| Summary: | 碩士 === 國立臺灣大學 === 臨床牙醫學研究所 === 101 === Aim： In tissue regeneration, growth factors and scaffolds play important roles in cell proliferation, differentiation, extracellular matrix production and dentin formation. The purpose of this study is to investigate the effects of TGF-β1 and collagen on behaviors of stem cells from apical papilla (SCAP). We hypothesize that collagen can affect cell attachment, viability, and alkaline phosphatase (ALP) expression in SCAP. Besides, culturing SCAP in scaffold with combination of TGF-β1 can regenerate tissues successfully in vitro.
Materials and Methods： SCAP were cultured on the surface coated with different densities of collagen. Then, SCAP were evaluated for cell attachment ability, cell viability by MTT test and cell differentiation by ALP activity. Besides, SCAP were treated with TGF-β1 with different concentrations. CTGF and α1-integrin expression were determined by reverse-transcription polymerase chain reaction（RT-PCR）. Finally, we cultured SCAP in collagen scaffolds and dentin disc scaffolds with combination of TGF-β1 to observe the morphological changes of cells and tissue regeneration under histological analysis.
Results： SCAP cultured on surfaces coated with collagen showed better attachment ability. SCAP viability elevated when cultured on the surface coated with lower densities of collagen. However, decreased cell viability and ALP activity were observed when cultured SCAP on the surfaces coated with high densities of collagen. TGF-β1 could promote CTGF and α1-integrin gene expression of SCAP in a dose-dependent manner. Furthermore, we cultured SCAP in collagen scaffold with TGF-β1 stimulation and observed the prominent cell proliferation and matrix-like tissue regeneration. When SCAP were cultured in dentin disc scaffold with TGF-β1 stimulation, regenerated dentin-like tissues containing dentinal tubules and calcified nodules were found.
Conclusion： SCAP cultured on the collagen matrices with various concentrations showed different biological characteristics like cell attachment, proliferation, and differentiation. Besides, collagen and dentin disc scaffolds with combination of SCAP and TGF-β1 may be practical models in regeneration of dentin-pulp tissues. Further research may be engaged in applying the models in animal studies, and even in clinical practice to repair the teeth with pulp necrosis or immature root formation.
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