本文選題：皮膚缺損 + 脫細(xì)胞真皮基質(zhì)　； 參考：《瀘州醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的： 腫瘤、外傷等病變造成口腔頜面部皮膚及粘膜缺損是目前較為常見的疾病，臨床中最常使用的修復(fù)方式是周圍游離皮瓣轉(zhuǎn)移修復(fù)術(shù)，但大多數(shù)適用于小面積軟組織缺損，對于較大范圍的皮膚、粘膜缺損，由于受到周圍組織張力及組織量多少的限制，臨近游離皮瓣轉(zhuǎn)移修復(fù)常會導(dǎo)致較多的并發(fā)癥。目前在較大面積的缺損中多需使用修復(fù)材料。修復(fù)材料可分為人工合成材料及生物修復(fù)材料，臨床廣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、疼痛等癥狀。生物材料是近幾年國內(nèi)外頜面部皮膚、粘膜缺損修復(fù)的新趨勢，它包括動物源性材料如豬小腸粘膜下層（Porcine small intestinalsubmucosa，P-SIS）、豬脫細(xì)胞真皮基質(zhì)（Porcine acellular derualmatrix，P-ADM）、人體尸源性材料等。研究表明，生物材料不僅具有合成材料的優(yōu)點(diǎn)，也具有更好的生物相容性等特點(diǎn)，術(shù)后感染、疼痛、粘連等并發(fā)癥較合成材料低。 理想的皮膚修復(fù)材料不僅要具有一定強(qiáng)度及物理性質(zhì)穩(wěn)定，而且應(yīng)具有組織相容性高，排斥反應(yīng)低，促進(jìn)組織再生等特征。目前臨床上還未發(fā)現(xiàn)上述理想的修復(fù)材料，多使用的是生物相容性較好的材料，因此不斷的尋找合適的材料一直是臨床研究的熱點(diǎn)。 本研究通過將牛跟腱制取的材料及目前國內(nèi)常用的豬脫細(xì)胞真皮基質(zhì)材料，應(yīng)用于兔子皮膚全層缺損修復(fù)實(shí)驗(yàn)中，對術(shù)后一般情況、皮膚愈合率、病理組織切片、免疫組化等多方面進(jìn)行對比，研究兩種生物材料的生物相容性，并發(fā)癥，炎性反應(yīng)及組織愈合情況，提供實(shí)驗(yàn)數(shù)據(jù)為以后在臨床中更好的選用材料修復(fù)軟組織缺損。 材料和方法 實(shí)驗(yàn)用新西蘭兔16只，體重2-3.5kg，口腔修復(fù)膜（北京天新福），，豬脫細(xì)胞真皮基質(zhì)。于新西蘭兔雙側(cè)背部均切除三處3cm×2cm的皮膚全層，每側(cè)分為空白對照、口腔修復(fù)膜及豬脫細(xì)胞真皮基質(zhì)修復(fù)，并將兔子隨機(jī)分為四組，四組分別于術(shù)后7、14、21、28天進(jìn)行取材，取材后的標(biāo)本進(jìn)行大體觀察、組織學(xué)HE染色、免疫組化及Masson染色觀察，并運(yùn)用圖像分析軟件測量皮膚愈合面積的百分比及炎性細(xì)胞的數(shù)量，將得到的數(shù)據(jù)進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果 一、大體觀察 16只實(shí)驗(yàn)用新西蘭兔術(shù)區(qū)軟組織均未見明顯炎癥反應(yīng)，實(shí)驗(yàn)組及對照組材料均成活未脫落，實(shí)驗(yàn)組于21天左右傷口愈合，對照組于23天左右傷口愈合，空白組于26天左右傷口愈合，兩種材料傷口愈合時間明顯快于空白組，28天時三組傷口均已完全愈合�？瞻滋庱：劬^口腔修復(fù)膜處及豬脫細(xì)胞真皮基質(zhì)處大，口腔修復(fù)膜材料修復(fù)后的瘢痕略小于豬脫細(xì)胞真皮基質(zhì)。 二、HE染色 空白處明顯炎癥反應(yīng)，術(shù)后7天空白組炎性細(xì)胞最多，明顯多于同一時期的兩種材料修復(fù)后的組織，炎性反應(yīng)可能空白組修復(fù)較兩種材料修復(fù)高；兩種修復(fù)材料處早期均形成大量成纖維細(xì)胞及毛細(xì)血管，相互之間差別不大，但均明顯多于空白處；鏡下見修復(fù)后（28天左右）空白組瘢痕最厚，實(shí)驗(yàn)組及對照組形成的瘢痕較空白處小，兩者肉眼觀察相差不大。 三、Masson染色 術(shù)后7天時實(shí)驗(yàn)組及對照組均出現(xiàn)少量膠原面積，對照組膠原新生不明顯。修復(fù)完成時（28天左右）實(shí)驗(yàn)組中真皮新生膠原粗大，呈條索狀，排列較為規(guī)律，三維層次感明顯，對照組中膠原排列相對于實(shí)驗(yàn)組疏松，纖細(xì)，方向紊亂。 四、免疫組化 7天時選取空白組、實(shí)驗(yàn)組和對照組的組織制成切片行免疫組化觀察，結(jié)果顯示：鏡下觀察空白組形成少量血管，兩種材料修復(fù)后的組織中血管形成較多。 五、創(chuàng)面愈合率 創(chuàng)面愈合率=原始創(chuàng)面面積-未愈合創(chuàng)面面積/原始創(chuàng)面面積×100%。通過Image Pro Plus軟件測量不同時期內(nèi)創(chuàng)面面積的大小進(jìn)行統(tǒng)計(jì)學(xué)分析，實(shí)驗(yàn)組及對照組愈合率均高于空白組，相互差別不大，各組總體相比（P＜0.05）具有統(tǒng)計(jì)學(xué)差異。 六、炎性細(xì)胞數(shù)量的統(tǒng)計(jì)觀察 術(shù)后7天空白組炎性細(xì)胞達(dá)到高峰，實(shí)驗(yàn)組及對照組炎性細(xì)胞數(shù)量均較空白組少，14天時空白組炎性細(xì)胞數(shù)量下降，但仍高于兩組修復(fù)材料，各組總體相比（P＜0.05）具有統(tǒng)計(jì)學(xué)差異。 結(jié)論： 1.使用兩種材料均較好的修復(fù)兔皮膚軟組織，具有良好的抗感染能力及生物相容性，可以明顯的縮短兔皮膚傷口的愈合時間，促進(jìn)皮膚傷口的愈合時間、血管化和膠原層積。 2．口腔修復(fù)膜材料在術(shù)后形成瘢痕較豬脫細(xì)胞真皮基質(zhì)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.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R782.2

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