浓缩生长因子与1,25二羟基维生素D3对骨髓间充质干细胞增殖与成骨分化的影响
发布时间：2019-02-22 11:09
【摘要】：目的:研究浓缩生长因子(concentrated growth factor,CGF)及其与1,25二羟基维生素D3(1,25(OH)_2VD_3)联合应用时,对骨髓间充质干细胞(BMSCs)增殖及成骨分化的影响。方法:全骨髓培养法提取、分离SD大鼠股骨BMSCs,分别采用6种不同培养液而分为6组。使用SD大鼠血液在Medifuge离心机中制备CGF,并收集CGF浓缩液。实验A、B、C组,分别为采用含5%、10%和20%浓度CGF的L-DMEM培养液组,单独使用1,25(OH)_2VD_3(10~(-10)mol/L)的为D组。B组与D组混合为E组。F组为空白对照组。采用上述6组培养基孵育BMSCs 7 d。采用CCK-8实验评价大鼠BMSCs在不同条件下的生长增殖情况;ALP活性染色实验检测各组ALP的生成,实时定量PCR检测成骨分化标志物基因OCN、Col-1和Runx2共3种成骨分化标志物基因的表达情况。结果:5%和10%的CGF浓缩液能促进大鼠BMSCs增殖,且10%浓度优于5%浓度。而20%CGF和1,25(OH)_2VD_3(10~(-10)mol/L)都抑制了大鼠BMSCs的增殖。1,25(OH)_2VD_3(10~(-10)mol/L)与CGF(10%)浓缩液联合应用时,细胞增殖介于两者之间。A、B、C组ALP活性及OCN、Col-1和Runx2的mRNA表达均受到了抑制。D组和E组均促进了大鼠BMSCs的ALP活性,并提高了OCN、Col-1和Runx2的mRNA表达水平。结论:CGF浓缩液(10%)与1,25(OH)_2VD_3(10~(-10)mol/L)的联合应用,可促进大鼠BMSCs的增殖,并对其成骨分化具有促进作用。
[Abstract]:Aim: to study the effects of concentrated growth factor (concentrated growth factor,CGF) and its combination with 1O25 (OH) _ 2VD_3 on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods: whole bone marrow culture method was used to extract and separate femoral BMSCs, from SD rats into 6 groups. SD rat blood was used to prepare CGF, and collect CGF concentrate in Medifuge centrifuge. Experiment A, group B, group C were treated with L-DMEM medium containing 5%, 10% and 20% CGF, respectively. Group D was treated with 25 (OH) _ 2VD_3 (10 ~ (-10) mol/L), group B was mixed with group D as group E and group F as blank control group. BMSCs was incubated in the above 6 groups for 7 days. CCK-8 test was used to evaluate the growth and proliferation of rat BMSCs under different conditions. ALP activity staining assay was used to detect the formation of ALP in each group, and real-time quantitative PCR was used to detect the expression of three osteogenic marker genes, OCN,Col-1 and Runx2. Results: the concentration of 5% and 10% CGF could promote the proliferation of BMSCs in rats, and the concentration of 10% was better than that of 5%. The proliferation of BMSCs in rats was inhibited by both 20%CGF and 1o 25 (OH) _ 2VD_3 (10 ~ (-10) mol/L). The combination of 1o 25 (OH) _ 2VD_3 (10 ~ (-10) mol/L) and CGF (10%) concentrated solution could inhibit the proliferation of BMSCs. The activity of ALP and the expression of mRNA of OCN,Col-1 and Runx2 in group A were inhibited. Group D and group E promoted the ALP activity of BMSCs and increased the expression of mRNA in OCN,Col-1 and Runx2. Conclusion: the combination of CGF concentrated solution (10%) and 1o 25 (OH) _ 2VD_3 (10 ~ (-10) mol/L) can promote the proliferation of rat BMSCs and promote its osteogenic differentiation.
【作者单位】： 同济大学附属口腔医院种植科 上海牙组织修复与再生工程技术研究中心;
【基金】：国家科技支撑计划项目(201413AI041300) 国家自然科学基金项目81271110 中央高校基本科研业务费专项资金项目20152957
【分类号】：R329.2
[Abstract]:Aim: to study the effects of concentrated growth factor (concentrated growth factor,CGF) and its combination with 1O25 (OH) _ 2VD_3 on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods: whole bone marrow culture method was used to extract and separate femoral BMSCs, from SD rats into 6 groups. SD rat blood was used to prepare CGF, and collect CGF concentrate in Medifuge centrifuge. Experiment A, group B, group C were treated with L-DMEM medium containing 5%, 10% and 20% CGF, respectively. Group D was treated with 25 (OH) _ 2VD_3 (10 ~ (-10) mol/L), group B was mixed with group D as group E and group F as blank control group. BMSCs was incubated in the above 6 groups for 7 days. CCK-8 test was used to evaluate the growth and proliferation of rat BMSCs under different conditions. ALP activity staining assay was used to detect the formation of ALP in each group, and real-time quantitative PCR was used to detect the expression of three osteogenic marker genes, OCN,Col-1 and Runx2. Results: the concentration of 5% and 10% CGF could promote the proliferation of BMSCs in rats, and the concentration of 10% was better than that of 5%. The proliferation of BMSCs in rats was inhibited by both 20%CGF and 1o 25 (OH) _ 2VD_3 (10 ~ (-10) mol/L). The combination of 1o 25 (OH) _ 2VD_3 (10 ~ (-10) mol/L) and CGF (10%) concentrated solution could inhibit the proliferation of BMSCs. The activity of ALP and the expression of mRNA of OCN,Col-1 and Runx2 in group A were inhibited. Group D and group E promoted the ALP activity of BMSCs and increased the expression of mRNA in OCN,Col-1 and Runx2. Conclusion: the combination of CGF concentrated solution (10%) and 1o 25 (OH) _ 2VD_3 (10 ~ (-10) mol/L) can promote the proliferation of rat BMSCs and promote its osteogenic differentiation.
【作者单位】： 同济大学附属口腔医院种植科 上海牙组织修复与再生工程技术研究中心;
【基金】：国家科技支撑计划项目(201413AI041300) 国家自然科学基金项目81271110 中央高校基本科研业务费专项资金项目20152957
【分类号】：R329.2
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