摘要
背景:基于医工结合理念和中医药治疗的优势,将中药有效活性成分单体负载于组织工程材料构建新型复合材料在损伤局部递送药物,是修复脊髓损伤的研究热点,有望成为解决这一难题的有效手段。目的:观察携川芎嗪超分子导电水凝胶修复大鼠脊髓损伤的效果。方法:制备携川芎嗪超分子导电水凝胶,表征其微观结构、电导率、流变性、溶胀率及体外缓释性能。采用随机数字表法将45只SD大鼠分为3组,每组15只:假手术组不损伤脊髓,模型组建立脊髓损伤模型,水凝胶组建立脊髓损伤模型后向脊髓损伤区域注射携川芎嗪超分子导电水凝胶。造模前及造模后1,7,14,21,28 d,采用BBB评分评估大鼠后肢运动功能恢复情况;造模后28 d取损伤处脊髓组织,分别进行苏木精-伊红染色、免疫组化染色及Western blot检测。结果与结论:①扫描电镜下可见,携川芎嗪超分子导电水凝胶呈现出三维微米尺度的多孔网络结构,具有较高的孔隙率,孔径约为100μm;该水凝胶的电导率为7.66 S/m、溶胀率为3764.42%,具备一定的力学稳定性能,同时具有可注射性;体外缓释实验显示,携川芎嗪超分子导电水凝胶可持续释放川芎嗪达800 h以上,且随着时间延长,药物累计释放量呈上升趋势。②随着造模时间的延长,模型组、导电水凝胶组大鼠后肢运动功能逐渐恢复,水凝胶组造模后14,21,28 d的后肢运动功能优于模型组(P<0.05)。苏木精-伊红染色显示,模型组脊髓组织出现空洞,残端可见大量炎性细胞浸润;水凝胶组脊髓损伤区可见部分炎性细胞浸润,损伤区域空洞面积缩小,交接区出现神经元细胞,组织排列较为整齐。免疫组化染色及Western blot检测显示,水凝胶组大鼠脊髓组织中肿瘤坏死因子α、白细胞介素6蛋白表达低于模型组(P<0.05),神经元核抗原蛋白表达高于模型组(P<0.05)。③结果显示,携川芎嗪超分子导电水凝胶可促进损伤脊髓组织的修复。
BACKGROUND:Based on the concept of the combination of medicine and industry and the advantages of traditional Chinese medicine treatment,the construction of a new composite material loaded with the effective active ingredient of traditional Chinese medicine is a hot research spot in the repair of spinal cord injury,and is expected to become an effective means to solve this problem.OBJECTIVE:To observe the effect of supramolecular conducting hydrogel carrying ligustrazine in repairing spinal cord injury in rats.METHODS:The supramolecular conducting hydrogel carrying ligustrazine was prepared and its microstructure,conductivity,rheology,swelling rate and in vitro release performance were characterized.45 SD rats were divided into 3 groups by random number table method,with 15 rats in each group:no spinal cord injury in the sham operation group;spinal cord injury model was established in the model group;and supramolecular conducting hydrogel carrying ligustrazine was injected into the spinal cord injury area after model establishment in hydrogel group.BBB score was used to evaluate the recovery of hind limb motor function of each group before and 1,7,14,21 and 28 days after modeling,respectively.28 days after the model establishment,the spinal cord tissues were collected and analyzed by hematoxylin-eosin staining,immunohistochemical staining and western blot assay.RESULTS AND CONCLUSION:(1)Under scanning electron microscopy,the supramolecular conducting hydrogel with ligustrazine displayed a three-dimensional micrometer-scale porous network structure with high porosity and a pore size of approximately 100μm.The conductivity of the hydrogel was 7.66 S/m;the swelling rate was 3764.42%,and it had certain mechanical stability and injection property.In vitro sustained release experiments demonstrated that the supramolecular conducting hydrogel with ligustrazine sustainably released ligustrazine for more than 800 hours.With the extension of time,the cumulative release of ligustrazine exhibited an increasing trend.(2)With the extension of modeling time,the hind limb motor function gradually recovered in the model group and the hydrogel group,and the hind limb motor function of the hydrogel group was better than that of the model group on 14,21,and 28 days after modeling(P<0.05).Hematoxylin-eosin staining demonstrated that the spinal cord tissue of the model group had cavities and a large number of inflammatory cells could be seen at the stump.In the hydrogel group,some inflammatory cells were infiltrated in the injured area of the spinal cord;the void area of the injured area was reduced;neuron cells appeared in the junction area,and the tissue arrangement was relatively neat.Immunohistochemical staining and western blot assay exhibited that the expression of tumor necrosis factorαand interleukin-6 protein in the rat spinal cord of the hydrogel group was lower than that in the model group(P<0.05),and the expression of neuronal nuclear antigen protein was higher than that in the model group(P<0.05).(3)These findings confirm that the supramolecular conducting hydrogel carrying ligustrazine can promote the repair of spinal cord injury.
作者
吴杨鹏
杨晓慧
劳克诚
戴世友
范筱
Wu Yangpeng;Yang Xiaohui;Lao Kecheng;Dai Shiyou;Fan Xiao(Department of Traditional Chinese Medicine Rehabilitation,The 909 Hospital of Joint Logistic Support Force(Dongnan Hospital of Xiamen University),Zhangzhou 363000,Fujian Province,China;Qingdao Hospital(Qingdao Municipal Hospital),University of Health and Rehabilitation Sciences,Qingdao 266011,Shandong Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2024年第10期1505-1511,共7页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金课题(82205149)
项目负责人:范筱
中国博士后科学基金(2022M710472)
项目负责人:范筱。
关键词
脊髓损伤
川芎嗪
可注射导电水凝胶
炎症
组织工程支架
spinal cord injury
ligustrazine
injectable conducting hydrogel
inflammation
tissue engineering scaffold
作者简介
第一作者:吴杨鹏,男,1990年生,福建省漳州市人,汉族,硕士,医师,主要从事脊髓损伤修复方面的研究,https://orcid.org/0009-0009-2372-2122;通讯作者:范筱,主治医师。
