α-半水硫酸钙/磷酸八钙/RGD多肽复合材料的生物相容性研究  

Biocompatibility ofα-calcium sulfate hemihydrate/octacalcium phosphate/RGD polypeptide composite

作　　者：韩先顺 张之涵 柳辉 胡祥[1] 陶圣祥[1] Han Xianshun;Zhang Zhihan;Liu Hui;Hu Xiang;Tao Shengxiang(Department of Trauma and Micro-orthopedics,Zhongnan Hospital of Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院创伤与显微骨科,430071

出　　处：《中华实验外科杂志》2022年第5期912-915,共4页Chinese Journal of Experimental Surgery

摘　　要：目的将α-半水硫酸钙(α-CSH)、磷酸八钙(OCP)与RGD多肽(RGD)复合,探究该复合材料的生物相容性。方法制备材料浸提液备用并作为实验组,α-CSH组、α-CSH/OCP组、α-CSH/OCP/RGD组。CCK-8实验将材料浸提液与前期实验保存的第3代新西兰大白兔骨髓间充质干细胞(BMSCs)共培养,以完全培养液、0.64%苯酚溶液作为阴性及阳性对照,使用细胞计数试剂盒(CCK-8)检测细胞增殖状态,评估材料的细胞毒性。急性毒性实验:将材料浸提液注入购自武汉万千佳兴生物科技有限公司的MK小鼠(6个月龄,18~22 g)腹腔内,以生理盐水作为阴性对照,通过观察小鼠的生物学行为及死亡情况评估材料的急性毒性作用。皮内刺激实验将材料浸提液注入新西兰大白兔(3个月龄,2.0~2.5 kg)背部脊柱两侧皮肤,以生理盐水为阴性对照,通过观察注射部位皮肤出现红斑、水肿的情况评估材料的皮内刺激反应。以P<0.05为差异有统计学意义。结果CCK-8实验中α-CSH组1、3、5、7 d时BMSCs的相对增殖率(RGR)分别为85.87%、91.49%、91.30%、91.20%;α-CSH/OCP组分别为86.99%、92.53%、94.11%、89.98%;α-CSH/OCP/RGD组分别为91.45%、97.40%、97.66%、95.10%,3种材料浸提液共培养的BMSCs的RGR均>80%,按照细胞毒性分级其为0~1级,说明材料无细胞毒性。急性毒性实验中各组实验小鼠术后一般情况、行为活动、神经反应方面均未见异常,说明材料无急性毒性作用。皮内刺激实验中α-CSH组、α-CSH/OCP组、α-CSH/OCP/RGD组材料的原发刺激指数(PII)分别为0.125、0.250、0,均在0.0~0.4范围内,其反应类型为极轻微型。结论α-CSH/OCP/RGD多肽复合材料具有良好的生物相容性。Objective To explore the biocompatibility ofα-calcium sulfate hemihydrate(α-CSH),octacalcium phosphate(OCP)and RGD polypeptide(RGD)composites.Methods The material extracts were prepared and used as the experimental groups:α-CSH group,α-CSH/OCP group andα-CSH/OCP/RGD group.For the cell counting kit-8(CCK-8)assay,the extracts were co-cultured with the third generation of bone marrow mesenchymal stem cells(BMSCs)of New Zealand white rabbits preserved in previous experiments.The complete culture medium and 0.64%phenol solution were used as negative and positive controls.The cell proliferation was examined by CCK-8 Kit to evaluate the cytotoxicity of the materials.For acute toxicity test,the material extracts were injected into the abdominal cavity of MK mice(6 months old,18-22 g)purchased from Wuhan Qianqianjiaxing Biotechnology Co.,LTD,and normal saline was used as the negative control.The acute toxicity of the materials were evaluated by observing the biological behavior and death of mice.For the intradermal stimulation experiment,the material extracts were injected into the skin on both sides of the back spine of New Zealand white rabbits(3 months old,2.0-2.5 kg)purchased from Wuhan Wanwanjiaxing Biotechnology Co.,LTD,and the normal saline was used as the negative control.The intradermal stimulation reaction of the materials was evaluated by observing the erythema and edema of the skin at the injection site.P<0.05 was considered as statistically significant difference.Results In the CCK-8 assay,the relative growth rate(RGR)of BMSCs inα-CSH group at day 1,3,5 and 7 was 85.87%,91.49%,91.30%and 91.20%respectively;Inα-CSH/OCP group,the RGR of BMSCs was 86.99%,92.53%,94.11%and 89.98%respectively;Inα-CSH/OCP/RGD group,the RGR of BMSCs was 91.45%,97.40%,97.66%and 95.10%respectively.The RGR of BMSCs co-cultured with the extracts of the three materials was greater than 80%.According to the cytotoxicity grade,they were 0-1,indicating that the materials had no cytotoxicity.In the acute toxicity test,there was no abno

关 键 词：α-半水硫酸钙/磷酸八钙/RGD多肽 复合材料 生物相容性 

分 类 号：R318.08[医药卫生—生物医学工程]