Abstract
Purpose: The development of new biomaterials and the application of growth factors are expected for osseoustissue regeneration. Therefore, we developed a high-molecular complex material as a carrier for variousgrowth factors, and evaluated its effects on bone marrow-derived undifferentiated mesenchymal cells. Methods: We developed DAC 50-TYPE I collagen as a completely new gel by clone reaction of Type I-Acollagen with deacetylated chitin 50 (DAC 50), and performed three-dimensional culture of bone marrow-derivedundifferentiated mesenchymal cells in this complex material. In terms of cell proliferation and differentiation, cell proliferation tests and morphological observation by staining with hematoxylin and eosin (H-Estaining) were performed, alkaline phosphatase (ALP) activity was observed by ALP staining, and the expressionsof bone sialoprotein (BSP), ALP, osteopontin, osteocalcin, and glyceraldehyde-3-phosphatedehydrogenase (GAPDH) genes were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Results: Cell proliferation tests showed cell proliferation but no marked proliferation. H-E staining revealedthree-dimensional cell expansion. ALP staining was positive. RT-PCR confirmed the expression of all targetgenes except the osteocalcin gene.Conclusion: This new material did not damage bone marrow-derived mesenchymal cells and allowed theirthree-dimensional proliferation. This material may be useful as a new biomaterial for osseous tissue regeneration.
