【摘要】：目的:研究濃縮生長因子(concentrated growth factor,CGF)及其與1,25二羥基維生素D3(1,25(OH)_2VD_3)聯(lián)合應(yīng)用時,對骨髓間充質(zhì)干細(xì)胞(BMSCs)增殖及成骨分化的影響。方法:全骨髓培養(yǎng)法提取、分離SD大鼠股骨BMSCs,分別采用6種不同培養(yǎng)液而分為6組。使用SD大鼠血液在Medifuge離心機(jī)中制備CGF,并收集CGF濃縮液。實驗A、B、C組,分別為采用含5%、10%和20%濃度CGF的L-DMEM培養(yǎng)液組,單獨使用1,25(OH)_2VD_3(10~(-10)mol/L)的為D組。B組與D組混合為E組。F組為空白對照組。采用上述6組培養(yǎng)基孵育BMSCs 7 d。采用CCK-8實驗評價大鼠BMSCs在不同條件下的生長增殖情況;ALP活性染色實驗檢測各組ALP的生成,實時定量PCR檢測成骨分化標(biāo)志物基因OCN、Col-1和Runx2共3種成骨分化標(biāo)志物基因的表達(dá)情況。結(jié)果:5%和10%的CGF濃縮液能促進(jìn)大鼠BMSCs增殖,且10%濃度優(yōu)于5%濃度。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)與CGF(10%)濃縮液聯(lián)合應(yīng)用時,細(xì)胞增殖介于兩者之間。A、B、C組ALP活性及OCN、Col-1和Runx2的mRNA表達(dá)均受到了抑制。D組和E組均促進(jìn)了大鼠BMSCs的ALP活性,并提高了OCN、Col-1和Runx2的mRNA表達(dá)水平。結(jié)論:CGF濃縮液(10%)與1,25(OH)_2VD_3(10~(-10)mol/L)的聯(lián)合應(yīng)用,可促進(jìn)大鼠BMSCs的增殖,并對其成骨分化具有促進(jìn)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.
【作者單位】： 同濟(jì)大學(xué)附屬口腔醫(yī)院種植科 上海牙組織修復(fù)與再生工程技術(shù)研究中心;
【基金】：國家科技支撐計劃項目(201413AI041300) 國家自然科學(xué)基金項目81271110 中央高校基本科研業(yè)務(wù)費專項資金項目20152957
【分類號】：R329.2

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