[1]郭小伟,吴俊铮,崔 鑫,等.脂肪干细胞复合透明质酸支架对糖尿病小鼠皮肤创面愈合的影响实验研究[J].陕西医学杂志,2025,54(2):158-163,169.[doi:DOI:10.3969/j.issn.1000-7377.2025.02.003]
　GUO Xiaowei,WU Junzheng,CUI Xin,et al.Effect of adipose-derived stem cells combined with hyaluronic acid scffolad on skin wound healing in diabetic mice[J].,2025,54(2):158-163,169.[doi:DOI:10.3969/j.issn.1000-7377.2025.02.003]
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脂肪干细胞复合透明质酸支架对糖尿病小鼠皮肤创面愈合的影响实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
54
期数:
2025年2期
页码:
158-163,169
栏目:
基础研究
出版日期:
2025-02-05

文章信息/Info

Title:
Effect of adipose-derived stem cells combined with hyaluronic acid scffolad on skin wound healing in diabetic mice
作者:
郭小伟; 吴俊铮; 崔 鑫; 王文飞; 秦傲霜; 刘 宾
(西安交通大学附属西安市中心医院烧伤整形美容外科,陕西 西安 710004)
Author(s):
GUO Xiaowei; WU Junzheng; CUI Xin; WANG Wenfei; QIN Aoshuang; LIU Bin
(Department of Burn,Plastic and Cosmetic Surgery,Xi'an Central Hospital,Xi'an Jiaotong University,Xi'an 710004,China)
关键词:
糖尿病;  皮肤创面;  脂肪干细胞;  透明质酸;  微血管;  胶原纤维;  小鼠
Keywords:
Diabetes;  Skin wound;  Adipose-derived stem cells;  Hyaluronic acid;  Microvessels;  Collagen fibers;  Mice
分类号:
R -33
DOI:
DOI:10.3969/j.issn.1000-7377.2025.02.003
文献标志码:
A
摘要:
目的:探讨脂肪干细胞(ADSCs)复合透明质酸(HA)支架对糖尿病小鼠皮肤创面愈合的影响。方法:使用高蔗糖和高脂肪食物饲养30只小鼠10周,第11周于腹膜腔内注射50 mg/kg链脲佐菌素建立糖尿病小鼠模型。切取10只健康小鼠双侧腹股沟区域的脂肪组织,分离培养ADSCs,制备ADSCs-HA复合物。将30只糖尿病小鼠随机分为ADSCs-HA组、HA组和对照组,每组10只,每只小鼠背部制备2个创面模型。其中,ADSCs-HA组小鼠创面基底下及创周皮下注射0.3 ml ADSCs-HA复合物(ADSCs密度6×107个/ml,HA浓度10 mg/ml),HA组和对照组小鼠同部位分别注射0.3 ml同等浓度HA、0.3 ml 0.9%氯化钠溶液。于术后3、7、10、14 d,观察各组创面愈合情况,并计算创面愈合率。记录各组创面愈合时间。术后14 d,采用免疫组织化学染色法观察创面组织微血管数量,行Masson染色观察创面组织胶原纤维沉积情况,采用RT-qPCR法检测创面组织生长因子及胶原蛋白mRNA表达水平。结果:术后7、10、14 d,ADSCs-HA组小鼠创面较对照组、HA组明显缩小,且ADSCs-HA组创面愈合率高于HA组和对照组(均P<0.05)。ADSCs-HA组创面愈合时间较HA组和对照组缩短(均P<0.05)。免疫组织化学染色结果显示,ADSCs-HA组术后14 d创面组织可见许多圆形或椭圆形微血管,数量多于HA组和对照组。Masson染色结果显示,与HA组和对照组比较,ADSCs-HA组术后14 d创面组织中的胶原纤维排列更整齐,胶原沉积以及胶原纤维更多。ADSCs-HA组术后14 d创面组织微血管数量、胶原纤维沉积百分比及血管内皮生长因子(VEGF)、成纤维细胞生长因子(FGF)、Ⅰ型胶原蛋白(COL Ⅰ)、COL Ⅲ mRNA表达水平高于HA组和对照组(均P<0.05)。而HA组与对照组上述指标比较差异无统计学意义(均P>0.05)。结论:ADSCs复合HA支架能够促进糖尿病小鼠皮肤创面愈合。
Abstract:
Objective:To investigate the effect of adipose-derived stem cells(ADSCs)combined with hyaluronic acid(HA)scaffold on skin wound healing in diabetic mice.Methods:Thirty mice were fed with high-sucrose and high-fat food for 10 weeks,and then injected with 50 mg/kg streptozotocin intraperitoneally in the 11th week to establish a diabetic mouse model.Adipose tissue was excised from the inguinal region of 10 