三紫创愈海绵敷料的药效学及质量评价研究

发布时间：2018-07-10 16:55

本文选题：中药海绵敷料 + 创面愈合　；参考：《南方医科大学》2016年硕士论文

【摘要】：研究背景：由于人口老龄化加剧,交通运输业迅速发展,车祸等创伤事故发生率不断增加,使得皮肤伤口这一问题越来越成为世界范围内非常严重的健康问题与负担。近年来,国内外在伤口愈合方面也提出了很多新的治疗理论、方法和技术。1974年诞生了全球第一块密闭性敷料安舒妥,提出湿性愈合的理论,并逐渐被广泛接受。近50年来,国内大量关于创面的基础研究和临床治疗证实了湿润环境有利于伤口的愈合,新型保湿敷料也不断进入医疗市场。但新型敷料存在功能单一、缺乏复合治疗功能、制备工艺复杂、价格昂贵、市场普及慢等缺点,且国内市场上的新型敷料主要来自欧美行业巨头,本土企业的产品很少。中药敷料在治疗伤口修复方面独具特色,能主动促进创面愈合,防止瘢痕增生,适合于各类急慢性伤口的治疗护理,且来源简单、安全、副作用小,费用不高,能提高伤口护理水平,降低医疗服务成本,提高医院工作效率。然而,中药敷料也存在不能维持密闭、微湿润的环境、使用不便等不足,其安全性问题也一直备受关注。医用敷料质量的好坏直接关系到患者的生命安全,发达国家从20世纪60年代就开始对医用敷料进行安全性监测。1976年,美国食品药物管理局(FDA)开始管理医用敷料,并实行售前审批制度,建立了一系列生物和质量评价标准。我国从70年代后期开始研究医用敷料的生物学评价,中国医疗器械注册的相关规定要求医用敷料正式进入市场前,在完成加工性能、外形设计等程序后,必须对其进行物理和化学性能评价,生物学评价、制定产品质量标准等,但是与发达国家相比还是有一定差距。如由于中药敷料成分复杂,且制剂形式多样,国家尚未建立统一化的中药敷料评价标准,制约着其产业化发展。因此,中药敷料的研究存在一些不足,研发体系存在着缺陷,需要进一步完善。本课题组在总结中草药和民间广为流传使用的伤科验方基础上,依据中药组方原则,选取功效不同的中草药创制出伤科验方“三七复方”。全验方由三七、紫珠草、白芨、儿茶、鸡矢藤、两面针、虎杖等9味中草药组成,具有良好的止血、抗菌、消肿、镇痛等作用,但原剂型存在使用不便、容易污染伤口等不足。本课题组己对该伤科验方进行改革,将该中药伤科验方的特色与新型保湿敷料的优势结合起来,研制开发出一种能够提供一个湿性愈合的环境,快速吸收渗出液,促进肉芽生长,减少感染,主动促进创面愈合的中药复合海绵敷料,即三紫创愈海绵敷料,适用于各类机械性或创伤性伤口、慢性感染伤口、糖尿病足溃疡等急慢性伤口。然而,其安全性和药效有待证实,质量有待评价。为此,我们对三紫创愈海绵敷料进行药效学和质量分析研究,确保海绵敷料安全有效、稳定可控,为医用敷料的注册审批提供重要科研资料,为进一步扩大生产提供科学的依据。研究目的：1.验证三紫创愈海绵敷料对大鼠创面修复的影响,探讨其治疗机理。2.评价三紫创愈海绵敷料的物理性能。3.鉴别三紫创愈海绵敷料中药材三七、虎杖。研究方法：1.三紫创愈海绵敷料的药效学研究本研究采用大鼠背部小面积皮肤切除伤模型,将大鼠随机分为3组,阴性对照组10只,阳性对照组10只,实验组10只。大鼠取俯卧位,脱去大鼠背部毛发,用3%的戊巴比妥钠腹腔注射麻醉,碘伏及70%乙醇常规消毒皮肤后,用利刀于大鼠背部左右对称制作2个直径为2cm的圆形全层缺损创面,醋酸洗必泰溶液充分清洗消毒创面。实验组用自制的三紫创愈海绵敷料覆盖创面,阳性对照组用空白海绵基质覆盖创面,阴性对照组用凡士林纱布覆盖创面,三组外层均用无菌纱布敷料包扎。前三天每日换药一次,三天后隔一天换药,一周后隔两天换药。每次换药时,观察三组大鼠性情和行为等生活情况以及创面大体情况,包括精神状况、摄食及饮水情况、活动情况、创面渗出、出血、感染迹象、肉芽组织生长情况,并选定创面面积、炎症积分、病理学情况作为修复指标。治疗后第3d、7d、14d,数码相机拍照,利用Image J软件测定刻度尺像素比例,测出创面面积,计算创面愈合率；治疗后第3d、7d测创面炎症积分,计算炎症积分值并进行统计学分析；于治疗后第3d、7d、14d随机抽取6只大鼠(每组2只)过量麻醉处死,立即切取创面皮肤,4%的多聚甲酵固定,用H.E染色,观察炎细胞浸润及坏死、纤维细胞及毛细血管增生、胶原纤维等情况,进行形态学描述。2.三紫创愈海绵敷料的质量研究本研究应用文献回顾的方法,参照《中国药典》2010版的相关要求及中国医疗器械注册的相关规定标准,咨询中药制剂学、护理学的临床及教学一线专家,采用薄层色谱法对三紫创愈海绵敷料进行定性鉴别,鉴别海绵敷料中人参皂苷Rb1、人参皂苷Rg1、虎杖苷。将对照品溶液、供试品溶液、阴性溶液点于同一薄层板上,在展开容器内用展开剂展开,使试样溶液所含成分分离,所得色谱图进行对比,进行制剂的鉴别。同时选取透气率、吸水率、保湿性、抗撕强度、重量差异、耐热性、耐寒性等7个指标作为海绵敷料的评价指标。为了验证工艺条件合理性,进行了三批三紫创愈海绵敷料的研究。研究结果：1.大鼠饮食和活动情况麻醉清醒后,各组大鼠均精神不佳,无任何主动活动及饮食；治疗后第3d,实验组和阳性对照组大鼠精神状态、主动活动、饮食均接近正常,阴性对照组有少量活动及饮食,精神状态好转；治疗后第7d、14d组各大鼠精神状态、主动活动、饮食均正常,组间无明显差别。各组均未见实验性的大鼠死亡。2.大鼠伤口愈合情况创面愈合率比较,治疗后第3天,实验组创面愈合率为(40.22±0.05)%,阳性对照组创面愈合率为(38.33±0.1)%,高于阴性对照组创面愈合率(22.02±0.03)%,比较有显著性差异(P0.05)；治疗后第7天,实验组创面愈合率为(71.32±0.08)%,高于阳性对照组创面愈合率(60.66±0.09)%和阴性对照组创面愈合率(50.01±0.10)%,比较有显著性差异(P0.05)；治疗后第14天,实验组创面愈合率为(90.44±0.07)%,高于阳性对照组创面愈合率(81.03±0.04)%和阴性对照组创面愈合率(71.65±0.10)%,比较有显著性差异(P0.05)。三紫创愈海绵敷料能缩小创面面积,缩短创面的愈合时间,提高创面愈合率。病理学观察,与对照组相比,治疗后第3d,实验组较早出现毛细血管,成纤维细胞开始增殖。治疗后第7d,实验照组创面纤维增生明显,胶原纤维明显多于对照组。治疗后第14d,与对照组相比实验组胶原纤维增生明显,与术后第7d相比没有明显改变,伤口未见异物组织,没有组织破坏和坏死现象。三紫创愈海绵敷料能促进毛细血管和胶原纤维的生长。炎症积分比较,治疗后第3天,实验组创面炎症积分为(3.20±0.42)分,低于阳性对照组创面炎症积分(4.18±0.60)分和阴性对照组创面炎症积分(5.67±0.50)分,比较有显著性差异(P0.05)；治疗后第7天,实验组创面炎症积分为(1.70±0.48)分,低于阳性对照组创面炎症积分(2.55±0.52)分和阴性对照组创面炎症积分(3.89±0.33)分,比较有显著性差异(P0.05)。三紫创愈海绵敷料能减少创面渗液的过多分泌。3.三紫创愈海绵敷料的质量评价实验结果显示三紫创愈海绵敷料的平均透气率为80.86%,故本品透气率暂定为不低于78%；平均保湿率为38.11%,保湿率暂定为不低于35%；平均吸水率为11.15倍,吸水率暂定为不低于10倍；平均抗撕强度为131.59g/cm3,抗撕强度标准暂定为不小于130g/cm3；最大片重差异为3.5%,最小片重差异为3.4%,重量差异限度在平均重量的±5%以内,符合药典评价标准。于冰箱中贮存后,敷料未见明显分层现象；放置烘箱后,敷料表面无渗油现象,有光泽。三紫创愈海绵敷料具有良好的透气率、吸水率和保湿性,耐热、耐寒、赋形性良好,抗撕强度高,重量差异差别小。薄层色谱鉴别三紫创愈海绵敷料中三七及虎杖的分离度好,样品色谱中,在与对照品色谱相应的位置上有相同颜色的斑点,且阴性无干扰。结论：1.通过对三紫创愈海绵敷料主要药效学的研究表明,三紫创愈海绵敷料具有快速止血、抗炎抑菌、活血化瘀等作用,能吸收伤口过多的渗出物,防止感染,为创面愈合创造良好的微环境,促进创面血管新生,促进肉芽组织生长,加速创面的愈合,并能减少瘢痕的产生,减少后遗症,降低死亡率。三紫创愈海绵敷料作为创面敷料具有安全可靠性。2.三紫创愈海绵敷料具有良好的透气率、吸水率和保湿性,耐热、耐寒性良好,抗撕强度高,重量差异限度在5%的标准范围内。薄层色谱法鉴别人参皂苷Rg1、人参皂苷Rb1、虎杖苷,操作方法简便、快捷、准确、可靠、重复性好,可以作为三紫创愈海绵敷料的定性鉴别方法,为后期产品敷料产业化扩大生产提供了实验依据。
