血小板伪装银纳米颗粒水凝胶加速1型糖尿病大鼠创面愈合  

Platelet-camouflaged silver nanoparticle hydrogel accelerates wound healing in type 1 diabetic rats

作　　者：王金磊 李科 赵亮 Wang Jinlei;Li Ke;Zhao Liang(School of Pharmacy,Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China;Life Science Institute,Jinzhou Medical University,Jinzhou 121001,Liaoning Province,China)

机构地区：[1]锦州医科大学药学院,辽宁省锦州市121001 [2]锦州医科大学生命科学研究院,辽宁省锦州市121001

出　　处：《中国组织工程研究》2024年第17期2659-2666,共8页Chinese Journal of Tissue Engineering Research

摘　　要：背景:细菌感染和血管生成功能受损一直是糖尿病创面愈合的障碍,多药耐药的问题同样不容忽视,因此迫切需要寻找一种新的治疗策略。目的:制备血小板伪装银纳米颗粒水凝胶,观察其对糖尿病创面的治疗作用。方法:①体外实验:通过回流加热的方式制备超小银纳米颗粒(ultra-small silver nanoparticles,usAgNPs),将其与血小板(platelets,PL)偶联后加入羧甲基纤维素(carboxymethyl cellulose,CMC)水凝胶中,制备usAgNPs-PL/CMC水凝胶,表征水凝胶的微观结构、溶胀性能及力学性能,采用二甲氧唑黄比色法、活性氧法、细菌生物膜法、细菌活性等方法检测该水凝胶的抗菌能力。②体内实验:将40只SD大鼠随机分为假手术组、模型组、CMC水凝胶组、usAgNPs/CMC水凝胶组和usAgNPs-PL/CMC水凝胶组,每组8只,除假手术组外,其余4组建立1型糖尿病模型,造模成功后在5组大鼠背部制作1个深达筋膜层的皮肤创面(直径20 mm),假手术组、模型组大鼠创面注射生理盐水,CMC水凝胶组、usAgNPs/CMC水凝胶组和usAgNPs-PL/CMC水凝胶组大鼠创面分别植入对应的水凝胶。治疗14 d后,观察创面愈合情况与愈合质量。结果与结论:①体外实验:usAgNPs-PL/CMC水凝胶具有均匀的三维网状结构、良好的力学性能与较强的吸水能力,可明显抑制大肠杆菌和金黄色葡萄球菌的生长,并且对两细菌生物膜形成具有良好的抑制和清除作用,具有良好的抗菌能力;②体内实验:治疗14 d后,usAgNPs-PL/CMC水凝胶组创面基本闭合,创面愈合率最高,创面菌落数量明显少于模型组;治疗14 d的苏木精-伊红、Masson染色显示,usAgNPs-PL/CMC水凝胶组、usAgNPs/CMC水凝胶组和CMC水凝胶组创面出现再生真皮的组织结构,新生肉芽组织表皮均完整且厚,其中usAgNPs-PL/CMC水凝胶组可见堆积的平行的成熟胶原纤维;治疗7 d的CD31免疫组化及Ki67免疫荧光染色显示,usAgNPs-PL/CMC水凝胶组创面微血BACKGROUND:Bacterial infections and impaired angiogenesis have been obstacles to diabetic wound healing,and the problem of multidrug resistance cannot be ignored,so there is an urgent need to find a new therapeutic strategy.OBJECTIVE:To prepare platelet-camouflaged silver nanoparticle hydrogel and observe its therapeutic effect on diabetic wounds.METHODS:(1)In vitro test:The ultra-small silver nanoparticles(usAgNPs)were obtained by reflux heating.usAgNPs-PL/CMC hydrogel was prepared by coupling it with platelets(PL)and adding it to the carboxymethyl cellulose(CMC)hydrogel.The microstructures,swelling properties,and mechanical properties of the hydrogel were characterized.The antibacterial activity of the hydrogel was tested by 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)5[(phenylamino)carbonyl]-2H-tetrazolium hydroxide method,reactive oxygen species method,bacterial biofilm method,and bacterial activity.(2)In vivo test:Forty SD rats were randomly divided into sham operation group,model group,CMC hydrogel group,usAgNPs/CMC hydrogel group,and usAgNPs-PL/CMC hydrogel group,with eight rats in each group.The type 1 diabetes model was established in the other four groups except the sham operation group.After successful model establishment,a skin wound(20 mm in diameter)deep to the fascia layer was made on the back of rats of the five groups.Rats in the sham operation group and model group were injected with normal saline.The rats in the CMC hydrogel group,usAgNPs/CMC hydrogel group,and usAgNPs-PL/CMC hydrogel group were implanted with the corresponding hydrogel,separately.The wound healing condition and healing quality were observed after 14 days of treatment.RESULTS AND CONCLUSION:(1)In vitro test:The usAgNPs-PL/CMC hydrogel had a uniform three-dimensional network structure,good mechanical properties,and strong water absorption ability,which could remarkably inhibit the growth of Escherichia coli and Staphylococcus aureus,and had a good inhibition and clearance effect on the formation of biofilms of the two bacteria,and h

关 键 词：超小纳米银颗粒 血小板 羧甲基纤维素 水凝胶 糖尿病创面 创面愈合 抑菌 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R331