centre for Applied Human Toxicology (SCAHT), Basel, Switzerland
| | - David Pamies
- Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland.
- Swiss Centre for Applied Human Toxicology (SCAHT), Basel, Switzerland.
- Stem Cells and Organoid Facility, University of Lausanne, Rue du Bugnon 9, CH-1005, Lausanne, Switzerland.
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Akter MZ, Tufail F, Ahmad A, Oh YW, Kim JM, Kim S, Hasan MM, Li L, Lee DW, Kim YS, Lee SJ, Kim HS, Ahn Y, Choi YJ, Yi HG. Harnessing native blueprints for designing bioinks to bioprint functional cardiac tissue. iScience 2025; 28:111882. [PMID: 40177403 PMCID: PMC11964760 DOI: 10.1016/j.isci.2025.111882] [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: 04/05/2025] Open
Abstract
Cardiac tissue lacks regenerative capacity, making heart transplantation the primary treatment for end-stage heart failure. Engineered cardiac tissues developed through three-dimensional bioprinting (3DBP) offer a promising alternative. However, reproducing the native structure, cellular diversity, and functionality of cardiac tissue requires advanced cardiac bioinks. Major obstacles in CTE (cardiac tissue engineering) include accurately characterizing bioink properties, replicating the cardiac microenvironment, and achieving precise spatial organization. Optimizing bioink properties to closely mimic the extracellular matrix (ECM) is essential, as deviations may result in pathological effects. This review encompasses the rheological and electromechanical properties of bioinks and the function of the cardiac microenvironment in the design of functional cardiac constructs. Furthermore, it focuses on improving the rheological characteristics, printability, and functionality of bioinks, offering valuable perspectives for developing new bioinks especially designed for CTE.
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Affiliation(s)
- Mst Zobaida Akter
- Department of Convergence Biosystems Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
- Interdisciplinary Program in IT-Bio Convergence System, Chonnam National University, Gwangju 61186, Republic of Korea
- Advanced Medical Device Research Center for Cardiovascular Disease, Chonnam National University, Gwangju 61186, Republic of Korea
| | - Fatima Tufail
- Department of Convergence Biosystems Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
- Interdisciplinary Program in IT-Bio Convergence System, Chonnam National University, Gwangju 61186, Republic of Korea
- Advanced Medical Device Research Center for Cardiovascular Disease, Chonnam National University, Gwangju 61186, Republic of Korea
| | - Ashfaq Ahmad
- Department of Convergence Biosystems Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
- Interdisciplinary Program in IT-Bio Convergence System, Chonnam National University, Gwangju 61186, Republic of Korea
- Advanced Medical Device Research Center for Cardiovascular Disease, Chonnam National University, Gwangju 61186, Republic of Korea
| | - Yoon Wha Oh
- Department of Convergence Biosystems Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
| | - Jung Min Kim
- Department of Convergence Biosystems Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
| | - Seoyeon Kim
- Department of Convergence Biosystems Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
- Interdisciplinary Program in IT-Bio Convergence System, Chonnam National University, Gwangju 61186, Republic of Korea
| | - Md Mehedee Hasan
- Department of Convergence Biosystems Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
| | - Longlong Li
- Department of Mechanical Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
| | - Dong-Weon Lee
- Advanced Medical Device Research Center for Cardiovascular Disease, Chonnam National University, Gwangju 61186, Republic of Korea
- Department of Mechanical Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
- Center for Next-Generation Sensor Research and Development, Chonnam National University, Gwangju 61186, Republic of Korea
| | - Yong Sook Kim
- Biomedical Research Institute, Chonnam National University Hospital, Gwangju 61469, Republic of Korea
| | - Su-jin Lee
- Biomedical Research Institute, Chonnam National University Hospital, Gwangju 61469, Republic of Korea
- Department of Forensic Medicine, Chonnam National University Medical School, Gwangju 61469, Republic of Korea
| | - Hyung-Seok Kim
- Department of Forensic Medicine, Chonnam National University Medical School, Gwangju 61469, Republic of Korea
| | - Youngkeun Ahn
- Division of Cardiology, Department of Internal Medicine, Chonnam National University Hospital, Chonnam National University Medical School, Gwangju, 61469, Republic of Korea
| | - Yeong-Jin Choi
- Advanced Bio and Healthcare Materials Research Division, Korea Institute of Materials Science (KIMS), Changwon 51508, Republic of Korea
- Advanced Materials Engineering, Korea National University of Science and Technology (UST), Changwon, Republic of Korea
| | - Hee-Gyeong Yi
- Department of Convergence Biosystems Engineering, Chonnam National University, Gwangju 61186, Republic of Korea
- Interdisciplinary Program in IT-Bio Convergence System, Chonnam National University, Gwangju 61186, Republic of Korea
- Advanced Medical Device Research Center for Cardiovascular Disease, Chonnam National University, Gwangju 61186, Republic of Korea
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Zhong J, Li W, Li H, Zhang J, Hou Z, Wang X, Zhou E, Lu K, Zhuang W, Sang H. A self-forming bone membrane generated by periosteum-derived stem cell spheroids enhances the repair of bone defects. Acta Biomater 2025; 193:185-201. [PMID: 39742905 DOI: 10.1016/j.actbio.2024.12.058] [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: 09/19/2024] [Revised: 12/24/2024] [Accepted: 12/30/2024] [Indexed: 01/04/2025]
Abstract
The periosteum, a highly specialized thin tissue, is instrumental in contributing to as much as 70 % of early bone formation. Recognizing the periosteum's vital physiological roles, the fabrication of a biomimetic periosteum has risen as an auspicious strategy for addressing extensive bone defects. In the study, we obtained such biomimetic periosteum by utilizing periosteum-derived stem cells (PDSCs) spheroids. These spheroids are induced to spontaneously generate a bioactive membrane on a delicate 3D-printed polycaprolactone (PCL) substrate. This process yields a biomimetic periosteum rich in the resources needed for bone repair. The in vitro evaluations demonstrated that this membrane can act as a repository for growth factors and stem cells. The release kinetics confirmed a sustained delivery of BMP-2 and VEGF, which promoted enhanced osteogenesis and angiogenesis in vitro, respectively. The in vivo results further highlighted robust bone regeneration from critical cranial defects upon the application of this biomimetic periosteum. The biomimetic periosteum, easily harvested and potent in bioactivity, presents substantial clinical potential, particularly for the treatment of critical-sized bone defects. STATEMENT OF SIGNIFICANCE: PDSC theoretically demonstrates substantial potential in membrane construction, a value we've harnessed in this pioneering application. By employing cell spheroids, we've successfully integrated a substantial number of cells into the membrane framework. PDSC spheroids exhibit the remarkable ability to self-assemble into functional membranes, endowing them with robust biological capabilities that enhance their performance in biological systems. The in vitro evaluations demonstrated that this membrane can act as a repository for growth factors and stem cells. The in vivo bone repair facilitated by this membrane is notably effective, characterized by superior bone quality and accelerated formation rates. This process mirrors the natural intramembrane ossification, offering a promising approach to bone integration and regeneration.
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Affiliation(s)
- Jintao Zhong
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China; The Third School of Clinical Medicine, Southern Medical University, Guangzhou 510000, PR China.
| | - Wenhua Li
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China; The Third School of Clinical Medicine, Southern Medical University, Guangzhou 510000, PR China.
| | - Hetong Li
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China.
| | - Jin Zhang
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China; The Third School of Clinical Medicine, Southern Medical University, Guangzhou 510000, PR China.
| | - Zuoxu Hou
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China.
| | - Xiao Wang
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China.
| | - Enhui Zhou
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China.
| | - Ke Lu
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China.
| | - Weida Zhuang
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China.
| | - Hongxun Sang
- Department of Orthopedics, Shenzhen Hospital, Southern Medical University, Shenzhen 518000, PR China; The Third School of Clinical Medicine, Southern Medical University, Guangzhou 510000, PR China.
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Park CH, Park JH, Suh YJ. Perspective of 3D culture in medicine: transforming disease research and therapeutic applications. Front Bioeng Biotechnol 2024; 12:1491669. [PMID: 39749112 PMCID: PMC11693738 DOI: 10.3389/fbioe.2024.1491669] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/05/2024] [Accepted: 12/06/2024] [Indexed: 01/04/2025] Open
Abstract
3D cell culture is gaining momentum in medicine due to its ability to mimic real tissues (in vivo) and provide more accurate biological data compared to traditional methods. This review explores the current state of 3D cell culture in medicine and discusses future directions, including the need for standardization and simpler protocols to facilitate wider use in research. Purpose 3D cell culture develops life sciences by mimicking the natural cellular environment. Cells in 3D cultures grow in three dimensions and interact with a matrix, fostering realistic cell behavior and interactions. This enhanced model offers significant advantages for diverse research areas. Methods By mimicking the cellular organization and functionalities found in human tissues, 3D cultures provide superior platforms for studying complex diseases like cancer and neurodegenerative disorders. This enables researchers to gain deeper insights into disease progression and identify promising therapeutic targets with greater accuracy. 3D cultures also play a crucial role in drug discovery by allowing researchers to effectively assess potential drugs' safety and efficacy. Results 3D cell culture's impact goes beyond disease research. It holds promise for tissue engineering. By replicating the natural tissue environment and providing a scaffold for cell growth, 3D cultures pave the way for regenerating damaged tissues, offering hope for treating burns, organ failure, and musculoskeletal injuries. Additionally, 3D cultures contribute to personalized medicine. Researchers can use patient-derived cells to create personalized disease models and identify the most effective treatment for each individual. Conclusion With ongoing advancements in cell imaging techniques, the development of novel biocompatible scaffolds and bioreactor systems, and a deeper understanding of cellular behavior within 3D environments, 3D cell culture technology stands poised to revolutionize various aspects of healthcare and scientific discovery.
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Affiliation(s)
- Chan Hum Park
- Nano-Bio Regenerative Medical Institute, College of Medicine, Hallym University, Chuncheon, Republic of Korea
- Departments of Otorhinolaryngology-Head and Neck Surgery, Chuncheon Sacred Heart Hospital, School of Medicine, Hallym University, Chuncheon, Republic of Korea
| | - Jung Ho Park
- Department of Breast and Endocrine Surgery, Hallym University Sacred Heart Hospital, Anyang, Republic of Korea
| | - Yong Joon Suh
- Department of Breast and Endocrine Surgery, Hallym University Sacred Heart Hospital, Anyang, Republic of Korea
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Shen L, Zhang QL, Yao Y, Huang YL, Zheng Z, Li M, Xu H, Tan L, Liao X, Xia B, Li L, Redshaw C, Bai Y, Yang C. Alkyl chain length-regulated in situ intelligent nano-assemblies with AIE-active photosensitizers for photodynamic cancer therapy. Asian J Pharm Sci 2024; 19:100967. [PMID: 39640060 PMCID: PMC11617974 DOI: 10.1016/j.ajps.2024.100967] [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: 12/22/2023] [Revised: 03/08/2024] [Accepted: 07/03/2024] [Indexed: 12/07/2024] Open
Abstract
Photodynamic therapy (PDT) brings new hope for the treatment of breast cancer due to few side effects and highly effective cell killing; however, the low bioavailability of traditional photosensitizers (PSs) and their dependence on oxygen severely limits their application. Aggregation-induced emission (AIE) PSs can dramatically facilitate the photosensitization effect, which can have positive impacts on tumor PDT. To-date, most AIE PSs lack tumor targeting capability and possess poor cell delivery, resulting in their use in large quantities that are harmful to healthy tissues. In this study, a series of AIE PSs based on pyridinium-substituted triphenylamine salts ( TTPAs 1-6) with different alkyl chain lengths are synthesized. Results reveal that TTPAs 1-6 promote the generation of type I and II ROS, including ·OH and 1O2. In particular, the membrane permeability and targeting of TTPAs 4-6 bearing C8-C10 side-chains are higher than TTPAs 1-3 bearing shorter alkyl chains. Additionally, they can assemble with albumin, thereby forming nanoparticles (TTPA 4-6 NPs) in situ in blood, which significantly facilitates mitochondrial-targeting and strong ROS generation ability. Moreover, the TTPA 4-6 NPs are pH-responsive, allowing for increased accumulation or endocytosis of the tumor and enhancing the imaging or therapeutic effect. Therefore, the in vivo distributions of TTPA 4-6 NPs are visually enriched in tumor sites and exhibited excellent PDT efficacy. This work demonstrates a novel strategy for AIE PDT and has the potential to play an essential role in clinical applications using nano-delivery systems.
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Affiliation(s)
- Lingyi Shen
- School of Basic Medicine, Guizhou Medical University, Guiyang 550025, China
| | - Qi-Long Zhang
- School of Basic Medicine, Guizhou Medical University, Guiyang 550025, China
| | - Yongchao Yao
- Department of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China
| | - Ya-Li Huang
- School of Basic Medicine, Guizhou Medical University, Guiyang 550025, China
| | - Zhichang Zheng
- Department of Pharmacy, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
| | - Ming Li
- Department of Pharmacy, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
| | - Hong Xu
- School of Basic Medicine, Guizhou Medical University, Guiyang 550025, China
| | - Lin Tan
- Department of Pharmacy, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
| | - Xukun Liao
- Department of Pharmacy, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
| | - Binyi Xia
- Department of Pharmacy, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
| | - Lin Li
- Department of Pharmacy, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
| | - Carl Redshaw
- Chemistry, School of Natural Sciences, University of Hull, Yorkshire HU6 7RX, UK
| | - Yang Bai
- Department of Pharmacy, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
| | - Chengli Yang
- Department of Pharmacy, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
- Clinical Research Center, The Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China
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Hodaei H, Esmaeili Z, Erfani Y, Esnaashari SS, Geravand M, Adabi M. Preparation of biocompatible Zein/Gelatin/Chitosan/PVA based nanofibers loaded with vitamin E-TPGS via dual-opposite electrospinning method. Sci Rep 2024; 14:23796. [PMID: 39394234 PMCID: PMC11470087 DOI: 10.1038/s41598-024-74865-9] [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: 05/20/2024] [Accepted: 09/30/2024] [Indexed: 10/13/2024] Open
Abstract
Wound management is a critical aspect of healthcare, necessitating effective and innovative wound dressing materials. Many existing wound dressings lack effectiveness and exhibit limitations, including poor antimicrobial activity, toxicity, inadequate moisture regulation, and weak mechanical performance. The aim of this study is to develop a natural-based nanofibrous structure that possesses desirable characteristics for use as a wound dressing. The chemical analysis confirmed the successful creation of Zein (Ze) (25% w/v) /gelatin (Gel) (10% w/v) /chitosan (CS) (2% w/v) /Polyvinyl alcohol (PVA) (10% w/v) nanofibrous scaffolds loaded with vitamin E tocopheryl polyethylene glycol succinate (Vit E). The swelling percentages of nanofiber (NF), NF + Vit E, cross-linked nanofiber (CNF), and CNF + Vit E were 49%, 110%, 410%, and 676%, respectively; and the degradation rates of NF, NF + Vit E, CNF, and CNF + Vit E were 29.57 ± 5.06%, 33.78 ± 7.8%, 14.03 ± 7.52%, 43 ± 6.27%, respectively. The antibacterial properties demonstrated that CNF impregnated with antibiotics reduced Escherichia coli (E. coli) counts by approximately 27-28% and Staphylococcus aureus (S. aureus) counts by about 34-35% compared to negative control. In conclusion, cross-linked Ze/Gel/CS/PVA nanofibrous scaffolds loaded with Vit E have potential as suitable wound dressing materials because environmentally friendly materials contribute to sustainable wound care and controlled degradation ensures wound dressings breakdown harmlessly.
