芦荟苷氧化石墨烯凝胶的制备及其对创伤修复作用的研究

发布时间：2017-12-26 16:02

本文关键词：芦荟苷氧化石墨烯凝胶的制备及其对创伤修复作用的研究　出处：《浙江大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:考察芦荟苷对急性创伤的治疗效果,进一步制备以氧化石墨烯水凝胶为载体的芦荟苷局部给药体系,并探讨其对皮肤急性创伤治疗和疤痕抑制的作用及其相关机制。方法:1.通过体内外一系列实验考察芦荟苷对皮肤创伤的治疗效果。体外方面,考察了芦荟苷对人永生化角质形成细胞(HaCaT)、人皮肤成纤维细胞(HSF)以及血管内皮细胞(Endo)的增殖与迁移的影响,采用Tube formation实验考察芦荟苷对血管形成的作用。体内实验方面,通过建立大鼠急性创伤模型考察芦荟苷对伤口愈合率的影响,并对愈合后的皮肤进行扫描电镜观察、组织染色(HE、Masson)、免疫组化染色(CD31、EGF)考察愈合后的皮肤微观结构及组成。2.构建携载芦荟苷的氧化石墨烯水凝胶,对处方进行优化,并进一步对形态、药物释放等进行一系列研究。建立凝胶中芦荟苷的HPLC检测方法。3.对制备的芦荟苷氧化石墨烯凝胶进行体外抑菌作用的考察。在体内实验中,对芦荟苷氧化石墨烯凝胶的皮肤刺激性以及创伤修复实验做进一步的考察。采用扫描电镜、HE与Masson三色染色、CD31与α-SMA免疫组化染色以及测定修复皮肤内的VEGF的含量等方法验证芦荟苷氧化石墨烯凝胶的创伤修复效果。结果:1.芦荟苷对血管内皮细胞和皮肤成纤维细胞有一定的促进增殖作用,对HSF细胞有较强的迁移促进作用,同时还具有一定的抑菌作用,显示出一定的促进愈合的效果。体内实验结果显示芦荟苷溶液能提高伤口愈合率。治疗后形成的皮肤,新生血管数量较多,含有毛囊等附属器官,胶原蛋白排列较为整齐疏松,与正常皮肤呈现相似的形态。2.确定制剂处方中各个成分的含量:芦荟苷1mg/mL;卡波姆0.5%;氧化石墨烯125μg/mL;三乙醇胺调节pH到5。采用本处方制备的芦荟苷氧化石墨烯凝胶质地均匀细腻,具有合适的粘度,易于涂布,且具有一定的药物缓释功能,适合皮肤局部给药。3.按照处方量配制的芦荟苷氧化石墨烯凝胶具有良好的抑菌效果,皮肤刺激性实验证明其没有皮肤刺激性。同时,芦荟苷氧化石墨烯凝胶能提高皮肤创面的伤口愈合率,对愈合后的皮肤进行组织形态以及免疫组化等考察,结果表明芦荟苷氧化石墨烯凝胶能促进皮肤血管新生及部分皮肤附属器官的形成,显示了一定的疤痕抑制效果。结论:芦荟苷氧化石墨烯凝胶可以作为良好的局部给药体系,促进创面的愈合并减少疤痕的形成。
[Abstract]:Objective: To investigate the therapeutic effect of aloin on acute trauma, and further prepare the aloin local drug delivery system based on graphene oxide hydrogel, and to explore its effect on skin acute wound treatment and scar inhibition and its related mechanism. Methods: 1. through a series of experiments in vivo and in vivo, the therapeutic effect of aloin on skin trauma was investigated. In vitro, the effects of aloin on the proliferation and migration of human immortalized keratinocytes (HaCaT), human skin fibroblasts (HSF) and vascular endothelial cells (Endo) were investigated. The effect of aloin on angiogenesis was investigated by Tube formation. In vivo experiment, through the establishment of rat model to investigate the effect of acute trauma of aloin on wound healing rate, and after the healing of skin were observed by scanning electron microscope, tissue staining (HE, Masson), immunohistochemical staining (CD31, EGF) skin microstructure and composition of healing. 2. to construct the graphene oxide hydrogel carrying aloin, optimize the prescription, and further study a series of studies on morphology and drug release. To establish a HPLC method for the determination of aloin in gel. 3. the Bacteriostasis of aloin oxide graphene gel prepared in vitro was investigated. In vivo experiments, the skin irritation of aloin oxide gels and the experiment of trauma repair were further investigated. SEM, HE and Masson trichrome staining, CD31 and -SMA immunohistochemical staining and the content of VEGF in the repaired skin were used to verify the wound healing effect of aloin glucoside graphene gel. Results: 1., aloin has certain promoting effect on proliferation of vascular endothelial cells and dermal fibroblasts, and has a strong migration and promotion effect on HSF cells, and also has a certain bacteriostatic effect, showing a certain effect of promoting healing. The results of the experiment in vivo showed that the aloin solution could improve the healing rate of the wound. After treatment, there are many new skin vessels, including hair follicle and other accessory organs. Collagen is arranged orderly and loose, showing similar morphology with normal skin. 2. to determine the content of each component in the formulation: aloin 1mg/mL; carbomer 0.5%; graphene oxide 125 g/mL triethanolamine; adjust pH to 5. The aloe glucoside graphene oxide gel prepared by this prescription is homogeneous and exquisite in texture. It has the appropriate viscosity and is easy to coat, and has certain drug release function. It is suitable for topical administration of skin. 3. the aloin graphene oxide gel prepared according to the prescription has a good bacteriostasis effect, and the skin irritation test proved that it has no skin irritation. At the same time, aloin graphene oxide gel can improve the healing rate of skin wound wound tissue morphology and immunohistochemical study on the skin after healing, results show that aloin graphene oxide gel can promote the formation of skin angiogenesis and part of skin appendages, showed a certain inhibitory effect of scar. Conclusion: the aloin graphene oxide gel can be used as a good local drug delivery system to promote the healing of the wound and reduce the formation of scar.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R283.6;R285

【参考文献】