富血小板血浆凝胶联合脂肪干细胞促进大鼠创面修复的实验研究

目的：探讨富血小板血浆（PRP）凝胶联合脂肪干细胞（ADSCs）对大鼠创面修复的影响，为其临床应用提供理论基础。方法体外分离、培养、鉴定大鼠ADSCs。取16只SD大鼠，制备大鼠自体PRP凝胶，采用自身对照的方法，每只实验大鼠背部制作4个1 cm2的正方形皮肤全层缺损创面，分为AP组、A组、P组、O组，分别给予ADSCs＋PRP 处理、ADSCs处理、PRP处理以及空白对照处理，观察术后各组创面愈合情况。并于术后3、5、7、14 d分别随机处死4只实验大鼠，取创面组织，HE染色进行病理组织学观察，血小板?内皮细胞黏附分子（ CD31）免疫组化分析各时间点新生血管化情况，天狼猩红染色对Ⅰ型胶原含量和Ⅲ型胶原含量进行检测，并对创面组织的转化生长因子β1（ TGF?β1）表达情况进行逆转录聚合酶链反应（ RT?PCR）检测。结果大体观察AP组和P组较A组和O组结痂快、创缘收缩也较快；微血管计数CD31细胞阳性表达率及I型胶原蛋白含量AP组、A组、P组均高于O组，且AP组与O组差异更为显著，差异有统计学意义（ P＜0．05）；创面愈合过程中，AP组创面组织的TGF?β1表达也明显高于空白对照组，差异有统计学意义（ P＜0．05）。结论 ADSCs与PRP凝胶的应用能加速新生血管化及增加胶原蛋白合成，可有效加速创面愈合。

Thereseach of platelet rich plasma gel combined with adipose-derived stem cells in repairing soft tissue wounds in rats

Objective To explore the role of ADSCs and PRP in skin defect repairing, and to provide theoretical basis for clinical application. Methods Adipose tissues from inguinalis fat pad of SD rats were harvested, isolated, cultured, and identified. 16 SD rats were taken, and autologous PRP gel was prepared. 4 square shaped full?thickness skin defect ( a=1 cm2 ) was made on the rat back using self?control method. The wounds were divided into 4 groups randomly, of which group AP was treated with ADSCs and PRP; group A was treated with ADSCs;group P was treated with PRP and group O is blank control group. The wound healing process was observed. HE, sirius red, CD31 stain were observed at 3 d, 5 d,7 d, 14 d after surgery. TGF?beta 1 expression was detected by Rt?PCR in wound tissue. Results The wound contraction of group AP and group P were faster than the group A and O;CD31 positive expression rate and content of type I collagen of microvascular counts in group A, group P and group AP were higher than that of O group. There was significant difference between group AP and group O ( P<0.05) . In the process of wound healing, the expression of TGF ?beta 1 in wound tissue of group AP was significantly higher than the blank control group ( P<0.05) . Conclusion Application of ADSCs comnined with PRP gel by means of accelerating new blood vessels and increasing collagen synthesis can effectively accelerate wound healing.