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Naumann J, Singer K, Shukla S, Maurya A, Schlichter S, Szenti I, Kukovecz A, Rawal A, Zink M. Sustainable Nonwoven Scaffolds Engineered with Recycled Carbon Fiber for Enhanced Biocompatibility and Cell Interaction: From Waste to Health. ACS APPLIED BIO MATERIALS 2025; 8:1984-1996. [PMID: 39960631 PMCID: PMC11921018 DOI: 10.1021/acsabm.4c01475] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/09/2024] [Revised: 01/13/2025] [Accepted: 02/11/2025] [Indexed: 03/18/2025]
Abstract
Carbon fibers, driven by ever-increasing demand, are contributing to a continuous rise in the generation of waste and byproducts destined for landfills or incineration. Recycling carbon fibers presents a promising strategy for reducing carbon emissions and conserving resources, thus contributing to more sustainable waste management practices. Discovering applications of recycled carbon fibers (rCFs) would inevitably accelerate the targeted integration of sustainable materials, fostering a circular economy. Herein, we have engineered rCF-based needlepunched nonwoven scaffolds and their blends with polypropylene (PP) fibers, providing the first example of investigating their interactions with human lung epithelial cells (Calu-3) and murine fibroblast cells (NIH/3T3). To promote the adsorption of extracellular matrix proteins such as laminin, these three-dimensional (3D) nonwoven scaffolds are designed and developed to feature tunable porous characteristics and wetting properties. Although cell adhesion and laminin adsorption are minimal on PP fibers, cells are preferentially organized on the rCFs. These nonwovens, composed exclusively of rCFs or their blends with PP fibers, exhibit no cytotoxic effects, with both cell types showing proliferation on the scaffolds and a progressive increase in cell numbers over time. Cell viability and apoptosis assays are also employed to comprehensively evaluate biocompatibility. Thus, our study proves rCF-based nonwoven scaffolds as potential candidates for artificial lung tissue scaffolds.
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Affiliation(s)
- Jonas Naumann
- Research
Group Biotechnology and Biomedicine, Peter-Debye-Institute for Soft
Matter Physics, Leipzig University, Linnéstraße 5, 04103 Leipzig, Germany
| | - Kresten Singer
- Research
Group Biotechnology and Biomedicine, Peter-Debye-Institute for Soft
Matter Physics, Leipzig University, Linnéstraße 5, 04103 Leipzig, Germany
| | - Siddharth Shukla
- Department
of Textile and Fibre Engineering, Indian
Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India
| | - Alok Maurya
- Department
of Textile and Fibre Engineering, Indian
Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India
| | - Stefan Schlichter
- Faculty
of Mechanical and Process Engineering, Makers labs Recycling &
AI, Technische Hochschule Augsburg, University
of Applied Sciences, An der Hochschule 1, 86161 Augsburg, Germany
| | - Imre Szenti
- Interdisciplinary
Excellence Centre, Department of Applied and Environmental Chemistry, University of Szeged, Rerrich Béla tér 1., 6720 Szeged, Hungary
| | - Akos Kukovecz
- Interdisciplinary
Excellence Centre, Department of Applied and Environmental Chemistry, University of Szeged, Rerrich Béla tér 1., 6720 Szeged, Hungary
| | - Amit Rawal
- Department
of Textile and Fibre Engineering, Indian
Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India
| | - Mareike Zink
- Research
Group Biotechnology and Biomedicine, Peter-Debye-Institute for Soft
Matter Physics, Leipzig University, Linnéstraße 5, 04103 Leipzig, Germany
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2
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Esmaeili J, Barati A, Charelli LE. Discussing the final size and shape of the reconstructed tissues in tissue engineering. J Artif Organs 2022:10.1007/s10047-022-01360-1. [PMID: 36125581 DOI: 10.1007/s10047-022-01360-1] [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] [Accepted: 08/15/2022] [Indexed: 11/30/2022]
Abstract
Tissue engineering (TE) has made a revolution in repairing, replacing, or regenerating tissues or organs, but it has still a long way ahead. The mechanical properties along with suitable physicochemical and biological characteristics are the initial criteria for scaffolds in TE that should be fulfilled. This research will provide another point of view toward TE challenges concerning the morphological and geometrical aspects of the reconstructed tissue and which parameters may affect it. Based on our survey, there is a high possibility that the final reconstructed tissue may be different in size and shape compared to the original design scaffold. Thereby, the 3D-printed scaffold might not guarantee an accurate tissue reconstruction. The main justification for this is the unpredicted behavior of cells, specifically in the outer layer of the scaffold. It can also be a concern when the scaffold is implanted while cell migration cannot be controlled through the in vivo signaling pathways, which might cause cancer challenges. To sum up, it is concluded that more studies are necessary to focus on the size and geometry of the final reconstructed tissue.
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Affiliation(s)
- Javad Esmaeili
- Department of Chemical Engineering, Faculty of Engineering, Arak University, Arak, 38156-88349, Iran.,Tissue Engineering Department, TISSUEHUB Co., Tehran, Iran
| | - Aboulfazl Barati
- Department of Chemical Engineering, Faculty of Engineering, Arak University, Arak, 38156-88349, Iran.
| | - Letícia Emiliano Charelli
- Nanotechnology Engineering Program, Alberto Luiz Coimbra Institute for Graduate Studies and Research in Engineering, COPPE, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil
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3
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Machado-Paula MM, Corat MAF, de Vasconcellos LMR, Araújo JCR, Mi G, Ghannadian P, Toniato TV, Marciano FR, Webster TJ, Lobo AO. Rotary Jet-Spun Polycaprolactone/Hydroxyapatite and Carbon Nanotube Scaffolds Seeded with Bone Marrow Mesenchymal Stem Cells Increase Bone Neoformation. ACS APPLIED BIO MATERIALS 2022; 5:1013-1024. [PMID: 35171572 DOI: 10.1021/acsabm.1c00365] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/19/2022]
Abstract
Clinically, bone tissue replacements and/or bone repair are challenging. Strategies based on well-defined combinations of osteoconductive materials and osteogenic cells are promising to improve bone regeneration but still require improvement. Herein, we combined polycaprolactone (PCL) fibers, carbon nanotubes (CNT), and hydroxyapatite (nHap) nanoparticles to develop the next generation of bone regeneration material. Fibers formed by rotary jet spinning (RJS) instead of traditional electrospinning (ES) with embedded bone marrow mesenchymal stem cells (BMMSCs) showed the best outcomes to repair rat calvarial defects after 6 weeks. To understand this, it was observed that different morphologies were formed depending on the manufacturing method used. RJS fibers presented a particular topography with rough fibers, which allowed for better cellular growth and cell spreading in vitro around and into a three-dimensional (3D) mesh, while fibers made by ES were more smooth and cellular growth was only measured on the 3D mesh surface. The fibers with incorporated nHap/CNT nanoparticles enhanced in vitro cell performance as indicated by more cellular proliferation, alkaline phosphatase activity, proliferation, and deposition of calcium. Greater bone neoformation occurred by combining three characteristics: the presence of nHap and CNT nanoparticles, the topography of the RJS fibers, and the addition of BMMSCs. RJS fibers with nanoparticles and seeded with BMMSCs showed 10 136 mm3 of bone neoformation, meaning a 10-fold increase compared to using RJS only and BMMSCs (0.853 mm3) and a 5-fold increase from using ES only (2054 mm3) after 6 weeks of implantation. Conversely, none of these approaches used individually showed any significant difference for in vivo bone neoformation, suggesting that their combination is essential for optimizing bone formation. In summary, our work generated a potential material composed of well-defined combinations of suitable scaffolds seeded with BMMSCs for enhancing numerous orthopedic tissue engineering applications.