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Affiliation(s)
- Homa Hodaei
- Department of Medical Nanotechnology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
| | - Zahra Esmaeili
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
| | - Yousef Erfani
- Department of Medical Laboratory Sciences, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
| | - Seyedeh Sara Esnaashari
- Department of Medical Nanotechnology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
| | - Mahvash Geravand
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
| | - Mahdi Adabi
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
- Food Microbiology Research Center, Tehran University of Medical Sciences, Tehran, Iran.
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7
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Akhter F. Evaluation of the Biocompatibility of Periodontal Dressings: An In Vitro Study. JOURNAL OF PHARMACY AND BIOALLIED SCIENCES 2024; 16:S2500-S2502. [PMID: 39346369 PMCID: PMC11426820 DOI: 10.4103/jpbs.jpbs_335_24] [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: 04/01/2024] [Revised: 04/05/2024] [Accepted: 04/09/2024] [Indexed: 10/01/2024] Open
Abstract
Background Periodontal dressings play a crucial role in post-operative management of periodontal surgeries by protecting surgical sites and promoting wound healing. Biocompatibility evaluation is essential to ensure the safety and effectiveness of these materials. This study aims to assess the biocompatibility of periodontal dressings through an in vitro approach. Materials and Methods In this in vitro study, three commercially available periodontal dressings were evaluated for their biocompatibility. The Dressings were Dressing A: Coe-Pak™ Dressing B: Barricaid® and Dressing C: PeriAcryl®. Human gingival fibroblast cells were cultured and exposed to extracts of the dressings using the MTT assay to assess cell viability. Additionally, cell morphology and attachment were observed under a microscope. The experiment was conducted in triplicate for each dressing. Results The MTT assay revealed that Dressing A exhibited the highest cell viability with an absorbance value of 0.8 ± 0.1, followed by Dressing B with a value of 0.6 ± 0.2 and Dressing C with a value of 0.5 ± 0.1. Microscopic analysis showed normal cell morphology and attachment in cells exposed to Dressing A and Dressing B extracts, while cells exposed to Dressing C exhibited slight detachment and irregular morphology. Conclusion Our findings suggest that Dressing A and Dressing B demonstrate favorable biocompatibility profiles, as evidenced by higher cell viability and normal cell morphology.
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Affiliation(s)
- Fatema Akhter
- Department of Surgical and Diagnostic Sciences, College of Dentistry, Dar Al Uloom University, Riyadh, Saudi Arabia
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8
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Foreman RE, Lucey R, Leaney AR, Lee MY, Naseer H, Wilson A. Optimized LC-MS/MS methods for quantifying antibody-drug conjugate payloads in cell culture media containing phenol red. Bioanalysis 2024; 16:575-585. [PMID: 39185791 DOI: 10.1080/17576180.2024.2349422] [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: 02/02/2024] [Accepted: 04/26/2024] [Indexed: 08/27/2024] Open
Abstract
Aim: Phenol red is commonly used in cell culture media, but can be detrimental to bioanalysis of in vitro samples as it may impact instrument reliability. Many researchers do their final stage of culture in 'phenol red free' media, but in collaborative work this is not always feasible.Materials & methods: A comparison was made between typical extraction methods to reduce phenol red matrix interferences, including organic solvent precipitation and solid phase extraction.Results: The final method was demonstrated to be precise and accurate for the measurement of a target analyte by LC-MS/MS, and was applied to an in vitro ADC deconjugation study.Conclusion: This method allows for for continued bioanalytical support of in vitro models used in drug development.
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Affiliation(s)
- Rachel E Foreman
- Integrated Bioanalysis, Clinical Pharmacology & Safety Sciences, AstraZeneca, Cambridge Biomedical Campus, 1 Francis Crick Avenue, Cambridge, CB2 0AA
| | - Richard Lucey
- Integrated Bioanalysis, Clinical Pharmacology & Safety Sciences, AstraZeneca, Cambridge Biomedical Campus, 1 Francis Crick Avenue, Cambridge, CB2 0AA
| | - Adam R Leaney
- Integrated Bioanalysis, Clinical Pharmacology & Safety Sciences, AstraZeneca, Cambridge Biomedical Campus, 1 Francis Crick Avenue, Cambridge, CB2 0AA
| | - Mi-Young Lee
- Integrated Bioanalysis, Clinical Pharmacology & Safety Sciences, AstraZeneca, Cambridge Biomedical Campus, 1 Francis Crick Avenue, Cambridge, CB2 0AA
| | - Humaira Naseer
- Integrated Bioanalysis, Clinical Pharmacology & Safety Sciences, AstraZeneca, Cambridge Biomedical Campus, 1 Francis Crick Avenue, Cambridge, CB2 0AA
| | - Amanda Wilson
- Integrated Bioanalysis, Clinical Pharmacology & Safety Sciences, AstraZeneca, Cambridge Biomedical Campus, 1 Francis Crick Avenue, Cambridge, CB2 0AA
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9
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Lee D, Kim SM, Kim D, Baek SY, Yeo SJ, Lee JJ, Cha C, Park SA, Kim TD. Microfluidics-assisted fabrication of natural killer cell-laden microgel enhances the therapeutic efficacy for tumor immunotherapy. Mater Today Bio 2024; 26:101055. [PMID: 38693995 PMCID: PMC11061753 DOI: 10.1016/j.mtbio.2024.101055] [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: 02/05/2024] [Revised: 03/29/2024] [Accepted: 04/09/2024] [Indexed: 05/03/2024] Open
Abstract
Recently, interest in cancer immunotherapy has increased over traditional anti-cancer therapies such as chemotherapy or targeted therapy. Natural killer (NK) cells are part of the immune cell family and essential to tumor immunotherapy as they detect and kill cancer cells. However, the disadvantage of NK cells is that cell culture is difficult. In this study, porous microgels have been fabricated using microfluidic channels to effectively culture NK cells. Microgel fabrication using microfluidics can be mass-produced in a short time and can be made in a uniform size. Microgels consist of photo cross-linkable polymers such as methacrylic gelatin (GelMa) and can be regulated via controlled GelMa concentrations. NK92 cell-laden three-dimensional (3D) microgels increase mRNA expression levels, NK92 cell proliferation, cytokine release, and anti-tumor efficacy, compared with two-dimensional (2D) cultures. In addition, the study confirms that 3D-cultured NK92 cells enhance anti-tumor effects compared with enhancement by 2D-cultured NK92 cells in the K562 leukemia mouse model. Microgels containing healthy NK cells are designed to completely degrade after 5 days allowing NK cells to be released to achieve cell-to-cell interaction with cancer cells. Overall, this microgel system provides a new cell culture platform for the effective culturing of NK cells and a new strategy for developing immune cell therapy.
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Affiliation(s)
- Dongjin Lee
- Immunotherapy Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Republic of Korea
- Nano-Convergence Manufacturing Research Division, Korea Institute of Machinery and Materials (KIMM), Daejeon, 34103, Republic of Korea
| | - Seok Min Kim
- Immunotherapy Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Republic of Korea
| | - Dahong Kim
- Nano-Convergence Manufacturing Research Division, Korea Institute of Machinery and Materials (KIMM), Daejeon, 34103, Republic of Korea
- Department of Applied Bioengineering, Seoul National University, Seoul, 08826, Republic of Korea
| | - Seung Yeop Baek
- School of Materials Science and Engineering, Ulsan National Institute of Science and Technology, Ulsan 44919, Republic of Korea
| | - Seon Ju Yeo
- Nano-Convergence Manufacturing Research Division, Korea Institute of Machinery and Materials (KIMM), Daejeon, 34103, Republic of Korea
| | - Jae Jong Lee
- Nano-Convergence Manufacturing Research Division, Korea Institute of Machinery and Materials (KIMM), Daejeon, 34103, Republic of Korea
| | - Chaenyung Cha
- School of Materials Science and Engineering, Ulsan National Institute of Science and Technology, Ulsan 44919, Republic of Korea
- Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan, 44919, Republic of Korea
| | - Su A Park
- Nano-Convergence Manufacturing Research Division, Korea Institute of Machinery and Materials (KIMM), Daejeon, 34103, Republic of Korea
| | - Tae-Don Kim
- Immunotherapy Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Republic of Korea
- Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, 34113, Republic of Korea
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10
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Jamshidi N, Jamshidi N, Modarresi Chahardehi A, Shams E, Chaleshi V. A promising breakthrough in pancreatic cancer research: The potential of spheroids as 3D models. BIOIMPACTS : BI 2024; 15:30241. [PMID: 39963557 PMCID: PMC11830132 DOI: 10.34172/bi.30241] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 12/27/2023] [Revised: 01/29/2024] [Accepted: 02/06/2024] [Indexed: 02/20/2025]
Abstract
Pancreatic ductal adenocarcinoma (PDAC) stands as the fourth leading cause of cancer-related deaths, primarily attributable to its resistance to chemotherapy, resulting in a nearly universal fatality rate. Despite the promise exhibited by numerous drugs in preclinical studies, their subsequent failure in clinical trials underscores the inherent limitations of conventional two-dimensional cell culture models commonly employed in early drug screening endeavors. The inadequacies of two-dimensional (2D) models prompted the exploration of three-dimensional (3D) culture systems, which more faithfully recapitulate the native tumor microenvironment. These 3D systems have distinct advantages over 2D models in morphology, proliferation, drug response, and protein expression. Among these 3D platforms, tumor organoids and spheroids, generated through different methodologies, have emerged as next-generation models that closely mirror aspects of pancreatic tumor biology. This comprehensive review scrutinizes pancreatic cancer spheroids' techniques, tissue sources, and applications, offering a nuanced analysis of their advantages and limitations. By comparing these distinct 3D culture systems, researchers gain valuable insights to inform the selection of optimal model designs aligned with their specific experimental objectives. The utilization of these advanced models holds significant promise for enhancing the clinical relevance of both in vitro and in vivo cancer research, thereby contributing to the development of improved therapeutics against pancreatic cancer.
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Affiliation(s)
- Nazanin Jamshidi
- Kimia Andisheh Teb Medical and Molecular Laboratory Research Co, Tehran, Iran
| | - Negar Jamshidi
- Kimia Andisheh Teb Medical and Molecular Laboratory Research Co, Tehran, Iran
| | | | - Elahe Shams
- Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
| | - Vahid Chaleshi
- Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran
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11
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Nie HY, Ge J, Liu KG, Yue Y, Li H, Lin HG, Yan HF, Zhang T, Sun HW, Yang JW, Zhou JL, Cui Y. The effects of microgravity on stem cells and the new insights it brings to tissue engineering and regenerative medicine. LIFE SCIENCES IN SPACE RESEARCH 2024; 41:1-17. [PMID: 38670635 DOI: 10.1016/j.lssr.2024.01.001] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/01/2023] [Revised: 12/13/2023] [Accepted: 01/06/2024] [Indexed: 04/28/2024]
Abstract
Conventional two-dimensional (2D) cell culture techniques may undergo modifications in the future, as life scientists have widely acknowledged the ability of three-dimensional (3D) in vitro culture systems to accurately simulate in vivo biology. In recent years, researchers have discovered that microgravity devices can address many challenges associated with 3D cell culture. Stem cells, being pluripotent cells, are regarded as a promising resource for regenerative medicine. Recent studies have demonstrated that 3D culture in microgravity devices can effectively guide stem cells towards differentiation and facilitate the formation of functional tissue, thereby exhibiting advantages within the field of tissue engineering and regenerative medicine. Furthermore, We delineate the impact of microgravity on the biological behavior of various types of stem cells, while elucidating the underlying mechanisms governing these alterations. These findings offer exciting prospects for diverse applications.
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Affiliation(s)
- Hong-Yun Nie
- Department of General Surgery, The 306th Hospital of PLA-Peking University Teaching Hospital, Beijing 100101, China; Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China
| | - Jun Ge
- Department of General Surgery, The 306th Hospital of PLA-Peking University Teaching Hospital, Beijing 100101, China; Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China
| | - Kai-Ge Liu
- Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China
| | - Yuan Yue
- Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China
| | - Hao Li
- Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China.
| | - Hai-Guan Lin
- Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China
| | - Hong-Feng Yan
- Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China
| | - Tao Zhang
- Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China
| | - Hong-Wei Sun
- Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China
| | - Jian-Wu Yang
- Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China
| | - Jin-Lian Zhou
- Department of Pathology, Strategic Support Force Medical Center, Beijing 100101, China
| | - Yan Cui
- Department of General Surgery, The 306th Hospital of PLA-Peking University Teaching Hospital, Beijing 100101, China; Department of General Surgery, Strategic Support Force Medical Center, Beijing 100101, China.
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12
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Farhang Doost N, Srivastava SK. A Comprehensive Review of Organ-on-a-Chip Technology and Its Applications. BIOSENSORS 2024; 14:225. [PMID: 38785699 PMCID: PMC11118005 DOI: 10.3390/bios14050225] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/14/2024] [Revised: 04/09/2024] [Accepted: 04/23/2024] [Indexed: 05/25/2024]
Abstract
Organ-on-a-chip (OOC) is an emerging technology that simulates an artificial organ within a microfluidic cell culture chip. Current cell biology research focuses on in vitro cell cultures due to various limitations of in vivo testing. Unfortunately, in-vitro cell culturing fails to provide an accurate microenvironment, and in vivo cell culturing is expensive and has historically been a source of ethical controversy. OOC aims to overcome these shortcomings and provide the best of both in vivo and in vitro cell culture research. The critical component of the OOC design is utilizing microfluidics to ensure a stable concentration gradient, dynamic mechanical stress modeling, and accurate reconstruction of a cellular microenvironment. OOC also has the advantage of complete observation and control of the system, which is impossible to recreate in in-vivo research. Multiple throughputs, channels, membranes, and chambers are constructed in a polydimethylsiloxane (PDMS) array to simulate various organs on a chip. Various experiments can be performed utilizing OOC technology, including drug delivery research and toxicology. Current technological expansions involve multiple organ microenvironments on a single chip, allowing for studying inter-tissue interactions. Other developments in the OOC technology include finding a more suitable material as a replacement for PDMS and minimizing artefactual error and non-translatable differences.
