MEDLINE/PubMed Journal Browser Search

Pubmed for Handhelds

PUBMED FOR HANDHELDS

Search MEDLINE/PubMed

Title: [Effects of allogeneic mouse adipose-derived mesenchymal stem cell-microporous sheep acellular dermal matrix on healing of wound with full-thickness skin defect in mouse and the related mechanism].
Author: Cao SJ, Wang LF, Ba T, Fu X, Li F, Hao CG.
Journal: Zhonghua Shao Shang Za Zhi; 2018 Dec 20; 34(12):901-906. PubMed ID: 30585055.
Abstract:
Objective: To explore the allogeneic mouse adipose-derived mesenchymal stem cell (ADSC)-microporous sheep acellular dermal matrix (ADM) on healing of wound with full-thickness skin defect in mouse and the related mechanism. Methods: One Kunming mouse was sacrificed by cervical dislocation to collect adipose tissue from inguinal region. Mouse ADSCs were isolated from the adipose tissue and cultured in vitro. Cells of the third passage were identified by cell adipogenic and osteogenic differentiation. The expressions of CD73, CD90, CD105, and CD34 were analyzed by flow cytometry. After one sheep was sacrificed, microporous sheep ADM was prepared from sheep back using decellularization method and freezing-thawing method. A 12 mm diameter, round, full-thickness skin defect wound was made on the back of each one of 36 Kunming mice. The wounds were covered by microporous sheep ADM. The mice were divided into group ADSC and control (C) group with 18 mice in each group according to the random number table after surgery. A volume of 0.2 mL DMEM/F12 culture medium containing 1×10(6) ADSCs was injected between microporous sheep ADM and wound of mice in group ADSC. While 0.2 mL DMEM/F12 culture medium was injected between microporous sheep ADM and wound of mice in group C. On post surgery day (PSD) 12 and 17, wound healing rates of mice in the 2 groups were calculated. On PSD 7, 12, and 17, wound vascularization of mice in the 2 groups was observed under reverse irradiation of backlight. On PSD 7, 12, and 17, the wound granulation tissue of mice in group ADSC was observed by hematoxylin and eosin staining. On PSD 7, the thicknesses of granulation tissue of mice in the 2 groups was measured. On PSD 12 and 17, expressions of VEGF in wounds of mice in the 2 groups were detected by immunohistochemical method. The sample number was 6 in each group at each time point in the above experiments. Data were processed with t test and analysis of variance of factorial design. Results: (1) After 7 days of adipogenic induction, lipid droplet was observed in cytoplasm using oil red O staining. After 21 days of osteogenic induction, black deposits of calcium salts were detected using silver nitrate staining. Expression rates of CD73, CD90, CD105, and CD34 in cells were 97.82%, 99.32%, 97.35%, and 5.88% respectively. The cells were identified as ADSCs. (2) The wound healing rates of mice in group ADSC on PSD 12 and 17 [(78±6)%, (98±3)%] were significantly higher than those in group C [(60±9)%, (90±4)%, t=4.26, 4.46, P<0.01]. (3) On PSD 7, no vessel obviously grew into the center of wounds of mice in the 2 groups, while the granulation tissue has covered the wounds of mice in group ADSC. On PSD 12, the vessels were more abundant in wounds of mice in group ADSC than those in group C. On PSD 17, big vessels crossing the whole wounds was observed in wounds of mice in group ADSC, while big vessels were observed without crossing the whole wounds in wounds of mice in group C. (4) The wounds were covered with thin granulation tissue on PSD 7, and the granulation tissue began to thicken on PSD 12 and were covered by epidermis on PSD 17 in wounds of mice in group ADSC. On PSD 7, the granulation tissue in wounds of mice in group ADSC [(0.62±0.05) mm] was significantly thicker than that in group C [ (0.31±0.04) mm, t=12.27, P<0.01]. (5) On PSD 12 and 17, expressions of VEGF in wounds of mice in group ADSC [(80.7±2.2), (0.98±0.03)/mm(2)] were significantly than those in group C [(59.5±2.4), (81.5±2.6)/mm(2,) t=15.95, 14.14, P<0.01]. Conclusions: Allogeneic mouse ADSC-microporous sheep ADM can accelerate angiogenesis and growth of granulation tissue, thus promoting wound healing, which may be due to the increase of expression of VEGF. 目的： 探讨异体小鼠脂肪源性间充质干细胞(ADSC)-微孔化羊脱细胞真皮基质(ADM)对小鼠全层皮肤缺损创面愈合的影响及相关机制。 方法： 取1只昆明小鼠，断颈处死后取腹股沟处脂肪组织，体外分离培养小鼠ADSC，取第3代细胞行细胞成脂、成骨诱导分化鉴定，采用流式细胞仪检测CD34、CD73、CD90、CD105的表达。取1只绵羊，宰杀后取背部皮肤，脱细胞处理和冻融法制备微孔化羊ADM。取36只昆明小鼠，均在背部制备1个直径为12 mm的圆形全层皮肤缺损创面，以微孔化羊ADM覆盖创面。术后将小鼠按随机数字表法分为ADSC组和对照组，每组18只。将0.2 mL的悬浮有1×10(6)个ADSC的DMEM/F12培养液注入ADSC组小鼠微孔化羊ADM与创面之间，将0.2 mL的DMEM/F12培养液注入对照组小鼠微孔化羊ADM与创面之间。术后12、17 d，计算2组小鼠创面愈合率；术后7、12、17 d，背光反向照射下观察2组小鼠创面血管生长情况；术后7、12、17 d，行苏木素-伊红染色观察ADSC组小鼠创面肉芽组织，并于术后7 d测量2组小鼠创面肉芽组织厚度；术后12、17 d，采用免疫组织化学法检测2组小鼠创面血管内皮生长因子(VEGF)表达。以上实验中各组各时间点样本数均为6。对数据行t检验、析因设计方差分析。 结果： (1)成脂诱导7 d，经油红O染色细胞质可见红色小油滴；成骨诱导21 d，硝酸银染色培养基内可见黑色的钙盐沉积。细胞CD73、CD90、CD105、CD34表达率分别为97.82%、99.32%、97.35%、5.88%。细胞鉴定为ADSC。(2)术后12、17 d，ADSC组创面愈合率[(78±6)%、(98±3)%]均高于对照组[(60±9)%、(90±4)%，t＝4.26、4.46，P<0.01]。(3)术后7 d，2组小鼠都无明显的向创面中心生长的血管，但ADSC组的创面肉芽组织已全部覆盖创面；术后12 d，ADSC组小鼠创面比对照组血管更加丰富；术后17 d，ADSC组小鼠创面可见粗大的血管且贯穿整个创面，对照组也可见粗大的血管生成，但未贯穿创面。(4)ADSC组小鼠术后7 d创面已经覆盖肉芽组织但较薄，术后12 d肉芽组织明显增厚，术后17 d肉芽组织已被表皮覆盖。术后7 d，ADSC组小鼠创面肉芽组织厚度为(0.62±0.05)mm，明显厚于对照组的(0.31±0.04)mm，t＝12.27，P<0.01。(5)术后12、17 d，ADSC组小鼠创面VEGF的表达[(80.7±2.2)、(102.8±2.6)个/mm(2)]均高于对照组[(59.5±2.4)、(81.5±2.6)个/mm(2)，t＝15.95、14.14，P<0.01]。 结论： 异体小鼠ADSC-微孔化羊ADM可促进小鼠全层皮肤缺损创面的血管生成和肉芽组织生长，加速创面愈合，其机制可能与增加VEGF表达相关。.

[Abstract] [Full Text] [Related] [New Search]