葡甘聚糖一胶原蛋白一壳聚糖共混膜（I）

用溶液共混法制备了葡甘聚糖一胶原蛋白一壳聚糖（KCCS）共混膜。并用FT—IR，X—RD，SEM及透光率表征了膜的结构，同时测试了膜的抗张强度、断裂伸长率、吸水率、透水汽性、渗透性和吸附性。结果表明：共混膜中葡甘聚糖、胶原蛋白及壳聚糖之间存在着强烈的相互作用和良好的相容性，三者共混明显改善了纯聚合物和二元膜的性能。以共混膜为载体培养内皮细胞，发现共混膜具有良好的细胞相容性，预示着共混膜可作为潜在的组织工程支架材料。     

魔芋葡甘聚糖/透明质酸共混膜的体外细胞生物相容性

背景：高分子材料透明质酸与魔芋葡甘聚糖均可用于防治术后粘连。 目的：观察兔骨髓间充质干细胞与魔芋葡甘聚糖-透明质酸共混膜复合培养的生物相容性。 方法：取第3代兔骨髓间充质干细胞与魔芋葡甘聚糖-透明质酸共混膜体外复合培养,倒置显微镜和扫描电镜观察复合程度,MTT法检测细胞增殖。体外定向外诱导魔芋葡甘聚糖-透明质酸共混膜上的骨髓间充质干细胞向脂肪细胞分化,油红O染色检测其分化效果。 结果与结论：骨髓间充质干细胞与魔芋葡甘聚糖-透明质酸共混膜修复材料复合良好,倒置显微镜和扫面电镜观察均可见细胞在共混膜上良好黏附与增殖;体外定向诱导魔芋葡甘聚糖-透明质酸共混膜上的骨髓间充质干细胞可向脂肪细胞分化,具有成脂潜能。说明魔芋葡甘聚糖-透明质酸共混膜与骨髓间充质干细胞具有良好的生物相容性。     

胶原蛋白/壳聚糖复合纳米纤维膜在生物医学工程中的应用

背景：胶原蛋白-壳聚糖复合纳米纤维膜以其优异的力学性能和良好的组织细胞相容性而成为近年来科学研究的热点。 目的：总结胶原蛋白/壳聚糖复合纳米纤维膜在生物医学工程中的应用进展。 方法：以“胶原蛋白、壳聚糖、复合纳米纤维膜、胶原蛋白/壳聚糖复合纳米纤维膜、collagen/chitosan、compound Nanofiber membrane、collagen/chitosan compound nanofiber membrane、development of research” 为检索词,应用计算机检索Pubmed数据库、Elsevier数据库、万方数据库1993-01/2010-05关于胶原蛋白/壳聚糖复合纳米纤维膜研究的相关文章,对53篇文献进行分析。 结果与结论：研究表明,将胶原蛋白/壳聚糖共混,在不同条件下交联,其共混复合物在力学性能方面较单一的胶原蛋白有一定的改善,其共混膜可以作为较小软骨缺损的修复的支架材料。研究证实胶原蛋白/壳聚糖复合纳米纤维膜有着优异的力学性能、很好的组织细胞相容性和生物可降解性。文章从胶原蛋白和壳聚糖单一生物材料的缺陷性、复合纤维膜的优势及其在生物医药工程中的应用方面进行了探讨。     

壳聚糖-透明质酸共混膜对兔角膜基质细胞生长的影响

观察各种壳聚糖-透明质酸共混膜对角膜基质细胞生长的影响作用,研究透明质酸混入比例对共混膜与细胞相容性的影响.以共混膜为载体培养兔角膜基质细胞,通过光学和电子显微镜,观察细胞在膜上的生长情况;通过MTT法,检测细胞在共混膜上的贴附率和生长活性;通过检测培养基中乳酸脱氢酶的活性,预示壳聚糖-透明质酸共混膜与角膜基质细胞的相容性.以低于1:0.1的比例混入透明质酸.可以提高细胞在共混膜上的贴附率、生长速度,细胞在共混膜上的生长状态好于在壳聚糖膜上的生长状态.结果提示,以低于1:0.1的比例混入透明质酸,可以提高壳聚糖膜与角膜基质细胞的相容性,促进细胞生长;而以高于1:0.1的比例混入透明质酸,则不利于细胞在共混膜上的生长,降低了壳聚糖膜与角膜基质细胞的相容性.     

