基于細(xì)菌纖維素的皮膚組織修復(fù)材料的制備和評價
發(fā)布時間：2018-10-26 15:21
【摘要】：細(xì)菌纖維素(bacterial cellulose, BC)是一種具有獨(dú)特的納米結(jié)構(gòu)和優(yōu)異綜合性能的高分子材料,如優(yōu)良的生物相容性,有望成為促進(jìn)組織再生與修復(fù)的重要生物材料之一。但是,BC的產(chǎn)量和成本是制約BC的應(yīng)用和產(chǎn)業(yè)化發(fā)展的關(guān)鍵。BC用于皮膚損傷修復(fù)的安全性和有效性則是決定其臨床應(yīng)用的重要因素。本文研制了系列納米細(xì)菌纖維素及其復(fù)合膜材料,表征了其結(jié)構(gòu)和性能,通過紅外光譜、掃描電鏡、比表面積分析、接觸角分析、X-射線衍射、差示量熱掃描和紫外可見分光光度計等儀器和設(shè)備表征了基于細(xì)菌纖維素的皮膚組織修復(fù)材料的結(jié)構(gòu)和性能。采用成纖維細(xì)胞和干細(xì)胞對其進(jìn)行體外細(xì)胞評價,建立小鼠全層皮膚損傷模型,對細(xì)菌纖維素和其它材料進(jìn)行動物實驗評價。 使用木醋桿菌菌株Acetobacter xylinum Y05 (CCTCC M207163)和葡糖醋桿菌菌株Gluconacetobacter xylinus (ATCC 53582)生物合成了細(xì)菌纖維素。采用多孔培養(yǎng)板一次培養(yǎng)多個BC膜適用于細(xì)胞大規(guī)模培養(yǎng),培養(yǎng)板的形狀和大小可調(diào)控菌體行為和細(xì)菌纖維素的合成。在BC的合成中,培養(yǎng)環(huán)境的空間是決定BC的孔徑分布和比表面積的重要因素之一。建立了動靜結(jié)合的多層發(fā)酵法,制備品質(zhì)均一的多層細(xì)菌纖維素(multilayer BC)。該方法快速、簡便、可控、均一性和重現(xiàn)性好,有利于推動BC材料的工業(yè)化發(fā)展。 通過在培養(yǎng)基中原位加入海藻酸鈉和具有抗菌活性的殼聚糖,制備了細(xì)菌纖維素/殼聚糖/海藻酸鈉三元復(fù)合膜,改善了細(xì)菌纖維素膜的抗菌性能。在BC的原位復(fù)合修飾過程中,殼聚糖和海藻酸鈉均可在BC的分泌和合成中被其網(wǎng)絡(luò)原位包裹進(jìn)去。隨著培養(yǎng)基中殼聚糖和海藻酸鈉濃度的增加,BC/殼聚糖/海藻酸鈉原位復(fù)合膜的產(chǎn)量隨著濃度的升高總體呈下降的趨勢。殼聚糖的結(jié)晶度在培養(yǎng)基中的殼聚糖濃度為2%的時候最高,之后隨著濃度的增加反而下降。添加了海藻酸鈉的培養(yǎng)基中,BC/海藻酸鈉原位復(fù)合膜的結(jié)晶度在培養(yǎng)基含有0.05%濃度時最高。 多層BC和普通BC一樣,都是納米網(wǎng)絡(luò)結(jié)構(gòu)。細(xì)胞評價證明了BC膜不僅能夠促進(jìn)普通細(xì)胞如成纖維細(xì)胞NIH/3T3的增殖和粘附,也有利于干細(xì)胞如人脂肪干細(xì)胞(hASCs)的生長。采用BALB/c小鼠全層皮膚缺損模型,比較了多層BC和醫(yī)用創(chuàng)傷敷料、異種皮膚及同種異體皮膚的創(chuàng)傷修復(fù)效果。動物評價表明使用細(xì)菌纖維素可以有效保護(hù)創(chuàng)面,防止感染,有利于皮膚組織再生和傷口愈合,優(yōu)于異種和同種異體皮膚移植以及臨床常用的傷口敷料的修復(fù)效果。檢測了血清中的細(xì)胞生長因子TGF-β1和bFGF的表達(dá)水平,初步探討了細(xì)菌纖維素修復(fù)皮膚損傷的分子機(jī)制。 基于BC的皮膚組織修復(fù)材料,其合成可控,可以大批量高效率的生產(chǎn),且重現(xiàn)性好,材料形狀可塑,含水量高,透氣性好,具備良好的力學(xué)性能以及適用于皮膚組織修復(fù)材料的機(jī)械強(qiáng)度。分子水平、細(xì)胞水平和個體水平的評價結(jié)果表明其生物相容性良好,包括其細(xì)胞相容性和組織相容性,具有很高的應(yīng)用價值。
[Abstract]:Bacterial cellulose (BC) is a kind of high molecular material with unique nano-structure and excellent comprehensive properties, such as good biocompatibility, which is expected to be one of the important biological materials to promote tissue regeneration and repair. However, BC's output and cost are the key to the application of BC and the development of industrialization. The safety and effectiveness of BC for skin injury repair is an important factor to determine its clinical application. In this paper, a series of nano-bacterial cellulose and its composite membrane materials are developed, their structure and properties are characterized, the infrared spectrum, scanning electron microscope, specific surface area analysis, contact angle analysis, X-ray diffraction, The apparatus and apparatus, such as differential thermal scan and ultraviolet visible spectrophotometer, characterize the structure and performance of the bacterial cellulose-based skin tissue repair material. In vitro cell evaluation of fibroblasts and stem cells was carried out to establish a model of whole-layer skin injury in mice, and animal experiments were performed on bacterial cellulose and other materials. Bacterial fibers were synthesized using Acinetobacter xylinum Y05 (CCTCC M207163) and Gluconobacter xylinus (ATCC 53582). The method adopts a porous culture plate to culture a plurality of BC films at one time, is suitable for large-scale culture of cells, and the shape and the size of the culture plate can regulate the cell behavior and the combination of bacterial cellulose In the synthesis of BC, the space of culture environment is an important factor to determine the pore size distribution and