盐酸四环素缓释微球对大鼠成骨细胞活性的影响

Effect of Tetracycline-HCl sustained release microspheres on the activity of rat osteoblast in vitro

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 study the active effect of Tetracycline - HCl PLGA microspheres on cultured SD rat osteoblast in vitro. METHODS Osteoblasts were isolated from SD rats. Its morphological changes were observed and the cytobiological features were evaluated by ALP staining. MTT colorimetric assay were used to evaluate the proliferation of rat osteoblast by adding Tetracycline - HCl PLGA microspheres or only Tetracycline - HCl. Then, the ALP activity was measured. Expression of Col Ⅰ was examined with iinmunohistochemieal staining. RESULTS Statistical analysis showed that Tetracycline - HCl PLGA microspheres could significantly enhance proliferation and activity of rat osteoblast than that of Tetracycline - HCI and control group. Col Ⅰ was expressed in three groups. CONCLUSION Tetracycline - HCl PLGA microspheres enhance proliferation and ALP activity of rat osteoblast compared to Tetracycline - HCl.