胶原蛋白-壳聚糖复合纳米纤维膜用于皮肤缺损的修复

背景：体外实验证实：胶原蛋白-壳聚糖(80∶20)复合纳米纤维膜具有优异的生物力学性能、生物相容性和生物可降解性。但目前缺乏此材料在体内方面的研究。 目的：观察胶原蛋白-壳聚糖复合纳米纤维膜修复SD大鼠背部全层皮肤缺损创面的作用。 方法：30只SD大鼠制备背部全层皮肤缺损创面后随机抽签法分为2组,实验组用胶原蛋白-壳聚糖(80∶20)复合纳米纤维膜覆盖后,用油纱及干纱布包扎并创缘外缝线打包固定;对照组仅用油纱及干纱布包扎并创缘外打包固定。 结果与结论：修复后14 d实验组创面已基本对合,干痂;苏木精-伊红染色显示创面毛细血管数量减少,纤维含量增多。对照组创面对合不整齐,创面较实验组大;苏木精-伊红染色显示毛细血管数量较多,扩张明显,大量炎性细胞浸润。提示胶原蛋白-壳聚糖复合纳米纤维膜较普通纱布敷料能更好的促进创伤修复、愈合。     

壳聚糖-明胶-透明质酸薄膜的制备及其性能的研究

本实验采用表面修饰法和共混法两种不同的方法将透明质酸引入壳聚糖-明胶体系，制备了壳聚糖-明胶-透明质酸复合薄膜，考察了薄膜的表面形貌、亲水性和力学性能等理化性能以及生物相容性。实验结果表明，壳聚糖-明胶-透明质酸薄膜具有良好的亲水性，而共混法和表面修饰法制备的薄膜在力学性能上各有所长。体外细胞培养结果表明，成纤维细胞在共混法制备的薄膜上的黏附和生长情况要明显好于表面修饰法制备的薄膜。     

壳聚糖与硫酸软骨素共混膜性质的研究

以壳聚糖和硫酸软骨素按一定比例制备出共混膜，研究了膜片的透光性、含水量、渗透性、力学性质、表面结构、生物降解性、生物相容性等性质。结果表明该共混膜具有较好的透光性、通透性、生物降解性和生物相容性，膜表面较粗糙。以此共混膜为载体培养兔角膜基质细胞，发现细胞在此共混膜上生长良好。制备膜片随着加入CaSO4量的增加，膜的通透性也随之增加。     

壳聚糖／魔芋葡甘聚糖共混膜细胞毒性和机械性能研究

目的利用天然高分子材料壳聚糖（CS）和魔芋葡甘聚糖（KGM）制备两者含量不同比例的生物膜,测定其细胞毒性和机械性能。方法流延法制备CS与KGM含量不同比例的混合膜。并将其与人脐带间充质干细胞联合培养,观察其细胞毒性：测定其断裂伸长率、拉伸强度及吸水率。结果CS/KGM含量比例分别为3：0、2：1、1：1和1：2制备的8种共混膜,具有良好的细胞毒性：膜平均厚度0．110mm,拉伸强度为35．84～48．17MPa,断裂伸长率为4．84％-6．25％,吸水率均随着CS含量增加而增加。结论CS／KGM含量比为3：0、2：1、1：1和1：2制备的共混膜,都具有良好的细胞毒性,初步具备了引导组织再生膜的基本性能。     

