Background:Acute skin wounds may compromise the skin barrier,posing a risk of infection.Small intestinal submucosa(SIS)is widely used to treat acute and chronic wounds.However,the efficacy of SIS to accelerate wound h...Background:Acute skin wounds may compromise the skin barrier,posing a risk of infection.Small intestinal submucosa(SIS)is widely used to treat acute and chronic wounds.However,the efficacy of SIS to accelerate wound healing still needs to be improved to meet clinical demands.To tackle this problem,platelet-rich plasma(PRP)is used due to its potency to promote proliferation,migration and adhesion of target cells.In this study,we applied PRP and SIS to skin wounds to explore their effects on wound healing by evaluating re-epithelialization,collagen production,angiogenesis and the inflammatory response.Methods:A1×1-cm full-thickness skin defectwas established in mice.Sixty mice were divided into four treatment groups:PRP+SIS,PRP,SIS and control.On days 3,5,7,10 and 14 post-surgery,tissue specimens were harvested.Haematoxylin and eosin,Masson’s trichrome,immunohistochemical and immunofluorescence double staining were used to visualize epidermal thickness,collagen and vascular regeneration and inflammation.Results:Wound contraction in the PRP and PRP+SIS groups was significantly greater,compared with the other groups,on days 3 and 5 post-surgery.A histological analysis showed higher collagen expression in the PRP and PRP+SIS groups on day 7,whichwas associated with a thicker epidermal layer on day 14.In addition,immunohistochemical staining showed that CD31-positive blood vessels and vascular endothelial growth factor expression in the PRP+SIS and PRP groups were significantly higher,compared with the control group.Furthermore,immunofluorescence double staining showed that the number of M1 and M2 macrophages in the PRP+SIS and PRP groups was higher,compared with the control and SIS groups alone,on day 3.However,on day 7,the number of M1 macrophages dramatically decreased in the PRP+SIS and PRP groups.The ratio of M2 to M1 macrophages in the PRP+SIS and PRP groups was 3.97 and 2.93 times that of the control group and 4.56 and 3.37 times that of the SIS group,respectively.Conclusion:Co-administration of SIS and PRP has a better effect on promoting angiogenesis,reepithelialization and collagen regeneration in managing acute wound healing than either agent alone.展开更多
目的:通过体内外实验研究复层猪小肠黏膜下层(multi-laminated small intestinal submucosa,mSIS)可吸收膜的降解性能,并与应用最为广泛的Bio-Gide可吸收生物膜进行比较,为其进一步应用于临床提供实验依据。方法:体外模拟降解采用胶原...目的:通过体内外实验研究复层猪小肠黏膜下层(multi-laminated small intestinal submucosa,mSIS)可吸收膜的降解性能,并与应用最为广泛的Bio-Gide可吸收生物膜进行比较,为其进一步应用于临床提供实验依据。方法:体外模拟降解采用胶原酶配制降解液,对mSIS膜和Bio-Gide膜进行降解,分别于不同时间点观察二者在降解液中的形态并取出称重,计算降解率。体内皮下埋植采用9只新西兰兔,每只动物背部皮下制备6个皮囊,分别埋入mSIS膜和Bio-Gide膜。于术后4、8、12周取材,通过肉眼观察及HE染色观察不同时间二者的降解程度及组织相容性。结果:体外降解实验显示mSIS膜在第12天降解完全,而Bio-Gide膜在第7天降解完全,且mSIS在降解液中维持形状的时间更长。皮下埋植4周时,mSIS膜和Bio-Gide膜形态相对完整,镜下观二者胶原纤维连续,膜周围少许炎症细胞浸润,Bio-Gide膜部分胶原纤维与周围组织融合。术后8周,mSIS膜形态基本完整,镜下观部分区域与结缔组织融合;肉眼观Bio-Gide膜已破碎,镜下仅可见部分残留纤维与周围组织结合,无完整膜的形态。术后12周时肉眼仅见少量mSIS膜残留碎片,镜下可见mSIS膜残留纤维,与周围结缔组织基本融合;肉眼及镜下观Bio-Gide膜均已消失。结论:mSIS膜皮下埋植降解时间约为12周,Bio-Gide膜约为8周,植入体内生物相容性良好。体外降解mSIS比Bio-Gide膜降解时间延长,且空间维持能力更佳。展开更多
