【摘要】 目的研究氯磷酸二钠修饰羟基磷灰石(hydroxyapatite,HA)在体内外促进骨形成对骨缺损修复效果的影响。方法制备氯磷酸二钠-HA复合材料,以HA为对照;分离培养成骨细胞,接种于氯磷酸二钠-HA和HA支架,四唑盐(methyl thiazolyl tetrazolium,MTT)比色法检测细胞增殖活性,测定碱性磷酸酶(alkaline phosphatase,ALP)活性,实时荧光定量PCR检测成骨细胞骨保护蛋白(osteoprotegerin,OPG)、核因子-κβ受体活化因子配体(receptor activator of nuclear factor kappa beta ligand,RANKL)mRNA表达的变化;进而将氯磷酸二钠-HA和HA植入兔下颌骨缺损区域,改良Masson染色及生物力学检测观察氯磷酸二钠对HA修复颌骨缺损的影响。结果 MTT和ALP活性检测表明成骨细胞在氯磷酸二钠-HA和HA支架上生长的增殖活性差异无统计学意义(P>0.05)。实时荧光定量PCR结果表明,两组细胞RANKL mRNA表达无明显变化,而氯磷酸二钠-HA组OPGmRNA显著上调(P<0.05)。改良Masson染色表明,与HA相比,氯磷酸二钠-HA更有利于材料周围新骨形成。生物力学检测结果表明氯磷酸二钠-HA组的压缩强度、拉伸强度、剪切强度均大于HA组(P<0.05)。结论复合在HA表面的氯磷酸二钠可能通过上调成骨细胞OPG表达,促进新骨形成,提高骨缺损修复的生物力学性能。更多还原

【Abstract】 Objective To evaluate the effects of clodronate modifying hydroxyapatite(HA) on cultured osteoblasts in vitro,and to observe the function of restoring mandibular defect in vivo.Methods Clodronate-HA composites were prepared,and HA as control;osteoblasts were isolated and cultured,and inoculated on Clodronate-HA and HA scaffold.The proliferation of osteoblasts was detected by methyl thiazolyl tetrazolium( MTT) and alkaline phosphatasel(ALP) activity test,and the mRNA expression of osteoprotegerin(OPG) and receptor activator of nuclear factor kappa beta ligand(RANKL) was detected by quantitative real-time RT-PCR.Clodronate-HA was implanted into rabbit mandibular defects,and the repairing effects were evaluated using Masson staining and biomechanics test.Results MTT and ALP activity test indicated that there was no significant difference between the Clodronate-HA and HA scaffolds in the growth activity of osteoblasts.The results of quantitative real-time RT-PCR showed there was no significant difference between two groups in mRNA expression of RANKL gene,while the expression of OPG gene showed higher expression in Clodronate-HA group(P<0.05).Masson staining indicated Clodronate-HA was more beneficial to new bone formation around the material compared with HA.The result of biomechanics test showed the compressive strength,tensile strength and shear strength of Clodronate-HA group were greater than those of HA group(P<0.05).Conclusion Clodronate modifying HA could upregulate the expression of OPG in osteoblasts,and be beneficial for HA to repair bone defects.更多还原