基于琼脂糖/壳聚糖共混凝胶模型的壳聚糖生物相容性的机理研究

目的 以琼脂糖/壳聚糖共混凝胶为模型,研究壳聚糖材料生物相容性的可能机理.方法 通过共混法,制备出一系列不同壳聚糖含量的琼脂糖/壳聚糖共混凝胶.利用傅里叶变换红外光谱(FTIR)分析共混凝胶的化学基团,利用荧光素-4-异硫氰酸酯(FITC)标记法观察琼脂糖和壳聚糖之间的可共混性.通过Zeta电势测量共混凝胶的电荷,利用二喹啉甲酸(BCA)法分别测定胎牛血清(FBS)总蛋白和牛血清白蛋白(RSA)在共混凝胶上的吸附,利用酶联免疫吸附(ELISA)法测定纤黏连蛋白(FN)在共混凝胶上的吸附.细胞实验以人微血管内皮细胞系(HMEC-1)为模型,通过观测细胞的黏附、增殖和形态来评价共混凝胶的细胞相容性.结果 琼脂糖/壳聚糖共混凝胶含有壳聚糖特征性的化学基团.琼脂糖和壳聚糖之间存在着良好的可共混性,壳聚糖的氨基基团在共混凝胶中呈均匀分布.在pH酸性条件下(pH 3.0)共混凝胶带有较强的正电荷,然而在pH中性条件下(pH 7.4)所有共混凝胶的Zeta电势均降低至0 mV附近.各组共混凝胶之间对FBS总蛋白以及BSA的吸附差异无统计学意义,但是共混凝胶对FN的吸附却随着壳聚糖含量的升高而显著升高.细胞实验结果显示:随着壳聚糖含量的提高,共混凝胶的细胞相容性有明显改善,HMECs在壳聚糖含量较高的凝胶上表现出良好的黏附、铺展和增殖.结论 相对于血清中的其他蛋白,壳聚糖组分对FN存在优先吸附,从而能够促进细胞在共混凝胶表面的黏附铺展.与传统观点不同,本研究发现壳聚糖的生物相容性与其所携带的正电荷无关.     

胶原-壳聚糖支架材料与间充质干细胞的组织相容性

背景：近年来一些研究发现胶原蛋白-壳聚糖复合支架材料可作为神经组织工程的支架材料,但相关细胞相容性研究较少。 目的：观察兔骨髓间充质干细胞在胶原蛋白-壳聚糖复合支架材料表面生长及分化情况。 方法：分离培养兔骨髓间充质干细胞,无血清培养液培养,流式细胞仪检查细胞表型;然后,将其接种到凝胶支架材料表面(实验组)及多聚赖氨酸包被的盖玻片表面(对照组),神经诱导培养基内培养,倒置相差显微镜观察干细胞的生长及分化情况。 结果与结论：细胞表型为CD29⁺、CD44⁺、CD166⁺。倒置相差显微镜观察：实验组中,接种的骨髓间充质干细胞生长良好,7 d后可见有突起神经细胞,细胞生长情况与对照组未见有明显差别。证实胶原蛋白-壳聚糖复合支架材料对骨髓间充质干细胞有良好细胞相容性。     

Study on physical properties and nerve cell affinity of composite films from chitosan and gelatin solutions

A series of chitosan-gelatin composite films was prepared by varying the ratio of constituents. FT-IR and X-ray analysis showed good compatibility between these two biopolymers. Differential scanning calorimetry (DSC) analysis indicated that the water take-up of chitosan film increased when blended with gelatin. Composite film exhibited a lower Young's modulus and a higher percentage of elongation-at-break compared with chitosan film, especially in wet state. All composite films were hydrophilic materials with water contact angles ranging from 55 degrees to 65 degrees. The results obtained from ELISA indicated the adsorption amount of fibronectin on composite films was much higher than on chitosan film. PC12 cells culture was used to evaluate the nerve cell affinity of materials. The cells cultured on the composite film with 60wt% gelatin differentiated more rapidly and extended longer neurites than on chitosan film. The results suggest that the soft and elastic complex of chitosan and gelatin, which has better nerve cell affinity compared to chitosan, is a promising candidate biomaterial for nerve regeneration.     

三种角膜内皮细胞载体膜片的性质研究

以壳聚糖分别与硫酸软骨素、羧甲基壳聚糖、透明质酸按一定比例制备出共混膜，研究了膜片的透光性、吸水率、渗透性、力学性质、表面结构、红外图谱等物理性质。以共混膜为载体培养兔角膜内皮细胞，从而筛选出了透明度高，亲水性好，机械性能优异，最适合内皮细胞贴附生长的壳聚糖．硫酸软骨素共混膜片，并对其生物降解性、生物相容性等性质进行了研究。为了提高壳聚糖．硫酸软骨素共混膜片的通透性，在膜片制备过程中加入不同剂量的醋酸盐作为扩孔剂，结果表明当加入一定量的醋酸盐为扩孔剂时，膜片的通透性提高。     

