负载aPexos和EGCG的可溶微针用于大鼠烫伤伤口愈合的研究  

Study on soluble microneedles loaded with aPexos and EGCG for healing of burn wounds in rats

作　　者：张阿龙 李琬晶 王红[1] 刘嘉馨[1] 曹晔[1] 钟锐[1] ZHANG Along;LI Wanjing;WANG Hong;LIU Jiaxin;CAO Ye;ZHONG Rui(Institute of Blood Transfusion,Chinese Academy of Medical Science&Peking Union Medical College,Chengdu 610052,China)

机构地区：[1]中国医学科学院北京协和医学院输血研究所,四川成都610052

出　　处：《中国输血杂志》2025年第3期333-342,共10页Chinese Journal of Blood Transfusion

基　　金：中国医学科学院医学与健康科技创新工程项目“血液安全保障及新产品新技术研究”(2021-I2M-1-060)。

摘　　要：目的 采用负载人工血小板外泌体(artificial platelet-derived exosomes, aPexos)的甲基丙烯酰化壳聚糖(Chitosan Methacryloyl, CSMA)作为针体材料,负载表没食子儿茶素没食子酸酯(Epigallocatechin gallate,EGCG)的明胶和羧甲基壳聚糖(Carboxymethyl chitosan, CMCS)作为背衬材料制备微针,探究aPexos-EGCG微针在治疗烫伤伤口愈合的治疗效果以及作用机制。方法 采用超声和梯度超离的方式制备提取aPexos,使用纳米颗粒追踪分析(NTA),透射电镜,ELISA试剂分别检测aPexos的浓度、形态及其生长因子TGF-β1,PDGF-BB,VEGF的含量。再将aPexos和EGCG分别负载在针体和背衬内,考察微针的针体形貌和机械性能。通过动物实验评价aPexos-EGCG微针应用于大鼠烫伤伤口愈合的效果。结果 aPexos的尺寸分布主要在50~150 nm,粒径为(132.7±3.8)nm,浓度为(2.88±0.36)×10^(13)个/mL。TGF-β1浓度为(1 363.0±135.39)ng/mL,PDGF-BB浓度为(63.5±14.8)ng/mL,VEGF的浓度为(1 606.0±77.5)pg/mL。机械性能检测结果证明其具有足够的的机械强度刺穿皮肤角质层。大鼠烫伤伤口愈合实验结果显示,d3和d7 aPexos-EGCG微针组的伤口愈合率与对照组相比有统计学差异,分别是(47.64±12.5)%vs(18.11±6.40)%,(87.45±5.57)%vs(79.85±5.03)%(P<0.05)。HE和Masson染色显示aPexos-EGCG微针组可见更完整的表皮层,胶原沉积百分比高于对照组。免疫荧光染色结果显示,aPexos-EGCG微针组的M1型巨噬细胞的含量下降,M2型巨噬细胞含量上升,新生血管标志物CD31和α-SMA均表达上升,且与其它3组相比有统计学差异(P<0.05)。结论 aPexos-EGCG微针具有良好的机械性能,能够成功刺穿皮肤角质层并递送aPexos和EGCG,可调控创面的M1型和M2型巨噬细胞的比例并抑制炎症反应,有效地促进大鼠烫伤伤口的愈合,为组织修复领域提供一种新的思路。Objective To use Chitosan Methacryloyl(CSMA)loaded with artificial platelet-derived exosomes(aPexos)as the needle body material,and gelatin and carboxymethyl chitosan(CMCS)loaded with Epigallocatechin gal-late(EGCG)as the backing material to prepare microneedles,aims to investigate the therapeutic effect and mechanism of aPexos-EGCG microneedles in the treatment of burn wound healing.Methods First,aPexos were extracted using ultra-sound and gradient ultracentrifugation.The concentration,morphology,and growth factor content(TGF-β1,PDGF-BB,VEGF)of the exosomes were assessed using NTA,transmission electron microscopy,and ELISA kits.The aPexos and EGCG were then dissolved in the needle body and backing materials,respectively.The aPexos and EGCG were loaded into the needle body and backing layer,respectively.The morphology,mechanical properties,and puncture performance of the microneedles were examined,and the preparation conditions for the microneedles loaded with aPexos-EGCG were optimized.Finally,animal experiments and tissue staining were conducted to assess the efficacy of the aPexos-EGCG microneedles in promoting burn wound healing in rats.Results The size distribution of aPexos was mainly in the range of 50-150 nm,with an average diameter of 132.7±3.8 nm and an average concentration of approximately 2.88×10^(13)±3.62×10^(12) particles/mL.The concentrations of TGF-β1,PDGF-BB,and VEGF were approximately 1363±135.9 ng/mL,63.5±14.8 ng/mL,and 1606.0±77.5 pg/mL,respectively.Mechanical property testing of the microneedles indicated that,compared with the blank microneedles,the drug-loaded microneedles had sufficient piercing ability to penetrate the stratum corneum of the skin.The results of the rat burn wound healing experiment showed that the wound healing rates of the d 3,d 7,aPexos-EGCG microneedle group compared to the control group showed significant differences,with respective values of(47.64±12.5)%vs(18.11±6.40)%,(87.45±5.57)%vs(79.85±5.03)%(P<0.05).Hematoxylin-eosin(HE)staining re-vealed more int

关 键 词：人工血小板外泌体 表没食子儿茶素没食子酸酯 烫伤创面 可溶微针 组织修复 

分 类 号：R331.143[医药卫生—人体生理学] R457.1[医药卫生—基础医学] R644