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目的观察盐酸四环素缓释微球对体外培养的成骨细胞活性的影响。方法体外分离培养SD大鼠成骨细胞并通过形态学观察、碱性磷酸酶(ALP)钙-钴法染色鉴定其细胞生物学特征;将盐酸四环素聚乳酸-聚羟基乙酸共聚物(PLGA)微球、盐酸四环素分别与成骨细胞共培养,采用四甲基偶氮唑盐(MTT)法检测各实验组成骨细胞的增殖情况,通过碱性磷酸酶活性测定法检测各实验组成骨细胞中ALP的活性,免疫组化Ⅰ型胶原蛋白(ColⅠtype)染色观察各实验组成骨细胞中I型胶原的表达情况。结果盐酸四环素-PLGA微球能显著促进SD大鼠成骨细胞的增殖,同时增强了ALP的表达,其效应持续时间高于盐酸四环素组和空白对照组,Ⅰ型胶原在3组细胞中的表达无显著差异。结论制备的盐酸四环素-PLGA微球具有缓释促进成骨细胞增殖和增加ALP活性的作用,在骨创伤的修复重建治疗中具有潜在的应用价值。
Objective To observe the effect of tetracycline hydrochloride sustained-release microspheres on osteoblast activity in vitro. Methods SD rat osteoblasts were isolated and cultured in vitro. The cell biological characteristics were identified by alkaline phosphatase (ALP) calcium-cobalt staining and morphological observation. The effect of tetracycline polylactic acid-polyglycolic acid copolymer (PLGA) And tetracycline hydrochloride were respectively co-cultured with osteoblasts. The proliferation of osteoblasts in each experimental group was detected by MTT assay. The ALP levels in osteoblasts of each experimental group were measured by alkaline phosphatase activity assay The expression of type I collagen in osteoblasts of each experimental group was observed by immunohistochemistry with type I collagen. Results Tetracycline-PLGA microspheres could significantly promote the proliferation of osteoblasts and increase the expression of ALP in SD rats. The duration of effect was longer than that of tetracycline hydrochloride and blank control group. The expression of type Ⅰ collagen in 3 groups of cells No significant difference. Conclusion The prepared tetracycline-PLGA microspheres have the effects of slow release and promote the proliferation of osteoblasts and increase the activity of ALP, which has potential value in the repair and reconstruction of bone trauma.