摘要
背景:水凝胶作为一种基于湿性愈合理论开发的新型敷料,为创面提供适宜湿度环境、为细胞迁移和组织再生提供足够空间和机械支撑,还可作为药物、生长因子、细胞、纳米颗粒的载体,在生物医学领域应用广泛。葛根素具有抗氧化、抗炎、促血管生成等作用,但脂溶性和水溶性欠佳,临床应用受限。目的:制备负载葛根素的双网络水凝胶,评估水凝胶对细胞行为及动物创面愈合的影响。方法:①将葛根素溶液(0.5 mg/mL)加入羧甲基壳聚糖-苯硼酸溶液中,分别制备单网络的海藻酸钠-呋喃/双马来酰亚胺水凝胶、羧甲基壳聚糖-苯硼酸-表没食子儿茶素没食子酸酯水凝胶,将2种单网络水凝胶按照原料的不同体积比例混合制备负载葛根素的双网络水凝胶,未负载葛根素的H0水凝胶中,海藻酸钠-呋喃、羧甲基壳聚糖-苯硼酸、表没食子儿茶素没食子酸酯、马来酰亚胺-马来酰亚胺的体积比为25∶40∶8∶3,负载葛根素的H1、H2、H3、H4、H5水凝胶中,海藻酸钠-呋喃、羧甲基壳聚糖-苯硼酸(含葛根素)、表没食子儿茶素没食子酸酯、马来酰亚胺-马来酰亚胺的体积比分别为25∶30∶6∶3、25∶40∶8∶3、25∶50∶10∶3、25∶60∶12∶3、25∶70∶14∶3。表征水凝胶的微观形貌、力学性能、黏附性、溶胀性能、降解性能及药物释放性能,检测水凝胶对大肠杆菌的抑制作用,检测水凝胶对L929细胞增殖与迁移的影响。②在9只SD大鼠背部制备直径1 cm的圆形全层皮肤缺损创面,每只制作3个创面,对照组(n=3)创面注射生理盐水,H0组(n=3)创面注射H0水凝胶,H3组(n=3)创面注射H3水凝胶,观察创面愈合情况。创面造模后3,7d取材,分别进行苏木精-伊红、Masson与CD31免疫组化染色。结果与结论:①负载葛根素的双网络水凝胶可以成胶,扫描电镜扫描下可见水凝胶微孔密集、孔数多、孔径小,具有可注射性、组织黏附性和力学性能;负载葛根素的双网络水凝胶吸水能力较强,降解速度较缓慢,能够保持形态与体积的稳定,对pH值和葡萄糖有响应性,可稳定释放负载其中的葛根素;负载葛根素的双网络水凝胶可促进L929细胞的增殖及迁移,抑制大肠杆菌的生长繁殖。②相较于对照组、H0组,H3组创面愈合速率较快,造模后7 d的创面愈合率高达70%。造模后7 d的苏木精-伊红、Masson与CD31免疫组化染色结果显示,相较于对照组、H0组,H3组创面创面肉芽组织生长、胶原积累和血管新生最多。③结果表明,负载葛根素的双网络水凝胶具有良好的生物相容性与释药性能,可促进L929细胞的生长与迁移以及大鼠全层皮肤缺损的愈合。
BACKGROUND:As a new type of dressing developed based on the theory of moist healing,hydrogel provides a suitable humidity environment for the wound surface,sufficient space and mechanical support for cell migration and tissue regeneration,and can also be used as a carrier of drugs,growth factors,cells,and nanoparticles.It is widely used in the biomedical field.Puerarin has antioxidant,anti-inflammatory,and angiogenic effects,but its lipid solubility and water solubility are poor,which limits its clinical application.OBJECTIVE:To prepare a puerarin-loaded double-network hydrogel and evaluate its effects on cellular behavior and wound healing in animal models.METHODS:(1)Puerarin solution(0.5 mg/mL)was added to carboxymethyl chitosan-phenylboronic acid solution to prepare single-network sodium alginate-furan/bismaleimide hydrogel and carboxymethyl chitosan-phenylboronic acid-epigallocatechin gallate hydrogel,respectively.The two single-network hydrogels were mixed according to different volume ratios of the raw materials to prepare double-network hydrogel loaded with puerarin.In the H0 hydrogel without puerarin,the volume ratio of sodium alginate-furan,carboxymethyl chitosan-phenylboronic acid,epigallocatechin gallate and maleimide-maleimide was 25:40:8:3.In the puerarin-loaded H1,H2,H3,H4,and H5 hydrogels,the volume ratios of sodium alginate-furan,carboxymethyl chitosan-phenylboronic acid(containing puerarin),epigallocatechin gallate and maleimide-maleimide were 25:30:6:3,25:40:8:3,25:50:10:3,25:60:12:3,and 25:70:14:3,respectively.The micromorphology,mechanical properties,adhesion,swelling properties,degradation properties,and drug release properties of the hydrogels were characterized,and the inhibitory effect of the hydrogels on Escherichia coli and the effects of the hydrogels on the proliferation and migration of L929 cells were detected.(2)A circular full-thickness skin defect with a diameter of 1 cm on the back of 9 SD rats was made.Three wounds were made on each rat.The wounds of the control group(n=3)were injected with normal saline;the wounds of the H0 group(n=3)were injected with H0 hydrogel,and the wounds of the H3 group(n=3)were injected with H3 hydrogel.The wound healing was observed.The wounds were collected 3 and 7 days after the wound modeling.Hematoxylin-eosin staining,Masson staining,and CD31 immunohistochemical staining were performed respectively.RESULTS AND CONCLUSION:(1)The double-network hydrogel loaded with puerarin could gel.Scanning electron microscopy showed that the hydrogel had dense micropores,a large number of pores,and a small pore size.It had injectability,tissue adhesion and mechanical properties.The double-network hydrogel loaded with puerarin had strong water absorption capacity,slow degradation rate,could maintain the stability of morphology and volume,was responsive to pH and glucose,and could stably release the puerarin loaded therein.The double-network hydrogel loaded with puerarin could promote the proliferation and migration of L929 cells and inhibit the growth and reproduction of Escherichia coli.(2)Compared with the control group and H0 group,the wound healing rate of the H3 group was faster,and the wound healing rate was as high as 70%7 days after modeling.The results of hematoxylin-eosin staining,Masson staining,and CD31 immunohistochemical staining 7 days after modeling showed that the wounds of the H3 group had the most granulation tissue growth,collagen accumulation and angiogenesis compared with the control group and H0 group.(3)The results showed that the double-network hydrogel loaded with puerarin had good biocompatibility and drug release properties,which could promote the growth and migration of L929 cells and the healing of full-thickness skin defects in rats.
作者
姚茵璇
温素如
陈超盛
温鑫
冯可滢
邝枣园
张文
Yao Yinxuan;Wen Suru;Chen Chaosheng;Wen Xin;Feng Keying;Kuang Zaoyuan;Zhang Wen(School of Basic Medical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong Province,China)
出处
《中国组织工程研究》
北大核心
2026年第20期5201-5213,共13页
Chinese Journal of Tissue Engineering Research
基金
广东省中医药管理局项目(20251096),项目负责人:张文。
关键词
创面修复
三维多孔结构
双网络水凝胶
葛根素
药物释放
生物敷料
中药小分子
点击化学
wound repair
three-dimensional porous structure
double-network hydrogel
puerarin
drug release
biological dressing
small molecules of traditional Chinese medicine
click chemistry
