巴沙鱼皮胶原支架在兔颅骨缺损再生中的研究
发布时间：2019-06-04 19:52
【摘要】：研究目的对巴沙鱼皮的组成成分和性质进行分析,评估巴沙鱼皮是否可作为胶原蛋白原来来源的可能性。利用冻干巴沙鱼皮制备出一种主要成分为胶原蛋白的膜状支架材料。分析该材料的相关理化性能,观察该材料在兔颅骨缺损的引导骨再生术中的效果,分析其作为引导骨再生屏障膜的可行性,为巴沙鱼皮胶原蛋白支架作为引导骨再生屏障膜应用于临床提供理论依据和可能性,以探索巴沙鱼皮是否可以成为胶原支架材料的新来源。研究方法对来源于巴沙鱼皮所提取的胶原蛋白进行成分分析和性质的分析。对制备出的巴沙鱼皮胶原支架进行理化性能的检测,主要有肉眼观察材料的外形,扫描电镜观察和分析材料的结构以及孔隙的大小和分布等,采用液体置换法测定巴沙鱼皮胶原支架的孔隙率,使用万能试验机测定材料的机械强度。根据其在磷酸缓冲液中浸泡28d后的失重率来测定该材料的体外降解率。测定该材料的吸水率并观察该材料在湿润状态下的溶胀情况。通过随温度变化导致胶原蛋白黏度变化的特点来测定材料的变性温度。CCK-8细胞毒性试验检测材料是否对生物体有毒性。通过兔子颅骨缺损的引导骨再生实验初步判断该材料作为引导骨再生屏障膜的作用效果和潜能。结果巴沙鱼皮粗蛋白含量为95.20%;胶原蛋白类型主要为I型胶原;与其他鱼类相比,巴沙鱼皮胶原蛋白中所含的PrO和Hyp占全部氨基酸含量的比例最高;肉眼观察厚薄均匀。扫描电镜显示材料为双层结构,一面疏松多孔,孔径大小不一,分布均匀;另一面光滑,孔隙致密。孔隙率(55.50±1.94)%,厚度(0.66±0.10)mm,抗拉强度(18.82±0.94)MPa。该材料28d的降解率为(80.22%±2.49),48h的吸水率为(379.77%±77.81)。巴沙鱼皮胶原蛋白支架的变性温度为34℃左右,经过干热交联后的巴沙鱼皮变性温度可升高至35℃左右。在兔颅骨缺损再生实验中,巴沙鱼皮胶原支架能起到引导骨再生的作用,在缺损区与皮肤黏膜等软组织阻隔开,有效保护了缺损区域,阻挡了纤维细胞的长入,同时为成骨过程提供了支架,促进了再生组织内的血管化。最终结果经统计分析可以发现巴沙鱼皮胶原支架的成骨效果较空白对照组好,比Bio-Gide膜组稍差,证明巴沙鱼皮胶原支架具有一定的成骨能力。结论巴沙鱼皮胶原蛋白支架理化性能稳定,表面具有一定的孔隙率,且为双层结构,一面粗糙,有较多孔隙,一面光滑致密,符合作为屏障膜的基本结构要求。吸水性适中,吸水后未见有明显变形和溶解。稳定性良好,降解速率与Bio-Gide胶原膜相比偏高。无细胞毒性,具有良好的生物相容性。在兔颅骨缺损再生术中可起到引导骨再生的作用。该材料具有进一步研究开发的潜能,具有成为一种新型临床医用膜材料的可能性。
[Abstract]:Objective to analyze the composition and properties of Barsha fish skin and to evaluate the possibility that Barsha fish skin can be used as the original source of collagen. A kind of membrane scaffold material with collagen, which is the main component of lyophilized Barthard fish skin, was prepared. The physical and chemical properties of the material were analyzed, the effect of the material in guided bone regeneration of rabbit skull defect was observed, and the feasibility of the material as a guide bone regeneration barrier membrane was analyzed. It provides a theoretical basis and possibility for Barsha fish skin collagen scaffold to be used as a guiding bone regeneration barrier membrane in clinical practice, so as to explore whether Barsha fish skin can become a new source of collagen scaffold material. Methods the components and properties of collagen extracted from Barsha fish skin were analyzed. The physical and chemical properties of the prepared Barsha fish skin collagen scaffold were mainly observed by naked eye, the structure, pore size and distribution of the material were observed and analyzed by scanning electron microscope (SEM). The porosity of Barsha fish skin collagen scaffold was measured by liquid replacement method, and the mechanical strength of the material was measured by universal testing machine. The degradation rate of the material in vitro was measured according to the weight loss rate of the material after soaking in phosphate buffer for 28 days. The water absorption of the material was measured and the swelling of the material in wet state was observed. The denaturation temperature of the material was measured by changing the viscosity of collagen with the change of temperature. CCK-8 cytotoxicity test was used to detect whether the material was toxic to organisms. The effect and potential of the material as the barrier membrane of guided bone regeneration were preliminarily judged by the guided bone regeneration experiment of rabbit skull defect. Results the crude protein content of Barsha fish skin was 95.20%, and the collagen type was mainly type I collagen. Compared with other fish, the proportion of PrO and Hyp in Barsha fish skin collagen was the highest, and the thickness was uniform. Scanning electron microscope (SEM) shows that the material is double-layer structure, one side is loose and porous, the pore size is different, and the distribution is uniform, the other side is smooth and the pores are dense. Porosity (55. 