目的观察人骨髓间充质干细胞(BM-MSCs)与猪小肠黏膜下层(SIS)的生物相容性,并检测其对人骨髓间充质干细胞增殖和分泌能力的影响。方法检测人BM-MSCs表面标记物以及多向分化能力。将第三代人BM-MSCs接种于SIS上作为为实验组,培养板单纯...目的观察人骨髓间充质干细胞(BM-MSCs)与猪小肠黏膜下层(SIS)的生物相容性,并检测其对人骨髓间充质干细胞增殖和分泌能力的影响。方法检测人BM-MSCs表面标记物以及多向分化能力。将第三代人BM-MSCs接种于SIS上作为为实验组,培养板单纯培养作为对照组。计算两组细胞分别在2h、4h、6h的贴壁率。通过CCK8的实验方法分别检测共培养12h、24h、36h、48h的OD值测定细胞的增殖情况。采用Hoechst33258荧光法检测2d、4d、6d两组细胞内的DNA含量。采用RT-PCR测定2d,4d,6d两组细胞VEGF m RNA的表达情况。共培养7d后在扫瞄电镜下观察人BM-MSCs在SIS上的生长状态。结果共培养2h、4h时,实验组的细胞贴壁率与对照组贴壁率差异有统计学意义(P<0.05),共培养6h,两组贴壁率差异无统计学意义(P>0.05)。共培养12h,两组细胞增殖差异无统计学意义(P>0.05),其余各时间点两组细胞增殖差异有统计学意义(P<0.05)。各时间点实验组细胞DNA含量明显高于对照组,差异有统计学意义(P<0.05)。各时间点实验组VEGF m RNA的表达量明显高于对照组,差异有统计学意义(P<0.05)。扫描电镜观察人BM-MSCs在小肠黏膜下层上生长状态良好。结论人骨髓间充质干细胞与小肠黏膜下层具有良好的生物相容性,SIS能够促进人BM-MSCs的增殖和分泌。展开更多
目的在猪全层皮肤切除模型中,观察负载白藜芦醇的猪小肠黏膜下层(small intestine submucosa,SIS)脱细胞基质对创面炎性反应和瘢痕形成的影响,评价愈合质量。方法自2022年3月至2022年7月,新疆医科大学第一附属医院整形美容外科将SIS脱...目的在猪全层皮肤切除模型中,观察负载白藜芦醇的猪小肠黏膜下层(small intestine submucosa,SIS)脱细胞基质对创面炎性反应和瘢痕形成的影响,评价愈合质量。方法自2022年3月至2022年7月,新疆医科大学第一附属医院整形美容外科将SIS脱细胞基质胶和白藜芦醇(resveratrol,Rev)溶液配制成膜片状的负载白藜芦醇的SIS脱细胞基质(Rev/SIS)。取小型猪3只,在每只猪背建立4个4 cm×4 cm的全层皮肤缺损创面,按照不同治疗手段随机分为4组:生理盐水纱布组(对照组)、白藜芦醇组(Rev组)、SIS脱细胞基质组(SIS组)和负载白藜芦醇的SIS脱细胞基质组(Rev/SIS组)。分别记录术后第7、14、21、28天的创面愈合及术后第42天的瘢痕形成状况;采集创缘组织行组织学检查和免疫组化染色分析,评估愈合过程中的炎性浸润和再血管化水平,以及所形成瘢痕组织的皮肤厚度和胶原沉积情况;并分别在术后第3、7天检测各组创面中炎性因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、M1型巨噬细胞标志物i NOS和M2型巨噬细胞标志物ARG-1的m RNA相对表达量。结果术后第7、14、21天,SIS组和Rev/SIS组创面残留率均显著低于对照组(P<0.05),术后第21天Rev/SIS组创面残留率低于SIS组(P<0.001),术后第28天除Rev/SIS组外其余3组创面均未完全愈合;术后第42天Rev/SIS组瘢痕面积最小,质地柔软,扁平且无明显色沉。Rev/SIS组TNF-a、IL-6 m RNA表达量均低于对照组(P<0.05),3种治疗手段均能降低创面i NOS m RNA表达(P<0.001),但以Rev/SIS组最为显著(P<0.05),且Rev/SIS组的ARG-1 m RNA表达高于其余3组(P<0.05),该组创面炎性浸润程度最低,再血管化水平最高。其余3组瘢痕的表皮、真皮厚度均低于对照组(P<0.01),也以Rev/SIS组改善最为显著(P<0.0001),且该组胶原沉积与正常皮肤相似,排列有序。结论尽管SIS脱细胞基质和白藜芦醇单独使用也可能产生抗炎效应,在促进创面愈合的同时,减少瘢痕形成面积,有效提高愈合质量;但二者联合应用的效果更为显著,Rev/SIS脱细胞基质为开发新型皮肤创面敷料提供了一种潜在选择。展开更多
基金funded by National Natural Science Foundation of China,grant number 81671924,81272105National Key Research and Development Plan of China,grant number 2017YFC1103301+6 种基金Health and Medical Treatment Collaborative Innovation Major Special Projects of Guangzhou,grant number 201508020253Science and Technology Key Project of Guangdong Province,grant number 2014B020212010Science and Technology Planning Project of Guangdong Province of China,grant number 2015B020233012Military Medical Innovation Special Projects,grant number 18CXZ029Zhejiang Provincial Basic Public Welfare Research Project,grant number GJ19H140001and China’s National Key R&D Programs(NKPs)grant number 2018YFB0407204.