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Affiliation(s)
- Mirian M Machado-Paula
- Institute of Research and Development, University of Vale do Paraiba, São José dos Campos, SP 12244 - 000, Brazil.,Nanomedicine Laboratory, Department of Chemical Engineering, Northeastern University, Boston, Massachusetts 02115, United States.,Multidisciplinary Center for Biological Research, State University of Campinas, Campinas, SP 13083-877, Brazil
| | - Marcus A F Corat
- Multidisciplinary Center for Biological Research, State University of Campinas, Campinas, SP 13083-877, Brazil
| | - Luana M R de Vasconcellos
- Department of Bioscience and Oral Diagnosis, Institute of Science and Technology, Sao Paulo State University, Sao Jose dos Campos, Sao Paulo 12245000, Brazil
| | - Juliani C R Araújo
- Department of Bioscience and Oral Diagnosis, Institute of Science and Technology, Sao Paulo State University, Sao Jose dos Campos, Sao Paulo 12245000, Brazil
| | - Gujie Mi
- Nanomedicine Laboratory, Department of Chemical Engineering, Northeastern University, Boston, Massachusetts 02115, United States
| | - Paria Ghannadian
- Nanomedicine Laboratory, Department of Chemical Engineering, Northeastern University, Boston, Massachusetts 02115, United States
| | - Tatiane V Toniato
- Institute of Research and Development, University of Vale do Paraiba, São José dos Campos, SP 12244 - 000, Brazil
| | - Fernanda R Marciano
- Department of Physics, UFPI - Federal University of Piaui, 64049-550 Teresina, PI, Brazil
| | - Thomas J Webster
- Nanomedicine Laboratory, Department of Chemical Engineering, Northeastern University, Boston, Massachusetts 02115, United States
| | - Anderson O Lobo
- Nanomedicine Laboratory, Department of Chemical Engineering, Northeastern University, Boston, Massachusetts 02115, United States.,LIMAV-Interdisciplinary Laboratory for Advanced Materials, BioMatLab, UFPI - Federal University of Piaui, 64049-550 Teresina, PI, Brazil
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4
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Li Y, Qin Z, Zhang F, Yang ST. Two-color fluorescent proteins reporting survivin regulation in breast cancer cells for high throughput drug screening. Biotechnol Bioeng 2021; 119:1004-1017. [PMID: 34914099 DOI: 10.1002/bit.28006] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2021] [Revised: 11/25/2021] [Accepted: 12/09/2021] [Indexed: 02/06/2023]
Abstract
Reporter gene assay is widely used for high throughput drug screening and drug action mechanism evaluation. In this study, we developed a robust dual-fluorescent reporter assay to detect drugs repressing the transcription of survivin, a cancer biomarker from the inhibitor of apoptosis family, in breast cancer cells cultured in three-dimensional (3D) microbioreactors. Survivin is overexpressed in numerous malignancies but almost silent in normal tissue cells and is considered a lead target for cancer therapy. Breast cancer MCF-7 cells were engineered to express enhanced green fluorescent protein driven by a survivin promoter and red fluorescent protein driven by a cytomegalovirus promoter as internal control to detect changes in survivin expression in cells as affected by drugs. This 3D dual-fluorescent reporter assay was validated with YM155 and doxorubicin, which were known to downregulate survivin in cancer cells, and further evaluated with two widely used anticancer compounds, cisplatin, and epigallocatechin gallate, to evaluate their effects on survivin expression. The results showed that the 3D dual-fluorescent reporter assay was robust for high throughput screening of drugs targeting survivin in breast cancer cells.
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Affiliation(s)
- You Li
- William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA
| | - Zhen Qin
- William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA
| | - Fengli Zhang
- William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA
| | - Shang-Tian Yang
- William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA
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Yadav P, Chatterjee K, Saini DK. Senescent cells in 3D culture show suppressed senescence signatures. Biomater Sci 2021; 9:6461-6473. [PMID: 34582533 DOI: 10.1039/d1bm00536g] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
Abstract
Cellular senescence, an irreversible proliferation arrested but viable cellular state, has been implicated in the progression of several age-associated pathologies. A vast amount of information about senescence has been acquired in cultured cells; however, senescence in living organisms (in vivo) remains poorly understood, mainly because of technical limitations. Furthermore, it is now widely recognized that three-dimensional (3D) culture systems are a better mimic of the in vivo physiology. Herein, senescence was induced in HeLa cells by irradiation. Non-senescent or senescent cells were cultured in soft 3D polymer scaffolds and compared with cells in conventional two-dimensional (2D) culture. This work shows that the morphology of the senescent cells markedly varies between substrates/culture platforms, driving the differences in the cytoskeletal organization, cellular division, and nanomechanical properties. One characteristic feature of senescent cells on 2D culture systems is the enlarged and flattened morphology; however, such drastic changes are not seen in vivo. This is an artificial effect of the substrate, which renders such non-physiological morphology to senescent cells. In the 3D scaffolds, this artifact is reduced. Hence, it serves as a better mimic of tissues, leading to reduced expression of senescence-associated genes, implying that the 3D scaffolds suppress the senescence in cells.
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Affiliation(s)
- Parul Yadav
- Centre for BioSystems Science and Engineering, Indian Institute of Science, C.V Raman Avenue, Bangalore 560012, India
| | - Kaushik Chatterjee
- Centre for BioSystems Science and Engineering, Indian Institute of Science, C.V Raman Avenue, Bangalore 560012, India.,Department of Materials Engineering, Indian Institute of Science, C.V Raman Avenue, Bangalore 560012, India.
| | - Deepak Kumar Saini
- Centre for BioSystems Science and Engineering, Indian Institute of Science, C.V Raman Avenue, Bangalore 560012, India.,Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, C.V Raman Avenue, Bangalore 560012, India.
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6
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Capuana E, Lopresti F, Carfì Pavia F, Brucato V, La Carrubba V. Solution-Based Processing for Scaffold Fabrication in Tissue Engineering Applications: A Brief Review. Polymers (Basel) 2021; 13:2041. [PMID: 34206515 PMCID: PMC8271609 DOI: 10.3390/polym13132041] [Citation(s) in RCA: 29] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/01/2021] [Revised: 06/17/2021] [Accepted: 06/18/2021] [Indexed: 12/13/2022] Open
Abstract
The fabrication of 3D scaffolds is under wide investigation in tissue engineering (TE) because of its incessant development of new advanced technologies and the improvement of traditional processes. Currently, scientific and clinical research focuses on scaffold characterization to restore the function of missing or damaged tissues. A key for suitable scaffold production is the guarantee of an interconnected porous structure that allows the cells to grow as in native tissue. The fabrication techniques should meet the appropriate requirements, including feasible reproducibility and time- and cost-effective assets. This is necessary for easy processability, which is associated with the large range of biomaterials supporting the use of fabrication technologies. This paper presents a review of scaffold fabrication methods starting from polymer solutions that provide highly porous structures under controlled process parameters. In this review, general information of solution-based technologies, including freeze-drying, thermally or diffusion induced phase separation (TIPS or DIPS), and electrospinning, are presented, along with an overview of their technological strategies and applications. Furthermore, the differences in the fabricated constructs in terms of pore size and distribution, porosity, morphology, and mechanical and biological properties, are clarified and critically reviewed. Then, the combination of these techniques for obtaining scaffolds is described, offering the advantages of mimicking the unique architecture of tissues and organs that are intrinsically difficult to design.
