【摘要】：蠶絲蛋白作為一種優(yōu)異的天然高分子纖維蛋白已廣泛應(yīng)用于生物醫(yī)用材料領(lǐng)域。其來(lái)源可靠,獲取方便,具有良好的力學(xué)性能,良好的細(xì)胞相容性,組織相容性。本課題探討了透明絲素蛋白膜作為角膜修復(fù)材料的可行性。采用酰胺類小分子與絲素蛋白復(fù)合,研究絲素蛋白復(fù)合膜的機(jī)械性能,透光性能,以及細(xì)胞相容性。通過(guò)X-射線衍射法研究了絲素蛋白復(fù)合膜的結(jié)構(gòu),結(jié)果表明當(dāng)酰胺添加量較低時(shí),復(fù)合膜主要為Silk I結(jié)晶結(jié)構(gòu),當(dāng)酰胺添加量較多時(shí),復(fù)合膜轉(zhuǎn)變?yōu)镾ilk II結(jié)晶結(jié)構(gòu)。力學(xué)性能測(cè)試表明,復(fù)合膜的斷裂強(qiáng)度雖然比純絲素膜有所降低,但是酰胺的加入改善了其柔韌性,使得其斷裂伸長(zhǎng)率增大。溶失率以及溶脹度測(cè)定的結(jié)果表明,酰胺的加入極大地降低了復(fù)合膜的溶失率,并且具有較低的溶脹度。采用酶標(biāo)儀檢測(cè)了復(fù)合膜的透光性能,并對(duì)復(fù)合膜的透光性進(jìn)行了長(zhǎng)時(shí)間的追蹤檢測(cè),結(jié)果表明,酰胺/絲素蛋白膜具有較長(zhǎng)時(shí)間的透光穩(wěn)定性。提取家兔角膜基質(zhì)細(xì)胞,通過(guò)在絲素蛋白膜上的體外培養(yǎng),研究了酰胺/絲素蛋白復(fù)合膜與角膜基質(zhì)細(xì)胞的相容性,并與乙醇處理的純絲素蛋白膜進(jìn)行對(duì)比。采用血球計(jì)數(shù)板測(cè)定細(xì)胞在不同膜上的黏附率,使用阿爾瑪藍(lán)測(cè)定細(xì)胞在不同膜上生長(zhǎng)不同天后的熒光值,利用熒光倒置顯微鏡觀察角膜基質(zhì)細(xì)胞生長(zhǎng)形態(tài),Masson染色觀察細(xì)胞外膠原蛋白的分泌。結(jié)果表明,酰胺/絲素膜與純絲素膜之間并無(wú)顯著差異,說(shuō)明酰胺/絲素膜能夠支持角膜基質(zhì)細(xì)胞正常生長(zhǎng),酰胺/絲素復(fù)合膜有望用于角膜組織的修復(fù)。
[Abstract]:Silk protein as an excellent natural polymer fibrin has been widely used in the field of biomedical materials. Its source is reliable, easy to obtain, with good mechanical properties, good cytocompatibility and histocompatibility. In this paper, the feasibility of hyaline fibroin membrane as corneal repair material was discussed. The mechanical properties, transmittance and cytocompatibility of silk fibroin composite membrane were studied by using amides and silk fibroin. The structure of silk fibroin composite membrane was studied by X-ray diffraction. The results showed that the composite film was mainly Silk I crystalline structure when the amount of amide was low, and the composite film changed into Silk II crystal structure when the amount of amide was more. The mechanical properties test showed that the fracture strength of the composite membrane was lower than that of the pure silk fibroin film, but the addition of amide improved its flexibility and increased its elongation at break. The results of dissolution loss and swelling degree measurement showed that the addition of amide greatly reduced the dissolution rate of the composite membrane and had a lower swelling degree. The transmittance of the composite membrane was measured by enzyme labeling instrument, and the light transmittance of the composite membrane was measured for a long time. The results showed that the amide / silk fibroin film had a long time transmittance stability. Rabbit corneal stromal cells were extracted and cultured on fibroin membrane in vitro. The compatibility of amide-fibroin composite membrane with corneal stromal cells was studied and compared with the pure fibroin membrane treated with ethanol. The adhesion rate of cells on different membranes was measured by blood cell counter, the fluorescence value of cells growing on different membranes was measured by Alma blue, and the morphology of corneal stromal cells was observed by fluorescence inverted microscope. The secretion of extracellular collagen was observed by Masson staining. The results showed that there was no significant difference between the amide / silk fibroin membrane and the pure silk fibroin membrane, which indicated that the amide / silk fibroin membrane could support the normal growth of corneal stromal cells, and the amide / silk fibroin composite membrane could be used in corneal tissue repair.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R318.08

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本文編號(hào)：2382689