[Abstract]:Background: as the aging of the population, the rapid development of traffic and transportation, and the increasing incidence of accidents, such as traffic accidents, the problem of skin wound has become a very serious health problem and burden in the world. In recent years, many new theories, methods and methods have been put forward in the field of wound healing at home and abroad. Technology.1974 was born in.1974, the first closed dressing in the world, and put forward the theory of wet healing and was gradually accepted. In the past 50 years, a large number of basic research and clinical treatment of the wound have proved that the moist environment is beneficial to the healing of the wound, and the new moisturizing dressing has also entered the medical market. However, the new dressing has a good work. Single, lack of compound treatment function, complex preparation process, high price, slow market popularization, and the new dressings on the domestic market are mainly from the European and American industry giants, and the local enterprises have few products. The traditional Chinese medicine dressing has the unique features in the treatment of wound repair. It can actively promote wound healing and prevent scar hyperplasia, suitable for various kinds of wound. The treatment and nursing of acute and chronic wounds, with simple sources, safety, small side effects and low cost, can improve the level of wound care, reduce the cost of medical service and improve the efficiency of hospital work. However, the traditional Chinese medicine dressings have also been unable to maintain the closed, micro humid environment and use inadequacy, and the safety problems have been paid much attention to. The quality is directly related to the safety of the patient's life. The developed countries began to monitor the safety of medical dressings in 1960s for.1976 years. The US Food and Drug Administration (FDA) began to manage medical dressings and implemented the pre sale examination and approval system, and established a series of raw materials and quality evaluation standards. China began in the late 70s. The biological evaluation of medical dressings is studied. The relevant regulations of medical instruments registration in China require medical dressings to enter the market formally. After completing the processing performance and shape design procedures, the medical dressings must be evaluated in physical and chemical properties, biological evaluation, and product quality standards, but there is still a certain difference compared with the developed countries. For example, because of the complex composition of traditional Chinese medicine dressing and the variety of preparation forms, the country has not established a unified standard for evaluation of traditional Chinese medicine dressing, which restricts the development of its industrialization. Therefore, there are some deficiencies in the research of traditional Chinese medicine dressings. There are defects in the R & D system and need further improvement. On the basis of Department of Traumatology test, according to the principle of prescription of traditional Chinese medicine, select the Chinese herbal medicine of different efficacy of Chinese herbal medicine "37 compound". The whole prescription is composed of 9 herbs, including 37, the herb, the