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Affiliation(s)
- Elisa Capuana
- Department of Engineering, University of Palermo, RU INSTM, Viale delle Scienze, 90128 Palermo, Italy; (E.C.); (F.L.); (F.C.P.); (V.B.)
| | - Francesco Lopresti
- Department of Engineering, University of Palermo, RU INSTM, Viale delle Scienze, 90128 Palermo, Italy; (E.C.); (F.L.); (F.C.P.); (V.B.)
| | - Francesco Carfì Pavia
- Department of Engineering, University of Palermo, RU INSTM, Viale delle Scienze, 90128 Palermo, Italy; (E.C.); (F.L.); (F.C.P.); (V.B.)
| | - Valerio Brucato
- Department of Engineering, University of Palermo, RU INSTM, Viale delle Scienze, 90128 Palermo, Italy; (E.C.); (F.L.); (F.C.P.); (V.B.)
| | - Vincenzo La Carrubba
- Department of Engineering, University of Palermo, RU INSTM, Viale delle Scienze, 90128 Palermo, Italy; (E.C.); (F.L.); (F.C.P.); (V.B.)
- ATeN Center, University of Palermo, Viale delle Scienze, 90128 Palermo, Italy
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7
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Rawal A, Sharma S, Singh D, Jangir NK, Saraswat H, Sebők D, Kukovecz A, Hietel D, Dauner M, Onal L. Out-of-plane auxetic nonwoven as a designer meta-biomaterial. J Mech Behav Biomed Mater 2020; 112:104069. [PMID: 32957055 DOI: 10.1016/j.jmbbm.2020.104069] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/21/2020] [Revised: 08/17/2020] [Accepted: 08/25/2020] [Indexed: 11/17/2022]
Abstract
Biomaterials are porous and three-dimensional (3D) templates, which are used as biological substitutes in tissue engineering. Targeting the optimal design of biomaterials requires a synergy between mechanical, porous, mass transport, and biological properties. To address this challenge, we propose a non-periodic meta-biomaterial in the form of an out-of-plane auxetic nonwoven scaffold that possesses a 3D interconnected highly porous structure with remarkable mechanical properties corresponding to conventional nonwoven material. A design strategy of utilizing larger fiber diameters to enhance the porosity and permeability characteristics successfully devised the nonwoven scaffold with an extraordinary out-of-plane auxetic effect. In situ tensile-X-ray microcomputed tomography (microCT) analysis has been carried out to monitor the variation in the morphological characteristics.
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Affiliation(s)
- Amit Rawal
- Department of Textile and Fibre Engineering, Indian Institute of Technology Delhi, New Delhi, India.
| | - Sumit Sharma
- Department of Textile and Fibre Engineering, Indian Institute of Technology Delhi, New Delhi, India
| | - Danvendra Singh
- Department of Textile and Fibre Engineering, Indian Institute of Technology Delhi, New Delhi, India
| | | | | | - Dániel Sebők
- University of Szeged, Interdisciplinary Excellence Center, Department of Applied and Environmental Chemistry, Szeged, Hungary
| | - Akos Kukovecz
- University of Szeged, Interdisciplinary Excellence Center, Department of Applied and Environmental Chemistry, Szeged, Hungary
| | - Dietmar Hietel
- Fraunhofer Institute for Industrial Mathematics (ITWM), Kaiserslautern, Germany
| | - Martin Dauner
- Deutsche Institute für Textil- und Faserforschung (DITF), Denkendorf, Germany
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8
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Sionkowska A, Tuwalska A. Preparation and characterization of new materials based on silk fibroin, chitosan and nanohydroxyapatite. INTERNATIONAL JOURNAL OF POLYMER ANALYSIS AND CHARACTERIZATION 2020. [DOI: 10.1080/1023666x.2020.1786271] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 01/04/2023]
Affiliation(s)
- Alina Sionkowska
- Faculty of Chemistry, Nicolaus Copernicus University in Torun, Torun, Poland
| | - Anna Tuwalska
- Faculty of Chemistry, Nicolaus Copernicus University in Torun, Torun, Poland
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9
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Xin X, Yang ST. A Dual Fluorescent 3-D Multicellular Coculture of Breast Cancer MCF-7 and Fibroblast NIH-3T3 Cells for High Throughput Cancer Drug Screening. Biochem Eng J 2019. [DOI: 10.1016/j.bej.2019.05.007] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/15/2023]
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10
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11
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Xin X, Wu Y, Zang R, Yang ST. A fluorescent 3D cell culture assay for high throughput screening of cancer drugs down-regulating survivin. J Biotechnol 2019; 289:80-87. [DOI: 10.1016/j.jbiotec.2018.11.018] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/15/2018] [Revised: 11/09/2018] [Accepted: 11/21/2018] [Indexed: 12/13/2022]
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12
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Lewis PL, Green RM, Shah RN. 3D-printed gelatin scaffolds of differing pore geometry modulate hepatocyte function and gene expression. Acta Biomater 2018; 69:63-70. [PMID: 29317370 PMCID: PMC5831494 DOI: 10.1016/j.actbio.2017.12.042] [Citation(s) in RCA: 113] [Impact Index Per Article: 16.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/19/2017] [Revised: 12/23/2017] [Accepted: 12/29/2017] [Indexed: 01/12/2023]
Abstract
Three dimensional (3D) printing is highly amenable to the fabrication of tissue-engineered organs of a repetitive microstructure such as the liver. The creation of uniform and geometrically repetitive tissue scaffolds can also allow for the control over cellular aggregation and nutrient diffusion. However, the effect of differing geometries, while controlling for pore size, has yet to be investigated in the context of hepatocyte function. In this study, we show the ability to precisely control pore geometry of 3D-printed gelatin scaffolds. An undifferentiated hepatocyte cell line (HUH7) demonstrated high viability and proliferation when seeded on 3D-printed scaffolds of two different geometries. However, hepatocyte specific functions (albumin secretion, CYP activity, and bile transport) increases in more interconnected 3D-printed gelatin cultures compared to a less interconnected geometry and to 2D controls. Additionally, we also illustrate the disparity between gene expression and protein function in simple 2D culture modes, and that recreation of a physiologically mimetic 3D environment is necessary to induce both expression and function of cultured hepatocytes. STATEMENT OF SIGNIFICANCE Three dimensional (3D) printing provides tissue engineers the ability spatially pattern cells and materials in precise geometries, however the biological effects of scaffold geometry on soft tissues such as the liver have not been rigorously investigated. In this manuscript, we describe a method to 3D print gelatin into well-defined repetitive geometries that show clear differences in biological effects on seeded hepatocytes. We show that a relatively simple and widely used biomaterial, such as gelatin, can significantly modulate biological processes when fabricated into specific 3D geometries. Furthermore, this study expands upon past research into hepatocyte aggregation by demonstrating how it can be manipulated to enhance protein function, and how function and expression may not precisely correlate in 2D models.
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Affiliation(s)
- Phillip L Lewis
- Department of Biomedical Engineering, Northwestern University, Evanston, IL, United States; Simpson Querrey Institute for Bionanotechnology, Northwestern University, Chicago, IL, United States.
| | - Richard M Green
- Division of Gastroenterology and Hepatology, Feinberg School of Medicine, Northwestern University, Chicago, IL, United States.
| | - Ramille N Shah
- Department of Biomedical Engineering, Northwestern University, Evanston, IL, United States; Simpson Querrey Institute for Bionanotechnology, Northwestern University, Chicago, IL, United States; Department of Materials Science and Engineering, Northwestern University, Evanston, IL, United States; Department of Surgery - Organ Transplantation, Feinberg School of Medicine, Northwestern University, Chicago, IL, United States.
