PRF凝胶所含细胞因子对SD大鼠骨髓间充质干细胞体外成骨成脂诱导分化影响的研究
发布时间：2019-07-01 16:02
【摘要】：目的 探究富血小板纤维蛋白（PRF）凝胶中的细胞因子对SD大鼠骨髓间充质干细胞体外成骨成脂定向诱导分化的影响，为组织工程种子细胞体外培养和口腔种植临床应用提供理论依据和实践指导。 方法 取SD大鼠股骨和胫骨骨髓液，贴壁法培养。传代后进行增殖，，在实验组培养基中加入PRF凝胶，对照组不加。培养1-7天进行MTT检测细胞增殖活性。同时分别进行成骨细胞和脂肪细胞诱导，7d后去除残余的PRF凝胶，培养14d成骨诱导组进行茜素红染色、矿化结节半定量分析、ALP检测，骨钙素含量检测。成脂诱导组进行油红o染色和定量分析。 结果 MTT检测显示1~3天内，PRF组和对照组无显著性差异，4~7天开始，PRF组吸光值大于对照组。成骨诱导茜素红染色观察PRF组红色矿化结节数量较对照组为多。骨钙素检测显示PRF组OD值大于对照组（P0.05），ALP检测、半定量分析显示PRF组A值均大于对照组（P0.05）。油红O染色显示PRF组脂滴含量大于对照组，定量分析PRF组A值对照组（P0.05） 结论 PRF凝胶所含细胞因子可促进大鼠骨髓间充质干细胞增殖，对成骨分化和成脂分化同样有促进作用。PRF凝胶可作为骨组织工程种子细胞体外培养生长因子源。
[Abstract]:Purpose To investigate the effect of cytokines in platelet-rich fibrin (PRF) gel on the differentiation of bone marrow-derived mesenchymal stem cells in vitro and to induce differentiation of bone marrow-derived mesenchymal stem cells in vitro, and to provide the theoretical basis and practical means for the in vitro culture of the tissue engineering seed cells and the clinical application of oral planting. Conductors. Methods SD rat femur and tibia bone marrow fluid were obtained. the culture was carried out after passage, and the propagation was carried out after passage, and the PRF gel was added to the culture medium of the experimental group, The control group was not cultured for 1-7 days and the MTT assay was fine. Cell proliferation was induced by osteoblasts and adipocytes, and the residual PRF gel was removed at 7 days. After 7 days, the residual PRF gel was removed, and the 14-day osteogenic induction group was cultured for fluorescein red staining, semi-quantitative analysis of mineralized nodules, ALP detection, and bone calcium. The detection of the content of the pigment. The fat-forming induction group was stained with oil red o. and The results showed that there was no significant difference between the PRF group and the control group within 1 to 3 days after the MTT test. The light absorption value of the group was greater than that of the control group. The red mineralization of the PRF group was observed by osteogenic induction. The results showed that the OD value of the PRF group was greater than that of the control group (P0.05), the ALP test and the semi-quantitative analysis showed that the value of the group A in the PRF group was greater than that of the control group (P0.05). Group A (P0.05). The oil red O staining showed that the lipid drop in the PRF group was greater than that of the control group, and the value of the PRF group A was quantitatively analyzed. control Conclusion Cytokine in the PRF gel can promote the proliferation of rat bone marrow-derived mesenchymal stem cells. The PRF gel can be used as bone tissue engineering.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
本文编号：2508621
[Abstract]:Purpose To investigate the effect of cytokines in platelet-rich fibrin (PRF) gel on the differentiation of bone marrow-derived mesenchymal stem cells in vitro and to induce differentiation of bone marrow-derived mesenchymal stem cells in vitro, and to provide the theoretical basis and practical means for the in vitro culture of the tissue engineering seed cells and the clinical application of oral planting. Conductors. Methods SD rat femur and tibia bone marrow fluid were obtained. the culture was carried out after passage, and the propagation was carried out after passage, and the PRF gel was added to the culture medium of the experimental group, The control group was not cultured for 1-7 days and the MTT assay was fine. Cell proliferation was induced by osteoblasts and adipocytes, and the residual PRF gel was removed at 7 days. After 7 days, the residual PRF gel was removed, and the 14-day osteogenic induction group was cultured for fluorescein red staining, semi-quantitative analysis of mineralized nodules, ALP detection, and bone calcium. The detection of the content of the pigment. The fat-forming induction group was stained with oil red o. and The results showed that there was no significant difference between the PRF group and the control group within 1 to 3 days after the MTT test. The light absorption value of the group was greater than that of the control group. The red mineralization of the PRF group was observed by osteogenic induction. The results showed that the OD value of the PRF group was greater than that of the control group (P0.05), the ALP test and the semi-quantitative analysis showed that the value of the group A in the PRF group was greater than that of the control group (P0.05). Group A (P0.05). The oil red O staining showed that the lipid drop in the PRF group was greater than that of the control group, and the value of the PRF group A was quantitatively analyzed. control Conclusion Cytokine in the PRF gel can promote the proliferation of rat bone marrow-derived mesenchymal stem cells. The PRF gel can be used as bone tissue engineering.
【学位授予单位】：辽宁医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
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