PLGA仿生矿化人工骨对BMSCs体外成骨影响的实验研究

发布时间：2018-06-18 04:15

本文选题：仿生 + 聚乳酸-羟基乙酸　；参考：《中国矫形外科杂志》2014年15期

【摘要】：[目的]通过仿生矿化的方法构建聚乳酸-羟基乙酸共聚物(poly lactide-co-glycolide PLGA)人工骨,检测仿生矿化对PLGA生物学性能的影响。[方法]将三维多孔PLGA材料在模拟体液中矿化2周,构建仿生矿化人工骨,应用扫描电镜进行矿化物形貌观察,X射线能谱分析钙磷比值,X射线衍射仪和傅立叶转换红外光谱仪行矿化物物相分析。获取大鼠骨髓基质干细胞(bone mesenchymal stem cells,BMSCs),经传代培养后作为种子细胞与仿生矿化人工骨于体外联合培养,同时以未矿化的PLGA材料和煅烧松质骨为对比,应用RT-PCR的方法测定各组材料上大鼠BMSCs成骨基因表达情况,通过比较成骨基因表达时间先后次序和量的变化,表明各组材料对BMSCs向成骨细胞分化的影响。[结果]矿化后的PLGA材料网孔壁有磷灰石矿化层形成,矿化物主要成分为羟基磷灰石,钙磷比为1.53,类似于人骨无机质。大鼠BMSCs在三种材料上培养7 d后,碱性磷酸酶(alkaline phosphatase,ALP)相对表达水平煅烧松质骨组矿化组未矿化组(P0.05);骨涎蛋白(bonesialoprotein,BSP)和骨钙素(osteocalcin,OCN)在各组均没有表达。培养14 d后ALP相对表达水平煅烧松质骨组和矿化组无显著性差别(P0.05),均明显高于未矿化组(P0.05);煅烧松质骨组BSP相对表达水平明显高于矿化组(P0.05);OCN在煅烧松质骨组有少量表达。培养21 d后ALP相对表达水平未矿化组矿化组煅烧松质骨组(P0.05);BSP和OCN在表达水平显示矿化组和煅烧松质骨组没有明显差别(P0.05),均明显高于未矿化组(P0.05)。[结论]PLGA仿生矿化人工骨能够显著提高骨髓基质干细胞成骨基因的表达,促进其向成骨细胞分化。
[Abstract]:[objective] to construct poly lactide-co-glycolide PLGA artificial bone by biomimetic mineralization, and to detect the effect of biomimetic mineralization on the biological properties of poly (lactic acid-glycolic acid) copolymers. [methods] Three-dimensional porous PLGA material was mineralized in simulated body fluid for 2 weeks to construct biomimetic mineralized artificial bone. Scanning electron microscopy (SEM) was used to observe the morphology of mineralized matter. X-ray diffraction and Fourier transform infrared spectroscopy (FTIR) were used to analyze the phase of mineralized matter. Rat bone marrow stromal stem cells (BMSCs) were obtained and cultured as seed cells and biomimetic mineralized artificial bone in vitro, and compared with unmineralized mesenchymal stem materials and calcined cancellous bone. The expression of osteogenic genes in BMSCs was determined by RT-PCR. The effects of each group on the differentiation of BMSCs into osteoblasts were compared by comparing the time sequence and quantity of expression of osteogenic genes. [results] there was apatite mineralized layer in the porous wall of PLGA material after mineralization. The main component of the mineralized material was hydroxyapatite and the ratio of calcium to phosphorus was 1.53, which was similar to the inorganic substance of human bone. Rat BMSCs cultured on three materials for 7 days, the relative expression level of alkaline phosphatase (ALP) in calcined cancellous bone mineralization group was P0.05a, bone sialoprotein (BSPP) and osteocalcin OCNs were not expressed in each group. The relative expression level of ALP in calcined cancellous bone group and mineralized group was significantly higher than that in unmineralized bone group (P 0.05), and the relative expression level of BSP in calcined cancellous bone group was significantly higher than that in mineralization group (P 0.05 OCN). After 21 days of culture, the relative expression level of ALP in calcined cancellous bone group (P0.05BSP) and OCN (OCN) group showed no significant difference between the mineralized group and calcined cancellous bone group, which was significantly higher than that in the unmineralized group (P0.05). [conclusion] PLGA biomimetic mineralized artificial bone can significantly increase the expression of osteogenic gene in bone marrow stromal cells and promote its differentiation into osteoblasts.
【作者单位】：山东省泰安市中心医院创伤骨科;华中科技大学同济医学院附属协和医院骨科;
【分类号】：R318.08

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