Mesenchymal stem cell(MSC)-derived exosomes(Exos)were reported to a prospective candidate in accelerating diabetic wound healing due to their pro-angiogenic effect.MSCs pretreated with chemistry or biology factors wer...Mesenchymal stem cell(MSC)-derived exosomes(Exos)were reported to a prospective candidate in accelerating diabetic wound healing due to their pro-angiogenic effect.MSCs pretreated with chemistry or biology factors were reported to advance the biological activities of MSC-derived exosomes.Hence,this study was designed to explore whether exosomes derived from human umbilical cord MSCs(hucMSCs)preconditioned with Nocardia rubra cell wall skeleton(Nr-CWS)exhibited superior proangiogenic effect on diabetic wound repair and its underlying molecular mechanisms.The results showed that Nr-CWS-Exos facilitated the proliferation,migration and tube formation of endothelial cells in vitro.In vivo,Nr-CWS-Exos exerted great effect on advancing wound healing by facilitating the angiogenesis of wound tissues compared with Exos.Furthermore,the expression of circIARS1 increased after HUVECs were treated with Nr-CWS-Exos.CircIARS1 promoted the pro-angiogenic effects of Nr-CWS-Exos on endothelial cells via the miR-4782-5p/VEGFA axis.Taken together,those data reveal that exosomes derived from Nr-CWS-pretreated MSCs might serve as an underlying strategy for diabetic wound treatment through advancing the biological function of endothelial cells via the circIARS1/miR-4782-5p/VEGFA axis.展开更多
目的观察人脂肪来源干细胞胶(SVF-GEL)中细胞因子含量及其体外对表皮、真皮细胞生物学行为的影响和临床疗效。方法(1)收集笔者单位行脂肪抽吸术获取的女性腹部脂肪抽吸液制备SVF-GEL。取1 mL SVF-GEL和1 mL DMEM高糖培养基,均用含体积分...目的观察人脂肪来源干细胞胶(SVF-GEL)中细胞因子含量及其体外对表皮、真皮细胞生物学行为的影响和临床疗效。方法(1)收集笔者单位行脂肪抽吸术获取的女性腹部脂肪抽吸液制备SVF-GEL。取1 mL SVF-GEL和1 mL DMEM高糖培养基,均用含体积分数10%胎牛血清、10 g/L青霉素、10 g/L链霉素的DMEM高糖培养基培养24 h,分别设为SVF-GEL组和阴性对照组,样本数均为6,采用酶联免疫吸附测定法检测上清液中表皮生长因子(EGF)、血管内皮生长因子(VEGF)含量。(2)选取处于对数生长期的5×105个人皮肤成纤维细胞(HSF)、HaCaT细胞分别接种于Transwell小室常规培养12 h,将所有培养细胞的Transwell小室分为2组,SVF-GEL处理组在Transwell小室中加入0.5 mL SVF-GEL与细胞共培养,对照组在Transwell小室中加入0.5 mL DMEM高糖培养基与细胞共培养,2种细胞各组样本数均为9。培养24 h后行划痕试验,观察划痕后0(即刻)、24、48 h划痕剩余宽度变化,测量划痕后24、48 h的细胞迁移距离;培养24、48、72 h采用细胞计数仪计数活细胞。(3)2018年6月—2019年6月,笔者单位应用SVF-GEL填充15例患者面部凹陷性瘢痕,其中男2例、女13例,年龄19~42岁。术后6个月,记录SVF-GEL存活情况、有无并发症。术前及术后6个月,对比患者瘢痕凹陷程度、色泽、柔软性,采用温哥华瘢痕量表进行色泽、血管、柔软性评分并计算总分。术后6个月记录十分满意、满意的患者数量。对数据行析因设计方差分析、配对样本t检验及Wilcoxon秩和检验。结果(1)SVF-GEL组、阴性对照组EGF含量分别为(316.6±12.8)、(3.4±0.6)pg/mL,VEGF含量分别为(568.67±12.19)、(4.93±0.16)pg/mL,组间比较,差异均有统计学意义(t=48.777、92.485,P<0.01)。(2)HSF、HaCaT各组细胞划痕剩余宽度随时间延长逐渐缩小,其中SVF-GEL处理组划痕后24、48 h的划痕剩余宽度均小于相应对照组。SVF-GEL处理组划痕后24、48 h HSF、HaCaT细胞迁移距离明显多于相应对照组(tHSF=-20.304、-43.516,tHaCaT=-15.060、-8.684,P<0.01);培养24、48、72 h,SVF-GEL处理组HSF、HaCaT活细胞数量明显多于相应对照组(tHSF=-3.374、-6.809、-18.036,tHaCaT=-4.793、-6.028、-8.141,P<0.05或P<0.01)。(3)术后6个月,患者移植的SVF-GEL均存活良好,无并发症,瘢痕凹陷程度较术前显著改善,色素沉着较术前显著减轻,瘢痕质地较术前柔软。术后6个月,15例患者面部凹陷性瘢痕的色泽、血管、柔软性评分及总分均明显低于术前(Z=-2.06,-2.07,-2.07,t=-15.811,P<0.05或P<0.01)。14例患者对疗效感到十分满意,1例患者对疗效感到满意。结论SVF-GEL含有细胞因子EGF、VEGF,可增强HSF、HaCaT细胞迁移和增殖能力,临床治疗凹陷性瘢痕的效果明显。展开更多
基金the National Natural Science Foundation of Jiangsu Province(No.BK20201155)the Foundation of Xuzhou Key Medical Talents(No.XWRCHT-20220067).
