氯磷酸二钠表面改性羟基磷灰石对成骨细胞的影响

Effect of hydroxyapatite with clodronate modified surface on osteoblast

背景:羟基磷灰石本身的骨诱导活性有限,植入体内难与组织发生键合,结合不牢固,尚不能用于较大骨缺损的修复,限制了其临床应用。目的:观察羟基磷灰石经氯磷酸二钠表面改性后对成骨细胞的影响。设计、时间及地点:随机对照实验,于2007-09/11在口腔疾病研究国家重点实验室(四川大学)完成。材料:致密型块状羟基磷灰石,压缩强度40.0MPa,由四川大学纳米生物材料研究中心提供,氯磷酸二钠由美国TRM公司提供。方法:将羟基磷灰石制成10mm×10mm×0.5mm大小的块状,共48块,按随机数字表法分为实验组与对照组,每组24块。实验组羟基磷灰石与氯磷酸二钠复合制成氯磷酸二钠-羟基磷灰石;将未经特殊处理的羟基磷灰石作为对照组。分别将两组材料与成骨细胞复合培养。主要观察指标:培养1,3,7dMTT法检测材料上细胞增殖活性,同时检测细胞碱性磷酸酶活性。结果:培养1,3,7d,实验组与对照组细胞增殖活性相近,差异无显著性意义(P>0.05)。培养3,7d,实验组碱性磷酸酶活性略高于对照组,差异无显著性意义(P>0.05)。结论:用二磷酸盐对羟基磷灰石进行表面改性后不影响成骨细胞的生长、增殖和活性。

BACKGROUND： Hydroxyapatite leads to local osteogenesis in an osseous site. However, the osteogenic potential of hydroxyapatite is not adequate for filling the large bone defect, which limits its clinical application. OBJECTIVE： To investigate the effect on the osteoblasts of hydroxyapatite with modified surface by clodronate. DESIGN, TIME AND SETTING： A randomized controlled experiment was carried out in the State Key Laboratory of Oral Disease （Sichuan University） from September to November in 2007. MATERIALS： Dense hydroxyapatite （compressive strength： 40.0 MPa） was provided by Research Center for Nano-Biomaterials, Sichuan University. Clodronate was purchased from TRM Company （USA）. METHODS： The hydroxyapatite was made into 48 pieces （10 mm×10 mm×0.5 mm） and then randomly divided into experimental group and control group with 24 pieces in each. Samples in the experimental group were combined with the clodronate to form compound material, while samples in the control group were only cleaned with alcohol and distilled water under ultrasonic wave, and then dried for using. Samples in both groups were co-cultured with osteoblasts. MAIN OUTCOME MEASURES： MTT assay was used to evaluate the proliferation of the cells attached on the materials at days 1, 3, 7. Alkaline phosphatase activity of cells was also tested. RESULTS： The results of both MTT assay and alkaline phosphatase activity assay indicated that the proliferation activity was similar in the experimental group to the control group at days 1, 3, 7 （P 〉 0.05）. Alkaline phosphatase activity of cells in the experimental group was slightly higher than that in the control group, but there was no significant difference （P 〉 0.05）. CONCLUSION： The growth, proliferation and activity of osteoblasts are not influenced by the hydroxyapatite with modified surface by clodronate.