温敏水凝胶负载脂源性干细胞对糖尿病大鼠皮肤创面血管新生影响的观察  

Effect of adipose-derived stem cell-thermosensitive hydrogel composite on angiogenesis of skin wounds in diabetic rate

作　　者：林凯桑 范丽君[1] 王思妤 潘道延 沈洁[1] 

机构地区：[1]南方医科大学第三附属医院内分泌代谢科,广州510630 [2]澳门大学健康科学学院

出　　处：《中国糖尿病杂志》2017年第5期449-453,共5页Chinese Journal of Diabetes

基　　金：广州市健康医疗协同创新重大专项(201604020007)

摘　　要：目的观察和探讨普兰尼克F127(PF127)温敏水凝胶负载脂源性干细胞(ADSCs)移植对糖尿病大鼠皮肤创面血管新生的影响。方法胶原酶消化法分离培养健康SD大鼠ADSCs并流式鉴定细胞表型。建立SD大鼠模型24只,在背部对称制作2个直径9 mm的全层皮肤切口,随机分为4组,创面注射ADSCs-PF127复合物(ADSCs-·PF127)组、ADSCs组、PF127组及PBS组,每组6只。术后评估各组创面愈合率,CD31免疫组织化学染色观察新生血管情况,qPCR检测血管内皮生长因子(VEGF)的表达。结果 ADSCs-PF127组愈合较快,术后14 d时愈合率为(97.8±1.8)%,高于其他3组(P=0.000)。免疫组织化学结果显示,术后7 d,ADSCs-PF127组和ADSCs组的血管数直观较PF127组、PBS组丰富,对术后14 d进行微血管计数,ADSCs-PF127组微脉管数高于ADSCs组[(21.0±2.4)vs(15.7±1.4)个,P=0.000]。qPCR结果显示,ADSCs-PF127组在7 d、14 d时,VEGF均高于其他组(P<0.05)。结论PF127水凝胶负载ADSCs可通过促进糖尿病大鼠创面新生毛细血管形成,从而加速创面愈合。Objective To observe the effect of transplanting adipose-derived stem cells （ADSCs） loaded on thermo-sensitive hydrogel pluronic F127 （PF127） on the angiogenesis of skin wounds in diabetic rats. Methods ADSCs were harvested from fat tissue of normal SD rats and were isolated and cultured by enzyme digestion. 24 diabetic rats were modeled. Two full-thickness 9 mm diameter skin incisions were made symmetrically in the back of each rat. These rats were randomly divided into four groups （n=6） and treated with ADSCs-PF127 composite,ADSCs, PF127 and PBS, respectively. The rates of wound healing among different groups were compared at day 3,7, 10 and 14. CD31 immunohistochemical staining was used to evaluate the degree of neovascularization and VEGF expression levels were detected via qPCR analysis. Results The healing rate of ADSCs-PF127 group （97. 8 ± 1.8%） was significantly higher than those in other groups on day 14 （P= 0. 000）. Histological observation showed that the number of visual vessels in ADSCs-PF127 group and ADSCs group was greater than PF127 group and PBS group on day 7. Microvessel count （MVD） of ADSCs-PF127 group was higher than ADSCs group on day 14 [（21.0 ±2. 4） vs （15.7±1.4） ,P=0. 000]. qPCR showed that VEGF levels were higher in ADSCs-PF127 group than those in other groups both on day 7 and 14 （P〈0.05）. Conclusion PF127 hydrogel loading ADSCs can accelerate the healing of diabetic wound by inducing angiogenesis.

关 键 词：脂源性干细胞 水凝胶 创面愈合 血管新生 

分 类 号：R587.2[医药卫生—内分泌]