PURPOSE: The purpose of this paper was to determine the ability of a mixture consisting of mesenchymal stem cells, beta-tricalcium phosphate β-TCP), and hydrogel, to support cells and form new tissue. MATERIALS AND METHODS: A composite was produced by adding β-TCP to hydrogel, and mesenchymal stem cells were cultivated in the composite. Then, reverse transcription polymerase chain reaction (RT-PCR) was conducted to measure the level of gene expression for the new bone formation in the cells. Moreover, a composite in which the mesenchymal stem cells were added was injected into the subcutaneous fat of sprague-dawley rats. After four weeks, H&E, Masson trichrome, silver nitrate staining, and osterix immunohistochemical staining were conducted by taking the tissue to evaluate whether the composite supported mesenchymal stem cells and formed new tissue. RESULTS: By using RT-PCR, we found that the level of gene expression became significantly higher in 3-dimensional gel culture with RUNX2 by 1.26 times, with osteopontin by 1.23 times, transforming growth factor-β by 2.12 times, osterix by 1.07 times, type I collagen by 1.3 times, and fibronectin by 1.3 times. In the animal experiment in which a composite was transplanted into the subcutaneous fat, newly formed tissue was observed. Also, it was found that the composite prevented mesenchymal stem cells from leaving and formed new tissue. Osteogenic differentiation cells in the tissue was observed through osterix immunostaining. CONCLUSION: It was identified that the composite prevented mesenchymal stem cells dispersal and contributed to the formation of neogenic tissue. Therefore we conclude that the composite plays a role of a scaffold to support the implanted cells and form neogenic tissue more effectively.
Animal Experimentation ; Collagen Type I ; Fibronectins ; Gene Expression ; Hydrogel ; Mesenchymal Stromal Cells* ; Osteogenesis* ; Osteopontin ; Polymerase Chain Reaction ; Rats, Sprague-Dawley ; Reverse Transcription ; Silver Staining ; Subcutaneous Fat