含絲蛋白肝臟組織工程支架材料的制備及體外評(píng)價(jià)
發(fā)布時(shí)間：2018-10-14 14:31
【摘要】：各類肝病引發(fā)的肝功能衰竭已嚴(yán)重影響人類健康。雖然肝臟組織工程技術(shù)是當(dāng)今最具潛力的治療手段，但是尋找理想的支架材料一直是該領(lǐng)域的技術(shù)難題。家蠶絲膠蛋白(silk siricin, SS)是一種寶貴的天然高分子材料，是人類利用很早的動(dòng)物蛋白質(zhì)之一，在絲綢工業(yè)中，產(chǎn)生的大量絲膠常作為廢物隨廢水直接排放，對(duì)其開(kāi)發(fā)利用可節(jié)約資源、保護(hù)環(huán)境。作為天然蛋白，絲膠具備特異性分子識(shí)別信號(hào)，可促進(jìn)細(xì)胞粘附和增殖，誘導(dǎo)細(xì)胞分化及為其提供支架�；诮z膠良好的凝膠化性能、低免疫原性，，特殊的生物相容性和可降解性等，逐漸在組織工程支架等醫(yī)學(xué)領(lǐng)域得到了廣泛的重視和研究。但是絲膠在低溫冷凍時(shí)為凝膠，加熱到室溫左右即成為溶膠，調(diào)控較為困難，對(duì)其改性調(diào)制必不可少。當(dāng)前報(bào)道較多的殼聚糖/甘油磷酸鈉(CS/GP)溫敏性復(fù)合凝膠材料，具有低溫溶膠，體溫范圍內(nèi)呈凝膠狀態(tài)的特點(diǎn)，可作為對(duì)絲膠的改性物質(zhì)之一。同時(shí)，玉米淀粉(Maize Starth, MS)是一類可完全生物降解的多糖類天然高分子，來(lái)源廣泛、價(jià)格低廉；聚乙烯醇(PolyvinylAlcohol, PVA)具有優(yōu)良成膜性、力學(xué)性能，以及良好的親水性、生物相容性和降解性，二者均可作為安全醫(yī)用生物材料用于對(duì)絲膠性質(zhì)的完善。 本課題分為兩個(gè)大部分，先后嘗試制備了溶膠狀態(tài)經(jīng)注射后可在體內(nèi)成型和直接在體外成型兩類可調(diào)控型肝臟組織工程支架材料，即SS/CS/GP溫敏性復(fù)合凝膠和SS/PVA/MS凝膠膜，從結(jié)構(gòu)、性能及應(yīng)用方面，分析了二者用于肝臟組織工程支架的可行性。首先進(jìn)行了對(duì)SS/CS/GP溫敏性復(fù)合凝膠的研究，即將廢棄蠶繭中提取的絲膠蛋白(SS)，外加高脫乙酰度的殼聚糖(CS)、α、β-甘油磷酸鈉(GP)，三者經(jīng)冰浴下的特定方式混合，得到不同配比SS/CS/GP復(fù)合物，該復(fù)合物可在20℃以下長(zhǎng)時(shí)間保持液態(tài)，在體溫環(huán)境(37±5℃)附近、大范圍生理pH值(6.9-7.45)下，快速凝膠化；對(duì)其結(jié)構(gòu)性能研究顯示SS/CS/GP凝膠具備貫通的三維多孔結(jié)構(gòu)、適當(dāng)?shù)牧W(xué)強(qiáng)度、良好的生物降解性且降解速率可調(diào)控，無(wú)細(xì)胞毒性且具備一定的促BRL大鼠肝細(xì)胞生長(zhǎng)性能，并可通過(guò)注射實(shí)現(xiàn)體內(nèi)成型。然后進(jìn)行了SS/PVA/MS凝膠膜材料的研究，即將廢棄蠶繭中提取的SS，與MS和PVA經(jīng)水相共混和反復(fù)凍融可實(shí)現(xiàn)體外不同配比的SS/PVA/MS凝膠膜材料成型。研究其力學(xué)性能和降解機(jī)制表明，SS/PVA質(zhì)量比為3/7，MS添加量為18%時(shí)，凝膠膜整體力學(xué)性能最優(yōu)，不同配比SS/PVA/MS凝膠膜均具備降解性，且降解速率可通過(guò)SS/PVA比例及MS含量得到調(diào)控，該凝膠膜材料通過(guò)體外成型且具備可調(diào)控的降解性能，有望作為體外肝臟組織工程支架使用。
[Abstract]:Liver failure caused by various liver diseases has seriously affected human health. Although liver tissue engineering technology is the most potential treatment, finding ideal scaffold materials has always been a technical problem in this field. Silk gelatin (silk siricin, SS) is a valuable natural polymer material, which is one of the early animal proteins used by human beings. In the silk industry, a large amount of sericin is often discharged directly with the waste water as waste water. It can save resources and protect environment. As a natural protein sericin has a specific molecular recognition signal which can promote cell adhesion and proliferation induce cell differentiation and provide scaffolds. Sericin with good gelation property, low immunogenicity, special biocompatibility and biodegradability has been paid more and more attention in the field of tissue engineering scaffolds. However, sericin is gel at low temperature and becomes sol when heated to room temperature, which is difficult to control and is necessary for modification and modulation of sericin. At present, more chitosan / sodium glycerophosphate (CS/GP) thermo-sensitive composite gel materials are reported, which have the characteristics of low temperature sol and gel state in the temperature range, and can be used as one of the modified materials for sericin. At the same time, corn starch (Maize Starth, MS) is a kind of completely biodegradable polysaccharides, which has a wide range of sources and low price. (PolyvinylAlcohol, PVA) has good film formation, mechanical properties and good hydrophilicity. Biocompatibility and biodegradability can be used as safe