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Title: In vitro study of stem cell communication via gap junctions for fibrocartilage regeneration at entheses
Authors: Nayak, B.P.
Hong Goh, J.C. 
Toh, S.L. 
Satpathy, G.R.
Keywords: Bone marrow-derived stem cells
Fibrocartilagen gap junction
Type 2 collagen
Issue Date: Mar-2010
Citation: Nayak, B.P., Hong Goh, J.C., Toh, S.L., Satpathy, G.R. (2010-03). In vitro study of stem cell communication via gap junctions for fibrocartilage regeneration at entheses. Regenerative Medicine 5 (2) : 221-229. ScholarBank@NUS Repository.
Abstract: Background: Entheses are fibrocartilaginous organs that bridge ligament with bone at their interface and add significant insertional strength. To replace a severely damaged ligament, a tissue-engineered graft preinstalled with interfacial fibrocartilage, which is being regenerated from stem cells, appears to be more promising than ligament-alone graft. Such a concept can be realized by a biomimetic approach of establishing a dynamic communication of stem cells with bone cells and/or ligament fibroblasts in vitro. Aim: The current study has two objectives. The first objective is to demonstrate functional coculture of bone marrow-derived stem cells (BMSCs) with mature bone cells/ligament fibroblasts as evidenced by gap-junctional communication in vitro. The second objective is to investigate the role of BMSCs in the regeneration of fibrocartilage within the coculture. Materials & methods: Rabbit bone/ligament fibroblasts were dual-stained with DiI-Red and calcein (gap-junction permeable dye), and cocultured with unlabeled BMSCs at fixed ratio (1:10). The functional gap junction was demonstrated by the transfer of calcein from donor to recipient cells that was confirmed and quantified by flow cytometry. Type 2 collagen (cartilage extracellular matrix-specific protein) expressed by the mixed cell lines in the cocultures were estimated by real-time reverse transcription PCR and compared with that of the ligament-bone coculture (control). Results: Significant transfer of calcein into BMSCs was observed and flow cytometry analyses showed a gradual increase in the percentage of BMSCs acquiring calcein with time. Cocultures that included BMSCs expressed significantly more type 2 collagen compared with the control. Conclusion: The current study, for the first time, reported the expression of gap-junctional communication of BMSCs with two adherent cell lines of musculoskeletal system in vitro and also confirmed that incorporation of stem cells augments fibrocartilage regeneration. The results open up a path to envisage a composite graft preinstalled with enthesial fibrocartilage using a stem cell-based coculture system. © 2010 Future Medicine Ltd.
Source Title: Regenerative Medicine
ISSN: 17460751
DOI: 10.2217/rme.09.86
Appears in Collections:Staff Publications

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