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Affiliation(s)
| | - Soumya K. Srivastava
- Department of Chemical and Biomedical Engineering, West Virginia University, Morgantown, WV 26506, USA;
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13
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Ferronato GDA, Vit FF, da Silveira JC. 3D culture applied to reproduction in females: possibilities and perspectives. Anim Reprod 2024; 21:e20230039. [PMID: 38510565 PMCID: PMC10954237 DOI: 10.1590/1984-3143-ar2023-0039] [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: 03/20/2023] [Accepted: 12/13/2023] [Indexed: 03/22/2024] Open
Abstract
In vitro cell culture is a well-established technique present in numerous laboratories in diverse areas. In reproduction, gametes, embryos, and reproductive tissues, such as the ovary and endometrium, can be cultured. These cultures are essential for embryo development studies, understanding signaling pathways, developing drugs for reproductive diseases, and in vitro embryo production (IVP). Although many culture systems are successful, they still have limitations to overcome. Three-dimensional (3D) culture systems can be close to physiological conditions, allowing greater interaction between cells and cells with the surrounding environment, maintenance of the cells' natural morphology, and expression of genes and proteins such as in vivo. Additionally, three-dimensional culture systems can stimulated extracellular matrix generating responses due to the mechanical force produced. Different techniques can be used to perform 3D culture systems, such as hydrogel matrix, hanging drop, low attachment surface, scaffold, levitation, liquid marble, and 3D printing. These systems demonstrate satisfactory results in follicle culture, allowing the culture from the pre-antral to antral phase, maintaining the follicular morphology, and increasing the development rates of embryos. Here, we review some of the different techniques of 3D culture systems and their applications to the culture of follicles and embryos, bringing new possibilities to the future of assisted reproduction.
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Affiliation(s)
| | - Franciele Flores Vit
- Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, SP, Brasil
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14
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Bodio C, Milesi A, Lonati PA, Chighizola CB, Mauro A, Pradotto LG, Meroni PL, Borghi MO, Raschi E. Fibroblasts and Endothelial Cells in Three-Dimensional Models: A New Tool for Addressing the Pathogenesis of Systemic Sclerosis as a Prototype of Fibrotic Vasculopathies. Int J Mol Sci 2024; 25:2780. [PMID: 38474040 DOI: 10.3390/ijms25052780] [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: 12/29/2023] [Revised: 02/22/2024] [Accepted: 02/26/2024] [Indexed: 03/14/2024] Open
Abstract
Two-dimensional in vitro cultures have represented a milestone in biomedical and pharmacological research. However, they cannot replicate the architecture and interactions of in vivo tissues. Moreover, ethical issues regarding the use of animals have triggered strategies alternative to animal models. The development of three-dimensional (3D) models offers a relevant tool to investigate disease pathogenesis and treatment, modeling in vitro the in vivo environment. We aimed to develop a dynamic 3D in vitro model for culturing human endothelial cells (ECs) and skin fibroblasts, simulating the structure of the tissues mainly affected in systemic sclerosis (SSc), a prototypical autoimmune fibrotic vasculopathy. Dermal fibroblasts and umbilical vein ECs grown in scaffold or hydrogel, respectively, were housed in bioreactors under flow. Fibroblasts formed a tissue-like texture with the deposition of a new extracellular matrix (ECM) and ECs assembled tube-shaped structures with cell polarization. The fine-tuned dynamic modular system allowing 3D fibroblast/EC culture connection represents a valuable model of the in vivo interplay between the main players in fibrotic vasculopathy as SSc. This model can lead to a more accurate study of the disease's pathogenesis, avoiding the use of animals, and to the development of novel therapies, possibly resulting in improved patient management.
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Affiliation(s)
- Caterina Bodio
- Experimental Laboratory of Immunological and Rheumatologic Researches, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Istituto Auxologico Italiano, 20095 Cusano Milanino, Italy
| | - Alessandra Milesi
- Laboratory of Clinical Neurobiology, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Istituto Auxologico Italiano, 28824 Piancavallo, Italy
| | - Paola Adele Lonati
- Experimental Laboratory of Immunological and Rheumatologic Researches, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Istituto Auxologico Italiano, 20095 Cusano Milanino, Italy
| | - Cecilia Beatrice Chighizola
- Department of Clinical Sciences and Community Health, University of Milan, 20122 Milan, Italy
- U.O.C. Clinica Reumatologica Pediatrica, ASST G. Pini-CTO, 20122 Milan, Italy
| | - Alessandro Mauro
- Laboratory of Clinical Neurobiology, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Istituto Auxologico Italiano, 28824 Piancavallo, Italy
- Department of Neuroscience, University of Turin, 10124 Turin, Italy
| | - Luca Guglielmo Pradotto
- Laboratory of Clinical Neurobiology, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Istituto Auxologico Italiano, 28824 Piancavallo, Italy
- Department of Neuroscience, University of Turin, 10124 Turin, Italy
| | - Pier Luigi Meroni
- Experimental Laboratory of Immunological and Rheumatologic Researches, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Istituto Auxologico Italiano, 20095 Cusano Milanino, Italy
| | - Maria Orietta Borghi
- Experimental Laboratory of Immunological and Rheumatologic Researches, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Istituto Auxologico Italiano, 20095 Cusano Milanino, Italy
- Department of Clinical Sciences and Community Health, University of Milan, 20122 Milan, Italy
| | - Elena Raschi
- Experimental Laboratory of Immunological and Rheumatologic Researches, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), Istituto Auxologico Italiano, 20095 Cusano Milanino, Italy
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15
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Hahn F, Ferrandez-Montero A, Queri M, Vancaeyzeele C, Plesse C, Agniel R, Leroy-Dudal J. Electroactive 4D Porous Scaffold Based on Conducting Polymer as a Responsive and Dynamic In Vitro Cell Culture Platform. ACS APPLIED MATERIALS & INTERFACES 2024; 16:5613-5626. [PMID: 38278772 PMCID: PMC10859895 DOI: 10.1021/acsami.3c16686] [Citation(s) in RCA: 3] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 11/07/2023] [Revised: 01/15/2024] [Accepted: 01/15/2024] [Indexed: 01/28/2024]
Abstract
In vivo, cells reside in a 3D porous and dynamic microenvironment. It provides biochemical and biophysical cues that regulate cell behavior in physiological and pathological processes. In the context of fundamental cell biology research, tissue engineering, and cell-based drug screening systems, a challenge is to develop relevant in vitro models that could integrate the dynamic properties of the cell microenvironment. Taking advantage of the promising high internal phase emulsion templating, we here designed a polyHIPE scaffold with a wide interconnected porosity and functionalized its internal 3D surface with a thin layer of electroactive conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) to turn it into a 4D electroresponsive scaffold. The resulting scaffold was cytocompatible with fibroblasts, supported cellular infiltration, and hosted cells, which display a 3D spreading morphology. It demonstrated robust actuation in ion- and protein-rich complex culture media, and its electroresponsiveness was not altered by fibroblast colonization. Thanks to customized electrochemical stimulation setups, the electromechanical response of the polyHIPE/PEDOT scaffolds was characterized in situ under a confocal microscope and showed 10% reversible volume variations. Finally, the setups were used to monitor in real time and in situ fibroblasts cultured into the polyHIPE/PEDOT scaffold during several cycles of electromechanical stimuli. Thus, we demonstrated the proof of concept of this tunable scaffold as a tool for future 4D cell culture and mechanobiology studies.
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Affiliation(s)
- Franziska Hahn
- Equipe
de Recherche sur les Relations Matrice Extracellulaire-Cellules (ERRMECe),
Groupe Matrice Extracellulaire et Physiopathologie (MECuP), I-Mat, CY Cergy Paris Université, 95000 Neuville
sur Oise, France
- Laboratoire
de Physicochimie des Polymères et des Interfaces (LPPI), I-Mat, CY Cergy Paris Université, 95000 Neuville sur Oise, France
| | - Ana Ferrandez-Montero
- Equipe
de Recherche sur les Relations Matrice Extracellulaire-Cellules (ERRMECe),
Groupe Matrice Extracellulaire et Physiopathologie (MECuP), I-Mat, CY Cergy Paris Université, 95000 Neuville
sur Oise, France
- Laboratoire
de Physicochimie des Polymères et des Interfaces (LPPI), I-Mat, CY Cergy Paris Université, 95000 Neuville sur Oise, France
- Instituto
de Ceramica y Vidrio (ICV), CSIC, Campus Cantoblanco, Kelsen 5., 28049 Madrid, Spain
| | - Mélodie Queri
- Equipe
de Recherche sur les Relations Matrice Extracellulaire-Cellules (ERRMECe),
Groupe Matrice Extracellulaire et Physiopathologie (MECuP), I-Mat, CY Cergy Paris Université, 95000 Neuville
sur Oise, France
- Laboratoire
de Physicochimie des Polymères et des Interfaces (LPPI), I-Mat, CY Cergy Paris Université, 95000 Neuville sur Oise, France
| | - Cédric Vancaeyzeele
- Laboratoire
de Physicochimie des Polymères et des Interfaces (LPPI), I-Mat, CY Cergy Paris Université, 95000 Neuville sur Oise, France
| | - Cédric Plesse
- Laboratoire
de Physicochimie des Polymères et des Interfaces (LPPI), I-Mat, CY Cergy Paris Université, 95000 Neuville sur Oise, France
| | - Rémy Agniel
- Equipe
de Recherche sur les Relations Matrice Extracellulaire-Cellules (ERRMECe),
Groupe Matrice Extracellulaire et Physiopathologie (MECuP), I-Mat, CY Cergy Paris Université, 95000 Neuville
sur Oise, France
| | - Johanne Leroy-Dudal
- Equipe
de Recherche sur les Relations Matrice Extracellulaire-Cellules (ERRMECe),
Groupe Matrice Extracellulaire et Physiopathologie (MECuP), I-Mat, CY Cergy Paris Université, 95000 Neuville
sur Oise, France
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16
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Oliveira A, Dias C, Oliveira R, Almeida C, Fuciños P, Sillankorva S, Oliveira H. Paving the way forward: Escherichia coli bacteriophages in a One Health approach. Crit Rev Microbiol 2024; 50:87-104. [PMID: 36608263 DOI: 10.1080/1040841x.2022.2161869] [Citation(s) in RCA: 6] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/17/2022] [Accepted: 12/19/2022] [Indexed: 01/07/2023]
Abstract
Escherichia coli is one of the most notorious pathogens for its ability to adapt, colonize, and proliferate in different habitats through a multitude of acquired virulence factors. Its presence affects the food-processing industry and causes food poisoning, being also a major economic burden for the food, agriculture, and health sectors. Bacteriophages are emerging as an appealing strategy to mitigate bacterial pathogens, including specific E. coli pathovars, without exerting a deleterious effect on humans and animals. This review globally analyzes the applied research on E. coli phages for veterinary, food, and human use. It starts by describing the pathogenic E. coli pathotypes and their relevance in human and animal context. The idea that phages can be used as a One Health approach to control and interrupt the transmission routes of pathogenic E. coli is sustained through an exhaustive revision of the recent literature. The emerging phage formulations, genetic engineering and encapsulation technologies are also discussed as a means of improving phage-based control strategies, with a particular focus on E. coli pathogens.
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Affiliation(s)
- Ana Oliveira
- CEB - Centre of Biological Engineering, University of Minho, Braga, Portugal
- LABBELS - Associate Laboratory, Braga, Guimarães, Portugal
| | - Carla Dias
- CEB - Centre of Biological Engineering, University of Minho, Braga, Portugal
- LABBELS - Associate Laboratory, Braga, Guimarães, Portugal
| | - Ricardo Oliveira
- INIAV, IP-National Institute for Agrarian and Veterinary Research, Vairão, Vila do Conde, Portugal
- LEPABE - Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, Porto, Portugal
- ALiCE - Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, Porto, Portugal
| | - Carina Almeida
- INIAV, IP-National Institute for Agrarian and Veterinary Research, Vairão, Vila do Conde, Portugal
- LEPABE - Laboratory for Process Engineering, Environment, Biotechnology and Energy, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, Porto, Portugal
- ALiCE - Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, Porto, Portugal
| | - Pablo Fuciños
- ALiCE - Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, Rua Dr. Roberto Frias, Porto, Portugal
| | - Sanna Sillankorva
- INL - International Iberian Nanotechnology Laboratory, Avenida Mestre José Veiga, Braga, Portugal
| | - Hugo Oliveira
- CEB - Centre of Biological Engineering, University of Minho, Braga, Portugal
- LABBELS - Associate Laboratory, Braga, Guimarães, Portugal
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17
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Dragic H, Chaveroux C, Cosset E, Manie SN. Modelling cancer metabolism in vitro: current improvements and future challenges. FEBS J 2024; 291:402-411. [PMID: 36516350 DOI: 10.1111/febs.16704] [Citation(s) in RCA: 2] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/09/2022] [Revised: 12/05/2022] [Accepted: 12/13/2022] [Indexed: 12/15/2022]
Abstract
Advances in cancer biology over the past decades have revealed that metabolic adaptation of cancer cells is an essential aspect of tumorigenesis. However, recent insights into tumour metabolism in vivo have revealed dissimilarities with results obtained in vitro. This is partly due to the reductionism of in vitro cancer models that struggle to reproduce the complexity of tumour tissues. This review describes some of the discrepancies in cancer cell metabolism between in vitro and in vivo conditions, and presents current methodological approaches and tools used to bridge the gap with the clinically relevant microenvironment. As such, these approaches should generate new knowledge that could be more effectively translated into therapeutic opportunities.
