【摘要】：目的:1、制備絲素蛋白-殼聚糖(SFCS)新型高分子材料,通過(guò)表征觀察,降解實(shí)驗(yàn),研究符合組織工程要求的兩者最優(yōu)配置比例。2、使用絲素蛋白-殼聚糖復(fù)合材料進(jìn)行生物相容性實(shí)驗(yàn),初步評(píng)估其作為高分子醫(yī)用材料應(yīng)用到肛瘺栓上的可行性。方法:1、絲素蛋白 殼聚糖三維多孔支架的制做;使用氯化鈣 醇 水體系(Ca CL2:C2H5OH:H2O=1:2:8)溶解,經(jīng)透析、濃縮、離心獲得2%的絲素蛋白溶液,殼聚糖溶解于醋酸溶液中配制出3%的殼聚糖溶液,將二者以不同的比例混合,裝模、冷凍干燥、交聯(lián),獲得絲素蛋白 殼聚糖三維支架產(chǎn)品。2、材料表征實(shí)驗(yàn);采用掃描電子顯微鏡測(cè)試法,觀察支架材料的特征,計(jì)算出材料的溶脹率、吸水率、孔隙率。3、材料靜態(tài)降解實(shí)驗(yàn);在磷酸鹽緩沖溶液(PBS溶液)中放入SFCS復(fù)合材料,降解溫度為37℃的恒溫烤箱中,觀察絲素蛋白 殼聚糖的降解速率及穩(wěn)定性。4、絲素蛋白 殼聚糖復(fù)合材料的細(xì)胞生物相容性評(píng)價(jià)。將SFCS進(jìn)行溶血實(shí)驗(yàn),將SFCS支架和橡膠的樣條分別植入SD大鼠大腿肌肉,在第7、14、21、28天,四個(gè)時(shí)間點(diǎn),分別處死大鼠,分別在創(chuàng)面組織進(jìn)行HE染色、Masson三色染色。結(jié)果:1、通過(guò)絲素蛋白與殼聚糖溶液不同質(zhì)量比相混合后,可以得到不同成分組成的SFCS支架。2、不同配比的SFCS孔隙率值均高于95%。材料的溶脹率和吸水率是隨著殼聚糖比例的增加而變大的。3、體外降解實(shí)驗(yàn)表明在同一時(shí)間點(diǎn),隨著絲素蛋白含量的增高復(fù)合材料的降解率依次遞減。3、絲素蛋白 殼聚糖復(fù)合材料的溶血率小于5%,符合國(guó)家相關(guān)標(biāo)準(zhǔn)。4、動(dòng)物實(shí)驗(yàn)表明,4周末,SFCS支架相比對(duì)照組孔內(nèi)部炎癥細(xì)胞幾乎完全消失,被正常細(xì)胞填滿,形成了完整地纖維囊壁結(jié)構(gòu),膠原含量明顯高于對(duì)照組,膠原纖維較粗大、緊密,趨向更加明顯。結(jié)論:1、將絲素蛋白和殼聚糖相混合,成功地制備出了擁有穩(wěn)定構(gòu)造的三維多孔支架,其中40%SF 60%CS組可作為生物支架研究的首選配比。2、動(dòng)物埋植實(shí)驗(yàn)中,顯示SFCS復(fù)合支架材料具有良好的生物相容性,是進(jìn)行肛瘺栓前期材料研究中比較好的一個(gè)選擇參考。
[Abstract]:Objective to prepare a new polymer material of silk fibroin chitosan (SFCS). In order to evaluate the feasibility of using fibroin-chitosan composite as a high molecular material for anal fistula suppository, the biocompatibility experiment was carried out with silk fibroin-chitosan composite. Methods 1: 1, preparation of silk fibroin / chitosan three-dimensional porous scaffold, 2% silk fibroin solution was obtained by dialysis, concentration, centrifugation and dissolution with calcium chloride CL2:C2H5OH:H2O=1:2:8 system (Ca CL2:C2H5OH:H2O=1:2:8). Chitosan was dissolved in acetic acid solution and 3% chitosan solution was prepared. The chitosan solution was mixed in different proportion, mold, freeze-drying, crosslinking, silk fibroin protein chitosan three-dimensional scaffold product. The characteristics of the scaffold material were observed by scanning electron microscopy (SEM). The swelling rate, water absorption, porosity, static degradation of the material were calculated, and the SFCS composite was added to the phosphate buffer solution (PBS solution). The degradation rate and stability of fibroin / chitosan were observed in a constant temperature oven at 37 鈩，

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