目的探討臍帶血富血小板血漿（CB-PRP）對糖尿病性難愈合創(chuàng)口修復(fù)的促進(jìn)作用。方法 ELISA試劑盒法檢測CB-PRP和外周血PRP（PB-PRP）的血管內(nèi)皮生長因子（VEGF）和血小板衍生生長因子BB（PDGF-BB）的含量；SD大鼠隨機(jī)分為對照組、模型組、PB-PRP組和CB-PRP組，除對照組外，按65 mg/kg劑量1次性ip鏈脲佐菌素（STZ）溶液誘發(fā)糖尿病模型；所有大鼠均建立全層皮膚缺損創(chuàng)傷模型。測定創(chuàng)傷再上皮化速率；術(shù)后10 d，采集組織學(xué)標(biāo)本，進(jìn)行蘇木精伊紅（HE）染色、天狼星紅染色和CD31免疫熒光染色。結(jié)果 CB-PRP中VEGF、PDGF-BB含量高于PB-PRP，其中VEGF差異顯著（P<0.05）。術(shù)后10、14、21 d，PB-PRP和CB-PRP組創(chuàng)口再上皮化速率顯著快于模型組（P<0.05），術(shù)后21 d時，CB-PRP組的再上皮化率（92.8%）顯著快于PB-PRP組（84.6%）（P<0.05）。HE染色結(jié)果顯示，與模型組比較，PB-PRP和CB-PRP組肉芽組織生長更為旺盛，可見膠原纖維存在較多，新生的毛細(xì)血管較多。天狼猩紅染色結(jié)果顯示，PB-PRP和CB-PRP組創(chuàng)面增生區(qū)膠原纖維豐富，以Ⅰ型膠原為主，其間見Ⅲ型膠原。CD31免疫熒光染色結(jié)果顯示，與模型組比較，CB-PRP和PB-PRP組毛細(xì)血管數(shù)量顯著增加（P<0.05）；與PB-PRP組比較，CB-PRP組的毛細(xì)血管數(shù)量顯著增加（P<0.05）。結(jié)論 CB-PRP具有促進(jìn)糖尿病性難愈合皮膚創(chuàng)口修復(fù)的作用，有望成為組織工程治療的新方法。

Objective To investigate the promoting effect of cord blood-derived PRP (CB-PRP) on the diabetic refractory wound repair. Methods We detected the content of Vascular Endothelial Growth Factor (VEGF) and Platelet-Derived Growth Factor BB (PDGF-BB) in CB-PRP and peripheral blood-derived PRP (PB-PRP) by ELISA kit method. SD rats were randomly divided into control group, model group, PB-PRP group and CB-PRP group. Except for the control group, diabetic model was induced by once ip streptozotocin (STZ) solution at a dose of 65 mg/kg. A full-thickness skin defect model was established with all rats. The effects of refractory skin wound healing were compared including rate of re-epithelialization and histological examination. Hematoxylin eosin (HE) staining, Sirius red staining and CD31 immunofluorescence staining were performed respectively to detect the wound healing process in histology. Results The level of VEGF in CB-PRP group was significantly higher than that of PB-PRP (P < 0.05). CBPRP group showed higher level of PDGF-BB than PB-PRP, but there was no statistically significant difference between two groups. The re-epithelization rate of PB-PRP and CB-PRP group was significantly faster than that of model group at 10, 14 and 21 days after operation (P < 0.05). At 21 days after operation, the re-epithelization rate of CB-PRP group (92.8%) was significantly faster than that of PB-PRP group (84.6%) (P < 0.05). HE staining results showed that compared with model group, the granulation tissue of PB-PRP and CB-PRP groups grew more vigorously, and there were more collagen fibers and new capillaries. The results of Sirius red staining showed that there were abundant collagen fibers in the proliferative area of the wounds in PB-PRP and CB-PRP groups, mainly type I collagen, with type III collagen in between. CD31 immunofluorescence staining results showed that, compared with model group, the number of capillaries in CB-PRP and PB-PRP groups was significantly increased (P < 0.05); Compared with PBPRP group, the number of capillaries in CB-PRP group was significantly increased (P < 0.05). Conclusions CB-PRP can accelerate diabetic refractory wound repair. It is expected to be a novel therapeutic method in tissue engineering.

R965

天津市科技計劃創(chuàng)新平臺專項（18PTSYJC00070）；天津市博士后擇優(yōu)資助計劃項目（TJQYBSH2018030）