biomaterials for the improvement of sericin properties. This thesis is divided into two parts. Two kinds of controllable scaffold materials for liver tissue engineering, SS/CS/GP thermo-sensitive composite gel and SS/PVA/MS gel membrane, have been prepared after injection of sol-state in vivo and directly in vitro. The feasibility of the two scaffolds for liver tissue engineering was analyzed in terms of performance and application. Firstly, the SS/CS/GP thermo-sensitive composite gel was studied. The sericin protein (SS), extracted from the abandoned cocoon and chitosan (CS), 偽, 尾 -glycerophosphate sodium (GP), with high deacetylation degree were mixed in a special way under ice bath to obtain the SS/CS/GP complex with different proportions. The complex can be rapidly gelatinized under 20 鈩
本文編號(hào)：2270758
[Abstract]:Liver failure caused by various liver diseases has seriously affected human health. Although liver tissue engineering technology is the most potential treatment, finding ideal scaffold materials has always been a technical problem in this field. Silk gelatin (silk siricin, SS) is a valuable natural polymer material, which is one of the early animal proteins used by human beings. In the silk industry, a large amount of sericin is often discharged directly with the waste water as waste water. It can save resources and protect environment. As a natural protein sericin has a specific molecular recognition signal which can promote cell adhesion and proliferation induce cell differentiation and provide scaffolds. Sericin with good gelation property, low immunogenicity, special biocompatibility and biodegradability has been paid more and more attention in the field of tissue engineering scaffolds. However, sericin is gel at low temperature and becomes sol when heated to room temperature, which is difficult to control and is necessary for modification and modulation of sericin. At present, more chitosan / sodium glycerophosphate (CS/GP) thermo-sensitive composite gel materials are reported, which have the characteristics of low temperature sol and gel state in the temperature range, and can be used as one of the modified materials for sericin. At the same time, corn starch (Maize Starth, MS) is a kind of completely biodegradable polysaccharides, which has a wide range of sources and low price. (PolyvinylAlcohol, PVA) has good film formation, mechanical properties and good hydrophilicity. Biocompatibility and biodegradability can be used as safe biomaterials for the improvement of sericin properties. This thesis is divided into two parts. Two kinds of controllable scaffold materials for liver tissue engineering, SS/CS/GP thermo-sensitive composite gel and SS/PVA/MS gel membrane, have been prepared after injection of sol-state in vivo and directly in vitro. The feasibility of the two scaffolds for liver tissue engineering was analyzed in terms of performance and application. Firstly, the SS/CS/GP thermo-sensitive composite gel was studied. The sericin protein (SS), extracted from the abandoned cocoon and chitosan (CS), 偽, 尾 -glycerophosphate sodium (GP), with high deacetylation degree were mixed in a special way under ice bath to obtain the SS/CS/GP complex with different proportions. The complex can be rapidly gelatinized under 20 鈩
本文編號(hào)：2270758
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