specific surface area of BC. firstly, a multi-layer fermentation method combining static and static combining is established, and multi-layer bacterial cellulose (multi-layer BC) with uniform quality is prepared. The method is rapid, simple, controllable, homogeneous and reproducible, and is favorable for promoting the industrialized production of the BC material. The bacterial cellulose/ chitosan/ sodium alginate ternary composite membrane is prepared by adding sodium alginate and chitosan with antibacterial activity in situ in the culture medium, so that the resistance of the bacterial cellulose membrane is improved. In situ compound modification of BC, chitosan and sodium alginate can be packaged in situ in the secretion and synthesis of BC. As the concentration of chitosan and sodium alginate in the medium increased, the yield of BC/ chitosan/ sodium alginate in-situ composite membrane decreased with increasing concentration. The chitosan has the highest crystallinity in the medium at 2%, followed by the increase of the concentration. In the culture medium supplemented with sodium alginate, the crystallinity of BC/ sodium alginate in-situ composite membrane contained 0.05% concentration in the medium. It's the highest. Multilayer BC is the same as normal BC, all of them. The cell evaluation demonstrated that BC films not only promote proliferation and adhesion of normal cells, such as fibroblasts NIH/ 3T3, but also facilitate stem cells such as human adipose-derived stem cells (hASC). S) growth. A BALB/ c mouse whole-layer skin defect model was used to compare the creation of multi-layer BC and medical wound dressing, xenogeneic skin and allogeneic skin. The animal evaluation indicates that the use of bacterial cellulose can effectively protect the wound, prevent infection, facilitate skin tissue regeneration and wound healing, is superior to xenogeneic and allogenic skin transplantation and clinical commonly used wound dressing. The effects of bacterial cellulose on skin injury were studied. The skin tissue repairing material based on BC is controllable in synthesis, can be produced in large batch and high efficiency, has good reproducibility, is plastic in shape, high in water content, good in air permeability, good in mechanical property and suitable for skin tissue repair. The mechanical strength, molecular level, cell level and individual level of the material showed good biocompatibility, including their cell compatibility and histocompatibility.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2012
【分類號】：R318.08
本文編號：2296152
[Abstract]:Bacterial cellulose (BC) is a kind of high molecular material with unique nano-structure and excellent comprehensive properties, such as good biocompatibility, which is expected to be one of the important biological materials to promote tissue regeneration and repair. However, BC's output and cost are the key to the application of BC and the development of industrialization. The safety and effectiveness of BC for skin injury repair is an important factor to determine its clinical application. In this paper, a series of nano-bacterial cellulose and its composite membrane materials are developed, their structure and properties are characterized, the infrared spectrum, scanning electron microscope, specific surface area analysis, contact angle analysis, X-ray diffraction, The apparatus and apparatus, such as differential thermal scan and ultraviolet visible spectrophotometer, characterize the structure and performance