丁寅佳 崔磊 赵旭 司婷婷 赵启明

[摘要]目的：評价富血小板纤维蛋白（platelet-rich fibrin，PRF）和脂肪干细胞（adipose derived stem cells，ADSCs）对于糖尿病小鼠皮肤创面愈合的作用。方法：在18只糖尿病裸鼠和6只正常裸鼠上制成48个皮肤创面，将糖尿病裸鼠创面随机分为PRF-ADSCs组、PRF组、糖尿病对照组，观察各组创面愈合情况，并行组织病理学检查及CD31表达检测。结果：PRF存在大量纤维蛋白聚集并形成疏松的立体网络结构，在交界处有大量血小板和白细胞聚集。在接种ADSCs并培养1周后的PRF中，可见大量ADSCs细胞粘附在PRF的纤维蛋白网络中。伤后3、7、10、14d，PRF-ADSCs组创面愈合率分别为（35.2±4.2）%、（76.2±5.0）%、（88.3±5.0）%、（97.6±2.7）%，PRF组为（28.0±5.0）%、（62.0±5.2）%、（81.0±3.7）%、（92.3±3.2）%，糖尿病对照组为（16.0±2.7）%、（26.1±4.8）%、（60.9±7.7）%、（72.0±8.1）%，正常对照组为（31.0±4.0）%、（64.8±4.2）%、（83.0±3.5）%、（98.5±2.7）%，PRF-ADSCs组愈合率优于其它3组，差异均具有统计学意义（P<0.01）。PRF-ADSCs组创面愈合时间为（14.75±0.45）d，PRF组为（15.33±0.49）d，糖尿病对照组创面愈合时间为（17.67±0.49）d，正常对照组创面愈合时间为（14.42±0.51）d，PRF-ADSCs组和正常对照组的愈合时间优于其余2组（P<0.01）。伤后14d PRF-ADSCs组、PRF组和正常对照组创面毛细血管和组CD31阳性细胞计数多于糖尿病对照组（P<0.01）。结论：应用PRF联合ADSCs治疗糖尿病小鼠皮肤创面，可以显著促进创面愈合，且效果优于单纯使用PRF。

[关键词]富血小板纤维蛋白;血小板;脂肪干细胞;糖尿病小鼠;创面;愈合

[中图分类号]R622    [文献标志码]A    [文章编号]1008-6455（2019）02-0008-06

Abstract： Objective  To evaluate the effects of PRF and ADSCs on skin wound healing in diabetic mice. Methods  48 skin wounds were made on 18 diabetic nude mice and 6 normal nude mice. Diabetic nude mice wounds were randomly divided into PRF-ADSCs group， PRF group， and diabetes control group. Wound healing was observed in each group and was observed. Pathological examination and detection of CD31 expression of the specimen was made. Results  PRF has a large amount of fibrin aggregation and forms a loose three-dimensional network structure， with a large number of platelet and leukocyte aggregation at the junction. In PRF inoculated with ADSCs and cultured for one week， a large number of ADSCs cells were seen adhering to the PRF's fibrin network. At 3， 7， 10， and 14d after injury， the wound healing rate in the PRF-ADSCs group was （35.2±4.2）%， （76.2±5.0）%， （88.3±5.0）% and （97.6±2.7）% respectively， in the PRF group was （28.0±5.0）%， （62.0±5.2）%， （81.0±3.7）% and （92.3±3.2）% respectively， in diabetic control group was （16.0±2.7）%， （26.1±4.8）%，（60.9±7.7）% and （72.0±8.1）% respectively， and in normal control group was （31.0±4.0）%， （64.8±4.2）%， （83.0±3.5）% and （98.5±2.7）% respectively. The healing rate of PRF-ADSCs group was better than the other three groups， and the differences were statistically significant （P<0.01）. The wound healing time in PRF-ADSCs group was （14.75±0.45）d， in PRF group was （15.33±0.49）d， in diabetic control group was （17.67±0.49）d， and in normal control group was （14.42±0.51）d， The healing time of PRF-ADSCs group and normal control group were better than other two groups （P<0.01）. The counts of capillary and CD31 positive cells in wounds of PRF-ADSCs group， PRF group and normal control group were higher than those of diabetic control group at 14d after injury （P<0.01）. Conclusion  This study found that the use of PRF in combination with ADSCs in the treatment of diabetic skin wounds， can significantly promote wound healing， and the effect is better than simply using PRF.

Key words： platelet-rich fibrin; platelet; adipose derived stem cells; diabetic mice; wound; healing

目前，糖尿病已经成为全球范围内的一种常见疾病。目前针对糖尿病足（diabetic foot ulcers，DFU）患者的治疗，除了控制患者全身血糖水平外，在局部创面所采用的传统治疗方法均需要较长的治疗时间，加重了患者的经济负担，甚至最终也未能达到创面愈合。富血小板纤维蛋白（platelet-rich fibrin，PRF）主要由血浆及血小板构成，其包含有纤维蛋白支架及聚集于其中的白细胞和各类生长因子、细胞因子、趋化因子等，被广泛应用口腔、颌面以及整形美容外科等多个医学领域[1-3]。目前已有初步研究表明，PRF对于创面愈合具有促进作用[4-5]，应用PRF治疗糖尿病创面也具有积极作用[6-7]。随着干细胞研究的深入，也为治疗创面损伤提供了新的方法。……