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Krucińska I, Żywicka B, Komisarczyk A, Szymonowicz M, Kowalska S, Zaczyńska E, Struszczyk M, Czarny A, Jadczyk P, Umińska-Wasiluk B, Rybak Z, Kowalczuk M. Biological Properties of Low-Toxicity PLGA and PLGA/PHB Fibrous Nanocomposite Implants for Osseous Tissue Regeneration. Part I: Evaluation of Potential Biotoxicity. Molecules 2017; 22:molecules22122092. [PMID: 29186078 PMCID: PMC6149750 DOI: 10.3390/molecules22122092] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/24/2017] [Revised: 11/17/2017] [Accepted: 11/27/2017] [Indexed: 01/11/2023] Open
Abstract
In response to the demand for new implant materials characterized by high biocompatibility and bioresorption, two prototypes of fibrous nanocomposite implants for osseous tissue regeneration made of a newly developed blend of poly(l-lactide-co-glycolide) (PLGA) and syntheticpoly([R,S]-3-hydroxybutyrate), PLGA/PHB, have been developed and fabricated. Afibre-forming copolymer of glycolide and l-lactide (PLGA) was obtained by a unique method of synthesis carried out in blocksusing Zr(AcAc)4 as an initiator. The prototypes of the implants are composed of three layers of PLGA or PLGA/PHB, nonwoven fabrics with a pore structure designed to provide the best conditions for the cell proliferation. The bioactivity of the proposed implants has been imparted by introducing a hydroxyapatite material and IGF1, a growth factor. The developed prototypes of implants have been subjected to a set of in vitro and in vivobiocompatibility tests: in vitro cytotoxic effect, in vitro genotoxicity and systemic toxicity. Rabbitsshowed no signs of negative reactionafter implantation of the experimental implant prototypes.
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Affiliation(s)
- Izabella Krucińska
- Department of Material and Commodity Sciences and Textile Metrology, Technical University of Lodz, Zeromskiego 116, 90-924 Lodz, Poland.
| | - Bogusława Żywicka
- Department of Experimental Surgery and Biomaterials Research, Wroclaw Medical University, Pasteura 1, 50-367 Wroclaw, Poland.
| | - Agnieszka Komisarczyk
- Department of Material and Commodity Sciences and Textile Metrology, Technical University of Lodz, Zeromskiego 116, 90-924 Lodz, Poland.
| | - Maria Szymonowicz
- Department of Experimental Surgery and Biomaterials Research, Wroclaw Medical University, Pasteura 1, 50-367 Wroclaw, Poland.
| | - Stanisława Kowalska
- Department of Material and Commodity Sciences and Textile Metrology, Technical University of Lodz, Zeromskiego 116, 90-924 Lodz, Poland.
| | - Ewa Zaczyńska
- Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, RudollfaWeigla 12, 53-114 Wroclaw, Poland.
| | - Marcin Struszczyk
- Department of Material and Commodity Sciences and Textile Metrology, Technical University of Lodz, Zeromskiego 116, 90-924 Lodz, Poland.
| | - Anna Czarny
- Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, RudollfaWeigla 12, 53-114 Wroclaw, Poland.
| | - Piotr Jadczyk
- Department of Sanitary Biology and Ecotechnics, Wroclaw University of Technology, Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland.
| | - Barbara Umińska-Wasiluk
- Department of Sanitary Biology and Ecotechnics, Wroclaw University of Technology, Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland.
| | - Zbigniew Rybak
- Department of Experimental Surgery and Biomaterials Research, Wroclaw Medical University, Pasteura 1, 50-367 Wroclaw, Poland.
| | - Marek Kowalczuk
- Centre of Polymer and Carbon Materials of the Polish Academy of Sciences, MariiSkłodowskiej-Curie 34, 41-819 Zabrze, Poland.
- Faculty of Science and Engineering, University of Wolverhampton, Wulfruna Street, WV1 1SB Wolverhampton, UK.
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14
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Biological Properties of Low-Toxic PLGA and PLGA/PHB Fibrous Nanocomposite Scaffolds for Osseous Tissue Regeneration. Evaluation of Potential Bioactivity. Molecules 2017; 22:molecules22111852. [PMID: 29143781 PMCID: PMC6150223 DOI: 10.3390/molecules22111852] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/05/2017] [Revised: 10/19/2017] [Accepted: 10/24/2017] [Indexed: 11/18/2022] Open
Abstract
The aim of the study was to evaluate the biocompatibility and bioactivity of two new prototype implants for bone tissue regeneration made from biodegradable fibrous materials. The first is a newly developed poly(l-lactide-co-glycolide), (PLGA), and the second is a blend of PLGA with synthetic poly([R,S]-3-hydroxybutyrate) (PLGA/PHB). The implant prototypes comprise PLGA or PLGA/PHB nonwoven fabrics with designed pore structures to create the best conditions for cell proliferation. The bioactivity of the proposed implants was enhanced by introducing a hydroxyapatite material and a biologically active agent, namely, growth factor IGF1, encapsulated in calcium alginate microspheres. To assess the biocompatibility and bioactivity, allergenic tests and an assessment of the local reaction of bone tissue after implantation were performed. Comparative studies of local tissue response after implantation into trochanters for a period of 12 months were performed on New Zealand rabbits. Based on the results of the in vivo evaluation of the allergenic effects and the local tissue reaction 12 months after implantation, it was concluded that the two implant prototypes, PLGA + IGF1 and PLGA/PHB + IGF1, were characterized by high biocompatibility with the soft and bone tissues of the tested animals.
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Drobnik J, Pietrucha K, Kudzin M, Mader K, Szymański J, Szczepanowska A. Comparison of various types of collagenous scaffolds applied for embryonic nerve cell culture. Biologicals 2017; 46:74-80. [PMID: 28108210 DOI: 10.1016/j.biologicals.2017.01.001] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2015] [Revised: 12/28/2016] [Accepted: 01/04/2017] [Indexed: 11/30/2022] Open
Abstract
The purpose of the study was to confirm whether collagen-based scaffolds using different cross-linking methods are suitable elaborate environments for embryonic nerve cell culture. Three 3D sponge-shaped porous scaffolds were composed using collagen alone, collagen with chondroitin sulphate modified by 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride, and collagen cross-linked by 2,3-dialdehyde cellulose (DAC). Embryonic nerve cells from rats were applied to the scaffolds and stained with bisbenzimide to study cell entrapment within the scaffolds. The metabolic activity of the cells cultured in the scaffolds was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The majority of cells were differentiated into neurocytes or oligodendrocytes. Collagen and collagen-chondroitin sulphate scaffolds entrapped a low number of cells. The highest cell density was found in the collagen-DAC scaffold. Moreover, in collagen-DAC scaffolds, the metabolic activity was markedly higher than in the other samples. Although all used scaffolds are suitable for the culture of embryonic nerve cells, the collagen-DAC scaffold properties are the most favorable. This scaffold entraps the highest number of cells and constitutes a favorable environment for their culture. Hence, the Col-DAC scaffold is recommended as an effective carrier for embryonic nerve cells.
