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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 and investigate the effects of Pluronic F127 (PF127) hydrogel-loaded adipose-derived stem cells (ADSCs) transplantation on angiogenesis in diabetic wounds. Methods Collagenase digestion method was used to isolate and culture ADSCs of healthy SD rats and parallel flow cytometry. Twenty-four SD rat models were established, and two full-thickness skin incisions of 9 mm in diameter were made on the back. They were randomly divided into 4 groups: ADSCs-PF127 group (ADSCs- PF127), ADSCs group, PF127 group PBS group, 6 in each group. The wound healing rate was evaluated after operation. CD31 immunohistochemical staining was used to observe the angiogenesis. The expression of vascular endothelial growth factor (VEGF) was detected by qPCR. Results The healing rate of ADSCs-PF127 group was faster than that of the other three groups (P <0.0001) after (14 days) and (97.8 ± 1.8)% respectively. The results of immunohistochemistry showed that the numbers of blood vessels in ADSCs-PF127 group and ADSCs group were more obvious than those in PF127 group and PBS group on the 7th day after operation. Microvessel counts were observed on the 14th day after operation. The number of microvessels in ADSCs-PF127 group was higher than that in ADSCs Group [(21.0 ± 2.4) vs (15.7 ± 1.4), P = 0.000]. qPCR results showed that the VEGF levels of ADSCs-PF127 group were higher than those of the other groups (P <0.05) on the 7th day and the 14th day. Conclusion PF127 hydrogel-loaded ADSCs can accelerate the wound healing by promoting the formation of capillaries in the wounds of diabetic rats.