冷冻干燥牙本质对牙周膜干细胞影响的研究

The effects of freeze-dried dentin matrix on periodontal ligament stem cells

目的:制作冷冻干燥牙本质生物支架(FDDM)并研究其对牙周膜干细胞(PDLSCs)的影响。方法:制作FDDM生物支架;将PDLSCs分别接种在FDDM、普通牙本质(D)、羟基磷灰石/磷酸三钙(HA)、盖玻片/α-MEM表面或浸提液中。MTT法检测细胞增殖;粘胶纤维红检测胶原分泌量;扫描电镜检测细胞形态;共聚焦显微镜观察各组细胞骨架形态。结果:研制出FDDM;MTT结果显示FDDM组细胞增殖活性高于HA组(P<0.05);胶原检测显示FDDM组细胞分泌的胶原多于HA组(P<0.05);电镜结果显示FDDM组细胞紧密粘附,细胞突起的数目和长度大于HA与盖玻片组;共聚焦结果显示,FDDM组细胞骨架较HA、盖玻片组粗大、清晰。结论:FDDM促进PDLSCs的增殖、胶原分泌,促进细胞粘附及细胞应力纤维形成。

Objective： To fabricate freeze-dried dentin matrix（FDDM） and to evaluate its effects on the biological behavior of periodontal ligament stem cells（PDLSCs）. Methods： FDDM was fabricated. PDLSCs were cultured and identified. Then, PDLSCs were respectively cultured on FDDM, dentin（ D）, hydroxyapatite/tricalcium phosphate （HA） and coverslip or their liquid extracts of α-MEM respectively. The proliferation, collagen secretion of PDLSCs were tested by MTY assay, collagen staining; cell morphology was observed by SEM; cytoskeleton was observed by confocal laser scanning microscopy（CLSM）. Results： FDDM was successfully fabri-cated. FDDM increased the proliferation and collagen secretion of PDLSCs compared with HA （P 〈 0. 05 ）. PDLSCs cultured on FDDM demonstrated superior attachment, cell morphology, growth viability, collagen secretion and cytoskeleton structure compared with those on HA. Conclusion： FDDM has superior biocompatibility and can promote the viability, collagen secretion and better cytoskeleton structure of PDLSCs in vitro.