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Affiliation(s)
- Jacek Drobnik
- Laboratory of Connective Tissue Metabolism, Department of Neuropeptides Research, Medical University of Lodz, Poland.
| | - Krystyna Pietrucha
- Department of Material and Commodity Sciences and Textile Metrology, Lodz University of Technology, Poland
| | | | | | - Jacek Szymański
- Central Scientific Laboratory, Medical University of Lodz, Poland
| | - Alicja Szczepanowska
- Laboratory of Connective Tissue Metabolism, Department of Neuropeptides Research, Medical University of Lodz, Poland
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Ghanavati Z, Neisi N, Bayati V, Makvandi M. The influence of substrate topography and biomaterial substance on skin wound healing. Anat Cell Biol 2015; 48:251-7. [PMID: 26770875 PMCID: PMC4701698 DOI: 10.5115/acb.2015.48.4.251] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2015] [Revised: 05/25/2015] [Accepted: 11/27/2015] [Indexed: 11/27/2022] Open
Abstract
Tissue engineering is a new field of which the main purpose is to regenerate and repair the damaged tissues. Scaffolds serve as three dimensional matrices for neo-organogenesis and their substance can be biologic or synthetic. Natural polymers have good interactions with the cells and synthetic biomaterials are also highly useful in biomedical application because of their biocompatible properties. In addition to scaffold substance, surface properties of biomaterials have an important role in tissue engineering. In this study, we examined whether substrate substance is important for wound healing or its surface topography. Therefore, we fabricated two matrices, electrospun polycaprolactone (PCL) nanofibers and collagen/chitosan film, and implanted them to the same rat models. After 2 weeks, the sizes of healing wounds were measured and their cellular structures were evaluated by histochemistry and mmunohistochemistry. Histological staining showed a good level of cellularization and epidermis-dermis formation in PCL implant while no determinable epithelium was observed after 2 weeks in collagen-chitosan graft. Immunohistochemical study demonstrated the highly expressed pancytokeratin in PCL graft while its expression was weak in underdeveloped epidermis of collagen-chitosan implantation. In conclusion, this study suggested that PCL nanofibers with high surface area had a more ideal property than natural collagen-chitosan film, therefore the structure and topography of a matrix seemed to be more important in wound healing than its material substance.
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Affiliation(s)
- Zeinab Ghanavati
- Cellular and Molecular Research Center, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.; Department of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Niloofar Neisi
- Department of Medical Virology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Vahid Bayati
- Cellular and Molecular Research Center, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.; Department of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Manoochehr Makvandi
- Department of Medical Virology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
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Prabhakaran MP, Vatankhah E, Kai D, Ramakrishna S. Methods for Nano/Micropatterning of Substrates: Toward Stem Cells Differentiation. INT J POLYM MATER PO 2014. [DOI: 10.1080/00914037.2014.945207] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/19/2022]
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DeRosa ME, Hong Y, Faris RA, Rao H. Microtextured polystyrene surfaces for three-dimensional cell culture made by a simple solvent treatment method. J Appl Polym Sci 2013. [DOI: 10.1002/app.40181] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/05/2022]
Affiliation(s)
- Michael E. DeRosa
- Science and Technology Division; Corning Incorporated; Corning New York 14831-0001 United States
| | - Yulong Hong
- Science and Technology Division; Corning Incorporated; Corning New York 14831-0001 United States
| | - Ronald A. Faris
- Science and Technology Division; Corning Incorporated; Corning New York 14831-0001 United States
| | - Hongwei Rao
- Science and Technology Division; Corning Incorporated; Corning New York 14831-0001 United States
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Burkhardt B, Martinez-Sanchez JJ, Bachmann A, Ladurner R, Nüssler AK. Long-term culture of primary hepatocytes: new matrices and microfluidic devices. Hepatol Int 2013. [PMID: 26202403 DOI: 10.1007/s12072-013-9487-3] [Citation(s) in RCA: 17] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
Abstract
Prediction of in vivo drug-induced hepatotoxicity by in vitro cell culture systems is still one of the main challenges in drug development. To date, most in vitro approaches are based on monolayer cultures of primary hepatocytes, although it is known that they rapidly lose their morphology and liver-specific functions, such as activities of drug-metabolizing enzymes and transporters. Hepatocyte dedifferentiation can be delayed by culturing cells in a 3D environment. Combination with continuous medium flow, which creates a more physiological situation, further improves the maintenance of hepatic functions. Here, we present recently developed hydrogels and scaffolds for 3D culture of hepatocytes, which aim at preserving hepatic morphology and functionality for up to 4 weeks in culture. Furthermore, major benefits and drawbacks of microfluidic devices for in vitro hepatotoxicity screening are discussed. Although promising advances have been made regarding the preservation of hepatic functions in 3D flow culture, major issues, such as expensive equipment, large cell numbers and low throughput, are still hampering their use in drug toxicity screening. For these devices to be applied and accepted in the drug-developing industry, it is necessary to combine easily accessible matrices that highly preserve the activities of drug-metabolizing enzymes with a user-friendly microfluidic platform, thereby finding the right balance between reflecting the in vivo situation and enabling satisfying throughput for drug candidate screening.
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Affiliation(s)
- Britta Burkhardt
- BG Trauma Center, Siegfried Weller Institut, Eberhard Karls University Tübingen, Schnarrenbergstr. 95, 72076, Tübingen, Germany.
| | - Juan José Martinez-Sanchez
- BG Trauma Center, Siegfried Weller Institut, Eberhard Karls University Tübingen, Schnarrenbergstr. 95, 72076, Tübingen, Germany
| | - Anastasia Bachmann
- BG Trauma Center, Siegfried Weller Institut, Eberhard Karls University Tübingen, Schnarrenbergstr. 95, 72076, Tübingen, Germany
| | - Ruth Ladurner
- Clinic for General, Visceral and Transplantation Surgery, Eberhard Karls University Tübingen, Hoppe-Seyler-Str. 3, 72076, Tübingen, Germany
| | - Andreas K Nüssler
- BG Trauma Center, Siegfried Weller Institut, Eberhard Karls University Tübingen, Schnarrenbergstr. 95, 72076, Tübingen, Germany.
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Liu M, Liu N, Zang R, Li Y, Yang ST. Engineering stem cell niches in bioreactors. World J Stem Cells 2013; 5:124-35. [PMID: 24179601 PMCID: PMC3812517 DOI: 10.4252/wjsc.v5.i4.124] [Citation(s) in RCA: 26] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/15/2013] [Revised: 06/05/2013] [Accepted: 07/04/2013] [Indexed: 02/06/2023] Open
Abstract
Stem cells, including embryonic stem cells, induced pluripotent stem cells, mesenchymal stem cells and amniotic fluid stem cells have the potential to be expanded and differentiated into various cell types in the body. Efficient differentiation of stem cells with the desired tissue-specific function is critical for stem cell-based cell therapy, tissue engineering, drug discovery and disease modeling. Bioreactors provide a great platform to regulate the stem cell microenvironment, known as "niches", to impact stem cell fate decision. The niche factors include the regulatory factors such as oxygen, extracellular matrix (synthetic and decellularized), paracrine/autocrine signaling and physical forces (i.e., mechanical force, electrical force and flow shear). The use of novel bioreactors with precise control and recapitulation of niche factors through modulating reactor operation parameters can enable efficient stem cell expansion and differentiation. Recently, the development of microfluidic devices and microbioreactors also provides powerful tools to manipulate the stem cell microenvironment by adjusting flow rate and cytokine gradients. In general, bioreactor engineering can be used to better modulate stem cell niches critical for stem cell expansion, differentiation and applications as novel cell-based biomedicines. This paper reviews important factors that can be more precisely controlled in bioreactors and their effects on stem cell engineering.
