[关键词]
[摘要]
目的探讨脐带血富血小板血浆(CB-PRP)对糖尿病性难愈合创口修复的促进作用。方法 ELISA试剂盒法检测CB-PRP和外周血PRP(PB-PRP)的血管内皮生长因子(VEGF)和血小板衍生生长因子BB(PDGF-BB)的含量;SD大鼠随机分为对照组、模型组、PB-PRP组和CB-PRP组,除对照组外,按65 mg/kg剂量1次性ip链脲佐菌素(STZ)溶液诱发糖尿病模型;所有大鼠均建立全层皮肤缺损创伤模型。测定创伤再上皮化速率;术后10 d,采集组织学标本,进行苏木精伊红(HE)染色、天狼星红染色和CD31免疫荧光染色。结果 CB-PRP中VEGF、PDGF-BB含量高于PB-PRP,其中VEGF差异显著(P<0.05)。术后10、14、21 d,PB-PRP和CB-PRP组创口再上皮化速率显著快于模型组(P<0.05),术后21 d时,CB-PRP组的再上皮化率(92.8%)显著快于PB-PRP组(84.6%)(P<0.05)。HE染色结果显示,与模型组比较,PB-PRP和CB-PRP组肉芽组织生长更为旺盛,可见胶原纤维存在较多,新生的毛细血管较多。天狼猩红染色结果显示,PB-PRP和CB-PRP组创面增生区胶原纤维丰富,以Ⅰ型胶原为主,其间见Ⅲ型胶原。CD31免疫荧光染色结果显示,与模型组比较,CB-PRP和PB-PRP组毛细血管数量显著增加(P<0.05);与PB-PRP组比较,CB-PRP组的毛细血管数量显著增加(P<0.05)。结论 CB-PRP具有促进糖尿病性难愈合皮肤创口修复的作用,有望成为组织工程治疗的新方法。
[Key word]
[Abstract]
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.
[中图分类号]
R965
[基金项目]
天津市科技计划创新平台专项(18PTSYJC00070);天津市博士后择优资助计划项目(TJQYBSH2018030)