Bletilla striata, the catechu, the chicken Yato, the two needles and the Polygonum cuspidatum, which have good hemostatic, antibacterial, swelling and analgesic effects, but the original dosage forms are inconvenient and easy to use We have made a reform in this department of Traumatology, combined the characteristics of the Department of Traumatology laboratory prescription with the advantages of the new moisturizing dressing, and developed a kind of traditional Chinese medicine that can provide a wet healing environment, quickly absorb exudative fluid, promote the growth of granulation, reduce infection and promote wound healing. Sponge dressing, which is three purple wound healing sponges, is suitable for all kinds of mechanical or traumatic wounds, chronic infection wounds, diabetic foot ulcers and other acute and chronic wounds. However, its safety and efficacy need to be confirmed and the quality needs to be evaluated. Therefore, we study the pharmacodynamics and quality analysis of three purple healing sponge compress to ensure the sponge dressing. It is safe, effective, stable and controllable to provide important scientific data for the registration and approval of medical dressings, and provide scientific basis for further expansion of production. 1. the purpose of this study is to verify the effect of three purple wound healing sponges on the wound healing of rats, and to explore the therapeutic mechanism of.2. for the evaluation of the physical properties of the three purple healing sponges for the identification of the three purple Chuang Yu sea. Chinese herbal medicine 37, Radix Polygonum cuspidatum. Research methods: 1. three the pharmacodynamic study of three purple healing sponges. This study adopted a small area skin excision model on the back of rats. The rats were randomly divided into 3 groups, 10 negative control group, 10 positive control group and 10 experimental group. The rats were prone position, removed the back hair of rats, and 3% pentobarbital sodium. After intraperitoneal injection of anaesthesia, iodophor and 70% ethanol in the routine disinfection of the skin, 2 round full layer defect wounds with diameter of 2cm were made with a sharp knife on the back of the rat, and the surface of the wound was fully cleaned by chlorhexidine acetate solution. The experimental group covered the wound with a self-made three purple healing spongy dressing, and the positive control group covered the wound with a blank sponge matrix. The negative control group covered the wound with vaseline gauze. The outer layer of the three groups was bandaged with sterile gauze dressing. The first three days, the medicine was changed one day, after three days, the medicine was changed one day, and the other two days after the other was changed. Every change of medicine, the sex and behavior of the three groups of rats and the large body condition, including mental condition, feeding and drinking water, were observed. Activities, wound exudation, bleeding, infection signs, granulation tissue growth, and selected wound area, inflammation integral, pathological condition as repair index. After treatment, 3D, 7d, 14d, digital camera were photographed, Image J software was used to measure scale pixel ratio, surface area of wound surface was measured, wound healing rate was calculated, and 3D, 7d measurement of wound surface after treatment. After the treatment, 6 rats (2 rats in each group) were randomly selected to die at 3D, 7d, and 14d after the