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Affiliation(s)
- Helena Dragic
- Centre de Recherche en Cancérologie de Lyon, INSERM U1052, CNRS 5286, Centre Léon Bérard, Univ Lyon, Université Claude Bernard Lyon 1, France
| | - Cedric Chaveroux
- Centre de Recherche en Cancérologie de Lyon, INSERM U1052, CNRS 5286, Centre Léon Bérard, Univ Lyon, Université Claude Bernard Lyon 1, France
| | - Erika Cosset
- Centre de Recherche en Cancérologie de Lyon, INSERM U1052, CNRS 5286, Centre Léon Bérard, Univ Lyon, Université Claude Bernard Lyon 1, France
| | - Serge N Manie
- Centre de Recherche en Cancérologie de Lyon, INSERM U1052, CNRS 5286, Centre Léon Bérard, Univ Lyon, Université Claude Bernard Lyon 1, France
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18
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Choinière W, Petit È, Monfette V, Pelletier S, Godbout-Lavoie C, Lauzon MA. Dynamic three-dimensional coculture model: The future of tissue engineering applied to the peripheral nervous system. J Tissue Eng 2024; 15:20417314241265916. [PMID: 39139455 PMCID: PMC11320398 DOI: 10.1177/20417314241265916] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/03/2024] [Accepted: 06/18/2024] [Indexed: 08/15/2024] Open
Abstract
Traumatic injuries to the peripheral nervous system (PNI) can lead to severe consequences such as paralysis. Unfortunately, current treatments rarely allow for satisfactory functional recovery. The high healthcare costs associated with PNS injuries, worker disability, and low patient satisfaction press for alternative solutions that surpass current standards. For the treatment of injuries with a deficit of less than 30 mm to bridge, the use of synthetic nerve conduits (NGC) is favored. However, to develop such promising therapeutic strategies, in vitro models that more faithfully mimic nerve physiology are needed. The absence of a clinically scaled model with essential elements such as a three-dimension environment and dynamic coculture has hindered progress in this field. The presented research focuses on the development of an in vitro coculture model of the peripheral nervous system (PNS) involving the use of functional biomaterial which microstructure replicates nerve topography. Initially, the behavior of neuron-derived cell lines (N) and Schwann cells (SC) in contact with a short section of biomaterial (5 mm) was studied. Subsequent investigations, using fluorescent markers and survival assays, demonstrated the synergistic effects of coculture. These optimized parameters were then applied to longer biomaterials (30 mm), equivalent to clinically used NGC. The results obtained demonstrated the possibility of maintaining an extended coculture of SC and N over a 7-day period on a clinically scaled biomaterial, observing some functionality. In the long term, the knowledge gained from this work will contribute to a better understanding of the PNS regeneration process and promote the development of future therapeutic approaches while reducing reliance on animal experimentation. This model can be used for drug screening and adapted for personalized medicine trials. Ultimately, this work fills a critical gap in current research, providing a transformative approach to study and advance treatments for PNS injuries.
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Affiliation(s)
- William Choinière
- Department of Chemical Engineering and Biotechnological Engineering, Université de Sherbrooke, Sherbrooke, QC, Canada
| | - Ève Petit
- Department of Chemical Engineering and Biotechnological Engineering, Université de Sherbrooke, Sherbrooke, QC, Canada
| | - Vincent Monfette
- Department of Chemical Engineering and Biotechnological Engineering, Université de Sherbrooke, Sherbrooke, QC, Canada
| | - Samuel Pelletier
- Department of Electrical and Informatics Engineering, Université de Sherbrooke, Sherbrooke, QC, Canada
| | - Catherine Godbout-Lavoie
- Department of Chemical Engineering and Biotechnological Engineering, Université de Sherbrooke, Sherbrooke, QC, Canada
| | - Marc-Antoine Lauzon
- Department of Chemical Engineering and Biotechnological Engineering, Université de Sherbrooke, Sherbrooke, QC, Canada
- Research Center on Aging, CIUSS de l’ESTRIE-CHUS, Sherbrooke, QC, Canada
- The Quebec Network for Research on Protein Function, Engineering, and Applications, Montréal, QC, Canada
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19
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Fujisawa Y, Takao T, Yamada D, Takarada T. Development of cartilage tissue using a stirred bioreactor and human iPSC-derived limb bud mesenchymal cells. Biochem Biophys Res Commun 2023; 687:149146. [PMID: 37956599 DOI: 10.1016/j.bbrc.2023.149146] [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: 08/25/2023] [Accepted: 10/23/2023] [Indexed: 11/15/2023]
Abstract
Production of cartilaginous particles for regenerative medicine requires a large supply of chondrocytes and development of suitable production techniques. Previously, we successfully produced human induced pluripotent stem cell (hiPSC)-derived limb bud mesenchymal cells (ExpLBM cells) with a high chondrogenic differentiation potential that stably proliferate. It may be possible to use these cells in combination with a stirred bioreactor to develop a tissue-engineered cell culture technology with potential for scale-up to facilitate production of large amounts of cartilaginous particles. ExpLBM cells derived from 414C2 and Ff-I 14s04 (human leukocyte antigen homozygous) hiPSCs were seeded into a stirred bioreactor containing cartilage induction medium. To characterize the cartilaginous particles produced, we performed real-time quantitative reverse transcription-polymerase chain reaction and histological analyses. Additionally, we transplanted the cartilage tissue into osteochondral defects of immunocompromised rats to assess its functionality, and evaluated engraftment of the grafted tissue. We successfully produced large amounts of cartilaginous particles via cartilage induction culture in a stirred bioreactor. This tissue exhibited significantly increased expression levels of type II collagen (COL2), aggrecan (ACAN), and SRY-box transcription factor 9 (SOX9), as well as positive Safranin O and Toluidine blue staining, indicating that it possesses characteristics of hyaline cartilage. Furthermore, engrafted tissues in osteochondral knee defects of immunodeficient rats were positively stained for human vimentin, COL2, and ACAN as well as with Safranin O. In this study, we successfully generated large amounts of hiPSC-derived cartilaginous particles using a combination of tissue engineering techniques. This method is promising as a cartilage regeneration technology with potential for scale-up.
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Affiliation(s)
- Yuki Fujisawa
- Department of Regenerative Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, 700-8558, Japan
| | - Tomoka Takao
- Department of Regenerative Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, 700-8558, Japan
| | - Daisuke Yamada
- Department of Regenerative Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, 700-8558, Japan
| | - Takeshi Takarada
- Department of Regenerative Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, 700-8558, Japan.
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Flores-Espinoza AI, Garcia-Contreras R, Guzman-Rocha DA, Aranda-Herrera B, Chavez-Granados PA, Jurado CA, Alfawaz YF, Alshabib A. Gelatin-Chitosan Hydrogel Biological, Antimicrobial and Mechanical Properties for Dental Applications. Biomimetics (Basel) 2023; 8:575. [PMID: 38132514 PMCID: PMC10742194 DOI: 10.3390/biomimetics8080575] [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: 09/12/2023] [Revised: 11/26/2023] [Accepted: 11/28/2023] [Indexed: 12/23/2023] Open
Abstract
Chitosan, a natural polysaccharide sourced from crustaceans and insects, is often used with hydrogels in wound care. Evaluating its cytotoxicity and antimicrobial properties is crucial for its potential use in dentistry. OBJECTIVE To investigate the mechanical properties of gelatin hydrogels based on decaethylated chitosan and antimicrobial activity against Streptococcus mutans and their biological effects with stem cells from apical papilla (SCAPs). MATERIAL AND METHODS Gelatin-chitosan hydrogels were synthesized at concentrations of 0%, 0.2% and 0.5%. Enzymatic and hydrolytic degradation, along with swelling capacity, was assessed. Fourier transform infrared spectroscopy (FTIR) analysis was employed to characterize the hydrogels. The interaction between hydrogels and SCAPs was examined through initial adhesion and cell proliferation at 24 and 48 h, using the Thiazolyl Blue Tetrazolium Bromide (MTT assay). The antimicrobial effect was evaluated using agar diffusion and a microdilution test against S. mutans. Uniaxial tensile strength (UTS) was also measured to assess the mechanical properties of the hydrogels. RESULTS The hydrogels underwent hydrolytic and enzymatic degradation at 30, 220, 300 min and 15, 25, 30 min, respectively. Significantly, (p < 0.01) swelling capacity occurred at 20, 40, 30 min, respectively. Gelatin-chitosan hydrogels' functional groups were confirmed using vibrational pattern analysis. SCAPs proliferation corresponded to 24 h = 73 ± 2%, 82 ± 2%, 61 ± 6% and 48 h = 83 ± 11%, 86 ± 2%, 44 ± 2%, respectively. The bacterial survival of hydrogel interaction was found to be 96 ± 1%, 17 ± 1.5% (p < 0.01) and 1 ± 0.5% (p < 0.01), respectively. UTS showed enhanced (p < 0.05) mechanical properties with chitosan presence. CONCLUSION Gelatin-chitosan hydrogels displayed favorable degradation, swelling capacity, mild dose-dependent cytotoxicity, significant proliferation with stem cells from apical papilla (SCAPs), substantial antimicrobial effects against S. mutans and enhanced mechanical properties. These findings highlight their potential applications as postoperative care dressings.
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Affiliation(s)
- Andrea Itzamantul Flores-Espinoza
- Interdisciplinary Research Laboratory (LII), Nanostructures and Biomaterials Area, National School of Higher Studies (ENES), Leon Unit, National Autonomous University of Mexico (UNAM), Leon 37689, Mexico; (A.I.F.-E.); (R.G.-C.); (D.A.G.-R.); (B.A.-H.); (P.A.C.-G.)
| | - Rene Garcia-Contreras
- Interdisciplinary Research Laboratory (LII), Nanostructures and Biomaterials Area, National School of Higher Studies (ENES), Leon Unit, National Autonomous University of Mexico (UNAM), Leon 37689, Mexico; (A.I.F.-E.); (R.G.-C.); (D.A.G.-R.); (B.A.-H.); (P.A.C.-G.)
| | - Dulce Araceli Guzman-Rocha
- Interdisciplinary Research Laboratory (LII), Nanostructures and Biomaterials Area, National School of Higher Studies (ENES), Leon Unit, National Autonomous University of Mexico (UNAM), Leon 37689, Mexico; (A.I.F.-E.); (R.G.-C.); (D.A.G.-R.); (B.A.-H.); (P.A.C.-G.)
| | - Benjamin Aranda-Herrera
- Interdisciplinary Research Laboratory (LII), Nanostructures and Biomaterials Area, National School of Higher Studies (ENES), Leon Unit, National Autonomous University of Mexico (UNAM), Leon 37689, Mexico; (A.I.F.-E.); (R.G.-C.); (D.A.G.-R.); (B.A.-H.); (P.A.C.-G.)
| | - Patricia Alejandra Chavez-Granados
- Interdisciplinary Research Laboratory (LII), Nanostructures and Biomaterials Area, National School of Higher Studies (ENES), Leon Unit, National Autonomous University of Mexico (UNAM), Leon 37689, Mexico; (A.I.F.-E.); (R.G.-C.); (D.A.G.-R.); (B.A.-H.); (P.A.C.-G.)
| | - Carlos A. Jurado
- Department of Prosthodontics, The University of Iowa College of Dentistry and Dental Clinics, Iowa City, IA 52242, USA;
| | - Yasser F. Alfawaz
- Department of Restorative Dentistry, King Saud University College of Dentistry, Riyadh 11545, Saudi Arabia;
| | - Abdulrahman Alshabib
- Department of Restorative Dentistry, King Saud University College of Dentistry, Riyadh 11545, Saudi Arabia;
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Zein-based 3D tubular constructs with tunable porosity for 3D cell culture and drug delivery. BIOMEDICAL ENGINEERING ADVANCES 2023. [DOI: 10.1016/j.bea.2022.100059] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022] Open
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22
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Li N, Dai X, Yang F, Sun Y, Wu X, Zhou Q, Chen K, Sun J, Bi W, Shi L, Yu Y. Spontaneous spheroids from alveolar bone-derived mesenchymal stromal cells maintain pluripotency of stem cells by regulating hypoxia-inducible factors. Biol Res 2023; 56:17. [PMID: 37016436 PMCID: PMC10074860 DOI: 10.1186/s40659-023-00421-w] [Citation(s) in RCA: 6] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/06/2022] [Accepted: 02/27/2023] [Indexed: 04/06/2023] Open
Abstract
BACKGROUND Spontaneous spheroid culture is a novel three-dimensional (3D) culture strategy for the rapid and efficient selection of progenitor cells. The objectives of this study are to investigate the pluripotency and differentiation capability of spontaneous spheroids from alveolar bone-derived mesenchymal stromal cells (AB-MSCs); compare the advantages of spontaneous spheroids to those of mechanical spheroids; and explore the mechanisms of stemness enhancement during spheroid formation from two-dimensional (2D) cultured cells. METHODS AB-MSCs were isolated from the alveolar bones of C57BL/6 J mice. Spontaneous spheroids formed in low-adherence specific culture plates. The stemness, proliferation, and multi-differentiation capacities of spheroids and monolayer cultures were investigated by reverse transcription quantitative polymerase chain reaction (RT-qPCR), immunofluorescence, alkaline phosphatase (ALP) activity, and oil-red O staining. The pluripotency difference between the spontaneous and mechanical spheroids was analyzed using RT-qPCR. Hypoxia-inducible factor (HIFs) inhibition experiments were performed to explore the mechanisms of stemness maintenance in AB-MSC spheroids. RESULTS AB-MSCs successfully formed spontaneous spheroids after 24 h. AB-MSC spheroids were positive for MSC markers and pluripotency markers (Oct4, KLF4, Sox2, and cMyc). Spheroids showed higher Ki67 expression and lower Caspase3 expression at 24 h. Under the corresponding conditions, the spheroids were successfully differentiated into osteogenic and adipogenic lineages. AB-MSC spheroids can induce neural-like cells after neurogenic differentiation. Higher expression of osteogenic markers, adipogenic markers, and neurogenic markers (NF-M, NeuN, and GFAP) was found in spheroids than in the monolayer. Spontaneous spheroids exhibited higher stemness than mechanical spheroids did. HIF-1α and HIF-2α were remarkably upregulated in spheroids. After HIF-1/2α-specific inhibition, spheroid formation was significantly reduced. Moreover, the expression of the pluripotency genes was suppressed. CONCLUSIONS Spontaneous spheroids from AB-MSCs enhance stemness and pluripotency. HIF-1/2α plays an important role in the stemness regulation of spheroids. AB-MSC spheroids exhibit excellent multi-differentiation capability, which may be a potent therapy for craniomaxillofacial tissue regeneration.
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Affiliation(s)
- Ni Li
- Shanghai University of Medicine and Health Sciences Affiliated Zhoupu Hospital, Shanghai, China, 201318
- The College of Medical Technology, Shanghai University of Medicine and Health Sciences, Shanghai, China, 201318
- Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China, 180 Fenglin Road, 200032
| | - Xiaofeng Dai
- Department of Stomatology, Shanghai Jing'an District Dental Clinic, Shanghai, China, 15 Pingxingguan Road, 200040
| | - Fei Yang
- Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China, 180 Fenglin Road, 200032
| | - Yang Sun
- Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China, 180 Fenglin Road, 200032
| | - Xingwen Wu
- Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China, 180 Fenglin Road, 200032
| | - Qianrong Zhou
- Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China, 180 Fenglin Road, 200032
| | - Kai Chen
- Department of Stomatology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai, China, 200072
| | - Jian Sun
- Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China, 180 Fenglin Road, 200032
| | - Wei Bi
- Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China, 180 Fenglin Road, 200032
| | - Le Shi
- Department of Stomatology, Shanghai Jing'an District Dental Clinic, Shanghai, China, 15 Pingxingguan Road, 200040.
| | - Youcheng Yu
- Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China, 180 Fenglin Road, 200032.