文摘Mesenchymal stem cell(MSC)-derived exosomes(Exos)were reported to a prospective candidate in accelerating diabetic wound healing due to their pro-angiogenic effect.MSCs pretreated with chemistry or biology factors were reported to advance the biological activities of MSC-derived exosomes.Hence,this study was designed to explore whether exosomes derived from human umbilical cord MSCs(hucMSCs)preconditioned with Nocardia rubra cell wall skeleton(Nr-CWS)exhibited superior proangiogenic effect on diabetic wound repair and its underlying molecular mechanisms.The results showed that Nr-CWS-Exos facilitated the proliferation,migration and tube formation of endothelial cells in vitro.In vivo,Nr-CWS-Exos exerted great effect on advancing wound healing by facilitating the angiogenesis of wound tissues compared with Exos.Furthermore,the expression of circIARS1 increased after HUVECs were treated with Nr-CWS-Exos.CircIARS1 promoted the pro-angiogenic effects of Nr-CWS-Exos on endothelial cells via the miR-4782-5p/VEGFA axis.Taken together,those data reveal that exosomes derived from Nr-CWS-pretreated MSCs might serve as an underlying strategy for diabetic wound treatment through advancing the biological function of endothelial cells via the circIARS1/miR-4782-5p/VEGFA axis.
文摘目的观察人脂肪来源干细胞胶(SVF-GEL)中细胞因子含量及其体外对表皮、真皮细胞生物学行为的影响和临床疗效。方法(1)收集笔者单位行脂肪抽吸术获取的女性腹部脂肪抽吸液制备SVF-GEL。取1 mL SVF-GEL和1 mL DMEM高糖培养基,均用含体积分数10%胎牛血清、10 g/L青霉素、10 g/L链霉素的DMEM高糖培养基培养24 h,分别设为SVF-GEL组和阴性对照组,样本数均为6,采用酶联免疫吸附测定法检测上清液中表皮生长因子(EGF)、血管内皮生长因子(VEGF)含量。(2)选取处于对数生长期的5×105个人皮肤成纤维细胞(HSF)、HaCaT细胞分别接种于Transwell小室常规培养12 h,将所有培养细胞的Transwell小室分为2组,SVF-GEL处理组在Transwell小室中加入0.5 mL SVF-GEL与细胞共培养,对照组在Transwell小室中加入0.5 mL DMEM高糖培养基与细胞共培养,2种细胞各组样本数均为9。培养24 h后行划痕试验,观察划痕后0(即刻)、24、48 h划痕剩余宽度变化,测量划痕后24、48 h的细胞迁移距离;培养24、48、72 h采用细胞计数仪计数活细胞。(3)2018年6月—2019年6月,笔者单位应用SVF-GEL填充15例患者面部凹陷性瘢痕,其中男2例、女13例,年龄19~42岁。术后6个月,记录SVF-GEL存活情况、有无并发症。术前及术后6个月,对比患者瘢痕凹陷程度、色泽、柔软性,采用温哥华瘢痕量表进行色泽、血管、柔软性评分并计算总分。术后6个月记录十分满意、满意的患者数量。对数据行析因设计方差分析、配对样本t检验及Wilcoxon秩和检验。结果(1)SVF-GEL组、阴性对照组EGF含量分别为(316.6±12.8)、(3.4±0.6)pg/mL,VEGF含量分别为(568.67±12.19)、(4.93±0.16)pg/mL,组间比较,差异均有统计学意义(t=48.777、92.485,P<0.01)。(2)HSF、HaCaT各组细胞划痕剩余宽度随时间延长逐渐缩小,其中SVF-GEL处理组划痕后24、48 h的划痕剩余宽度均小于相应对照组。SVF-GEL处理组划痕后24、48 h HSF、HaCaT细胞迁移距离明显多于相应对照组(tHSF=-20.304、-43.516,tHaCaT=-15.060、-8.684,P<0.01);培养24、48、72 h,SVF-GEL处理组HSF、HaCaT活细胞数量明显多于相应对照组(tHSF=-3.374、-6.809、-18.036,tHaCaT=-4.793、-6.028、-8.141,P<0.05或P<0.01)。(3)术后6个月,患者移植的SVF-GEL均存活良好,无并发症,瘢痕凹陷程度较术前显著改善,色素沉着较术前显著减轻,瘢痕质地较术前柔软。术后6个月,15例患者面部凹陷性瘢痕的色泽、血管、柔软性评分及总分均明显低于术前(Z=-2.06,-2.07,-2.07,t=-15.811,P<0.05或P<0.01)。14例患者对疗效感到十分满意,1例患者对疗效感到满意。结论SVF-GEL含有细胞因子EGF、VEGF,可增强HSF、HaCaT细胞迁移和增殖能力,临床治疗凹陷性瘢痕的效果明显。