healthy mice,and ADSCs were isolated and cultured to prepare ADSCs-HA complexes.Thirty diabetic mice were randomly divided into ADSCs-HA group,HA group and control group,with 10 mice in each group.Each mouse had two wound models prepared on the back.The ADSCs-HA group mice were injected with 0.3 ml ADSCs-HA complex(ADSCs density 6×107 cells/ml,HA concentration 10 mg/ml)under the wound base and surrounding subcutaneous tissue,while the HA group and control group mice were injected with 0.3 ml of the same concentration HA and 0.3 ml of 0.9% sodium chloride solution,respectively.The wound healing situation of each group was observed at 3,7,10 and 14 days postoperatively,and the wound healing rate was calculated.The wound healing time of each group was recorded.At 14 days postoperatively,immunohistochemical staining was used to observe the number of microvessels in the wound tissue,Masson's staining was used to observe the deposition of collagen fibers in the wound tissue,and RT-qPCR was used to detect the mRNA expression levels of growth factors and collagen in the wound tissue.Results:At 7,10 and 14 days postoperatively,the wounds in the ADSCs-HA group mice were significantly smaller than those in the control and HA groups,and the wound healing rate in the ADSCs-HA group was higher than that in the HA and control groups(all P<0.05).The wound healing time in the ADSCs-HA group was shorter than that in the HA and control groups(all P<0.05).Immunohistochemical staining results showed that at 14 days postoperatively,many round or oval microvessels were visible in the wound tissue of the ADSCs-HA group,with a greater number than in the HA and control groups.Masson's staining results showed that compared with the HA and control groups,the collagen fibers in the wound tissue of the ADSCs-HA group at 14 days postoperatively were more neatly arranged,with more collagen deposition and collagen fibers.The number of microvessels,percentage of collagen fiber deposition,and mRNA expression levels of vascular endothelial growth factor(VEGF),fibroblast growth factor(FGF),type Ⅰ collagen(COL Ⅰ)and COL Ⅲ in the wound tissue at 14 days postoperatively in the ADSCs-HA group were higher than those in the HA and control groups(all P<0.05).However,there was no statistically significant difference in the above indicators between the HA group and the control group(all P>0.05).Conclusion:ADSCs combined with HA scaffold can promote skin wound healing in diabetic mice.