treatment. The wound skin was immediately cut and 4% of the polymethylene was fixed. H.E staining was used to observe the infiltration and necrosis of the inflammatory cells, fibrous cells and capillary hyperplasia, collagen fibers and other conditions. Study on the quality of.2. three purple Chuang Yu sponge dressing, the method of literature review, reference to the related requirements of Chinese Pharmacopoeia >2010 and the relevant standards of Chinese medical instruments registration, consulting Chinese medicine preparation, nursing clinical and teaching frontline experts, using TLC to determine three purple healing sponges. Identifying the ginsenoside Rb1, ginsenoside Rg1 and polygonin in the spongy dressing, the control solution, the test solution, the negative solution on the same thin plate, the expansion agent in the unfolding container, the component of the sample solution separated, the chromatogram comparison, the identification of the preparation, and the selection of air permeability and water absorption. 7 indexes such as moisture, tear strength, weight difference, heat resistance and cold resistance were used as evaluation indexes of sponge dressing. In order to verify the rationality of the process conditions, three batches of three purple wound dressing were studied. The results were as follows: after the diet and activity situation of the 1. rats were awake, the rats in each group were inactive, without any active activity and Diet: the mental state of the rats in 3D, the experimental group and the positive control group, the active activity, the diet were all close to normal, the negative control group had a small amount of activity and diet, and the mental state was improved. After treatment, the mental state, active activity and diet were normal in group 7d and 14d, and there was no significant difference between the groups. No experimental rats were dead. The healing rate of wound healing in.2. rats was compared. The wound healing rate in the experimental group was (40.22 + 0.05)% after treatment and the wound healing rate in the positive control group was (38.33 + 0.1)%, which was higher than that of the negative control group (22.02 + 0.03)% (22.02 + 0.03), with a significant difference (P0.05), and the wound healing rate of the experimental group was (71.32 + 0.08)% after the treatment at the seventh day after treatment. The wound healing rate (60.66 + 0.09)% and the negative control group were (50.01 + 0.10)%, compared with the negative control group (P0.05). The wound healing rate in the experimental group was (90.44 + 0.07)% after fourteenth days after treatment, which was higher than that in the positive control group (81.03 + 0.04)% and the negative control group (71.65 + 0.10)%, compared with the negative control group (71.65 + 0.10)%. Significant difference (P0.05). Three purple wound healing sponger dressing can reduce wound area, shorten the wound healing time, improve wound healing rate. Pathological observation, compared with the control group, after treatment 3D, the experimental group earlier appeared capillary, fibroblasts began to proliferate. After treatment, 7d, experimental group of wound fiber hyperplasia, collagen fiber bright More than the control group. After treatment, 14d, compared with the control group, the collagen fibers proliferated obviously, and there was no significant change compared with the postoperative 7d. No tissue, tissue destruction and necrosis were not found in the wound. Three