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目的:研究多肽水凝胶复合骼金(nano-hydroxyapatite/collagen,nHAC)对大鼠脂肪干细胞(adipose-derived stemcells,ADSCs)体外增殖和早期骨向分化的作用。方法:用1%的多肽水凝胶与nHAC制备复合支架,未复合水凝胶的nHAC为对照组,使用扫描电子显微镜(field emission scanning electron microscope,FESEM)和共聚焦显微镜(Confocal laser scanningmicroscope,CLSM)进行表征,将ADSCs接种到两种支架材料上,以CCK-8检测1、3、5、7 d的增殖情况,以碱性磷酸酶(ALP)活性为早期骨向分化指标,检测其3、5、7 d的活性。结果:多肽水凝胶在nHAC表面形成涂层,ADSCs在有涂层的nHAC上黏附、铺展得更好,两种支架上的ADSCs在1、3、5、7 d都持续增殖,其中两组间在1、3 d无差异,5、7 d有差异,实验组增殖更快(P<0.05);ALP值在3、5、7 d都持续增高,实验组更高,两组间有差异(P<0.05),但在3、5、7 d时两两比较差异不显著(P>0.05)。结论:多肽水凝胶复合支架材料能显著促进ADSCs的黏附增殖,对细胞早期骨向分化有一定的促进作用,但不如对细胞增殖的促进作用显著。
OBJECTIVE: To study the effect of nHAC on the proliferation and early osteogenic differentiation of rat adipose-derived stem cells (ADSCs) in vitro. METHODS: The composite scaffolds were prepared with 1% peptide hydrogel and nHAC, and nHAC without the hydrogel as a control. The fluorescence intensity was measured using a field emission scanning electron microscope (FESEM) and a confocal laser scanning microscope (CLSM ). ADSCs were seeded on the two kinds of scaffolds. Proliferation of cells at 1, 3, 5 and 7 days was detected by CCK-8. Alkaline phosphatase (ALP) , 5, 7 d activity. RESULTS: Peptide hydrogel formed a coating on the surface of nHAC. ADSCs adhered and spread better on coated nHAC. ADSCs on both scaffolds continued to proliferate on the 1st, 3rd, 5th and 7th day. Two groups There was no difference between the two groups at 1 and 3 days, and there was a difference at 5 and 7 days. The proliferation of experimental group was faster (P <0.05). The ALP value increased continuously on the 3rd, 5th, 7th day in the experimental group with higher difference (P <0.05), but there was no significant difference between the two groups on the 3rd, 5th, 7th day (P> 0.05). CONCLUSION: The polypeptide hydrogel composite scaffold material can significantly promote the adhesion and proliferation of ADSCs and promote the differentiation of bone cells at early stage, but not as good as promoting proliferation of cells.