50 卤1. 94)%, thickness (0. 66 卤0. 10) mm, tensile strength (18. 82 卤0. 94) MPa. The degradation rate of the material was (80.22% 卤2.49) for 28 days and (379.77% 卤77.81) for 48 h. The denaturation temperature of Barsha fish skin collagen scaffold is about 34 鈩，
本文编号：2492959
[Abstract]:Objective to analyze the composition and properties of Barsha fish skin and to evaluate the possibility that Barsha fish skin can be used as the original source of collagen. A kind of membrane scaffold material with collagen, which is the main component of lyophilized Barthard fish skin, was prepared. The physical and chemical properties of the material were analyzed, the effect of the material in guided bone regeneration of rabbit skull defect was observed, and the feasibility of the material as a guide bone regeneration barrier membrane was analyzed. It provides a theoretical basis and possibility for Barsha fish skin collagen scaffold to be used as a guiding bone regeneration barrier membrane in clinical practice, so as to explore whether Barsha fish skin can become a new source of collagen scaffold material. Methods the components and properties of collagen extracted from Barsha fish skin were analyzed. The physical and chemical properties of the prepared Barsha fish skin collagen scaffold were mainly observed by naked eye, the structure, pore size and distribution of the material were observed and analyzed by scanning electron microscope (SEM). The porosity of Barsha fish skin collagen scaffold was measured by liquid replacement method, and the mechanical strength of the material was measured by universal testing machine. The degradation rate of the material in vitro was measured according to the weight loss rate of the material after soaking in phosphate buffer for 28 days. The water absorption of the material was measured and the swelling of the material in wet state was observed. The denaturation temperature of the material was measured by changing the viscosity of collagen with the change of temperature. CCK-8 cytotoxicity test was used to detect whether the material was toxic to organisms. The effect and potential of the material as the barrier membrane of guided bone regeneration were preliminarily judged by the guided bone regeneration experiment of rabbit skull defect. Results the crude protein content of Barsha fish skin was 95.20%, and the collagen type was mainly type I collagen. Compared with other fish, the proportion of PrO and Hyp in Barsha fish skin collagen was the highest, and the thickness was uniform. Scanning electron microscope (SEM) shows that the material is double-layer structure, one side is loose and porous, the pore size is different, and the distribution is uniform, the other side is smooth and the pores are dense. Porosity (55. 50 卤1. 94)%, thickness (0. 66 卤0. 10) mm, tensile strength (18. 82 卤0. 94) MPa. The degradation rate of the material was (80.22% 卤2.49) for 28 days and (379.77% 卤77.81) for 48 h. The denaturation temperature of Barsha fish skin collagen scaffold is about 34 鈩，
本文编号：2492959
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