外用红色诺卡氏菌细胞壁骨架提高脂肪间充质干细胞活性修复糖尿病创面  

Nocardia rubra cell wall skeleton for extemal use improves the viability of adipogenic mesenchymal stem cells and promotes diabetes wound repair

作　　者：闻丹丹 李强 沈才齐 纪哲 金培生 Wen Dandan;Li Qiang;Shen Caiqi;Ji Zhe;Jin Peisheng(Graduate School of Xuzhou Medical University,Xuzhou 221000,Jiangsu Province,China;Department of Plastic Surgery,Affiliated Hospital of Xuzhou Medical University,Xuzhou 221002,Jiangsu Province,China)

机构地区：[1]徐州医科大学研究生院,江苏省徐州市221000 [2]徐州医科大学附属医院整形外科,江苏省徐州市221002

出　　处：《中国组织工程研究》2022年第7期1038-1044,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:根据目前临床上外用红色诺卡氏菌细胞壁骨架对宫颈糜烂的治疗应用及其免疫作用,考虑到其对慢性创面治疗的可能性。目的:探讨外用红色诺卡氏菌细胞壁骨架对脂肪间充质干细胞体外增殖、凋亡的影响,以及对脂肪间充质干细胞体内移植存活率和皮肤愈合的影响。方法:以质量浓度为10 mg/L外用红色诺卡氏菌细胞壁骨架作用于脂肪间充质干细胞,采用CCK-8法检测细胞活力,EDU试剂盒检测细胞增殖能力;在脂肪间充质干细胞中加入50%蔗糖诱导细胞凋亡,给予外用红色诺卡氏菌细胞壁骨架干预后采用FITC-PI流式细胞凋亡试剂盒、TUNEL凋亡试剂盒检测细胞凋亡情况。第3代脂肪间充质干细胞中加入10 mg/L外用红色诺卡氏菌细胞壁骨架5 mL,共培养48 h,用荧光染料CM-Dil标记后以皮内注射方式注入糖尿病小鼠创缘皮肤,LB983活体成像系统检测细胞存活率,苏木精-伊红染色、Masson染色和免疫荧光染色验证慢性创面第14天愈合情况。结果与结论:外用红色诺卡氏菌细胞壁骨架可提高脂肪间充质干细胞的活力及增殖能力,并抑制其凋亡;在体内条件下外用红色诺卡氏菌细胞壁骨架可以增加脂肪间充质干细胞的存活率,糖尿病小鼠创面愈合速度更快。BACKGROUND:According to the current clinical application of Nocardia rubra cell wall skeleton to the treatment of cervical erosion and its immune effect,considering the possibility of its treatment of chronic wounds.OBJECTIVE:To study the effect of Nocardia rubra cell wall skeleton for extemal use on the proliferation and apoptosis of adipogenic mesenchymal stem cells in vitro,and the impact on the survival rate and skin healing of adipogenic mesenchymal stem cells transplantation in vivo.METHODS:Adipogenic mesenchymal stem cells were treated with 10 mg/L Nocardia rubra cell wall skeleton for extemal use.CCK-8 and EDU kits were used to detect the viability and proliferation of adipogenic mesenchymal stem cells.Adipogenic mesenchymal stem cells were treated with 50%sucrose to induce apoptosis,and then with Nocardia rubra cell wall skeleton for extemal use.Afterwards,flow cytometry and TUNEL assay were used to detect the apoptosis.Nocardia rubra cell wall skeleton for extemal use(10 mg/L,5 mL)was added to passage 3 adipogenic mesenchymal stem cells for 48 hours of culture.The pretreated cells labeled with the fluorescent dye CM-Dil were injected intracutaneously into the wound skin of diabetic mice.The cell survival rate was detected by the LB983 in vivo imaging system.Hematoxylin-eosin staining,Masson staining,and immunofluorescence were used to verify the healing of chronic wounds on day 14.RESULTS AND CONCLUSION:Nocardia rubra cell wall skeleton for extemal use could promote the viability and proliferation of adipogenic mesenchymal stem cells,and inhibit their apoptosis.Nocardia rubra cell wall skeleton for extemal use increased the survival rate of adipogenic mesenchymal stem cells under in vivo conditions.The wound healing rate is faster in diabetic mice.

关 键 词：红色诺卡氏菌细胞壁骨架 脂肪间充质干细胞 细胞活性 凋亡 慢性创面 糖尿病创面 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]