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Jankovic MG, Stojkovic M, Bojic S, Jovicic N, Kovacevic MM, Ivosevic Z, Juskovic A, Kovacevic V, Ljujic B. Scaling up human mesenchymal stem cell manufacturing using bioreactors for clinical uses. Curr Res Transl Med 2023; 71:103393. [PMID: 37163885 DOI: 10.1016/j.retram.2023.103393] [Citation(s) in RCA: 7] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/20/2022] [Revised: 03/13/2023] [Accepted: 04/26/2023] [Indexed: 05/12/2023]
Abstract
Human mesenchymal stem cells (hMSCs) are multipotent cells and an attractive therapeutic agent in regenerative medicine and intensive clinical research. Despite the great potential, the limitation that needs to be overcome is the necessity of ex vivo expansion because of insufficient number of hMSCs presented within adult organs and the high doses required for a transplantation. As a result, numerous research studies aim to provide novel expansion methods in order to achieve appropriate numbers of cells with preserved therapeutic quality. Bioreactor-based cell expansion provide high-level production of hMSCs in accordance with good manufacturing practice (GMP) and quality standards. This review summarizes current knowledge about the hMSCs manufacturing platforms with a main focus to the application of bioreactors for large-scale production of GMP-grade hMSCs.
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Affiliation(s)
- Marina Gazdic Jankovic
- University of Kragujevac, Serbia, Faculty of Medical Sciences, Department of Genetics, Serbia.
| | | | - Sanja Bojic
- Newcastle University, School of Computing, Newcastle upon Tyne, UK
| | - Nemanja Jovicic
- University of Kragujevac, Serbia, Faculty of Medical Sciences, Department of Histology and Embryology, Serbia
| | - Marina Miletic Kovacevic
- University of Kragujevac, Serbia, Faculty of Medical Sciences, Department of Histology and Embryology, Serbia
| | - Zeljko Ivosevic
- University of Kragujevac, Serbia, Faculty of Medical Sciences, Department of Genetics, Serbia
| | - Aleksandar Juskovic
- Department of Orthopaedic Surgery, Clinical Centre of Montenegro, 81110 Podgorica, Montenegro
| | - Vojin Kovacevic
- University of Kragujevac, Serbia, Faculty of Medical Sciences, Department of Surgery, Serbia
| | - Biljana Ljujic
- University of Kragujevac, Serbia, Faculty of Medical Sciences, Department of Genetics, Serbia
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Chipangura JK, Ntamo Y, Mohr B, Chellan N. A review of challenges and prospects of 3D cell-based culture models used for studying drug induced liver injury during early phases of drug development. Hum Exp Toxicol 2023; 42:9603271221147884. [PMID: 36879529 DOI: 10.1177/09603271221147884] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 03/08/2023]
Abstract
Drug-induced liver injury (DILI) is the leading cause of compound attrition during drug development. Over the years, a battery of in-vitro cell culture toxicity tests is being conducted to evaluate the toxicity of compounds prior to testing in laboratory animals. Two-dimensional (2D) in-vitro cell culture models are commonly used and have provided a great deal of knowledge; however, these models often fall short in mimicking natural structures of tissues in-vivo. Testing in humans is the most logical method, but unfortunately there are ethical limitations associated with human tests. To overcome these limitations better human-relevant, predictive models are required. The past decade has witnessed significant efforts towards the development of three-dimensional (3D) in-vitro cell culture models better mimicking in-vivo physiology. 3D cell culture has advantages in being representative of the interactions of cells in-vivo and when validated can act as an interphase between 2D cell culture models and in-vivo animal models. The current review seeks to provide an overview of the challenges that make biomarkers used for detection of DILI not to be sensitive enough during drug development and explore how 3D cell culture models can be used to address the gap with the current models.
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Affiliation(s)
- John K Chipangura
- Faculty of Health Science, University of Cape Town Research Animal Facility, South Africa
| | - Yonela Ntamo
- Biomedical Research and Innovation Platform, South African Medical Research Council, Tygerberg, South Africa
| | - Bert Mohr
- Faculty of Health Science, University of Cape Town Research Animal Facility, South Africa
| | - Nireshni Chellan
- Biomedical Research and Innovation Platform, South African Medical Research Council, Tygerberg, South Africa
- Centre for Cardio-metabolic Research in Africa (CARMA), Division of Medical Physiology, Faculty of Health Sciences, Stellenbosch University, South Africa
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25
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Chari D, Basit R, Wiseman J, Chowdhury F. Simulating traumatic brain injury in vitro: developing high throughput models to test biomaterial based therapies. Neural Regen Res 2023; 18:289-292. [PMID: 35900405 PMCID: PMC9396524 DOI: 10.4103/1673-5374.346465] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022] Open
Abstract
Traumatic brain injuries are serious clinical incidents associated with some of the poorest outcomes in neurological practice. Coupled with the limited regenerative capacity of the brain, this has significant implications for patients, carers, and healthcare systems, and the requirement for life-long care in some cases. Clinical treatment currently focuses on limiting the initial neural damage with long-term care/support from multidisciplinary teams. Therapies targeting neuroprotection and neural regeneration are not currently available but are the focus of intensive research. Biomaterial-based interventions are gaining popularity for a range of applications including biomolecule and drug delivery, and to function as cellular scaffolds. Experimental investigations into the development of such novel therapeutics for traumatic brain injury will be critically underpinned by the availability of appropriate high throughput, facile, ethically viable, and pathomimetic biological model systems. This represents a significant challenge for researchers given the pathological complexity of traumatic brain injury. Specifically, there is a concerted post-injury response mounted by multiple neural cell types which includes microglial activation and astroglial scarring with the expression of a range of growth inhibitory molecules and cytokines in the lesion environment. Here, we review common models used for the study of traumatic brain injury (ranging from live animal models to in vitro systems), focusing on penetrating traumatic brain injury models. We discuss their relative advantages and drawbacks for the developmental testing of biomaterial-based therapies.
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Krolinski A, Sommer K, Wiesner J, Friedrich O, Vielreicher M. Optimized Method of 3D Scaffold Seeding, Cell Cultivation, and Monitoring Cell Status for Bone Tissue Engineering. Methods Mol Biol 2023; 2644:467-480. [PMID: 37142941 DOI: 10.1007/978-1-0716-3052-5_30] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 05/06/2023]
Abstract
The cultivation of cells in 3D systems is commonly regarded to be more physiological than in 2D as it comes much closer to the natural situation in tissues in many different aspects. However, 3D cell culture is much more complex. Cells within the pores of a printed 3D scaffold face a special situation concerning cell-material interaction and cell adhesion, cell proliferation, and supply of medium and oxygen into the core of the scaffolds. Biological assays (for cell proliferation, viability, and activity) have been validated primarily for 2D cell cultures and need to be adapted for 3D cultures. Likewise, in imaging, a number of points need to be taken into account in order to get a clear picture of the cells in 3D scaffolds, preferably with the method of multiphoton microscopy. Here, we describe a method for pretreatment and cell seeding of porous inorganic composite scaffolds (α-TCP/HA) for bone tissue engineering and for cultivation of the cell-scaffold constructs. The analytical methods described are the cell proliferation assay and the ALP activity assay. A step-by-step protocol is provided here that safely tackles typical difficulties that arise with this 3D cell-scaffold setting. In addition, MPM imaging of cells is described both with and without labeling. The combination of biochemical assays and imaging provides valuable insights into the possibilities of analysis with this 3D cell-scaffold system.
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Affiliation(s)
- Adrian Krolinski
- Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander University (FAU) Erlangen-Nürnberg, Erlangen, Germany
| | - Kai Sommer
- Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander University (FAU) Erlangen-Nürnberg, Erlangen, Germany
| | - Johanna Wiesner
- Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander University (FAU) Erlangen-Nürnberg, Erlangen, Germany
| | - Oliver Friedrich
- Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander University (FAU) Erlangen-Nürnberg, Erlangen, Germany
| | - Martin Vielreicher
- Institute of Medical Biotechnology, Department of Chemical and Biological Engineering, Friedrich-Alexander University (FAU) Erlangen-Nürnberg, Erlangen, Germany.
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Rojas-Rojas L, Espinoza-Álvarez ML, Castro-Piedra S, Ulloa-Fernández A, Vargas-Segura W, Guillén-Girón T. Muscle-like Scaffolds for Biomechanical Stimulation in a Custom-Built Bioreactor. Polymers (Basel) 2022; 14:polym14245427. [PMID: 36559794 PMCID: PMC9781371 DOI: 10.3390/polym14245427] [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: 10/14/2022] [Revised: 11/21/2022] [Accepted: 11/23/2022] [Indexed: 12/14/2022] Open
Abstract
Tissue engineering aims to develop in-vitro substitutes of native tissues. One approach of tissue engineering relies on using bioreactors combined with biomimetic scaffolds to produce study models or in-vitro substitutes. Bioreactors provide control over environmental parameters, place and hold a scaffold under desired characteristics, and apply mechanical stimulation to scaffolds. Polymers are often used for fabricating tissue-engineering scaffolds. In this study, polycaprolactone (PCL) collagen-coated microfilament scaffolds were cell-seeded with C2C12 myoblasts; then, these were grown inside a custom-built bioreactor. Cell attachment and proliferation on the scaffolds were investigated. A loading pattern was used for mechanical stimulation of the cell-seeded scaffolds. Results showed that the microfilaments provided a suitable scaffold for myoblast anchorage and that the custom-built bioreactor provided a qualified environment for the survival of the myoblasts on the polymeric scaffold. This PCL-based microfilament scaffold located inside the bioreactor proved to be a promising structure for the study of skeletal muscle models and can be used for mechanical stimulation studies in tissue engineering applications.
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Affiliation(s)
- Laura Rojas-Rojas
- Materials Science and Engineering School, Instituto Tecnológico de Costa Rica, Cartago 30101, Costa Rica
- Physics School, Instituto Tecnológico de Costa Rica, Cartago 30101, Costa Rica
- Correspondence: ; Tel.: +506-25502284
| | - María Laura Espinoza-Álvarez
- Materials Science and Engineering School, Instituto Tecnológico de Costa Rica, Cartago 30101, Costa Rica
- Biology School, Instituto Tecnológico de Costa Rica, Cartago 30101, Costa Rica
| | | | | | | | - Teodolito Guillén-Girón
- Materials Science and Engineering School, Instituto Tecnológico de Costa Rica, Cartago 30101, Costa Rica
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Immohr MB, Dos Santos Adrego F, Teichert HL, Schmidt V, Sugimura Y, Bauer S, Barth M, Lichtenberg A, Akhyari P. 3D-bioprinting of aortic valve interstitial cells: impact of hydrogel and printing parameters on cell viability. Biomed Mater 2022; 18. [PMID: 36322974 DOI: 10.1088/1748-605x/ac9f91] [Citation(s) in RCA: 6] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/05/2022] [Accepted: 11/02/2022] [Indexed: 11/05/2022]
Abstract
Calcific aortic valve disease (CAVD) is a frequent cardiac pathology in the aging society. Although valvular interstitial cells (VICs) seem to play a crucial role, mechanisms of CAVD are not fully understood. Development of tissue-engineered cellular models by 3D-bioprinting may help to further investigate underlying mechanisms of CAVD. VIC were isolated from ovine aortic valves and cultured in Dulbecco's modified Eagle's Medium (DMEM). VIC of passages six to ten were dissolved in a hydrogel consisting of 2% alginate and 8% gelatin with a concentration of 2 × 106VIC ml-1. Cell-free and VIC-laden hydrogels were printed with an extrusion-based 3D-bioprinter (3D-Bioplotter®Developer Series, EnvisionTec, Gladbeck, Germany), cross-linked and incubated for up to 28 d. Accuracy and durability of scaffolds was examined by microscopy and cell viability was tested by cell counting kit-8 assay and live/dead staining. 3D-bioprinting of scaffolds was most accurate with a printing pressure ofP< 400 hPa, nozzle speed ofv< 20 mm s-1, hydrogel temperature ofTH= 37 °C and platform temperature ofTP= 5 °C in a 90° parallel line as well as in a honeycomb pattern. Dissolving the hydrogel components in DMEM increased VIC viability on day 21 by 2.5-fold compared to regular 0.5% saline-based hydrogels (p< 0.01). Examination at day 7 revealed dividing and proliferating cells. After 21 d the entire printed scaffolds were filled with proliferating cells. Live/dead cell viability/cytotoxicity staining confirmed beneficial effects of DMEM-based cell-laden VIC hydrogel scaffolds even 28 d after printing. By using low pressure printing methods, we were able to successfully culture cell-laden 3D-bioprinted VIC scaffolds for up to 28 d. Using DMEM-based hydrogels can significantly improve the long-term cell viability and overcome printing-related cell damage. Therefore, future applications 3D-bioprinting of VIC might enable the development of novel tissue engineered cellular 3D-models to examine mechanisms involved in initiation and progression of CAVD.
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Affiliation(s)
- Moritz Benjamin Immohr
- Department of Cardiac Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany
| | - Fabió Dos Santos Adrego
- Department of Cardiac Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany
| | - Helena Lauren Teichert
- Department of Cardiac Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany
| | - Vera Schmidt
- Department of Cardiac Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany
| | - Yukiharu Sugimura
- Department of Cardiac Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany
| | - Sebastian Bauer
- Department of Cardiac Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany
| | - Mareike Barth
- Department of Cardiac Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany
| | - Artur Lichtenberg
- Department of Cardiac Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany.,CARID-Cardiovascular Research Institute Düsseldorf, Heinrich Heine University Düsseldorf, Düsseldorf, Germany
| | - Payam Akhyari
- Department of Cardiac Surgery, Medical Faculty and University Hospital Düsseldorf, Heinrich-Heine-University Düsseldorf, Düsseldorf, Germany
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Fattahi E, Taheri S, Schilling AF, Becker T, Pörtner R. Generation and evaluation of input values for computational analysis of transport processes within tissue cultures. Eng Life Sci 2022; 22:681-698. [PMID: 36348656 PMCID: PMC9635004 DOI: 10.1002/elsc.202100128] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2021] [Revised: 01/27/2022] [Accepted: 02/11/2022] [Indexed: 11/15/2022] Open
Abstract
Techniques for tissue culture have seen significant advances during the last decades and novel 3D cell culture systems have become available. To control their high complexity, experimental techniques and their Digital Twins (modelling and computational tools) are combined to link different variables to process conditions and critical process parameters. This allows a rapid evaluation of the expected product quality. However, the use of mathematical simulation and Digital Twins is critically dependent on the precise description of the problem and correct input parameters. Errors here can lead to dramatically wrong conclusions. The intention of this review is to provide an overview of the state-of-the-art and remaining challenges with respect to generating input values for computational analysis of mass and momentum transport processes within tissue cultures. It gives an overview on relevant aspects of transport processes in tissue cultures as well as modelling and computational tools to tackle these problems. Further focus is on techniques used for the determination of cell-specific parameters and characterization of culture systems, including sensors for on-line determination of relevant parameters. In conclusion, tissue culture techniques are well-established, and modelling tools are technically mature. New sensor technologies are on the way, especially for organ chips. The greatest remaining challenge seems to be the proper addressing and handling of input parameters required for mathematical models. Following Good Modelling Practice approaches when setting up and validating computational models is, therefore, essential to get to better estimations of the interesting complex processes inside organotypic tissue cultures in the future.