文摘Background:Acute skin wounds may compromise the skin barrier,posing a risk of infection.Small intestinal submucosa(SIS)is widely used to treat acute and chronic wounds.However,the efficacy of SIS to accelerate wound healing still needs to be improved to meet clinical demands.To tackle this problem,platelet-rich plasma(PRP)is used due to its potency to promote proliferation,migration and adhesion of target cells.In this study,we applied PRP and SIS to skin wounds to explore their effects on wound healing by evaluating re-epithelialization,collagen production,angiogenesis and the inflammatory response.Methods:A1×1-cm full-thickness skin defectwas established in mice.Sixty mice were divided into four treatment groups:PRP+SIS,PRP,SIS and control.On days 3,5,7,10 and 14 post-surgery,tissue specimens were harvested.Haematoxylin and eosin,Masson’s trichrome,immunohistochemical and immunofluorescence double staining were used to visualize epidermal thickness,collagen and vascular regeneration and inflammation.Results:Wound contraction in the PRP and PRP+SIS groups was significantly greater,compared with the other groups,on days 3 and 5 post-surgery.A histological analysis showed higher collagen expression in the PRP and PRP+SIS groups on day 7,whichwas associated with a thicker epidermal layer on day 14.In addition,immunohistochemical staining showed that CD31-positive blood vessels and vascular endothelial growth factor expression in the PRP+SIS and PRP groups were significantly higher,compared with the control group.Furthermore,immunofluorescence double staining showed that the number of M1 and M2 macrophages in the PRP+SIS and PRP groups was higher,compared with the control and SIS groups alone,on day 3.However,on day 7,the number of M1 macrophages dramatically decreased in the PRP+SIS and PRP groups.The ratio of M2 to M1 macrophages in the PRP+SIS and PRP groups was 3.97 and 2.93 times that of the control group and 4.56 and 3.37 times that of the SIS group,respectively.Conclusion:Co-administration of SIS and PRP has a better effect on promoting angiogenesis,reepithelialization and collagen regeneration in managing acute wound healing than either agent alone.
