巴沙魚皮膠原支架在兔顱骨缺損再生中的研究
發(fā)布時(shí)間：2019-06-04 19:52
【摘要】：研究目的對(duì)巴沙魚皮的組成成分和性質(zhì)進(jìn)行分析,評(píng)估巴沙魚皮是否可作為膠原蛋白原來(lái)來(lái)源的可能性。利用凍干巴沙魚皮制備出一種主要成分為膠原蛋白的膜狀支架材料。分析該材料的相關(guān)理化性能,觀察該材料在兔顱骨缺損的引導(dǎo)骨再生術(shù)中的效果,分析其作為引導(dǎo)骨再生屏障膜的可行性,為巴沙魚皮膠原蛋白支架作為引導(dǎo)骨再生屏障膜應(yīng)用于臨床提供理論依據(jù)和可能性,以探索巴沙魚皮是否可以成為膠原支架材料的新來(lái)源。研究方法對(duì)來(lái)源于巴沙魚皮所提取的膠原蛋白進(jìn)行成分分析和性質(zhì)的分析。對(duì)制備出的巴沙魚皮膠原支架進(jìn)行理化性能的檢測(cè),主要有肉眼觀察材料的外形,掃描電鏡觀察和分析材料的結(jié)構(gòu)以及孔隙的大小和分布等,采用液體置換法測(cè)定巴沙魚皮膠原支架的孔隙率,使用萬(wàn)能試驗(yàn)機(jī)測(cè)定材料的機(jī)械強(qiáng)度。根據(jù)其在磷酸緩沖液中浸泡28d后的失重率來(lái)測(cè)定該材料的體外降解率。測(cè)定該材料的吸水率并觀察該材料在濕潤(rùn)狀態(tài)下的溶脹情況。通過(guò)隨溫度變化導(dǎo)致膠原蛋白黏度變化的特點(diǎn)來(lái)測(cè)定材料的變性溫度。CCK-8細(xì)胞毒性試驗(yàn)檢測(cè)材料是否對(duì)生物體有毒性。通過(guò)兔子顱骨缺損的引導(dǎo)骨再生實(shí)驗(yàn)初步判斷該材料作為引導(dǎo)骨再生屏障膜的作用效果和潛能。結(jié)果巴沙魚皮粗蛋白含量為95.20%;膠原蛋白類型主要為I型膠原;與其他魚類相比,巴沙魚皮膠原蛋白中所含的PrO和Hyp占全部氨基酸含量的比例最高;肉眼觀察厚薄均勻。掃描電鏡顯示材料為雙層結(jié)構(gòu),一面疏松多孔,孔徑大小不一,分布均勻;另一面光滑,孔隙致密�？紫堵�(55.50±1.94)%,厚度(0.66±0.10)mm,抗拉強(qiáng)度(18.82±0.94)MPa。該材料28d的降解率為(80.22%±2.49),48h的吸水率為(379.77%±77.81)。巴沙魚皮膠原蛋白支架的變性溫度為34℃左右,經(jīng)過(guò)干熱交聯(lián)后的巴沙魚皮變性溫度可升高至35℃左右。在兔顱骨缺損再生實(shí)驗(yàn)中,巴沙魚皮膠原支架能起到引導(dǎo)骨再生的作用,在缺損區(qū)與皮膚黏膜等軟組織阻隔開,有效保護(hù)了缺損區(qū)域,阻擋了纖維細(xì)胞的長(zhǎng)入,同時(shí)為成骨過(guò)程提供了支架,促進(jìn)了再生組織內(nèi)的血管化。最終結(jié)果經(jīng)統(tǒng)計(jì)分析可以發(fā)現(xiàn)巴沙魚皮膠原支架的成骨效果較空白對(duì)照組好,比Bio-Gide膜組稍差,證明巴沙魚皮膠原支架具有一定的成骨能力。結(jié)論巴沙魚皮膠原蛋白支架理化性能穩(wěn)定,表面具有一定的孔隙率,且為雙層結(jié)構(gòu),一面粗糙,有較多孔隙,一面光滑致密,符合作為屏障膜的基本結(jié)構(gòu)要求。吸水性適中,吸水后未見有明顯變形和溶解。穩(wěn)定性良好,降解速率與Bio-Gide膠原膜相比偏高。無(wú)細(xì)胞毒性,具有良好的生物相容性。在兔顱骨缺損再生術(shù)中可起到引導(dǎo)骨再生的作用。該材料具有進(jìn)一步研究開發(fā)的潛能,具有成為一種新型臨床醫(yī)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 鹵1. 94)%, thickness (0. 66 鹵0. 10) mm, tensile strength (18. 82 鹵0. 94) MPa. The degradation rate of the material was (80.22% 鹵2.49) for 28 days and (379.77% 鹵77.81) for 48 h. The denaturation temperature of Barsha fish skin collagen scaffold is about 34 鈩，
本文編號(hào)：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 鹵1. 94)%, thickness (0. 66 鹵0. 10) mm, tensile strength (18. 82 鹵0. 94) MPa. The degradation rate of the material was (80.22% 鹵2.49) for 28 days and (379.77% 鹵77.81) for 48 h. The denaturation temperature of Barsha fish skin collagen scaffold is about 34 鈩，
本文編號(hào)：2492959
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