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Affiliation(s)
- Ehsan Fattahi
- Chair of Brewing and Beverage TechnologyTUM School of Life SciencesTechnische Universität MünchenFreisingGermany
| | - Shahed Taheri
- Department of Trauma SurgeryOrthopaedics and Plastic SurgeryUniversity Medical Center GöttingenGöttingenGermany
| | - Arndt F. Schilling
- Department of Trauma SurgeryOrthopaedics and Plastic SurgeryUniversity Medical Center GöttingenGöttingenGermany
| | - Thomas Becker
- Chair of Brewing and Beverage TechnologyTUM School of Life SciencesTechnische Universität MünchenFreisingGermany
| | - Ralf Pörtner
- Institute of Bioprocess and Biosystems EngineeringHamburg University of TechnologyHamburgGermany
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Xu H, Su Y, Liao Z, Liu Z, Huang X, Zhao L, Duan R, Hu Y, Wei Y, Lian X, Huang D. Coaxial bioprinting vascular constructs: A review. Eur Polym J 2022. [DOI: 10.1016/j.eurpolymj.2022.111549] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
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Microfluidic Technology for Evaluating and Preserving Islet Function for Islet Transplant in Type 1 Diabetes. CURRENT TRANSPLANTATION REPORTS 2022. [DOI: 10.1007/s40472-022-00377-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/27/2022]
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Aljadi Z, Aval NA, Kumar T, Qin T, Ramachandraiah H, Pettersson T, Russom A. Layer-by-Layer Cellulose Nanofibrils: A New Coating Strategy for Development and Characterization of Tumor Spheroids as a Model for In-Vitro Anti-Cancer Drug Screening. Macromol Biosci 2022; 22:e2200137. [PMID: 35899862 DOI: 10.1002/mabi.202200137] [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: 04/05/2022] [Revised: 07/21/2022] [Indexed: 11/09/2022]
Abstract
Three-dimensional multicellular spheroids (MCSs) are complex structure of cellular aggregates and cell-to-matrix interaction that emulates the in-vivo microenvironment. This research field has progressively grown to develop and improve spheroid generation techniques. Here, we present a new platform for spheroid generation using Layer-by-Layer (LbL) technology. Layer-by-Layer (LbL) containing cellulose nanofibrils (CNF) assemble on a standard 96 well plate. Various LbL assembly parameters, multiple cell seeding concentration, and two tumor cell lines (HEK 293 T, HCT 116) are utilized to generate and characterize spheroids. The number and the proliferation of generated spheroids in correlation to the number of LbL-CNF bi-layers, the viability, and the response to the anti-cancer drug are examined. The spheroids are formed and proliferated on the LbL-CNF coated wells with no significant difference in connection to the number of LbL-CNF bi-layers; however, the number of formed spheroids correlates positively with the cell seeding concentration (122 ± 17) for HCT 116 and (42 ± 8) for HEK 293T cell lines at 700 cells ml-1 . The generated spheroids proliferate progressively up to (309, 663) μm of HCT 116 and HEK 293T cell lines on the 5 bi-layers coated wells respectively overtime with maintaining viability. The (HCT 116) spheroids react to the anti-cancer drug. We demonstrate a new platform (LbL-CNF) coating strategy for spheroids generation, with high performance and efficiency to test anti-cancer drugs. This article is protected by copyright. All rights reserved.
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Affiliation(s)
- Zenib Aljadi
- School of Engineering Sciences in Chemistry, Biotechnology and Health CBH, Division of Nanobiotechnology, Department of Protein Science, Science for Life Laboratory, KTH Royal Institute of Technology, Stockholm, Sweden
| | - Negar Abbasi Aval
- Fibre and Polymer Technology, KTH Royal Institute of Technology, Stockholm, Sweden
| | - Tharagan Kumar
- School of Engineering Sciences in Chemistry, Biotechnology and Health CBH, Division of Nanobiotechnology, Department of Protein Science, Science for Life Laboratory, KTH Royal Institute of Technology, Stockholm, Sweden
| | - Taoyu Qin
- School of Engineering Sciences in Chemistry, Biotechnology and Health CBH, Division of Nanobiotechnology, Department of Protein Science, Science for Life Laboratory, KTH Royal Institute of Technology, Stockholm, Sweden
| | - Harisha Ramachandraiah
- School of Engineering Sciences in Chemistry, Biotechnology and Health CBH, Division of Nanobiotechnology, Department of Protein Science, Science for Life Laboratory, KTH Royal Institute of Technology, Stockholm, Sweden
| | - Torbjörn Pettersson
- Fibre and Polymer Technology, KTH Royal Institute of Technology, Stockholm, Sweden
| | - Aman Russom
- School of Engineering Sciences in Chemistry, Biotechnology and Health CBH, Division of Nanobiotechnology, Department of Protein Science, Science for Life Laboratory, KTH Royal Institute of Technology, Stockholm, Sweden
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Joshi A, Kaur T, Singh N. 3D Bioprinted Alginate-Silk-Based Smart Cell-Instructive Scaffolds for Dual Differentiation of Human Mesenchymal Stem Cells. ACS APPLIED BIO MATERIALS 2022; 5:2870-2879. [PMID: 35679315 DOI: 10.1021/acsabm.2c00251] [Citation(s) in RCA: 13] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
Designing smart bioinks, which can provide multifunctionality and instructive cues to cells, is a current need of the tissue engineering field. Addressing these parameters, this work aims at developing a smart dual 3D bioprinted scaffold that is capable of differentiating human mesenchymal stem cells into two different lineages within the same construct without providing any exogenous cues. Here, biocompatible alginate- and silk-based bioinks were developed to print self-standing structures with the ability of spatially controlled differentiation of the encapsulated hMSCs. We present this proof of concept and have demonstrated a smart design where the incorporation of phosphate groups enhanced the osteogenic differentiation, whereas the addition of silk promoted the chondrogenic differentiation. Altogether, the present work suggests the potential of the developed bioinks for use in creating clinically viable osteochondral grafts.
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Affiliation(s)
- Akshay Joshi
- Centre for Biomedical Engineering, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India
| | - Tejinder Kaur
- Centre for Biomedical Engineering, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India
| | - Neetu Singh
- Centre for Biomedical Engineering, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.,Biomedical Engineering Unit, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India
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Bose A, Petsko GA, Studer L. Induced pluripotent stem cells: a tool for modeling Parkinson's disease. Trends Neurosci 2022; 45:608-620. [PMID: 35667922 PMCID: PMC9576003 DOI: 10.1016/j.tins.2022.05.001] [Citation(s) in RCA: 26] [Impact Index Per Article: 8.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/08/2021] [Revised: 04/04/2022] [Accepted: 05/09/2022] [Indexed: 12/26/2022]
Abstract
Parkinson's disease (PD) is the second most prevalent neurodegenerative disorder. Among its pathologies, progressive loss of dopaminergic (DA) neurons in the substantia nigra is characteristic and contributes to many of the most severe symptoms of PD. Recent advances in induced pluripotent stem cell (iPSC) technology have made it possible to generate patient-derived DA neuronal cell culture and organoid models of PD. These models have contributed to understanding disease mechanisms and the identification of novel targets and therapeutic candidates. Still needed are better ways to model the age-related aspects of PD, as well as a deeper understanding of the interactions among disease-modifying genes and between genetic and environmental contributions to the etiology and progression of PD.
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Affiliation(s)
- Anindita Bose
- Ann Romney Institute of Neurological Diseases, Harvard Medical School/Brigham and Women's Hospital, Boston, MA, USA; The Center for Stem Cell Biology, Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, New York, NY, USA.
| | - Gregory A Petsko
- Ann Romney Institute of Neurological Diseases, Harvard Medical School/Brigham and Women's Hospital, Boston, MA, USA; The Center for Stem Cell Biology, Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, New York, NY, USA
| | - Lorenz Studer
- The Center for Stem Cell Biology, Developmental Biology Program, Sloan-Kettering Institute for Cancer Research, New York, NY, USA
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Limongi T, Guzzi F, Parrotta E, Candeloro P, Scalise S, Lucchino V, Gentile F, Tirinato L, Coluccio ML, Torre B, Allione M, Marini M, Susa F, Fabrizio ED, Cuda G, Perozziello G. Microfluidics for 3D Cell and Tissue Cultures: Microfabricative and Ethical Aspects Updates. Cells 2022; 11:1699. [PMID: 35626736 PMCID: PMC9139493 DOI: 10.3390/cells11101699] [Citation(s) in RCA: 9] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/08/2022] [Revised: 05/16/2022] [Accepted: 05/18/2022] [Indexed: 12/10/2022] Open
Abstract
The necessity to improve in vitro cell screening assays is becoming ever more important. Pharmaceutical companies, research laboratories and hospitals require technologies that help to speed up conventional screening and therapeutic procedures to produce more data in a short time in a realistic and reliable manner. The design of new solutions for test biomaterials and active molecules is one of the urgent problems of preclinical screening and the limited correlation between in vitro and in vivo data remains one of the major issues. The establishment of the most suitable in vitro model provides reduction in times, costs and, last but not least, in the number of animal experiments as recommended by the 3Rs (replace, reduce, refine) ethical guiding principles for testing involving animals. Although two-dimensional (2D) traditional cell screening assays are generally cheap and practical to manage, they have strong limitations, as cells, within the transition from the three-dimensional (3D) in vivo to the 2D in vitro growth conditions, do not properly mimic the real morphologies and physiology of their native tissues. In the study of human pathologies, especially, animal experiments provide data closer to what happens in the target organ or apparatus, but they imply slow and costly procedures and they generally do not fully accomplish the 3Rs recommendations, i.e., the amount of laboratory animals and the stress that they undergo must be minimized. Microfluidic devices seem to offer different advantages in relation to the mentioned issues. This review aims to describe the critical issues connected with the conventional cells culture and screening procedures, showing what happens in the in vivo physiological micro and nano environment also from a physical point of view. During the discussion, some microfluidic tools and their components are described to explain how these devices can circumvent the actual limitations described in the introduction.
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Affiliation(s)
- Tania Limongi
- Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy; (T.L.); (B.T.); (M.A.); (M.M.); (F.S.); (E.D.F.)
| | - Francesco Guzzi
- Nanotechnology Research Centre, BioNEM Laboratory, Department of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy; (F.G.); (P.C.); (F.G.); (L.T.); (M.L.C.)
| | - Elvira Parrotta
- Department of Medical and Surgical Sciences, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy;
| | - Patrizio Candeloro
- Nanotechnology Research Centre, BioNEM Laboratory, Department of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy; (F.G.); (P.C.); (F.G.); (L.T.); (M.L.C.)
| | - Stefania Scalise
- Research Centre for Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, University Magna Graecia, 88100 Catanzaro, Italy; (S.S.); (V.L.); (G.C.)
| | - Valeria Lucchino
- Research Centre for Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, University Magna Graecia, 88100 Catanzaro, Italy; (S.S.); (V.L.); (G.C.)
| | - Francesco Gentile
- Nanotechnology Research Centre, BioNEM Laboratory, Department of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy; (F.G.); (P.C.); (F.G.); (L.T.); (M.L.C.)
| | - Luca Tirinato
- Nanotechnology Research Centre, BioNEM Laboratory, Department of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy; (F.G.); (P.C.); (F.G.); (L.T.); (M.L.C.)
| | - Maria Laura Coluccio
- Nanotechnology Research Centre, BioNEM Laboratory, Department of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy; (F.G.); (P.C.); (F.G.); (L.T.); (M.L.C.)
| | - Bruno Torre
- Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy; (T.L.); (B.T.); (M.A.); (M.M.); (F.S.); (E.D.F.)
| | - Marco Allione
- Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy; (T.L.); (B.T.); (M.A.); (M.M.); (F.S.); (E.D.F.)
| | - Monica Marini
- Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy; (T.L.); (B.T.); (M.A.); (M.M.); (F.S.); (E.D.F.)
| | - Francesca Susa
- Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy; (T.L.); (B.T.); (M.A.); (M.M.); (F.S.); (E.D.F.)
| | - Enzo Di Fabrizio
- Department of Applied Science and Technology, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Turin, Italy; (T.L.); (B.T.); (M.A.); (M.M.); (F.S.); (E.D.F.)
| | - Giovanni Cuda
- Research Centre for Advanced Biochemistry and Molecular Biology, Department of Experimental and Clinical Medicine, University Magna Graecia, 88100 Catanzaro, Italy; (S.S.); (V.L.); (G.C.)
| | - Gerardo Perozziello
- Nanotechnology Research Centre, BioNEM Laboratory, Department of Experimental and Clinical Medicine, University Magna Graecia of Catanzaro, 88100 Catanzaro, Italy; (F.G.); (P.C.); (F.G.); (L.T.); (M.L.C.)
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Rathore RS, R Ayyannan S, Mahto SK. Emerging three-dimensional neuronal culture assays for neurotherapeutics drug discovery. Expert Opin Drug Discov 2022; 17:619-628. [DOI: 10.1080/17460441.2022.2061458] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/18/2022]
Affiliation(s)
- Rahul S Rathore
- Pharmaceutical Chemistry Research Laboratory II, Department of Pharmaceutical Engineering & Technology, Indian Institute of Technology (Banaras Hindu University), Varanasi-221005, UP, India
| | - Senthil R Ayyannan
- Pharmaceutical Chemistry Research Laboratory II, Department of Pharmaceutical Engineering & Technology, Indian Institute of Technology (Banaras Hindu University), Varanasi-221005, UP, India
| | - Sanjeev K Mahto
- Tissue Engineering and Biomicrofluidics Laboratory, School of Biomedical Engineering, Indian Institute of Technology (Banaras Hindu University), Varanasi-221005, UP, India
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Langella A, Gadau SD, Serra E, Bebbere D, Ledda S. Microtubular Assessment of C6 Rat Glioma Cell Spheroids Developed in Transparent Liquid Marbles or Hanging Drops. BIOLOGY 2022; 11:biology11040492. [PMID: 35453692 PMCID: PMC9031767 DOI: 10.3390/biology11040492] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 02/09/2022] [Revised: 03/16/2022] [Accepted: 03/22/2022] [Indexed: 11/16/2022]
Abstract
Glioblastoma is a brain tumour frequently used as an experimental model to exploit innovative therapeutic approaches due to its high lethality and refractoriness to therapies. Part of these innovative anticancer therapies address cytoskeletal microtubules (MTs) since specific tubulin post-translational modifications (PTMs) are considered markers of tumour plasticity. In vitro studies, which traditionally employ two-dimensional (2D) culture systems, are now being replaced by three-dimensional (3D) systems that more closely mimic in vivo physiological conditions and allow a better understanding of the signalling between cells. In this work, we compared 2 liquid base 3D methods for the generation of spheroids from C6 rat glioma cells (RGCs) using 30 µL of liquid marble (LM) or the hanging drops (HDs), which contained 2 different cell numbers (5000 or 15,000). After 24 or 48 h of in vitro culture (IVC), the morphology of the spheroids was observed and the behaviour of the two main tubulin PTMs, tyrosinated α-tubulin (Tyr-T) and acetylated α-tubulin (Ac-T), was evaluated by fluorescence and Western blot (WB). RGCs spontaneously formed spherical agglomerates more rapidly in the LM than in the HD system. Cell density influenced the size of the spheroids, which reached a larger size (> of 300 µm Ø), with 15,000 cells compared to 5000 cells (150 µm Ø). Moreover, an increase in Tyr-T and Ac-T was observed in both the HD and LM system from 24 to 48 h, with the highest values shown in the 48 h/LM spheroids of 5000 cells (p < 0.05). In conclusion, by comparing the morphology and microtubular architecture of spheroids from C6 rat glioma cells developed by LM or HD methodology, our findings demonstrate that the use of a fumed silica microbioreactor boosts the induction and maintenance of a high plasticity state in glioma cells. RGCs cultured in LM express levels of tubulin PTMs that can be used to evaluate the efficacy of new anticancer therapies.
