艾灸与还原氧化石墨烯/二氧化铈纳米复合材料修复感染性创面

Moxibustion and reduced graphene oxide/cerium dioxide nanocomposites for repairing infectious wounds

背景:皮肤创伤修复过程复杂且易受感染,易导致预后不良,是目前创面修复研究的难点与热点,并受到中医药及组织工程研究领域的广泛关注。目的:观察艾灸和还原氧化石墨烯/二氧化铈纳米复合材料修复感染性创面的效果。方法:①采用水热法合成质量比分别为2∶1、1∶1和1∶2的还原氧化石墨烯/二氧化铈纳米复合材料,所得复合材料分别记为G2C1、G1C1和G1C2,检测3种材料的光热性能、细胞毒性及抗菌性能。取艾条,设置3种施灸距离(3.0-3.5 cm,记为灸1;2.5-3.0 cm,记为灸2;2.0-2.5 cm,记为灸3)对人体皮肤表面施灸10 min,检测光热性能;检测3种距离区间下实施艾灸的抗菌性能。同时,检测不同质量浓度G1C1材料、艾灸(3种施灸距离)及灸2+G1C1材料的大鼠背部体表红外热成像。②取60只成年SD大鼠,建立金黄色葡萄球菌感染创面模型,48 h后随机分10组干预,每组6只:对照组(不进行任何处理)、莫匹罗星组、灸2+G1C1组、灸1组、灸2组、灸3组及60,80,100,120μg/mL G1C1组(G1C1组给予808 nm近红外激光照射,10 min/次,每次治疗前在创面负载G1C1悬浮液;艾灸各组行原位悬起灸,干预时间10 min/次;灸2+G1C1组每次治疗前在在创面负载80μg/mL G1C1悬浮液,并用艾条行原位悬起灸,干预时间10 min/次),治疗频率2 d一次。干预7 d后,检测创面愈合情况、创面菌落计数及修复情况。结果与结论:①3种还原氧化石墨烯/二氧化铈纳米复合材料均具有良好的光热性能,复合材料质量浓度越高光热性能越好。灸2组施灸10 min温度能达到47.6℃,且不造成热损伤,更适合于动物实验。与NIH-3T3细胞共培养实验结果显示,60,80,100μg/mL G1C1具有良好的生物相容性。与金黄色葡萄球菌悬浮液共培养实验结果显示,G2C1、G1C1和G1C2均具有良好的抗菌作用,其中G1C1组表现出优异的抗菌性能,当其质量浓度为80μg/mL时抑菌率已达到100%。60-120μg/mL G1C1可有效清除金黄色葡萄球菌生物膜,材料质量浓度越高清除效果越好;艾灸也可有效清除金黄色葡萄球菌生物膜,且施灸距离越近清除效果越好。②莫匹罗星组、灸2组、灸2+G1C1组及80,100μg/mL G1C1组大鼠治疗第7天的创面面积相较于对照组明显减小,创面修复质量较好。莫匹罗星、G1C1、艾灸和灸2+G1C1均能有效清除创面细菌残留,且G1C1质量浓度越高细菌残留量越少,艾灸组施灸距离越近细菌残留量越少。其中80μg/mL G1C1组和灸2组的创面修复效率以及细菌残留量非常接近,且二者创面修复效率均优于莫匹罗星组。除此之外还观察到材料与艾灸联合后对创面细菌的清除能力要优于单独使用。③结果显示,艾灸、还原氧化石墨烯/二氧化铈纳米复合材料及其结合具有良好的抗感染和促创面愈合效果。

BACKGROUND:The repair process of skin trauma is complex and susceptible to infection,easy to lead to poor healing,is the current difficulty and hot spot in wound repair research,and has received extensive attention in the fields of traditional Chinese medicine and tissue engineering.OBJECTIVE:To investigate the effect of moxibustion and reduced graphene oxide/cerium oxide nanocomposite on promoting the healing of infectious wounds.METHODS:(1)Reduced graphene oxide/cerium dioxide nanocomposites with mass ratios of 2:1,1:1 and 1:2 were synthesized by hydrothermal method.The resulting composites were recorded as G2C1,G1C1 and G1C2,respectively.The photothermal properties,cytotoxicity and antibacterial properties of the three kinds of materials were tested.After taking moxa sticks,three kinds of moxibustion distances were set(3.0-3.5 cm,recorded as moxibustion 1;2.5-3.0 cm,recorded as moxibustion 2;2.0-2.5 cm,recorded as moxibustion 3).Moxibustion was applied to the surface of human skin for 10 minutes to detect the photothermal properties.The antibacterial properties of moxibustion were tested at three different distance intervals.Simultaneously,the back body surface infrared imaging of rats with different mass concentrations of G1C1 material,moxibustion(three kinds of moxibustion distances)and moxibustion 2+G1C1 material was detected.(2)Sixty male Sprague-Dawley rats were selected to model the wound of Staphylococcus aureus infection.48 hours later,they were randomly divided into 10 groups with 6 rats in each group:control group(did not receive any treatment),mupirocin group,moxibustion 2+G1C1 group,moxibustion 1 group,moxibustion 2 group,moxibustion 3 group and 60,80,100,and 120μg/mL G1C1 groups(The G1C1 group was given 808 nm near-infrared laser irradiation for 10 min/time,and the G1C1 suspension was loaded on the wound surface before each treatment.Each group of moxibustion underwent in-situ suspension moxibustion,and the intervention time was 10 min/time.Moxibustion 2+G1C1 group was loaded with G1C1 suspension on the wound surface before each treatment,and moxibustion was suspended in situ with moxa strips,and the intervention time was 10 min/time).The frequency of treatment was 2 days once.Wound healing,wound colony count and repair were detected after 7 days of intervention.RESULTS AND CONCLUSION:(1)The three kinds of reduced graphene oxide/cerium dioxide nanocomposites had good photothermal properties,and the higher the mass concentration of the composites,the better the photothermal properties.The temperature of the moxibustion 2 group reached 47.6°C for 10 minutes without causing thermal damage,which was more suitable for animal experiments.The results of co-culture with NIH-3T3 cells exhibited that 60,80,and 100μg/mL G1C1 had good biocompatibility.The results of a co-culture experiment with Staphylococcus aureus suspension displayed that G2C1,G1C1 and G1C2 had good antibacterial activity,among which G1C1 group demonstrated excellent antibacterial performance,and the antibacterial rate reached 100%when its mass concentration was 80μg/mL.60-120μg/mL G1C1 could effectively remove Staphylococcus aureus biofilm,and the higher the material mass concentration,the better the removal effect.Moxibustion could also effectively remove Staphylococcus aureus biofilm,and the closer the moxibustion was,the better the removal effect.(2)Compared with the control group,the wound area of the mupirocin group,moxibustion 2 group,moxibustion 2+G1C1 group and 80,100μg/mL G1C1 groups was significantly reduced on day 7 of treatment,and the quality of wound repair was better.Mupirocin,G1C1,moxibustion and moxibustion 2+G1C1 could effectively remove the residual bacteria on the wound surface,and the higher the mass concentration of G1C1,the lower the residual bacteria.Among them,the wound repair efficiency and bacterial residue of 80μg/mL G1C1 group and moxibustion 2 group were very similar,and the wound repair efficiency of both was better than that of mupirocin group.In addition,it was also observed that the combination of materials and moxibustion had a better ability to clear wound bacteria than that used alone.(3)The results confirm that moxibustion,reduced graphene oxide/cerium dioxide nanocomposites and their combination have good anti-infection and wound healing effects.