of the bacterial cellulose-based skin tissue repair material. In vitro cell evaluation of fibroblasts and stem cells was carried out to establish a model of whole-layer skin injury in mice, and animal experiments were performed on bacterial cellulose and other materials. Bacterial fibers were synthesized using Acinetobacter xylinum Y05 (CCTCC M207163) and Gluconobacter xylinus (ATCC 53582). The method adopts a porous culture plate to culture a plurality of BC films at one time, is suitable for large-scale culture of cells, and the shape and the size of the culture plate can regulate the cell behavior and the combination of bacterial cellulose In the synthesis of BC, the space of culture environment is an important factor to determine the pore size distribution and specific surface area of BC. firstly, a multi-layer fermentation method combining static and static combining is established, and multi-layer bacterial cellulose (multi-layer BC) with uniform quality is prepared. The method is rapid, simple, controllable, homogeneous and reproducible, and is favorable for promoting the industrialized production of the BC material. The bacterial cellulose/ chitosan/ sodium alginate ternary composite membrane is prepared by adding sodium alginate and chitosan with antibacterial activity in situ in the culture medium, so that the resistance of the bacterial cellulose membrane is improved. In situ compound modification of BC, chitosan and sodium alginate can be packaged in situ in the secretion and synthesis of BC. As the concentration of chitosan and sodium alginate in the medium increased, the yield of BC/ chitosan/ sodium alginate in-situ composite membrane decreased with increasing concentration. The chitosan has the highest crystallinity in the medium at 2%, followed by the increase of the concentration. In the culture medium supplemented with sodium alginate, the crystallinity of BC/ sodium alginate in-situ composite membrane contained 0.05% concentration in the medium. It's the highest. Multilayer BC is the same as normal BC, all of them. The cell evaluation demonstrated that BC films not only promote proliferation and adhesion of normal cells, such as fibroblasts NIH/ 3T3, but also facilitate stem cells such as human adipose-derived stem cells (hASC). S) growth. A BALB/ c mouse whole-layer skin defect model was used to compare the creation of multi-layer BC and medical wound dressing, xenogeneic skin and allogeneic skin. The animal evaluation indicates that the use of bacterial cellulose can effectively protect the wound, prevent infection, facilitate skin tissue regeneration and wound healing, is superior to xenogeneic and allogenic skin transplantation and clinical commonly used wound dressing. The effects of bacterial cellulose on skin injury were studied. The skin tissue repairing material based on BC is controllable in synthesis, can be produced in large batch and high efficiency, has good reproducibility, is plastic in shape, high in water content, good in air permeability, good in mechanical property and suitable for skin tissue repair. The mechanical strength, molecular level, cell level and individual level of the material showed good biocompatibility, including their cell compatibility and histocompatibility.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2012
【分類號】：R318.08
【引證文獻(xiàn)】
相關(guān)期刊論文 前1條
1 汪麗粉;李政;賈士儒;張健飛;;細(xì)菌纖維素性質(zhì)及應(yīng)用的研究進(jìn)展[J];微生物學(xué)通報;2014年08期
，本文編號：2296152
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