purple wound healing sponger dressing could promote the growth of capillary and collagen fibers. The score of inflammation was compared, and the experiment was third days after treatment. The score of inflammation in the wound of the group was (3.20 + 0.42), which was lower than that of the positive control group (4.18 + 0.60) and the score of inflammation (5.67 + 0.50) in the negative control group (5.67 + 0.50). The score of the wound inflammation in the experimental group was (1.70 + 0.48) after seventh days after the treatment, which was lower than that of the positive control group (2.55 + 0.52). The score of wound inflammation (3.89 + 0.33) in the negative control group was 3.89 + 0.33, and there was a significant difference (P0.05). The experimental results showed that the average air permeability of three purple wound dressing was 80.86%, so the permeability rate of this product was not less than 78%. The average moisture retention rate is 38.11%, the moisture retention rate is not less than 35%, the average water absorption rate is 11.15 times, the water absorption rate is not less than 10 times, the average anti tearing strength is 131.59g/cm3, the tearing strength standard is not less than less than 130g/cm3, the maximum slice weight difference is 3.5%, the minimum slice weight difference is 3.4%, the weight difference limit is within the average weight of + 5%, According to the standard of Pharmacopoeia evaluation. After storage in the refrigerator, there is no obvious stratification in the dressing. After placing the oven, the surface of the dressings has no seepage and luster. The three purple wound dressing has good air permeability, water absorption and moisture retention, heat resistance, cold resistance, good shape, high tear strength, and small difference in weight difference. TLC identification of three purple The separation degree of 37 and the Polygonum cuspidatum was good in Chuang Yue sponge dressing. In the sample chromatography, there was the same color spot in the corresponding position of the control substance and negative and no interference. Conclusion: 1. through the study of the main pharmacodynamics of three purple healing sponges, the three purple wound healing spongy compress has rapid hemostasis, anti-inflammatory and bacteriostasis, blood activating and removing stasis. It can absorb the excessive exudation of the wound, prevent the infection, create a good microenvironment for the wound healing, promote the angiogenesis of the wound, promote the growth of the granulation tissue, accelerate the healing of the wound, reduce the production of scar, reduce the sequelae and reduce the death rate. The three purple wound healing sponger dressing has the safety and reliability of the wound dressing with the safety and reliability of three purple. Chuang Yu sponge dressing has good air permeability, water absorption and moisture retention, good heat resistance, good cold resistance, high tearing strength and weight difference limit in the standard range of 5%. TLC identification of ginsenoside Rg1, ginsenoside Rb1 and Polygonum Polygonum are simple, quick, accurate, reliable, reproducible, and can be used as three purple healing sponges. The qualitative identification method provides an experimental basis for the industrialization of the product dressings and the expansion of production in the later stage.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R285

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