本文選題：脂肪干細(xì)胞 + 細(xì)胞膜片��； 參考：《第三軍醫(yī)大學(xué)學(xué)報(bào)》2017年23期

【摘要】：目的初步研究犬脂肪干細(xì)胞(adipose-derived stem cell,ADSCs)膜片的構(gòu)建及其生物學(xué)特性對(duì)膜片復(fù)合富血小板纖維蛋白(platelet-rich fibrin,PRF)后成骨能力的影響。方法取犬腹股溝處的脂肪進(jìn)行消化,分離培養(yǎng)成脂肪干細(xì)胞進(jìn)行相關(guān)檢測(cè)。將P3代細(xì)胞加入含抗壞血酸的成膜培養(yǎng)液培養(yǎng)2周以上,構(gòu)建脂肪干細(xì)胞膜片,采用HE染色及掃描電鏡對(duì)膜片進(jìn)行形態(tài)學(xué)檢測(cè)。提取靜脈血PRF,分為細(xì)胞膜片復(fù)合PRF的實(shí)驗(yàn)組和膜片對(duì)照組,2組分別成骨誘導(dǎo)后進(jìn)行ALP染色和茜素紅染色,并于誘導(dǎo)后第3、5天,采用RT-PCR檢測(cè)成骨相關(guān)基因ALP、Runx2的表達(dá)。結(jié)果體外分離培養(yǎng)脂肪干細(xì)胞,成功構(gòu)建干細(xì)胞膜片。光鏡下可見細(xì)胞極性排列緊密,細(xì)胞外基質(zhì)分泌。掃描電鏡可見膜片表面光滑,細(xì)胞緊密排列。HE染色可見藍(lán)染細(xì)胞緊密相連,細(xì)胞外大量基質(zhì)。成骨誘導(dǎo)后,ADSCs膜片復(fù)合PRF的實(shí)驗(yàn)組ALP表達(dá)及茜素紅染色鈣結(jié)節(jié)形成量明顯優(yōu)于ADSCs膜片對(duì)照組,實(shí)驗(yàn)組ALP、Runx2基因m RNA表達(dá)高于對(duì)照組。結(jié)論成功構(gòu)建犬脂肪干細(xì)胞膜片,膜片復(fù)合PRF后具有更強(qiáng)的成骨能力。
[Abstract]:Objective to study the construction of adipose-derived stem cell (ADSCs) membrane and its biological characteristics on the osteogenic ability of dog adipose-derived stem cell (ADSCs) composite with platelet-rich fibrin (PRF).Methods the fat of dog groin was digested and the adipose stem cells were isolated and cultured.P3 generation cells were cultured in the medium containing ascorbic acid for more than 2 weeks to construct the adipose stem cell membrane. The morphology of the membrane was detected by HE staining and scanning electron microscope.PRFs were extracted from venous blood and divided into two groups: the experimental group with membrane membrane combined with PRF and the control group with ALP staining and alizarin red staining after osteogenesis induction. The expression of osteoblast-associated gene ALPnRunx2 was detected by RT-PCR 5 days after induction.Results Adipose stem cells were isolated and cultured in vitro.Under the light microscope, the cell polarity was closely arranged and extracellular matrix secreted.Scanning electron microscopy showed that the surface of the membrane was smooth and the cells were closely arranged. He staining showed that the blue stained cells were closely connected and a large amount of extracellular matrix was found.The expression of ALP and alizarin red stained calcium nodules in the experimental group was significantly higher than that in the ADSCs membrane control group after osteogenic induction, and the expression of ALP Runx2 gene m RNA in the experimental group was higher than that in the control group.Conclusion the canine adipose stem cell membrane was successfully constructed, and the membrane combined with PRF has stronger osteogenic ability.
【作者單位】： 重慶醫(yī)科大學(xué)附屬口腔醫(yī)院正畸科口腔疾病與生物醫(yī)學(xué)重慶市重點(diǎn)實(shí)驗(yàn)室重慶市高校市級(jí)口腔生物醫(yī)學(xué)工程重點(diǎn)實(shí)驗(yàn)室;重慶醫(yī)科大學(xué)附屬口腔醫(yī)院兒童口腔科口腔疾病與生物醫(yī)學(xué)重慶市重點(diǎn)實(shí)驗(yàn)室重慶市高校市級(jí)口腔生物醫(yī)學(xué)工程重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金面上項(xiàng)目(81470772) 重慶市衛(wèi)計(jì)委醫(yī)學(xué)科研項(xiàng)目(20141013,2015HBRC009) 重慶市自然科學(xué)基金(CSTC2015jcyjA10028,CSTC2016jcyjA0238)~~
【分類號(hào)】：R781

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