低浓度万古霉素复合PMMA骨水泥的细胞毒性研究

Study on Cytotoxicity of Low Concentration Vancomycin/PMMA Composite Cements

目的:研究低浓度万古霉素(1%)复合聚甲基丙烯酸甲酯PMMA骨水泥对SD仔鼠原代成骨细胞的增殖以及凋亡的影响。方法:采用分离SD仔鼠颅骨与胰酶消化法获取SD仔鼠原代成骨细胞,通过细胞爬片碱性磷酸酶染色对原代成骨细胞进行鉴定;利用CCK-8法检测低浓度万古霉素复合PMMA及纯PMMA骨水泥浸提液对SD大鼠原代成骨细胞的增殖影响、流式细胞仪检测万古霉素复合PMMA及纯PMMA骨水泥浸提液对对SD大鼠原代成骨细胞的凋亡影响。结果:SD大鼠原代成骨细胞在低浓度万古霉素复合PMMA骨水泥浸提液中第1、2、3天的增殖更明显(P<0.05),在纯PMMA骨水泥浸提液中第1、5天凋亡显著增加(P<0.05)。结论:低浓度万古霉素复合PMMA骨水泥相比PMMA骨水泥具有较低的细胞毒性,在临床应用于骨缺损伴局部骨感染的治疗方面具有一定的优势。

Objective： To study the cytotoxicity of vancomycin （low concentration, 1% ）/polymethyl methacrylate （PMMA） cements on SD rat-derived primary osteoblasts. Methods： Primary osteoblasts were obtained by the calvarias trypsin-digestion method, which were isolated from newborn Sprague-Dawley rats. To identify the primary osteoblasts, alkaline phosphatase staining was used.The activity of cell proliferation in vancomycin （low concentration）/ polymethyl methacrylate （PMMA） and polymethyl methacrylate （PMMA） cements extract were assessed by CCK-8 assay. Apoptosis of osteoblasts induced by vancomycin （low concentration）/ polymethyl methacrylate （PMMA） and polymethyl methacrylate （PMMA） cements extract was observed by flow cytometry. Results： The activity of SD rat rat-derived osteoblasts proliferation in the vancomycin （low concentration）/polymethyl methacrylate （PMMA）cement extracts was significantly more than that in the group of PMMA cement extracts on day 1, 2, 3 （P 〈 0.05）. The apoptotic rate of primary osteoblasts in extracts from the vancomycin （low concentration）/PMMA cements was significantly lower than that in extract from the PMMA cement on day 1 and 5 （P 〈 0.05）. Conclusion： Compared with PMMA cements, the vancomycin （low concentration）/PMMA cements has a good cytocompatibility with primary osteoblasts. It will be useful inbone defect with infection repair.