新型复合人工骨的生物相容和成骨性能研究△

Biocompatibility and osteogenic properties of a novel composited artificial bone

摘要:

[目的] 探讨脱细胞骨基质/磷酸钙/半水硫酸钙 （ACBM/CPC/CSH） 复合人工骨材料的生物相容性及成骨性能。[方法] 物理混合法合成 ACBM/CPC/CSH 复合人工骨材料，制备材料浸提液，用于急性、亚急性毒理、热原和表皮刺激实验。材料肌肉埋植实验，第 7、14 和 21 d 取材组织病理学检查，观察肌肉炎症反应；第 14 d 取材行流式细胞检查，观察血液与组织 CD4⁺ 、CD8⁺T 淋巴细胞含量变化，CCK-8 法检测细胞毒性。建立大鼠股骨髁骨缺损模型，分别植入 ACBM/CPC/CSH 骨材料组 （材料组）、磷酸钙/半水硫酸钙 （CPC/CSH）（对照组） 和不植任何材料 （空白组），观察成骨情况。[结果]浸提液未引起小鼠急性与亚急性毒理反应，体重变化差异无统计学意义（P＞0.05）。热原实验体温正常。表皮注射实验 72 h 内皮肤未出现红斑。肌肉埋植实验，第 7 d 肌肉组织有轻微炎症反应，21 d 炎症消失；第 14 d 流式细胞学检查，材料组、对照组和空白组外周血 CD4⁺ [(9.6±1.8) vs (10.1±1.2) vs (10.7±1.4), P=0.470]，CD8⁺ [(9.5±1.1) vs (10.3±1.8) vs (10.5±1.7), P=0.249]; 脾脏 CD4⁺ [(18.1±1.5) vs (17.2± 7.3) vs (17.5±1.0), P=0.195], CD8⁺ [(8.8±7.2) vs (7.7±7.6) vs (7.8±7.2), P=0.359] 淋巴细胞含量差异无统计学意义（P＞0.05）。X 线片与 Micro-CT 结果显示，各组骨缺损均有不同程度修复，ACBM/CPC/CSH 骨材料修复效果最佳。[结论]ACBM/CPC/CSH 复合人工骨材料具有良好的生物相容性和体内成骨性能，有利于不规则骨缺损的修补。

Abstract:

[Objective] To investigate the biocompatibility and osteogenic properties of acellular bone matrix/calcium phosphate/calcium sulfate hemihydrate (ACBM/CPC/CSH) composite artificial bone. [Methods] ACBM/CPC/CSH composite was synthesized by physical mixing method, and the material extract was prepared for acute and subacute toxicity, pyrogen and epidermal irritation tests. The material was implanted in the muscle and histopathological examination was performed on the 7^th, 14^th and 21^d to observe the inflammatory reaction of the muscle. On the 14^th day, flow cytometry was taken to observe the changes of CD4⁺ and CD8⁺T lymphocyte content in blood and tissues, and the cytotoxicity was detected by CCK-8 method. Bone defect model was established in the rat femoral condyle, and the bone formation was observed by Xray and CT after implantation of ACBM/CPC/CSH group (material group), calcium phosphate/calcium sulfate hemihydrate (CPC/CSH) group (control group) and no material implantation (blank group). [Results] The extract did not cause acute or subacute toxicity in mice, and there was no significant difference in body weight (P＞0.05). In the pyrogen test, the body temperature remained normal. Skin erythema did not appear within 72 h after epidermal injection. In the muscle implantation experiment, there was slight inflammation in the muscle tissue on the 7th day, and the inflammation disappeared on the 21st day. As results of flow cytometry on the 14th day, there were no significant differences among the three groups in terms of the peripheral blood lymphocyte content of CD4⁺ [(9.6±1.8) vs (10.1±1.2) vs (10.7± 1.4), P=0.470], CD8⁺ [(9.5±1.1) vs (10.3±1.8) vs (10.5±1.7), P=0.249], as well as the spleen CD4⁺ [(18.1±1.5) vs (17.2±7.3) vs (17.5±1.0), P= 0.195], and CD8⁺ [(8.8±7.2) vs (7.7±7.6) vs (7.8±7.2), P=0.359]. The X-ray and Micro-CT images showed that bone defects in all groups were repaired to different degrees, and ACBM/CPC/CSH bone materials had the best repair effect. [Conclusion] The ACBM/CPC/CSH composite artificial bone material has good biocompatibility and in vivo osteogenic properties, which is beneficial to the repair of irregular bone defects