犬脂肪干细胞膜片的构建及其复合富血小板纤维蛋白后的成骨能力

发布时间：2018-04-16 08:09

本文选题：脂肪干细胞 + 细胞膜片　；参考：《第三军医大学学报》2017年23期

【摘要】：目的初步研究犬脂肪干细胞(adipose-derived stem cell,ADSCs)膜片的构建及其生物学特性对膜片复合富血小板纤维蛋白(platelet-rich fibrin,PRF)后成骨能力的影响。方法取犬腹股沟处的脂肪进行消化,分离培养成脂肪干细胞进行相关检测。将P3代细胞加入含抗坏血酸的成膜培养液培养2周以上,构建脂肪干细胞膜片,采用HE染色及扫描电镜对膜片进行形态学检测。提取静脉血PRF,分为细胞膜片复合PRF的实验组和膜片对照组,2组分别成骨诱导后进行ALP染色和茜素红染色,并于诱导后第3、5天,采用RT-PCR检测成骨相关基因ALP、Runx2的表达。结果体外分离培养脂肪干细胞,成功构建干细胞膜片。光镜下可见细胞极性排列紧密,细胞外基质分泌。扫描电镜可见膜片表面光滑,细胞紧密排列。HE染色可见蓝染细胞紧密相连,细胞外大量基质。成骨诱导后,ADSCs膜片复合PRF的实验组ALP表达及茜素红染色钙结节形成量明显优于ADSCs膜片对照组,实验组ALP、Runx2基因m RNA表达高于对照组。结论成功构建犬脂肪干细胞膜片,膜片复合PRF后具有更强的成骨能力。
[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.
【作者单位】：重庆医科大学附属口腔医院正畸科口腔疾病与生物医学重庆市重点实验室重庆市高校市级口腔生物医学工程重点实验室;重庆医科大学附属口腔医院儿童口腔科口腔疾病与生物医学重庆市重点实验室重庆市高校市级口腔生物医学工程重点实验室;
【基金】：国家自然科学基金面上项目(81470772) 重庆市卫计委医学科研项目(20141013,2015HBRC009) 重庆市自然科学基金(CSTC2015jcyjA10028,CSTC2016jcyjA0238)~~
【分类号】：R781

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