Reversible mitotic and metabolic inhibition following the encapsulation of fibroblasts in alginate hydrogels |
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| Authors: | Nicola C Hunt Richard M Shelton Liam M Grover |
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| Institution: | 1. School of Chemical Engineering, College of Engineering and Physical Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK;2. School of Dentistry, College of Medical and Dental Sciences, University of Birmingham, Birmingham B4 6NN, UK;2. Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, Leahurst Campus, Neston, Cheshire CH64 7TE, UK;3. Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, University Hospital, Aintree, Liverpool L9 7AL, UK;1. Cell Isolation and Transplantation Center, Department of Surgery, Geneva University Hospitals and Medical School, Geneva, Switzerland;2. Institut d’Ingénierie Biologique et Institut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland;3. Orthopedic Surgery, Department of Surgery, Geneva University Hospitals and Medical School, Geneva, Switzerland;1. Department of Bioengineering and IBB-Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal;2. The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology/Emory University, Atlanta, GA, USA;3. Gladstone Institute of Cardiovascular Disease, San Francisco, CA, USA;4. Department of Bioengineering & Therapeutic Sciences, University of California, San Francisco, USA;5. The Discoveries Centre for Regenerative and Precision Medicine, Lisbon Campus, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal;1. Biotechnology-Medical Science, KU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul 136-701, Republic of Korea;2. Department of Biomedical Engineering, College of Health Science, Korea University, Seoul 136-703, Republic of Korea |
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| Abstract: | Limiting cell proliferation without reducing cell viability for in vivo tissue engineering applications is important in co-culture applications where the growth of one cell type must be inhibited to prevent overgrowth of the scaffold at the expense of another cell type. Also, it is vital for maintaining viability of cells in large constructs before vascularisation occurs. In this study we have shown by means of the Thiazolyl blue (MTT) assay and immuno-staining for proliferating cell nuclear antigen (PCNA) that encapsulating fibroblasts in 2% and 5% w/v calcium-alginate at a density of 7.5 × 105 cells/ml as uniformly dispersed entities, enabled cells to maintain viability and caused a reversible mitotic inhibition. Alginate encapsulation also caused reversible metabolic inhibition as demonstrated by the MTT assay and fluorescent staining for mitochondrial membrane potential. Histological evaluation of the alginate constructs containing fibroblasts showed that mitotic and metabolic inhibition was possibly due to cell isolation during the first five weeks of culture. The alginate scaffold degraded with time releasing encapsulated fibroblasts. Upon implantation to a wound site this should ensure that encapsulated cells are able to replace the damaged tissue after sufficient proliferation of the co-cultured cell type or sufficient vascularisation of the construct. |
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