基于细菌纤维素的皮肤组织修复材料的制备和评价
发布时间：2018-10-26 15:21
【摘要】：细菌纤维素(bacterial cellulose, BC)是一种具有独特的纳米结构和优异综合性能的高分子材料,如优良的生物相容性,有望成为促进组织再生与修复的重要生物材料之一。但是,BC的产量和成本是制约BC的应用和产业化发展的关键。BC用于皮肤损伤修复的安全性和有效性则是决定其临床应用的重要因素。本文研制了系列纳米细菌纤维素及其复合膜材料,表征了其结构和性能,通过红外光谱、扫描电镜、比表面积分析、接触角分析、X-射线衍射、差示量热扫描和紫外可见分光光度计等仪器和设备表征了基于细菌纤维素的皮肤组织修复材料的结构和性能。采用成纤维细胞和干细胞对其进行体外细胞评价,建立小鼠全层皮肤损伤模型,对细菌纤维素和其它材料进行动物实验评价。 使用木醋杆菌菌株Acetobacter xylinum Y05 (CCTCC M207163)和葡糖醋杆菌菌株Gluconacetobacter xylinus (ATCC 53582)生物合成了细菌纤维素。采用多孔培养板一次培养多个BC膜适用于细胞大规模培养,培养板的形状和大小可调控菌体行为和细菌纤维素的合成。在BC的合成中,培养环境的空间是决定BC的孔径分布和比表面积的重要因素之一。建立了动静结合的多层发酵法,制备品质均一的多层细菌纤维素(multilayer BC)。该方法快速、简便、可控、均一性和重现性好,有利于推动BC材料的工业化发展。 通过在培养基中原位加入海藻酸钠和具有抗菌活性的壳聚糖,制备了细菌纤维素/壳聚糖/海藻酸钠三元复合膜,改善了细菌纤维素膜的抗菌性能。在BC的原位复合修饰过程中,壳聚糖和海藻酸钠均可在BC的分泌和合成中被其网络原位包裹进去。随着培养基中壳聚糖和海藻酸钠浓度的增加,BC/壳聚糖/海藻酸钠原位复合膜的产量随着浓度的升高总体呈下降的趋势。壳聚糖的结晶度在培养基中的壳聚糖浓度为2%的时候最高,之后随着浓度的增加反而下降。添加了海藻酸钠的培养基中,BC/海藻酸钠原位复合膜的结晶度在培养基含有0.05%浓度时最高。 多层BC和普通BC一样,都是纳米网络结构。细胞评价证明了BC膜不仅能够促进普通细胞如成纤维细胞NIH/3T3的增殖和粘附,也有利于干细胞如人脂肪干细胞(hASCs)的生长。采用BALB/c小鼠全层皮肤缺损模型,比较了多层BC和医用创伤敷料、异种皮肤及同种异体皮肤的创伤修复效果。动物评价表明使用细菌纤维素可以有效保护创面,防止感染,有利于皮肤组织再生和伤口愈合,优于异种和同种异体皮肤移植以及临床常用的伤口敷料的修复效果。检测了血清中的细胞生长因子TGF-β1和bFGF的表达水平,初步探讨了细菌纤维素修复皮肤损伤的分子机制。 基于BC的皮肤组织修复材料,其合成可控,可以大批量高效率的生产,且重现性好,材料形状可塑,含水量高,透气性好,具备良好的力学性能以及适用于皮肤组织修复材料的机械强度。分子水平、细胞水平和个体水平的评价结果表明其生物相容性良好,包括其细胞相容性和组织相容性,具有很高的应用价值。
[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.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R318.08
本文编号：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.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R318.08
【引证文献】
相关期刊论文 前1条
1 汪丽粉;李政;贾士儒;张健飞;;细菌纤维素性质及应用的研究进展[J];微生物学通报;2014年08期
，本文编号：2296152
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