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人脐带间充质干细胞联合自体Meek微型皮片移植在大面积烧伤患者中的应用前瞻性研究

[A prospective study on application of human umbilical cord mesenchymal stem cells combined with autologous Meek microskin transplantation in patients with extensive burns].

作者信息

Yan T T, Xiao R, Wang Y, Lin G A, Zheng Y, Zhao H, Li W J, Shang X Z, Meng J S, Hu D S, Li S, Wang C, Lin Z C, Chen H C, Zhao D Y, Tang D

机构信息

Military Burn Center, the 990th Hospital of PLA Joint Logistics Support Force, Zhumadian 463002, China.

Beijing Zhongjing Hi-Tech Biotechnology Co., Beijing 100089, China.

出版信息

Zhonghua Shao Shang Yu Chuang Mian Xiu Fu Za Zhi. 2023 Feb 20;39(2):114-121. doi: 10.3760/cma.j.cn501225-20220728-00319.

DOI:10.3760/cma.j.cn501225-20220728-00319
PMID:36878520
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11630450/
Abstract

To investigate the effects of human umbilical cord mesenchymal stem cells (hUCMSCs) combined with autologous Meek microskin transplantation on patients with extensive burns. The prospective self-controlled study was conducted. From May 2019 to June 2022, 16 patients with extensive burns admitted to the 990 Hospital of PLA Joint Logistics Support Force met the inclusion criteria, while 3 patients were excluded according to the exclusion criteria, and 13 patients were finally selected, including 10 males and 3 females, aged 24-61 (42±13) years. A total of 20 trial areas (40 wounds, with area of 10 cm×10 cm in each wound) were selected. Two adjacent wounds in each trial area were divided into hUCMSC+gel group applied with hyaluronic acid gel containing hUCMSCs and gel only group applied with hyaluronic acid gel only according to the random number table, with 20 wounds in each group. Afterwards the wounds in two groups were transplanted with autologous Meek microskin grafts with an extension ratio of 1∶6. In 2, 3, and 4 weeks post operation, the wound healing was observed, the wound healing rate was calculated, and the wound healing time was recorded. The specimen of wound secretion was collected for microorganism culture if there was purulent secretion on the wound post operation. In 3, 6, and 12 months post operation, the scar hyperplasia in wound was assessed using the Vancouver scar scale (VSS). In 3 months post operation, the wound tissue was collected for hematoxylin-eosin (HE) staining to observe the morphological changes and for immunohistochemical staining to observe the positive expressions of Ki67 and vimentin and to count the number of positive cells. Data were statistically analyzed with paired samples test and Bonferronni correction. In 2, 3, and 4 weeks post operation, the wound healing rates in hUCMSC+gel group were (80±11)%, (84±12)%, and (92±9)%, respectively, which were significantly higher than (67±18)%, (74±21)%, and (84±16)% in gel only group (with values of 4.01, 3.52, and 3.66, respectively, <0.05). The wound healing time in hUCMSC+gel group was (31±11) d, which was significantly shorter than (36±13) d in gel only group (=-3.68, <0.05). The microbiological culture of the postoperative wound secretion specimens from the adjacent wounds in 2 groups was identical, with negative results in 4 trial areas and positive results in 16 trial areas. In 3, 6, and 12 months post operation, the VSS scores of wounds in gel only group were 7.8±1.9, 6.7±2.1, and 5.4±1.6, which were significantly higher than 6.8±1.8, 5.6±1.6, and 4.0±1.4 in hUCMSC+gel group, respectively (with values of -4.79, -4.37, and -5.47, respectively, <0.05). In 3 months post operation, HE staining showed an increase in epidermal layer thickness and epidermal crest in wound in hUCMSC+gel group compared with those in gel only group, and immunohistochemical staining showed a significant increase in the number of Ki67 positive cells in wound in hUCMSC+gel group compared with those in gel only group (=4.39, <0.05), with no statistically significant difference in the number of vimentin positive cells in wound between the 2 groups (>0.05). The application of hyaluronic acid gel containing hUCMSCs to the wound is simple to perform and is therefore a preferable route. Topical application of hUCMSCs can promote healing of the autologous Meek microskin grafted area in patients with extensive burns, shorten wound healing time, and alleviate scar hyperplasia. The above effects may be related to the increased epidermal thickness and epidermal crest, and active cell proliferation.

摘要

探讨人脐带间充质干细胞(hUCMSCs)联合自体Meek微型皮片移植对大面积烧伤患者的影响。进行前瞻性自身对照研究。2019年5月至2022年6月,解放军联勤保障部队第990医院收治的16例大面积烧伤患者符合纳入标准,3例根据排除标准被排除,最终入选13例患者,其中男10例,女3例,年龄24 - 61(42±13)岁。共选取20个试验区(40个创面,每个创面面积为10 cm×10 cm)。每个试验区相邻的两个创面根据随机数字表分为hUCMSC +凝胶组(应用含hUCMSCs的透明质酸凝胶)和单纯凝胶组(仅应用透明质酸凝胶),每组20个创面。之后两组创面均移植自体Meek微型皮片,扩展比例为1∶6。术后2、3、4周,观察创面愈合情况,计算创面愈合率,记录创面愈合时间。术后创面如有脓性分泌物,则采集创面分泌物标本进行微生物培养。术后3、6、12个月,采用温哥华瘢痕量表(VSS)评估创面瘢痕增生情况。术后3个月,采集创面组织进行苏木精 - 伊红(HE)染色观察形态学变化,进行免疫组织化学染色观察Ki67和波形蛋白的阳性表达并计数阳性细胞数量。数据采用配对样本检验和Bonferronni校正进行统计学分析。术后2、3、4周,hUCMSC +凝胶组创面愈合率分别为(80±11)%、(84±12)%、(92±9)%,显著高于单纯凝胶组的(67±18)%、(74±21)%、(84±16)%(t值分别为4.01、3.52、3.66,均<0.05)。hUCMSC +凝胶组创面愈合时间为(31±11)d,显著短于单纯凝胶组的(36±13)d(t = -3.68,<0.05)。两组相邻创面术后创面分泌物标本的微生物培养结果相同,4个试验区为阴性,16个试验区为阳性。术后3、6、12个月,单纯凝胶组创面VSS评分分别为7.8±1.9、6.7±2.1、5.4±1.6,显著高于hUCMSC +凝胶组的6.8±1.8、5.6±1.6、4.0±1.4(t值分别为 -4.79、 -4.37、 -5.47,均<0.05)。术后3个月,HE染色显示hUCMSC +凝胶组创面表皮层厚度和表皮嵴较单纯凝胶组增加,免疫组织化学染色显示hUCMSC +凝胶组创面Ki67阳性细胞数量较单纯凝胶组显著增加(t = 4.39,<0.05),两组创面波形蛋白阳性细胞数量差异无统计学意义(P >0.05)。应用含hUCMSCs的透明质酸凝胶于创面操作简便,因此是一种较好的途径。局部应用hUCMSCs可促进大面积烧伤患者自体Meek微型皮片移植区愈合,缩短创面愈合时间,并减轻瘢痕增生。上述作用可能与表皮厚度增加、表皮嵴增加及细胞增殖活跃有关。

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