保留软骨的去细胞全喉支架制备方法及在喉软骨缺损中的修复效果

发布时间：2018-06-23 15:10

  本文选题：去细胞全喉支架 + 喉软骨缺损　； 参考：《中国老年学杂志》2017年24期

【摘要】：目的探讨保留软骨的去细胞全喉支架的制备方法及在大鼠喉软骨缺损中的修复效果。方法取6只大鼠用于支架的制备,分离颈总动脉,结扎分支,经颈总动脉灌注去污剂(SDS、Triton X-100、PBS离子型),3只大鼠去除喉黏膜、肌肉、韧带的同时,保留细胞外基质和软骨细胞制备保留软骨的去细胞全喉支架,其余3只用于制备对照组支架。另取34只大鼠,建立0.5 cm×0.5 cm全层喉软骨缺损模型,根据处理方法不同随机分为对照组(n=17)和保留软骨组(n=17),对照组植入未经处理的异体喉骨支架,保留软骨组植入保留软骨的去细胞全喉支架,比较两组的修复效果。结果 (1)两种支架经过脱细胞灌注处理后,全喉大体结构完整,体积未发生明显的变化;(2)保留软骨的去细胞全喉支架未见表面细胞组织填充,可见胶原、细胞外基质成分组成的腔隙,细胞之间结构紧密,存在于会厌、声带、环状软骨等表面;对照组支架组织中的细胞成分被完全去除,但是依稀可见网状连接,结构之间不紧密;(3)免疫荧光结果显示:保留软骨的去细胞全喉支架除了有肌肉、软骨细胞外,主要组织相容性复合体(MHC)-Ⅰ、MHC-Ⅱ染色均为阴性;对照组支架免疫荧光染色下MHC-Ⅰ、MHC-Ⅱ染色均为阳性;(4)保留软骨组大鼠修复6 w后缺损部位存在少许不成熟的软骨细胞,极少的软骨形成陷窝,表面存在纤维组织及炎症细胞;修复12 w后缺损部位得到修复,形成软骨陷窝,炎症反应减少;对照组修复6 w后缺损部位周围软骨细胞较少,缺损部位与周围组织边界明显,存在大量炎症细胞;修复12 w后存在少量纤维组织,存在炎症因子。结论保留软骨组植入保留软骨的去细胞全喉支架在喉软骨缺损中具有良好的修复效果。
[Abstract]:Objective to investigate the preparation method of chondrogenic acellular whole larynx scaffold and its effect in repairing rat laryngeal cartilage defect. Methods six rats were used in the preparation of stent, the common carotid artery was isolated, the branches were ligated, and three rats were perfused through the common carotid artery to remove the laryngeal mucosa, muscle and ligaments. Extracellular matrix (ECM) and chondrocytes were retained to prepare chondrogenic laryngeal scaffolds. Another 34 rats were selected to establish a 0.5 cm 脳 0.5 cm full-thickness laryngeal cartilage defect model. According to the different treatment methods, the rats were randomly divided into two groups: the control group (nong17) and the cartilage preserving group (nong17). The untreated laryngeal bone scaffold was implanted in the control group. Cartilage-preserving group was implanted with chondrocyte-preserving laryngeal scaffold to compare the repair effect between the two groups. Results (1) after the two scaffolds were treated with acellular perfusion, the whole laryngeal structure was intact, and the volume of the whole larynx was not changed obviously. (2) the chondrogenic acellular laryngeal scaffold was not filled with surface cellular tissue, and collagen was found in the scaffold. The interstitial space composed of extracellular matrix components, with a tight cell structure, which exists on the surface of epiglottis, vocal cord, annular cartilage, etc. In the control group, the cellular components of the scaffold tissue were completely removed, but reticular connections were observed. (3) the results of immunofluorescence showed that the main histocompatibility complex (MHC)-鈪，

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