In this study, we investigated how rat bone marrow stromal cells (BMSCs) under a physically stable microenvironment influenced the subcutaneous fibroblasts. The model for this study involved setting up a space made up of a titanium mesh cage inserted into the subcutaneous region in rats and filled with a collagen matrix seeded with (1) BMSCs, (2) fibroblasts or (3) a combination of BMSCs and fibroblasts. Fibroblasts for transplantations were taken from enhanced green fluorescence protein (EGFP) transgenic "green rats" which enabled us to trace the fate of the cells in vivo. A series of X-ray computed tomographic (CT) images were taken of each implant over a period of 8 weeks, and the implants were then removed and examined histologically. As a result, while generated bone was observed in each case that included BMSCs (the BMSCs and combination group), there was no generated bone observed in the group using fibroblasts only. Interestingly, EGFP-positive osteocytes were observed in the generated bone of the combination group, indicating that the transplanted fibroblasts differentiated into osteocytes during the bone formation. Thus, we demonstrated that genuine intrinsic fibroblasts are able to become osteocytes as a result of the influence of BMSCs.
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