参考文献/References:

[1] LI Y,TENG D,SHI X,et al.Prevalence of diabetes recorded in mainland China using 2018 diagnostic criteria from the American Diabetes Association:National cross sectional study[J].BMJ,2020,369:m997.
[2] 中国老年医学学会烧创伤分会,中华医学会烧伤外科学分会,中国医师协会创面修复专业委员会.糖尿病足溃疡合并下肢血管病变的外科诊疗全国专家共识(2024版)[J].中华烧伤与创面修复杂志,2024,40(3):206-220.
[3] ABOULHODA B E,ABD-EL-FATTAH S.Bone marrow-derived versus adipose-derived stem cells in wound healing:Value and route of administration[J].Cell Tissue Res,2018,374(2):285-302.
[4] CHEN L,ZHENG Q,LIU Y,et al.Adipose-derived stem cells promote diabetic wound healing via the recruitment and differentiation of endothelial progenitor cells into endothelial cells mediated by the VEGF-PLCγ-ERK pathway[J].Arch Biochem Biophys,2020,692:108531.
[5] CHANDA A,ADHIKARI J,GHOSH A,et al.Electrospun chitosan/polycaprolactone-hyaluronic acid bilayered scaffold for potential wound healing applications[J].Int J Biol Macromol,2018,116:774-785.
[6] ARPAG O F,DAMLAR I,ALTAN A,et al.To what extent does hyaluronic acid affect healing of xenografts? A histomorphometric study in a rabbit model[J].J Appl Oral Sci,2018,26:e20170004.
[7] SONG J,LIU A,LIU B,et al.Natural biologics accelerate healing of diabetic foot ulcers by regulating oxidative stress[J].Front Biosci(Landmark Ed),2022,27(10):285.
[8] ARMSTRONG D G,TAN T W,BOULTON A J M,et al.Diabetic foot ulcers:A review[J].JAMA,2023,330(1):62-75.
[9] 中国糖尿病足细胞与介入治疗技术联盟,中国医师协会介入医师分会介入医学与生物工程技术委员会,国家放射与治疗临床医学研究中心.糖尿病足介入综合诊治临床指南(第九版)[J].介入放射学杂志,2024,33(4):341-354.
[10] CHEN Y,KUANG X,ZHOU J,et al.Proximal tibial cortex transverse distraction facilitating healing and limb salvage in severe and recalcitrant diabetic foot ulcers[J].Clin Orthop Relat Res,2020,478(4):836-851.
[11] 杨越,于文霞,何青敏,等.三黄血竭方外敷治疗糖尿病足溃疡疗效研究[J].陕西中医,2023,44(12):1740-1744.
[12] 孙玉苹,尹明亮,王文艳,等.四妙勇安汤对糖尿病足部溃疡患者血清lncRNA MALAT1表达量及足背血流动力学的影响[J].陕西中医,2023,44(8):1056-1060.
[13] 龙晨艳,程飚,田举富.血小板血浆促进创面愈合的细胞和分子机制[J].中国组织工程研究,2024,29(13):2793-2801.
[14] 尚玉,王丹霞.局部注射富血小板血浆联合水凝胶敷料对糖尿病足溃疡患者创面修复的影响及安全性研究[J].陕西医学杂志,2024,53(6):777-781.
[15] GADELKARIM M,ABUSHOUK A I,GHANEM E,et al.Adipose-derived stem cells:Effectiveness and advances in delivery in diabetic wound healing[J].Biomed Pharmacother,2018,107:625-633.
[16] BNUUELL B A,FLAAT M,GAGLIARDI C,et al.Adipose-derived stem cells:Isolation,expansion and differentiation[J].Methods,2008,45(2):115-120.
[17] ZHOU J,WEI T,HE Z.ADSCs enhance VEGFR3-mediated lymphangiogenesis via METTL3-mediated VEGF-C m6A modification to improve wound healing of diabetic foot ulcers[J].Mol Med,2021,27(1):146.
[18] SUN N,PANETTA N J,GUPTA D M,et al.Feeder-free derivation of induced pluripotent stem cells from adult human adipose stem cells[J].Proc Natl Acad Sci U S A,2009,106(37):15720-15725.
[19] ALTMAN A M,MATTHIAS N,YAN Y,et al.Dermal matrix as a carrier for in vivo delivery of human adipose-derived stem cells[J].Biomaterials,2008,29(10):1431-1442.
[20] HE J,SHAN S,JIANG T,et al.Mechanical stretch preconditioned adipose-derived stem cells elicit polarization of anti-inflammatory M2-like macrophages and improve chronic wound healing[J].FASEB J,2024,38(10):e23626.
[21] 左楠,崔莉,王爽.脂肪间充质干细胞对小鼠创面组织超氧化物歧化酶及血管内皮生长因子、表皮生长因子、碱性成纤维细胞生长因子表达的影响[J].陕西医学杂志,2022,51(1):7-10.
[22] KAWANO Y,PATRULEA V,SUBLET E,et al.Wound healing promotion by hyaluronic acid:Effect of molecular weight on gene expression and in vivo wound closure[J].Pharmaceuticals(Basel),2021,14(4):301-318.
[23] MARTIN P.Wound healing-aiming for perfect skin regeneration[J].Science,1997,276(5309):75-81.
[24] MOON K M,PARK Y H,LEE J S,et al.The effect of secretory factors of adipose-derived stem cells on human keratinocytes[J].Int J Mol Sci,2012,13(1):1239-1257.
[25] ZOGRAFOU A,TSIGRIA C,PAPADOPOULOS O,et al.Improvement of skin-graft survival after autologous transplantation of adipose-derived stem cells in rats[J].J Plast Reconstr Aesthet Surg,2011,64(12):1647-1656.
[26] LEE S H,JIN S Y,SONG J S,et al.Paracrine effects of adipose-derived stem cells on keratinocytes and dermal fibroblasts[J].Ann Dermatol,2012,24(2):136-143.

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备注/Memo

备注/Memo:
[基金项目]陕西省重点研发计划项目(2020SF-184)
更新日期/Last Update: 2025-02-04