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Affiliation(s)
- Meimei Liu
- Meimei Liu, Ning Liu, Ru Zang, Shang-Tian Yang, William G Lowrie Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH 43210, United States
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Liu N, Zang R, Yang ST, Li Y. Stem cell engineering in bioreactors for large-scale bioprocessing. Eng Life Sci 2013. [DOI: 10.1002/elsc.201300013] [Citation(s) in RCA: 47] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/22/2022] Open
Affiliation(s)
- Ning Liu
- William G. Lowrie Department of Chemical and Biomolecular Engineering; Ohio State University; Columbus OH USA
| | - Ru Zang
- William G. Lowrie Department of Chemical and Biomolecular Engineering; Ohio State University; Columbus OH USA
| | - Shang-Tian Yang
- William G. Lowrie Department of Chemical and Biomolecular Engineering; Ohio State University; Columbus OH USA
| | - Yan Li
- Department of Chemical and Biomedical Engineering; FAMU-FSU College of Engineering; Florida State University; Tallahassee FL USA
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Danilevicius P, Rekstyte S, Balciunas E, Kraniauskas A, Jarasiene R, Sirmenis R, Baltriukiene D, Bukelskiene V, Gadonas R, Malinauskas M. Micro-structured polymer scaffolds fabricated by direct laser writing for tissue engineering. JOURNAL OF BIOMEDICAL OPTICS 2012; 17:081405-1. [PMID: 23224166 DOI: 10.1117/1.jbo.17.8.081405] [Citation(s) in RCA: 22] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/01/2023]
Abstract
This work presents the latest results on direct laser writing of polymeric materials for tissue engineering applications. A femtosecond Yb:KGW laser (300 fs, 200 kHz, 515 nm) was used as a light source for non-linear lithography. Fabrication was implemented in various photosensitive polymeric materials, such as: hybrid organic-inorganic sol-gel based on silicon-zirconium oxides, commercial ORMOCER® class photoresins. These materials were structured via multi-photon polymerization technique with submicron resolution. Porous three-dimensional scaffolds for artificial tissue engineering were fabricated with constructed system and were up to several millimeters in overall size with 10 to 100 μm internal pores. Biocompatibility of the used materials was tested in primary rabbit muscle-derived stem cell culture in vitro and using laboratory rats in vivo. This interdisciplinary study suggests that proposed technique and materials are suitable for tissue engineering applications.
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Affiliation(s)
- Paulius Danilevicius
- Vilnius University, Faculty of Physics, Department of Quantum Electronics, Laser Research Center, Sauletekio Avenue 10, LT-10223 Vilnius, Lithuania
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Abstract
Having good mechanical strength, biocompatibility, and workability, polyethylene terephthalate (PET) is often used as a biomaterial. In this study, PET filaments with various deniers are made into plied yarn with various coefficients of twist. The plied yarn is then made into PET knitted fabrics. Mechanical property tests are performed to determine the differences among the various PET knitted fabrics. Finally, by using cell culture, the PET knitted fabrics are analyzed and evaluated with their cell attachment.
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Wang XH, Shi S, Guo G, Fu SZ, Fan M, Luo F, Zhao X, Wei YQ, Qian ZY. Preparation and Characterization of a Porous Scaffold Based on Poly(D,L-Lactide) and N-Hydroxyapatite by Phase Separation. JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION 2012; 22:1917-29. [PMID: 20961495 DOI: 10.1163/092050610x529155] [Citation(s) in RCA: 11] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Affiliation(s)
- Xiu Hong Wang
- a State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, P. R. China
| | - Shuai Shi
- b State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, P. R. China
| | - Gang Guo
- c State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, P. R. China
| | - Shao Zhi Fu
- d State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, P. R. China
| | - Min Fan
- e State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, P. R. China
| | - Feng Luo
- f State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, P. R. China
| | - Xia Zhao
- g State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, P. R. China
| | - Yu Quan Wei
- h State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, P. R. China
| | - Zhi Yong Qian
- i State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, P. R. China.
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Sabri F, Cole JA, Scarbrough MC, Leventis N. Investigation of polyurea-crosslinked silica aerogels as a neuronal scaffold: a pilot study. PLoS One 2012; 7:e33242. [PMID: 22448239 PMCID: PMC3308972 DOI: 10.1371/journal.pone.0033242] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/14/2011] [Accepted: 02/06/2012] [Indexed: 11/18/2022] Open
Abstract
BACKGROUND Polymer crosslinked aerogels are an attractive class of materials for future implant applications particularly as a biomaterial for the support of nerve growth. The low density and nano-porous structure of this material combined with large surface area, high mechanical strength, and tunable surface properties, make aerogels materials with a high potential in aiding repair of injuries of the peripheral nervous system. however, the interaction of neurons with aerogels remains to be investigated. METHODOLOGY In this work the attachment and growth of neurons on clear polyurea crosslinked silica aerogels (PCSA) coated with: poly-L-lysine, basement membrane extract (BME), and laminin1 was investigated by means of optical and scanning electron microscopy. After comparing the attachment and growth capability of neurons on these different coatings, laminin1 and BME were chosen for nerve cell attachment and growth on PCSA surfaces. The behavior of neurons on treated petri dish surfaces was used as the control and behavior of neurons on treated PCSA discs was compared against it. CONCLUSIONS/SIGNIFICANCE This study demonstrates that: 1) untreated PCSA surfaces do not support attachment and growth of nerve cells, 2) a thin application of laminin1 layer onto the PCSA discs adhered well to the PCSA surface while also supporting growth and differentiation of neurons as evidenced by the number of processes extended and b3-tubulin expression, 3) three dimensional porous structure of PCSA remains intact after fixing protocols necessary for preservation of biological samples and 4) laminin1 coating proved to be the most effective method for attaching neurons to the desired regions on PCSA discs. This work provides the basis for potential use of PCSA as a biomaterial scaffold for neural regeneration.
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Affiliation(s)
- Firouzeh Sabri
- Department of Physics, University of Memphis, Memphis, Tennessee, United States of America.
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Czolkos I, Hakonen B, Orwar O, Jesorka A. High-resolution micropatterned Teflon AF substrates for biocompatible nanofluidic devices. LANGMUIR : THE ACS JOURNAL OF SURFACES AND COLLOIDS 2012; 28:3200-3205. [PMID: 22204476 DOI: 10.1021/la2044784] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/31/2023]
Abstract
We describe a general photolithography-based process for the microfabrication of surface-supported Teflon AF structures. Teflon AF patterns primarily benefit from superior optical properties such as very low autofluorescence and a low refractive index. The process ensures that the Teflon AF patterns remain strongly hydrophobic in order to allow rapid lipid monolayer spreading and generates a characteristic edge morphology which assists directed cell growth along the structured surfaces. We provide application examples, demonstrating the well-controlled mixing of lipid films on Teflon AF structures and showing how the patterned surfaces can be used as biocompatible growth-directing substrates for cell culture. Chinese hamster ovary (CHO) cells develop in a guided fashion along the sides of the microstructures, selectively avoiding to grow over the patterned areas.
