成人成骨细胞与珊瑚羟基磷灰石的体外生物相容性

Biocompatibility of adult human osteoblasts with coral-derived hydroxyapatite in vitro

目的研究成人骨髓成骨细胞与珊瑚羟基磷灰石(carollinehydroxyapatite,CHA)在体外培养条件下的生物相容性。方法抽取健康成人骨髓组织,置于含体积分数为10%胎牛血清的DMEM培养基中培养,传代后改用含地塞米松β-甘油磷酸钠和维生素C的条件培养基培养,分为CHA复合细胞组和单纯细胞组,不同时间用倒置相差显微镜、HE染色光镜及扫描电镜观察。MTT法进行细胞增殖测定,并进行细胞微量蛋白含量和碱性磷酸酶的定量检测。结果成人骨髓成骨细胞体外培养时复合或不复合CHA均生长良好,表现出典型成骨细胞的形态特征和生物学特性,CHA利于细胞的贴附、生长与增殖,并对细胞的功能无不良影响。结论CHA是较理想的骨组织工程支架材料,成骨细胞复合CHA用于骨缺损的修复,具有广阔的临床应用前景。

O bj ective To study the biocompatibility of the osteoblasts from adult human bone ma rrow with coral-derived hy-droxyapatite (CHA) in in vitro culture. Methods Bone marrow was obtained from healthy adult subjects and cultured in Dul-becco's modi fied Eagle's medium (DMEM) containing 10% fetal bovine serum. The subsequent cel l passaging was conducted in conditioned medium containing dexamethasone, beta-sodium glycerophosphate and ascorbic acid, with the osteoblasts in culture then divided into CHA group (in which the cells were cultured with CHA) and osteobla sts group (without CHA). The proliferation and differentiation of all the cultu red cells were observed at different time points under inverted phase contrast m i-croscope, optical microscope with HE staining and scanning electron microscope respectively. Proliferation of the cultured cells were evaluated by MTT assay, and the activity of alkaline phosphatase and total micro-protein contents in th ese cultured os-teoblasts were quantitatively detected. Results The osteoblasts from adult human bone marrow grow well in vitro, regardless of the presence of CHA, with biological and morphological characteristics similar to those of norma l osteoblasts. CHA improved the adhesion, growth and proliferation of the cultu red cells, showing no adverse effects on the cell functions. Conclusion CHA is an optimal scaffold material for bone tissue engineering, which may potentiall y find clinical application for bone defect repair.