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Li K, Tharwat M, Larson EL, Felgendreff P, Hosseiniasl SM, Rmilah AA, Safwat K, Ross JJ, Nyberg SL. Re-Endothelialization of Decellularized Liver Scaffolds: A Step for Bioengineered Liver Transplantation. Front Bioeng Biotechnol 2022; 10:833163. [PMID: 35360393 PMCID: PMC8960611 DOI: 10.3389/fbioe.2022.833163] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/10/2021] [Accepted: 02/11/2022] [Indexed: 12/12/2022] Open
Abstract
Bioengineered livers (BELs) are an attractive therapeutic alternative to address the donor organ shortage for liver transplantation. The goal of BELs technology aims at replacement or regeneration of the native human liver. A variety of approaches have been proposed for tissue engineering of transplantable livers; the current review will highlight the decellularization-recellularization approach to BELs. For example, vascular patency and appropriate cell distribution and expansion are critical components in the production of successful BELs. Proper solutions to these components of BELs have challenged its development. Several strategies, such as heparin immobilization, heparin-gelatin, REDV peptide, and anti-CD31 aptamer have been developed to extend the vascular patency of revascularized bioengineered livers (rBELs). Other novel methods have been developed to enhance cell seeding of parenchymal cells and to increase graft functionality during both bench and in vivo perfusion. These enhanced methods have been associated with up to 15 days of survival in large animal (porcine) models of heterotopic transplantation but have not yet permitted extended survival after implantation of BELs in the orthotopic position. This review will highlight both the remaining challenges and the potential for clinical application of functional bioengineered grafts.
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Affiliation(s)
- Kewei Li
- Department of Surgery, Mayo Clinic, Rochester, MN, United States
- Department of Pediatric Surgery, West China Hospital of Sichuan University, Chengdu, China
| | - Mohammad Tharwat
- Department of Surgery, Mayo Clinic, Rochester, MN, United States
- General Surgery Department, Faculty of Medicine, Zagazig University, Zagazig, Egypt
| | - Ellen L. Larson
- Department of Surgery, Mayo Clinic, Rochester, MN, United States
| | - Philipp Felgendreff
- Department of Surgery, Mayo Clinic, Rochester, MN, United States
- Department for General, Visceral and Vascular Surgery, University Hospital Jena, Jena, Germany
| | | | - Anan Abu Rmilah
- Department of Surgery, Mayo Clinic, Rochester, MN, United States
| | - Khaled Safwat
- General Surgery Department, Faculty of Medicine, Zagazig University, Zagazig, Egypt
| | | | - Scott L. Nyberg
- Department of Surgery, Mayo Clinic, Rochester, MN, United States
- William J. von Liebig Center for Transplantation and Clinical Regeneration, Mayo Clinic, Rochester, MN, United States
- *Correspondence: Scott L. Nyberg,
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Marei I, Abu Samaan T, Al-Quradaghi MA, Farah AA, Mahmud SH, Ding H, Triggle CR. 3D Tissue-Engineered Vascular Drug Screening Platforms: Promise and Considerations. Front Cardiovasc Med 2022; 9:847554. [PMID: 35310996 PMCID: PMC8931492 DOI: 10.3389/fcvm.2022.847554] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/02/2022] [Accepted: 02/03/2022] [Indexed: 12/12/2022] Open
Abstract
Despite the efforts devoted to drug discovery and development, the number of new drug approvals have been decreasing. Specifically, cardiovascular developments have been showing amongst the lowest levels of approvals. In addition, concerns over the adverse effects of drugs to the cardiovascular system have been increasing and resulting in failure at the preclinical level as well as withdrawal of drugs post-marketing. Besides factors such as the increased cost of clinical trials and increases in the requirements and the complexity of the regulatory processes, there is also a gap between the currently existing pre-clinical screening methods and the clinical studies in humans. This gap is mainly caused by the lack of complexity in the currently used 2D cell culture-based screening systems, which do not accurately reflect human physiological conditions. Cell-based drug screening is widely accepted and extensively used and can provide an initial indication of the drugs' therapeutic efficacy and potential cytotoxicity. However, in vitro cell-based evaluation could in many instances provide contradictory findings to the in vivo testing in animal models and clinical trials. This drawback is related to the failure of these 2D cell culture systems to recapitulate the human physiological microenvironment in which the cells reside. In the body, cells reside within a complex physiological setting, where they interact with and respond to neighboring cells, extracellular matrix, mechanical stress, blood shear stress, and many other factors. These factors in sum affect the cellular response and the specific pathways that regulate variable vital functions such as proliferation, apoptosis, and differentiation. Although pre-clinical in vivo animal models provide this level of complexity, cross species differences can also cause contradictory results from that seen when the drug enters clinical trials. Thus, there is a need to better mimic human physiological conditions in pre-clinical studies to improve the efficiency of drug screening. A novel approach is to develop 3D tissue engineered miniaturized constructs in vitro that are based on human cells. In this review, we discuss the factors that should be considered to produce a successful vascular construct that is derived from human cells and is both reliable and reproducible.
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Affiliation(s)
- Isra Marei
- Department of Pharmacology, Weill Cornell Medicine-Qatar, Doha, Qatar
- National Heart and Lung Institute, Imperial College London, London, United Kingdom
- *Correspondence: Isra Marei
| | - Tala Abu Samaan
- Department of Pharmacology, Weill Cornell Medicine-Qatar, Doha, Qatar
| | | | - Asmaa A. Farah
- Department of Pharmacology, Weill Cornell Medicine-Qatar, Doha, Qatar
| | | | - Hong Ding
- Department of Pharmacology, Weill Cornell Medicine-Qatar, Doha, Qatar
| | - Chris R. Triggle
- Department of Pharmacology, Weill Cornell Medicine-Qatar, Doha, Qatar
- Chris R. Triggle
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Development of Intracorporeal Differentiation of Stem Cells to Induce One-Step Mastoid Bone Reconstruction during Otitis Media Surgeries. Polymers (Basel) 2022; 14:polym14050877. [PMID: 35267699 PMCID: PMC8912861 DOI: 10.3390/polym14050877] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/24/2022] [Revised: 02/20/2022] [Accepted: 02/20/2022] [Indexed: 11/21/2022] Open
Abstract
Mastoidectomy is a surgical procedure for the treatment of chronic otitis media. This study investigated the ability of rat stromal vascular fraction cells (rSVF) in combination with polycaprolactone (PCL) scaffolds and osteogenic differentiation-enhancing blood products to promote the regeneration of mastoid bone defect. Twenty male Sprague Dawley rats were randomly divided according to obliteration materials: (1) control, (2) PCL scaffold only, (3) rSVFs + PCL, (4) rSVFs + PCL + platelet-rich plasma, and (5) rSVFs + PCL + whole plasma (WP). At 7 months after transplantation, the rSVFs + PCL + WP group showed remarkable new bone formation in the mastoid. These results indicate that SVFs, PCL scaffolds, and blood products accelerate bone regeneration for mastoid reconstruction. Autologous SVF cells with PCL scaffolds and autologous blood products are promising composites for mastoid reconstruction which can be easily harvested after mastoidectomy. With this approach, the reconstruction of mastoid bone defects can be performed right after mastoidectomy as a one-step procedure which can offer efficiency in the clinical field.
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Truong TA, Nguyen TK, Zhao H, Nguyen NK, Dinh T, Park Y, Nguyen T, Yamauchi Y, Nguyen NT, Phan HP. Engineering Stress in Thin Films: An Innovative Pathway Toward 3D Micro and Nanosystems. SMALL (WEINHEIM AN DER BERGSTRASSE, GERMANY) 2022; 18:e2105748. [PMID: 34874620 DOI: 10.1002/smll.202105748] [Citation(s) in RCA: 8] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/20/2021] [Revised: 10/23/2021] [Indexed: 06/13/2023]
Abstract
Transformation of conventional 2D platforms into unusual 3D configurations provides exciting opportunities for sensors, electronics, optical devices, and biological systems. Engineering material properties or controlling and modulating stresses in thin films to pop-up 3D structures out of standard planar surfaces has been a highly active research topic over the last decade. Implementation of 3D micro and nanoarchitectures enables unprecedented functionalities including multiplexed, monolithic mechanical sensors, vertical integration of electronics components, and recording of neuron activities in 3D organoids. This paper provides an overview on stress engineering approaches to developing 3D functional microsystems. The paper systematically presents the origin of stresses generated in thin films and methods to transform a 2D design into an out-of-plane configuration. Different types of 3D micro and nanostructures, along with their applications in several areas are discussed. The paper concludes with current technical challenges and potential approaches and applications of this fast-growing research direction.
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Affiliation(s)
- Thanh-An Truong
- Queensland Micro and Nanotechnology Centre, Griffith University, Nathan, Queensland, 4111, Australia
| | - Tuan-Khoa Nguyen
- Queensland Micro and Nanotechnology Centre, Griffith University, Nathan, Queensland, 4111, Australia
| | - Hangbo Zhao
- Department of Aerospace and Mechanical Engineering, University of Southern California, Los Angeles, CA, 90089, USA
| | - Nhat-Khuong Nguyen
- Queensland Micro and Nanotechnology Centre, Griffith University, Nathan, Queensland, 4111, Australia
| | - Toan Dinh
- Centre for Future Materials, University of Southern Queensland, Ipswich, Queensland, 4305, Australia
| | - Yoonseok Park
- Querrey Simpson Institute for Bioelectronics, Northwestern University, Evanston, IL, 60208, USA
| | - Thanh Nguyen
- Centre for Future Materials, University of Southern Queensland, Ipswich, Queensland, 4305, Australia
| | - Yusuke Yamauchi
- Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland, 4072, Australia
| | - Nam-Trung Nguyen
- Queensland Micro and Nanotechnology Centre, Griffith University, Nathan, Queensland, 4111, Australia
| | - Hoang-Phuong Phan
- Queensland Micro and Nanotechnology Centre, Griffith University, Nathan, Queensland, 4111, Australia
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OUP accepted manuscript. J Pharm Pharmacol 2022; 74:1282-1295. [DOI: 10.1093/jpp/rgac017] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/28/2021] [Accepted: 02/23/2022] [Indexed: 11/13/2022]
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Drug-Induced Liver Injury: Clinical Evidence of N-Acetyl Cysteine Protective Effects. OXIDATIVE MEDICINE AND CELLULAR LONGEVITY 2021; 2021:3320325. [PMID: 34912495 PMCID: PMC8668310 DOI: 10.1155/2021/3320325] [Citation(s) in RCA: 32] [Impact Index Per Article: 8.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 08/12/2021] [Revised: 11/12/2021] [Accepted: 11/22/2021] [Indexed: 12/29/2022]
Abstract
Oxidative stress is a key pathological feature implicated in both acute and chronic liver diseases, including drug-induced liver injury (DILI). The latter describes hepatic injury arising as a direct toxic effect of administered drugs or their metabolites. Although still underreported, DILI remains a significant cause of liver failure, especially in developed nations. Currently, it is understood that mitochondrial-generated oxidative stress and abnormalities in phase I/II metabolism, leading to glutathione (GSH) suppression, drive the onset of DILI. N-Acetyl cysteine (NAC) has attracted a lot of interest as a therapeutic agent against DILI because of its strong antioxidant properties, especially in relation to enhancing endogenous GSH content to counteract oxidative stress. Thus, in addition to updating information on the pathophysiological mechanisms implicated in oxidative-induced hepatic injury, the current review critically discusses clinical evidence on the protective effects of NAC against DILI, including the reduction of patient mortality. Besides injury caused by paracetamol, NAC can also improve liver function in relation to other forms of liver injury such as those induced by excessive alcohol intake. The implicated therapeutic mechanisms of NAC extend from enhancing hepatic GSH levels to reducing biomarkers of paracetamol toxicity such as keratin-18 and circulating caspase-cleaved cytokeratin-18. However, there is still lack of evidence confirming the benefits of using NAC in combination with other therapies in patients with DILI.
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Bedford R, Perkins E, Clements J, Hollings M. Recent advancements and application of in vitro models for predicting inhalation toxicity in humans. Toxicol In Vitro 2021; 79:105299. [PMID: 34920082 DOI: 10.1016/j.tiv.2021.105299] [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: 06/22/2021] [Revised: 11/20/2021] [Accepted: 12/10/2021] [Indexed: 12/01/2022]
Abstract
Animals have been indispensable in testing chemicals that can pose a risk to human health, including those delivered by inhalation. In recent years, the combination of societal debate on the use of animals in research and testing, the drive to continually enhance testing methodologies, and technology advancements have prompted a range of initiatives to develop non-animal alternative approaches for toxicity testing. In this review, we discuss emerging in vitro techniques being developed for the testing of inhaled compounds. Advanced tissue models that are able to recreate the human response to toxic exposures alongside examples of their ability to complement in vivo techniques are described. Furthermore, technology being developed that can provide multi-organ toxicity assessments are discussed.
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Affiliation(s)
- R Bedford
- Labcorp Early Development Laboratories Limited, Harrogate, UK.
| | - E Perkins
- Labcorp Early Development Laboratories Limited, Harrogate, UK.
| | - J Clements
- Labcorp Early Development Laboratories Limited, Harrogate, UK.
| | - M Hollings
- Labcorp Early Development Laboratories Limited, Harrogate, UK.