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Affiliation(s)
- Ilja Czolkos
- Department of Chemical and Biological Engineering, Chalmers University of Technology, Kemivägen 10, 412 96 Göteborg, Sweden
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Ng R, Zang R, Yang KK, Liu N, Yang ST. Three-dimensional fibrous scaffolds with microstructures and nanotextures for tissue engineering. RSC Adv 2012. [DOI: 10.1039/c2ra21085a] [Citation(s) in RCA: 110] [Impact Index Per Article: 8.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/20/2023] Open
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Mou ZL, Zhao LJ, Zhang QA, Zhang J, Zhang ZQ. Preparation of porous PLGA/HA/collagen scaffolds with supercritical CO2 and application in osteoblast cell culture. J Supercrit Fluids 2011. [DOI: 10.1016/j.supflu.2011.07.003] [Citation(s) in RCA: 41] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022]
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Zhang X, Yang ST. High-throughput 3-D cell-based proliferation and cytotoxicity assays for drug screening and bioprocess development. J Biotechnol 2010; 151:186-93. [PMID: 21115074 DOI: 10.1016/j.jbiotec.2010.11.012] [Citation(s) in RCA: 34] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/05/2010] [Revised: 09/07/2010] [Accepted: 11/17/2010] [Indexed: 02/06/2023]
Abstract
We have designed, built and tested a three-dimensional (3-D) cell culture system on modified microplates for high-throughput, real-time, proliferation and cytotoxicity assays. In this 3-D culture system, cells expressing the enhanced green fluorescent protein (EGFP) were cultured in nonwoven polyethylene terephthalate (PET) fibrous scaffolds. Compared to 2-D cultures in conventional microplates, 3-D cultures gave more than 10-fold higher fluorescence signals with significantly increased signal-to-noise ratio (SNR), thus extending the application of conventional fluorescence microplate readers for online monitoring of culture fluorescence. The 3-D system was successfully used to demonstrate the effects of fetal bovine serum, fibronectin coating of PET fibers, and cytotoxicity of dexamethasone on recombinant murine embryonic stem D3 cells. The dosage effects of 5-fluorouracil and gemcitabine on high-density colon cancer HT-29 cells were also tested. These studies demonstrated that the 3-D culture microplate system with EGFP expressing cells can be used as a high-throughput system in drug discovery and bioprocess development.
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Affiliation(s)
- Xudong Zhang
- William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 West 19th Avenue, Columbus, OH 43210, USA.
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Ng R, Gurm JS, Yang ST. Centrifugal seeding of mammalian cells in nonwoven fibrous matrices. Biotechnol Prog 2010; 26:239-45. [PMID: 19785042 DOI: 10.1002/btpr.317] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/07/2022]
Abstract
Three-dimensional (3D) cell cultures have many advantages over two-dimensional cultures. However, seeding cells in 3D scaffolds such as nonwoven fibrous polyethylene terephthalate (PET) matrices has been a challenge task in tissue engineering and cell culture bioprocessing. In this study, a centrifugal seeding method was investigated to improve the cell seeding efficiency in PET matrices with two different porosities (93% and 88%). Both the centrifugal force and centrifugation time were found to affect the seeding efficiency. With an appropriate centrifugation speed, a high 80-90% cell seeding efficiency was achieved and the time to reach this high seeding efficiency was less than 5 min. The seeding efficiency was similar for matrices with different porosities, although the optimal seeding time was significantly shorter for the low-porosity scaffold. Post seeding cell viability was demonstrated by culturing colon cancer cells seeded in PET matrices for over 5 days. The centrifugal seeding method developed in this work can be used to efficiently and uniformly seed small fibrous scaffolds for applications in 3D cell-based assays for high-throughput screening.
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Affiliation(s)
- Robin Ng
- William G. Lowrie Dept. of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, OH 43210, USA
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Subramanian A, Krishnan UM, Sethuraman S. Development of biomaterial scaffold for nerve tissue engineering: Biomaterial mediated neural regeneration. J Biomed Sci 2009; 16:108. [PMID: 19939265 PMCID: PMC2790452 DOI: 10.1186/1423-0127-16-108] [Citation(s) in RCA: 352] [Impact Index Per Article: 22.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/26/2009] [Accepted: 11/25/2009] [Indexed: 01/27/2023] Open
Abstract
Neural tissue repair and regeneration strategies have received a great deal of attention because it directly affects the quality of the patient's life. There are many scientific challenges to regenerate nerve while using conventional autologous nerve grafts and from the newly developed therapeutic strategies for the reconstruction of damaged nerves. Recent advancements in nerve regeneration have involved the application of tissue engineering principles and this has evolved a new perspective to neural therapy. The success of neural tissue engineering is mainly based on the regulation of cell behavior and tissue progression through the development of a synthetic scaffold that is analogous to the natural extracellular matrix and can support three-dimensional cell cultures. As the natural extracellular matrix provides an ideal environment for topographical, electrical and chemical cues to the adhesion and proliferation of neural cells, there exists a need to develop a synthetic scaffold that would be biocompatible, immunologically inert, conducting, biodegradable, and infection-resistant biomaterial to support neurite outgrowth. This review outlines the rationale for effective neural tissue engineering through the use of suitable biomaterials and scaffolding techniques for fabrication of a construct that would allow the neurons to adhere, proliferate and eventually form nerves.
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Affiliation(s)
- Anuradha Subramanian
- Center for Nanotechnology & Advanced Biomaterials, SASTRA University, Thanjavur, India.
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Ng R, Zhang X, Liu N, Yang ST. Modifications of nonwoven polyethylene terephthalate fibrous matrices via NaOH hydrolysis: Effects on pore size, fiber diameter, cell seeding and proliferation. Process Biochem 2009. [DOI: 10.1016/j.procbio.2009.04.024] [Citation(s) in RCA: 31] [Impact Index Per Article: 1.9] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 10/20/2022]
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Seo YK, Youn HH, Park CS, Song KY, Park JK. Reinforced bioartificial dermis constructed with collagen threads. BIOTECHNOL BIOPROC E 2009. [DOI: 10.1007/s12257-008-0118-0] [Citation(s) in RCA: 12] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/21/2022]
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Stillaert FB, Di Bartolo C, Hunt JA, Rhodes NP, Tognana E, Monstrey S, Blondeel PN. Human clinical experience with adipose precursor cells seeded on hyaluronic acid-based spongy scaffolds. Biomaterials 2008; 29:3953-9. [PMID: 18635258 DOI: 10.1016/j.biomaterials.2008.06.005] [Citation(s) in RCA: 69] [Impact Index Per Article: 4.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/24/2008] [Accepted: 06/10/2008] [Indexed: 12/20/2022]
Abstract
Histioconductive approaches to soft-tissue defects use scaffolds seeded with lineage- and tissue-specific progenitors to generate tissue which should reside in equilibrium with adjacent tissue. Scaffolds guide histiogenesis by ensuring cell-cell and cell-matrix interactions. Hyaluronic acid-based (HA) preadipocyte-seeded scaffolds were evaluated for their adipo-conductive potential and efficacy in humans. Preadipocytes were isolated from lipoaspirate material and seeded on HA scaffolds. The cellular bio-hybrid (ADIPOGRAFT) and an acellular control scaffold (HYAFF11) were implanted subcutaneously. At specific time points (2, 8 and 16 weeks) explants were analyzed histopathologically with immunohistochemistry. No adverse tissue effects occurred. Volume loss and consistent degradation of the HYAFF11 scaffolds compared to the ADIPOGRAFT group indicated progressive tissue integration. No consistent histological differences between both groups were observed. By 8 weeks all void spaces within the scaffolds were filled with cells with pronounced matrix deposition in the ADIPOGRAFT bio-hybrids. Here we show that HA scaffolds were stable cell carriers and had the potential to generate volume-retaining tissue. However, no adipogenic differentiation was observed within the preadipocyte-seeded scaffolds.
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Affiliation(s)
- F B Stillaert
- Department of Plastic and Reconstructive Surgery, University Hospital Gent, De Pintelaan 185, Gent, Belgium.
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35
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Lee LJ. Polymer nano-engineering for biomedical applications. Ann Biomed Eng 2006; 34:75-88. [PMID: 16541328 DOI: 10.1007/s10439-005-9011-6] [Citation(s) in RCA: 42] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/06/2004] [Accepted: 05/20/2005] [Indexed: 10/24/2022]
Abstract
Polymeric materials possess many attractive properties such as high toughness and recyclability. Some possess excellent biocompatibility, are biodegradable, and can provide various bio-functionalities. Proper combination of functional polymers and biomolecules can offer tailored properties for various biomedical applications. This overview article covers three major sections: Applications of Polymeric Structures and Devices, Nanoscale Polymer Fabrication Technologies, and Conclusions and Future Directions.
