牛胶原蛋白膜应用于兔皮肤缺损的实验研究

发布时间：2018-06-12 21:42

本文选题：皮肤缺损 + 脱细胞真皮基质　；参考：《泸州医学院》2014年硕士论文

【摘要】：目的：肿瘤、外伤等病变造成口腔颌面部皮肤及粘膜缺损是目前较为常见的疾病，临床中最常使用的修复方式是周围游离皮瓣转移修复术，但大多数适用于小面积软组织缺损，对于较大范围的皮肤、粘膜缺损，由于受到周围组织张力及组织量多少的限制，临近游离皮瓣转移修复常会导致较多的并发症。目前在较大面积的缺损中多需使用修复材料。修复材料可分为人工合成材料及生物修复材料，临床广泛使用的是人工合成材料，但其存在生物相容性差、材料移位、感染、疼痛等症状。生物材料是近几年国内外颌面部皮肤、粘膜缺损修复的新趋势，它包括动物源性材料如猪小肠粘膜下层（Porcine small intestinalsubmucosa，P-SIS）、猪脱细胞真皮基质（Porcine acellular derualmatrix，P-ADM）、人体尸源性材料等。研究表明，生物材料不仅具有合成材料的优点，也具有更好的生物相容性等特点，术后感染、疼痛、粘连等并发症较合成材料低。理想的皮肤修复材料不仅要具有一定强度及物理性质稳定，而且应具有组织相容性高，排斥反应低，促进组织再生等特征。目前临床上还未发现上述理想的修复材料，多使用的是生物相容性较好的材料，因此不断的寻找合适的材料一直是临床研究的热点。本研究通过将牛跟腱制取的材料及目前国内常用的猪脱细胞真皮基质材料，应用于兔子皮肤全层缺损修复实验中，对术后一般情况、皮肤愈合率、病理组织切片、免疫组化等多方面进行对比，研究两种生物材料的生物相容性，并发症，炎性反应及组织愈合情况，提供实验数据为以后在临床中更好的选用材料修复软组织缺损。材料和方法实验用新西兰兔16只，体重2-3.5kg，口腔修复膜（北京天新福），，猪脱细胞真皮基质。于新西兰兔双侧背部均切除三处3cm×2cm的皮肤全层，每侧分为空白对照、口腔修复膜及猪脱细胞真皮基质修复，并将兔子随机分为四组，四组分别于术后7、14、21、28天进行取材，取材后的标本进行大体观察、组织学HE染色、免疫组化及Masson染色观察，并运用图像分析软件测量皮肤愈合面积的百分比及炎性细胞的数量，将得到的数据进行统计学分析。结果一、大体观察 16只实验用新西兰兔术区软组织均未见明显炎症反应，实验组及对照组材料均成活未脱落，实验组于21天左右伤口愈合，对照组于23天左右伤口愈合，空白组于26天左右伤口愈合，两种材料伤口愈合时间明显快于空白组，28天时三组伤口均已完全愈合。空白处瘢痕均较口腔修复膜处及猪脱细胞真皮基质处大，口腔修复膜材料修复后的瘢痕略小于猪脱细胞真皮基质。二、HE染色空白处明显炎症反应，术后7天空白组炎性细胞最多，明显多于同一时期的两种材料修复后的组织，炎性反应可能空白组修复较两种材料修复高；两种修复材料处早期均形成大量成纤维细胞及毛细血管，相互之间差别不大，但均明显多于空白处；镜下见修复后（28天左右）空白组瘢痕最厚，实验组及对照组形成的瘢痕较空白处小，两者肉眼观察相差不大。三、Masson染色术后7天时实验组及对照组均出现少量胶原面积，对照组胶原新生不明显。修复完成时（28天左右）实验组中真皮新生胶原粗大，呈条索状，排列较为规律，三维层次感明显，对照组中胶原排列相对于实验组疏松，纤细，方向紊乱。四、免疫组化 7天时选取空白组、实验组和对照组的组织制成切片行免疫组化观察，结果显示：镜下观察空白组形成少量血管，两种材料修复后的组织中血管形成较多。五、创面愈合率创面愈合率=原始创面面积-未愈合创面面积/原始创面面积×100%。通过Image Pro Plus软件测量不同时期内创面面积的大小进行统计学分析，实验组及对照组愈合率均高于空白组，相互差别不大，各组总体相比（P＜0.05）具有统计学差异。六、炎性细胞数量的统计观察术后7天空白组炎性细胞达到高峰，实验组及对照组炎性细胞数量均较空白组少，14天时空白组炎性细胞数量下降，但仍高于两组修复材料，各组总体相比（P＜0.05）具有统计学差异。结论： 1.使用两种材料均较好的修复兔皮肤软组织，具有良好的抗感染能力及生物相容性，可以明显的缩短兔皮肤伤口的愈合时间，促进皮肤伤口的愈合时间、血管化和胶原层积。 2．口腔修复膜材料在术后形成瘢痕较猪脱细胞真皮基质小，具有良好的抗瘢痕形成能力。
[Abstract]:Objective:
The defects of the oral and maxillofacial skin and mucosa caused by tumor and trauma are the most common diseases of the oral and maxillofacial skin. The most commonly used repair method in clinic is free flap transfer repair, but most of them are suitable for small area soft tissue defects. For larger skin, mucous membrane defects, due to the surrounding tissue tension and tissue volume. Many limitations, near free flap transfer and repair often lead to more complications. Repair materials need to be used in large area defects. Repair materials can be divided into synthetic materials and bioremediation materials, and synthetic materials are widely used in clinic, but they exist in poor biocompatibility, material displacement, infection, pain and so on. Biomaterials are a new trend in the repair of skin and mucous defects in the maxillofacial region in recent years. It includes animal derived materials such as Porcine small intestinalsubmucosa (P-SIS), porcine acellular dermal matrix (Porcine acellular derualmatrix, P-ADM), human body source materials and so on. It has the advantages of synthetic materials and better biocompatibility. Postoperative infection, pain, adhesions and other complications are lower than synthetic materials.
Ideal skin repair materials should not only have a certain strength and physical properties, but also have the characteristics of high tissue compatibility, low rejection and promoting tissue regeneration. At present, the ideal materials have not been found in clinic, and most of them are biocompatible materials. It is a hot spot in clinical research.
In this study, the material of the bovine Achilles tendon and the current domestic porcine acellular dermal matrix materials were applied to the repair experiment of the whole skin defect of rabbit skin. The general condition, the healing rate of the skin, the pathological tissue section and the immunohistochemistry were compared, and the biocompatibility, complications and inflammation of the two kinds of biological materials were studied. Response and tissue healing, provide experimental data for later clinical selection of materials to repair soft tissue defects.
Materials and methods