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Gao Q, Kim BS, Gao G. Advanced Strategies for 3D Bioprinting of Tissue and Organ Analogs Using Alginate Hydrogel Bioinks. Mar Drugs 2021; 19:708. [PMID: 34940707 PMCID: PMC8708555 DOI: 10.3390/md19120708] [Citation(s) in RCA: 44] [Impact Index Per Article: 11.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/30/2021] [Revised: 12/12/2021] [Accepted: 12/12/2021] [Indexed: 12/15/2022] Open
Abstract
Alginate is a natural polysaccharide that typically originates from various species of algae. Due to its low cost, good biocompatibility, and rapid ionic gelation, the alginate hydrogel has become a good option of bioink source for 3D bioprinting. However, the lack of cell adhesive moieties, erratic biodegradability, and poor printability are the critical limitations of alginate hydrogel bioink. This review discusses the pivotal properties of alginate hydrogel as a bioink for 3D bioprinting technologies. Afterward, a variety of advanced material formulations and biofabrication strategies that have recently been developed to overcome the drawbacks of alginate hydrogel bioink will be focused on. In addition, the applications of these advanced solutions for 3D bioprinting of tissue/organ mimicries such as regenerative implants and in vitro tissue models using alginate-based bioink will be systematically summarized.
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Affiliation(s)
- Qiqi Gao
- Institute of Engineering Medicine, Beijing Institute of Technology, No. 5, South Street, Zhongguancun, Haidian District, Beijing 100081, China;
| | - Byoung-Soo Kim
- School of Biomedical Convergence Engineering, Pusan National University, Yangsan 626841, Kyungnam, Korea;
| | - Ge Gao
- Institute of Engineering Medicine, Beijing Institute of Technology, No. 5, South Street, Zhongguancun, Haidian District, Beijing 100081, China;
- Department of Medical Technology, Beijing Institute of Technology, No. 5, South Street, Zhongguancun, Haidian District, Beijing 100081, China
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From Spheroids to Organoids: The Next Generation of Model Systems of Human Cardiac Regeneration in a Dish. Int J Mol Sci 2021; 22:ijms222413180. [PMID: 34947977 PMCID: PMC8708686 DOI: 10.3390/ijms222413180] [Citation(s) in RCA: 35] [Impact Index Per Article: 8.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/08/2021] [Revised: 12/02/2021] [Accepted: 12/05/2021] [Indexed: 12/12/2022] Open
Abstract
Organoids are tiny, self-organized, three-dimensional tissue cultures that are derived from the differentiation of stem cells. The growing interest in the use of organoids arises from their ability to mimic the biology and physiology of specific tissue structures in vitro. Organoids indeed represent promising systems for the in vitro modeling of tissue morphogenesis and organogenesis, regenerative medicine and tissue engineering, drug therapy testing, toxicology screening, and disease modeling. Although 2D cell cultures have been used for more than 50 years, even for their simplicity and low-cost maintenance, recent years have witnessed a steep rise in the availability of organoid model systems. Exploiting the ability of cells to re-aggregate and reconstruct the original architecture of an organ makes it possible to overcome many limitations of 2D cell culture systems. In vitro replication of the cellular micro-environment of a specific tissue leads to reproducing the molecular, biochemical, and biomechanical mechanisms that directly influence cell behavior and fate within that specific tissue. Lineage-specific self-organizing organoids have now been generated for many organs. Currently, growing cardiac organoid (cardioids) from pluripotent stem cells and cardiac stem/progenitor cells remains an open challenge due to the complexity of the spreading, differentiation, and migration of cardiac muscle and vascular layers. Here, we summarize the evolution of biological model systems from the generation of 2D spheroids to 3D organoids by focusing on the generation of cardioids based on the currently available laboratory technologies and outline their high potential for cardiovascular research.
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Dituri F, Centonze M, Berenschot EJW, Tas NR, Susarrey-Arce A, Krol S. Complex Tumor Spheroid Formation and One-Step Cancer-Associated Fibroblasts Purification from Hepatocellular Carcinoma Tissue Promoted by Inorganic Surface Topography. NANOMATERIALS (BASEL, SWITZERLAND) 2021; 11:3233. [PMID: 34947582 PMCID: PMC8706479 DOI: 10.3390/nano11123233] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 10/20/2021] [Revised: 11/24/2021] [Accepted: 11/26/2021] [Indexed: 12/12/2022]
Abstract
In vitro cell models play important roles as testbeds for toxicity studies, drug development, or as replacements in animal experiments. In particular, complex tumor models such as hepatocellular carcinoma (HCC) are needed to predict drug efficacy and facilitate translation into clinical practice. In this work, topographical features of amorphous silicon dioxide (SiO2) are fabricated and tested for cell culture of primary HCC cells and cell lines. The topographies vary from pyramids to octahedrons to structures named fractals, with increased hierarchy and organized in periodic arrays (square or Hexagonal). The pyramids were found to promote complex 2D/3D tissue formation from primary HCC cells. It was found that the 2D layer was mainly composed of cancer-associated fibroblasts (CAFs), while the 3D spheroids were composed of tumor cells enwrapped by a CAF layer. Compared with conventional protocols for 3D cultures, this novel approach mimics the 2D/3D complexity of the original tumor by invading CAFs and a microtumor. Topographies such as octahedrons and fractals exclude tumor cells and allow one-step isolation of CAFs even directly from tumor tissue of patients as the CAFs migrate into the structured substrate. Cell lines form spheroids within a short time. The presented inorganic topographical surfaces stimulate complex spheroid formation while avoiding additional biological scaffolds and allowing direct visualization on the substrate.
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Affiliation(s)
- Francesco Dituri
- Laboratory for Personalized Medicine, National Institute of Gastroenterology, “S. de Bellis” Research Hospital, Castellana Grotte Via Turi 27, 70013 Bari, Italy; (F.D.); (M.C.)
| | - Matteo Centonze
- Laboratory for Personalized Medicine, National Institute of Gastroenterology, “S. de Bellis” Research Hospital, Castellana Grotte Via Turi 27, 70013 Bari, Italy; (F.D.); (M.C.)
| | - Erwin J. W. Berenschot
- Mesoscale Chemical Systems, MESA+ Institute, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands; (E.J.W.B.); (N.R.T.)
| | - Niels R. Tas
- Mesoscale Chemical Systems, MESA+ Institute, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands; (E.J.W.B.); (N.R.T.)
| | - Arturo Susarrey-Arce
- Mesoscale Chemical Systems, MESA+ Institute, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands; (E.J.W.B.); (N.R.T.)
| | - Silke Krol
- Laboratory for Personalized Medicine, National Institute of Gastroenterology, “S. de Bellis” Research Hospital, Castellana Grotte Via Turi 27, 70013 Bari, Italy; (F.D.); (M.C.)
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Polat A, Göktürk D. An alternative approach to tracing the volumic proliferation development of an entire tumor spheroid in 3D through a mini-Opto tomography platform. Micron 2021; 152:103173. [PMID: 34785434 DOI: 10.1016/j.micron.2021.103173] [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: 08/10/2021] [Revised: 10/21/2021] [Accepted: 10/28/2021] [Indexed: 11/17/2022]
Abstract
Microscopy, which is listed among the major in-situ imaging applications, allows to derive information from a biological sample on the existing architectural structures of cells and tissues and their changes over time. Large biological samples such as tumor spheroids cannot be imaged within one field of view, regional imaging in different areas and subsequent stitching are required to attain the full picture. Microscopy is not typically used to produce full-size visualization of tumor spheroids measuring a few millimeters in size. In this study, we propose a 3D volume imaging technique for tracing the growth of an entire tumor spheroid measuring up to 10 mm using a miniaturized optical (mini-Opto) tomography platform. We performed a primary analysis of the 3D imaging for the MIA PaCa-2 pancreatic tumoroid employing its 2D images produced with the mini-Opto tomography from different angles ranging from -25 ° to +25 ° at six different three-day-apart time points of consecutive image acquisition. These 2D images were reconstructed by using a 3D image reconstruction algorithm that we developed based on the algebraic reconstruction technique (ART). We were able to reconstruct the 3D images of the tumoroid to achieve 800 × 800-pixel 50-layer images at resolutions of 5-25 μm. We also created its 3D visuals to understand more clearly how its volume changed and how it looked over weeks. The volume of the tumor was calculated to be 6.761 mm3 at the first imaging time point and 46.899 mm3 15 days after the first (at the sixth time point), which is 6.94 times larger in volume. The mini-Opto tomography can be considered more advantageous than commercial microscopy because it is portable, more cost-effective, and easier to use, and enables full-size visualization of biological samples measuring a few millimeters in size.
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Affiliation(s)
- Adem Polat
- Çanakkale Onsekiz Mart University, Faculty of Engineering, Department of Electronics Engineering, 17100, Çanakkale, Turkey.
| | - Dilek Göktürk
- Adana Alparslan Türkeş Science and Technology University, Faculty of Engineering, Department of Bioengineering, 01250, Adana, Turkey
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Habanjar O, Diab-Assaf M, Caldefie-Chezet F, Delort L. 3D Cell Culture Systems: Tumor Application, Advantages, and Disadvantages. Int J Mol Sci 2021; 22:12200. [PMID: 34830082 PMCID: PMC8618305 DOI: 10.3390/ijms222212200] [Citation(s) in RCA: 251] [Impact Index Per Article: 62.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/27/2021] [Revised: 11/05/2021] [Accepted: 11/07/2021] [Indexed: 01/09/2023] Open
Abstract
The traditional two-dimensional (2D) in vitro cell culture system (on a flat support) has long been used in cancer research. However, this system cannot be fully translated into clinical trials to ideally represent physiological conditions. This culture cannot mimic the natural tumor microenvironment due to the lack of cellular communication (cell-cell) and interaction (cell-cell and cell-matrix). To overcome these limitations, three-dimensional (3D) culture systems are increasingly developed in research and have become essential for tumor research, tissue engineering, and basic biology research. 3D culture has received much attention in the field of biomedicine due to its ability to mimic tissue structure and function. The 3D matrix presents a highly dynamic framework where its components are deposited, degraded, or modified to delineate functions and provide a platform where cells attach to perform their specific functions, including adhesion, proliferation, communication, and apoptosis. So far, various types of models belong to this culture: either the culture based on natural or synthetic adherent matrices used to design 3D scaffolds as biomaterials to form a 3D matrix or based on non-adherent and/or matrix-free matrices to form the spheroids. In this review, we first summarize a comparison between 2D and 3D cultures. Then, we focus on the different components of the natural extracellular matrix that can be used as supports in 3D culture. Then we detail different types of natural supports such as matrigel, hydrogels, hard supports, and different synthetic strategies of 3D matrices such as lyophilization, electrospiding, stereolithography, microfluid by citing the advantages and disadvantages of each of them. Finally, we summarize the different methods of generating normal and tumor spheroids, citing their respective advantages and disadvantages in order to obtain an ideal 3D model (matrix) that retains the following characteristics: better biocompatibility, good mechanical properties corresponding to the tumor tissue, degradability, controllable microstructure and chemical components like the tumor tissue, favorable nutrient exchange and easy separation of the cells from the matrix.
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Affiliation(s)
- Ola Habanjar
- Université Clermont-Auvergne, INRAE, UNH, Unité de Nutrition Humaine, CRNH-Auvergne, 63000 Clermont-Ferrand, France; (O.H.); (F.C.-C.)
| | - Mona Diab-Assaf
- Equipe Tumorigénèse Pharmacologie Moléculaire et Anticancéreuse, Faculté des Sciences II, Université Libanaise Fanar, Beyrouth 1500, Liban;
| | - Florence Caldefie-Chezet
- Université Clermont-Auvergne, INRAE, UNH, Unité de Nutrition Humaine, CRNH-Auvergne, 63000 Clermont-Ferrand, France; (O.H.); (F.C.-C.)
| | - Laetitia Delort
- Université Clermont-Auvergne, INRAE, UNH, Unité de Nutrition Humaine, CRNH-Auvergne, 63000 Clermont-Ferrand, France; (O.H.); (F.C.-C.)
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50
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Vernazza S, Tirendi S, Passalacqua M, Piacente F, Scarfì S, Oddone F, Bassi AM. An Innovative In Vitro Open-Angle Glaucoma Model (IVOM) Shows Changes Induced by Increased Ocular Pressure and Oxidative Stress. Int J Mol Sci 2021; 22:12129. [PMID: 34830007 PMCID: PMC8622817 DOI: 10.3390/ijms222212129] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/24/2021] [Revised: 10/26/2021] [Accepted: 11/08/2021] [Indexed: 02/07/2023] Open
Abstract
Primary Open-Angle Glaucoma (POAG) is a neurodegenerative disease, and its clinical outcomes lead to visual field constriction and blindness. POAG's etiology is very complex and its pathogenesis is mainly explained through both mechanical and vascular theories. The trabecular meshwork (TM), the most sensitive tissue of the eye anterior segment to oxidative stress (OS), is the main tissue involved in early-stage POAG, characterized by an increase in pressure. Preclinical assessments of neuroprotective drugs on animal models have not always shown correspondence with human clinical studies. In addition, intra-ocular pressure management after a glaucoma diagnosis does not always prevent blindness. Recently, we have been developing an innovative in vitro 3Dadvanced human trabecular cell model on a millifluidicplatform as a tool to improve glaucoma studies. Herein, we analyze the effects of prolonged increased pressure alone and, in association with OS, on such in vitro platform. Moreover, we verify whethersuch damaged TM triggers apoptosis on neuron-like cells. The preliminary results show that TM cells are less sensitive to pressure elevation than OS, and OS-damaging effects were worsened by the pressure increase. The stressed TM releases harmful signals, which increase apoptosis stimuli on neuron-like cells, suggesting its pivotal role in the glaucoma cascade.
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Affiliation(s)
- Stefania Vernazza
- Department of Experimental Medicine (DIMES), University of Genoa, 16132 Genoa, Italy; (S.V.); (S.T.); (F.P.); (A.M.B.)
| | - Sara Tirendi
- Department of Experimental Medicine (DIMES), University of Genoa, 16132 Genoa, Italy; (S.V.); (S.T.); (F.P.); (A.M.B.)
- Inter-University Center for the Promotion of the 3Rs Principles in Teaching & Research (Centro 3R), 56122 Pisa, Italy;
| | - Mario Passalacqua
- Department of Experimental Medicine (DIMES), University of Genoa, 16132 Genoa, Italy; (S.V.); (S.T.); (F.P.); (A.M.B.)
- Inter-University Center for the Promotion of the 3Rs Principles in Teaching & Research (Centro 3R), 56122 Pisa, Italy;
| | - Francesco Piacente
- Department of Experimental Medicine (DIMES), University of Genoa, 16132 Genoa, Italy; (S.V.); (S.T.); (F.P.); (A.M.B.)
| | - Sonia Scarfì
- Inter-University Center for the Promotion of the 3Rs Principles in Teaching & Research (Centro 3R), 56122 Pisa, Italy;
- Department of Earth, Environmental and Life Sciences (DISTAV), University of Genoa, 16132 Genoa, Italy
| | | | - Anna Maria Bassi
- Department of Experimental Medicine (DIMES), University of Genoa, 16132 Genoa, Italy; (S.V.); (S.T.); (F.P.); (A.M.B.)
- Inter-University Center for the Promotion of the 3Rs Principles in Teaching & Research (Centro 3R), 56122 Pisa, Italy;
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