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Affiliation(s)
- L James Lee
- Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 W. 19th Ave., Columbus, Ohio 43210, USA.
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Kang X, Xie Y, Kniss DA. Adipose tissue model using three-dimensional cultivation of preadipocytes seeded onto fibrous polymer scaffolds. ACTA ACUST UNITED AC 2005; 11:458-68. [PMID: 15869424 DOI: 10.1089/ten.2005.11.458] [Citation(s) in RCA: 62] [Impact Index Per Article: 3.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
Abstract
A better understanding of the mechanism of adipose tissue differentiation is of paramount importance in the development of therapeutic strategies for the treatment and prevention of obesity and type 2 diabetes mellitus. Optimal results using tissue culture models can be expected only when the in vitro adipocyte resembles adipose tissue in vivo as closely as possible. In this study, we used tissue-engineering principles to develop a three-dimensional (3-D) culture system to mimic the geometry of adipose tissue in vivo. Mouse preadipocyte 3T3-L1 cells were seeded onto nonbiodegradable fibrous polyethylene terephthalate scaffolds and differentiated with a hormone cocktail consisting of insulin, dexamethasone, isobutylmethylxanthine, and fetal calf serum. Cell morphology, growth, differentiation, and function were studied by immunocytochemistry, reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, enzyme-linked immunosorbent assay, and oil red O staining. Cells grown on 3-D fibrous scaffolds were differentiated in situ by hormone induction with high efficiency (approximately 90%) as shown by scanning electron microscopy. Immunocytochemistry, immunoblot analysis, and RT-PCR revealed that the 3-D constructs expressed adipocyte-specific genes, including peroxisome proliferator-activated receptor gamma, leptin, adipsin, aP2, adiponectin, GLUT4, and resistin. Adipocytes matured on 3-D constructs secreted leptin at levels even greater than that of fully differentiated adipocytes in 2-D conventional cell cultures. Finally, adipocyte-specific phenotypic function was demonstrated by accumulation of neutral lipids in larger fat droplets. In conclusion, preadipocytes grown on 3-D matrices acquire morphology and biological features of mature adipocytes. This new culture model should have significant utility for in vitro studies of adipocyte cell biology and development.
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Affiliation(s)
- Xihai Kang
- Department of Obstetrics and Gynecology, Laboratory of Perinatal Research, Ohio State University, College of Medicine and Public Health, Columbus, 43210, USA
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Basu S, Yang ST. Astrocyte Growth and Glial Cell Line-Derived Neurotrophic Factor Secretion in Three-Dimensional Polyethylene Terephthalate Fibrous Matrices. ACTA ACUST UNITED AC 2005; 11:940-52. [PMID: 15998233 DOI: 10.1089/ten.2005.11.940] [Citation(s) in RCA: 17] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/13/2022]
Abstract
The ability of human astrocytes grown in nonwoven fibrous matrices to produce glial cell line-derived neurotrophic factor (GDNF) was studied. GDNF has the ability to selectively nourish and regenerate dopaminergic neurons and thus can provide a new treatment of Parkinson's disease. Compressed polyethylene terephthalate (PET) fabrics (porosity, 88.8%; mean pore diameter, 64 microm), treated with boiling NaOH, was effective in supporting high-density growth of astrocytes with stable GDNF production over the entire period of 18 days studied. Treatment of PET with NaOH renders the fiber surface more hydrophilic, thereby facilitating attachment and spreading of cells, whereas matrix compression allows cells to grow along and also between the fibers of these matrices to a higher density. The average production of GDNF by cells grown in these matrices (approximately 2 cm in diameter) was 21.7 pg/mL x day, with an average high concentration of 64.6 pg/mL, which is well above the effective concentration of 40 pg/mL. This work shows promise in culturing astrocytes in PET matrices as the first step in developing a potential implantable tissue-engineering device for treating patients with Parkinson's disease.
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Affiliation(s)
- Shubhayu Basu
- Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, Ohio 43210, USA
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Yang Y, Basu S, Tomasko DL, Lee LJ, Yang ST. Fabrication of well-defined PLGA scaffolds using novel microembossing and carbon dioxide bonding. Biomaterials 2005; 26:2585-94. [PMID: 15585261 DOI: 10.1016/j.biomaterials.2004.07.046] [Citation(s) in RCA: 45] [Impact Index Per Article: 2.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/23/2004] [Accepted: 07/22/2004] [Indexed: 10/26/2022]
Abstract
A novel biologically benign technique was developed to produce three-dimensional tissue engineering scaffolds with well-defined structure. Photolithography was used to design and pattern a planar scaffold skeletal structure on a photoresist (SU-8), and a variety of microembossing processes including sacrificial layer embossing and bilayer embossing were developed to transfer the skeletal pattern to the poly(DL-lactide-co-glycolide) substrate as scaffold skeletons. Subcritical carbon dioxide was then introduced to assemble these skeletons to a three-dimensional scaffold at a low temperature. Compared with conventional scaffolds, which have a broad pore size distribution and varying pore geometry, these microfabricated scaffolds have a uniform and well-defined geometry and structure. This uniformity of structural parameters allows for the studies of cell attachment, spreading, and proliferation in scaffolds in a controlled and logical manner. The cytocompatibility of these microfabricated scaffolds was tested by seeding three different cell lines with different morphologies and growth patterns into these scaffolds. All three cell lines attached well to the scaffolds and grew to high densities as observed with scanning electron microscopy. This study demonstrates a controllable method to fabricate tissue scaffolds with a well-defined 3D architecture that can be used to better elucidate the effect of structure parameters such as pore geometry and pore size on tissue growth in 3D scaffolds.
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Affiliation(s)
- Yong Yang
- The Department of Chemical and Biomolecular Engineering, The Ohio State University, 140 West 19th Avenue, Columbus, OH 43210, USA
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Luo J, Yang ST. Effects of three-dimensional culturing in a fibrous matrix on cell cycle, apoptosis, and MAb production by hybridoma cells. Biotechnol Prog 2004; 20:306-15. [PMID: 14763857 DOI: 10.1021/bp034181v] [Citation(s) in RCA: 31] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/30/2022]
Abstract
The effects of culturing hybridoma cells in a three-dimensional (3-D) poly(ethylene terephthalate) (PET) fibrous matrix on cell cycle, apoptosis, metabolism, and monoclonal antibody (MAb) production were evaluated by comparing with two-dimensional (2-D) culturing on microcarrier and multiwell plate surfaces. The percentage of cells in the G1/G0 phase increased during the long-term culturing period of approximately 4 weeks. Compared to the 2-D culture systems, cells grown in 3-D matrices had higher MAb productivity for long-term culture. Decreasing serum content in the culture medium increased both MAb productivity and apoptosis. However, the 3-D culture had a greater increase in MAb productivity and a much lower apoptotic rate than the 2-D culture, especially at 0% serum. Most cells in the 3-D fibrous matrix formed large aggregates and were smaller than cells grown on a 2-D surface or in suspension. The smaller cell size allowed cells to survive better in the high-cell-density environment. The fibrous matrix also selectively retained healthy, nonapoptotic cells. These results suggested that the 3-D fibrous matrix contributed to growth arrest, protected cells to better resist low-serum environments, and reduced apoptosis, all of which contributed to the high viable cell density and volumetric MAb productivity in the long-term 3-D culture.
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Affiliation(s)
- Jun Luo
- Department of Chemical Engineering, The Ohio State University, 140 West 19th Avenue, Columbus, Ohio 43210, USA
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