16 New Zealand rabbits, body weight 2-3.5kg, oral repair membrane (Beijing Tian Xinfu) and porcine acellular dermal matrix were used to remove three 3cm x 2cm skin layers on the bilateral back of New Zealand rabbits. Each side was divided into blank control, oral repair membrane and porcine acellular dermal matrix, and rabbits were randomly divided into four groups, and the four groups were respectively 7,14,21 after operation. After 28 days, the specimens were collected, the specimens were observed, the histological HE staining, immunohistochemistry and Masson staining were observed, and the percentage of skin healing area and the number of inflammatory cells were measured by the image analysis software. The data were analyzed statistically.
Result
First, general observation
There was no obvious inflammatory reaction in the soft tissue of the 16 New Zealand rabbits. The experimental group and the control group were all survived, the experimental group was healed around 21 days, the control group healed around 23 days, the blank group was healed around 26 days. The healing time of the two materials was faster than the blank group, and the three wounds were all at 28 days. The cicatricial scar of the blank was larger than that of the oral repair membrane and the porcine acellular dermal matrix, and the scar of the repair membrane was slightly less than the porcine acellular dermal matrix.
Two, HE staining
There were obvious inflammatory reactions in the blank area. The most inflammatory cells in the 7 day blank group were more than the two kinds of tissue repaired at the same time. The inflammatory response may be higher than that of the two materials, and the two kinds of repair materials formed a large number of fibroblasts and capillary vessels at the early stage. The scars in the blank group were the thickest in the blank group after the repair (about 28 days). The scar formed in the experimental group and the control group was smaller than that in the blank area, and the difference between the two eyes was little.
Three, Masson staining
On the 7 day after the operation, the experimental group and the control group had a small amount of collagen area, and the control group had no obvious collagen freshmen. When the repair was completed (about 28 days), the new collagen in the experimental group was thick and large, the arrangement was more regular and the three-dimensional sense was obvious. The collagen permutation phase in the control group was loose, thin and disorderly.
Four, immunohistochemistry
In the 7 day, the blank group was selected. The tissue of the experimental group and the control group were made into immunohistochemical staining. The results showed that a small amount of blood vessels were formed in the blank group under the microscope, and the blood vessels in the two tissues were formed more.
Five, wound healing rate
The wound healing rate = original wound area - the area of the original wound surface / original wound area * 100%. measured the size of the wound area through the Image Pro Plus software for statistical analysis. The healing rate of the experimental group and the control group was higher than that of the blank group, and the difference was not significant. The statistical difference was found between the groups of each group (P < 0.05).
Six, statistical observation of the number of inflammatory cells
The number of inflammatory cells in the blank group reached the peak at 7 days after the operation. The number of inflammatory cells in the experimental group and the control group was less than that in the blank group. The number of inflammatory cells in the blank group decreased at the 14 day, but it was still higher than the two groups of repair materials. Compared with the group (P < 0.05), there was a statistical difference.
Conclusion:
1. the use of two kinds of materials can better repair the skin soft tissue of rabbit. It has good anti infection ability and biocompatibility. It can obviously shorten the healing time of rabbit skin wound, promote the healing time of skin wound, vascularization and collagenous layer.
2. the postoperative scar formation of prosthodontics membrane is smaller than that of porcine acellular dermal matrix.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R782.2

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