文摘目的:通过体内外实验研究复层猪小肠黏膜下层(multi-laminated small intestinal submucosa,mSIS)可吸收膜的降解性能,并与应用最为广泛的Bio-Gide可吸收生物膜进行比较,为其进一步应用于临床提供实验依据。方法:体外模拟降解采用胶原酶配制降解液,对mSIS膜和Bio-Gide膜进行降解,分别于不同时间点观察二者在降解液中的形态并取出称重,计算降解率。体内皮下埋植采用9只新西兰兔,每只动物背部皮下制备6个皮囊,分别埋入mSIS膜和Bio-Gide膜。于术后4、8、12周取材,通过肉眼观察及HE染色观察不同时间二者的降解程度及组织相容性。结果:体外降解实验显示mSIS膜在第12天降解完全,而Bio-Gide膜在第7天降解完全,且mSIS在降解液中维持形状的时间更长。皮下埋植4周时,mSIS膜和Bio-Gide膜形态相对完整,镜下观二者胶原纤维连续,膜周围少许炎症细胞浸润,Bio-Gide膜部分胶原纤维与周围组织融合。术后8周,mSIS膜形态基本完整,镜下观部分区域与结缔组织融合;肉眼观Bio-Gide膜已破碎,镜下仅可见部分残留纤维与周围组织结合,无完整膜的形态。术后12周时肉眼仅见少量mSIS膜残留碎片,镜下可见mSIS膜残留纤维,与周围结缔组织基本融合;肉眼及镜下观Bio-Gide膜均已消失。结论:mSIS膜皮下埋植降解时间约为12周,Bio-Gide膜约为8周,植入体内生物相容性良好。体外降解mSIS比Bio-Gide膜降解时间延长,且空间维持能力更佳。
文摘目的观察人骨髓间充质干细胞(BM-MSCs)与猪小肠黏膜下层(SIS)的生物相容性,并检测其对人骨髓间充质干细胞增殖和分泌能力的影响。方法检测人BM-MSCs表面标记物以及多向分化能力。将第三代人BM-MSCs接种于SIS上作为为实验组,培养板单纯培养作为对照组。计算两组细胞分别在2h、4h、6h的贴壁率。通过CCK8的实验方法分别检测共培养12h、24h、36h、48h的OD值测定细胞的增殖情况。采用Hoechst33258荧光法检测2d、4d、6d两组细胞内的DNA含量。采用RT-PCR测定2d,4d,6d两组细胞VEGF m RNA的表达情况。共培养7d后在扫瞄电镜下观察人BM-MSCs在SIS上的生长状态。结果共培养2h、4h时,实验组的细胞贴壁率与对照组贴壁率差异有统计学意义(P<0.05),共培养6h,两组贴壁率差异无统计学意义(P>0.05)。共培养12h,两组细胞增殖差异无统计学意义(P>0.05),其余各时间点两组细胞增殖差异有统计学意义(P<0.05)。各时间点实验组细胞DNA含量明显高于对照组,差异有统计学意义(P<0.05)。各时间点实验组VEGF m RNA的表达量明显高于对照组,差异有统计学意义(P<0.05)。扫描电镜观察人BM-MSCs在小肠黏膜下层上生长状态良好。结论人骨髓间充质干细胞与小肠黏膜下层具有良好的生物相容性,SIS能够促进人BM-MSCs的增殖和分泌。
文摘目的在猪全层皮肤切除模型中,观察负载白藜芦醇的猪小肠黏膜下层(small intestine submucosa,SIS)脱细胞基质对创面炎性反应和瘢痕形成的影响,评价愈合质量。方法自2022年3月至2022年7月,新疆医科大学第一附属医院整形美容外科将SIS脱细胞基质胶和白藜芦醇(resveratrol,Rev)溶液配制成膜片状的负载白藜芦醇的SIS脱细胞基质(Rev/SIS)。取小型猪3只,在每只猪背建立4个4 cm×4 cm的全层皮肤缺损创面,按照不同治疗手段随机分为4组:生理盐水纱布组(对照组)、白藜芦醇组(Rev组)、SIS脱细胞基质组(SIS组)和负载白藜芦醇的SIS脱细胞基质组(Rev/SIS组)。分别记录术后第7、14、21、28天的创面愈合及术后第42天的瘢痕形成状况;采集创缘组织行组织学检查和免疫组化染色分析,评估愈合过程中的炎性浸润和再血管化水平,以及所形成瘢痕组织的皮肤厚度和胶原沉积情况;并分别在术后第3、7天检测各组创面中炎性因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、M1型巨噬细胞标志物i NOS和M2型巨噬细胞标志物ARG-1的m RNA相对表达量。结果术后第7、14、21天,SIS组和Rev/SIS组创面残留率均显著低于对照组(P<0.05),术后第21天Rev/SIS组创面残留率低于SIS组(P<0.001),术后第28天除Rev/SIS组外其余3组创面均未完全愈合;术后第42天Rev/SIS组瘢痕面积最小,质地柔软,扁平且无明显色沉。Rev/SIS组TNF-a、IL-6 m RNA表达量均低于对照组(P<0.05),3种治疗手段均能降低创面i NOS m RNA表达(P<0.001),但以Rev/SIS组最为显著(P<0.05),且Rev/SIS组的ARG-1 m RNA表达高于其余3组(P<0.05),该组创面炎性浸润程度最低,再血管化水平最高。其余3组瘢痕的表皮、真皮厚度均低于对照组(P<0.01),也以Rev/SIS组改善最为显著(P<0.0001),且该组胶原沉积与正常皮肤相似,排列有序。结论尽管SIS脱细胞基质和白藜芦醇单独使用也可能产生抗炎效应,在促进创面愈合的同时,减少瘢痕形成面积,有效提高愈合质量;但二者联合应用的效果更为显著,Rev/SIS脱细胞基质为开发新型皮肤创面敷料提供了一种潜在选择。