胶原-壳聚糖复合材料作为组织工程支架的研究

目的 本研究考察壳聚糖对于胶原膜理化性质的影响,选择合适比例的胶原-壳聚糖复合膜作为骨髓间充质干细胞（BMSCs）载体.方法 胶原溶涨液中添加一定比例的壳聚糖,交联并冷冻干燥制备多孔组织工程支架,研究壳聚糖对胶原膜形态学、孔隙率、机械强度、降解特性等理化性质的影响,并初步探讨了胶原-壳聚糖材料作为组织工程三维支架材料与BMSCs的相容性.结果 制备的胶原-壳聚糖支架孔径分布均匀;相对于单纯胶原海绵支架,胶原-壳聚糖复合材料支架降低体外降解速度,提高支架材料的力学性能,稳定支架的结构.BMSCs种植于胶原-壳聚糖（mCol∶mCS=9∶1）支架14 d时,SEM观察到细胞通过微绒毛与支架纤维复合,细胞相容性好.结论 胶原-壳聚糖复合支架有良好的细胞相容性,作为BMSCs诱导体外支架材料具有好的研究前景.     

Efficient transfer of human adipose-derived stem cells by chitosan/gelatin blend films

Adipose-derived stem cells (ASCs) are a potential source of abundant mesenchymal stem cells and represent a promising cell-based therapy for tissue damage or degeneration conditions. Previous investigations have demonstrated enhanced therapeutic effects of ASCs in a three-dimensional spheroid culture formulation. In this study, we hypothesize that a composite membrane made of chitosan/gelatin (C/G) is beneficial to facilitate transfer of human ASCs in spheroids. Increasing chitosan content within the blends enhanced the mechanical properties of the sample, including tensile strength and elongation-at-break ratio. Although ASC spheroids developed shortly after seeding on pure chitosan films, increasing gelatin proportion in the C/G blends promoted cell adhesion onto the membranes. We also found that ASCs did not proliferate on chitosan films, but C/G blends of different ratios supported ASC proliferation in the first 4 days of culture. However, ASCs on all C/G blends started to detach from the films to form spheroids after day 4, while ASCs on pure gelatin films remained attached and continued to grow. Gradual gelatin release from the C/G blend films, leading to enriched chitosan content in the blends, probably encouraged ASC detachment and spheroid formation. We placed porous collagen matrix on ASC-seeded C/G blends to simulate the application of ASC-seeded C/G films onto injured tissue and found that a C/G film composed of 75% chitosan could facilitate significantly more cell transfer into the overlying collagen sponge. Therefore, a blend film containing 75% chitosan and 25% gelatin showed promising results to serve as a biomaterial for human ASC-based cell therapy.     

The study of improved controlled release of vincristine sulfate from collagen-chitosan complex film

A water-in-oil-in-oil double-emulsion solvent/evaporation method was used to prepare vincristine sulfate (VCR) loaded poly(lactide-co-glycolide) microspheres, and then VCR microspheres were mixed with collagen and (or) chitosan swelling solution and lyophilized to form polymeric films. The films were cross-linked by 0.3% glutaraldehyde (GA). Encapsulation efficiency and release kinetics of VCR microspheres were determined, as well as release kinetics and in vitro degradation of the film. The rate of VCR release from the film submerged in PBS (pH 6.8) and the content were measured by high-performance liquid chromatography (HPLC). The physichemical properties of the film, such as surface morphology, mechanical function, and differential scanning calorimetry, were also measured. VCR was released from the film in a prolonged period and the initial burst release of the film was less significant. In the degradation experiment, the film containing chitosan degraded more slowly than that without chitosan. The films comprising collagen and chitosan could achieve the release kinectics of a relatively constant release. It has a promising future.     

Surface modification of poly(L-lactic acid) by entrapment of chitosan and its derivatives to promote osteoblasts-like compatibility

Surface modification of biomaterials has been adopted over the years to improve their biocompatibility. In this study, aiming to promote hydrophilicity and to introduce natural recognition sites onto poly(L-lactic acid) (PLLA) films, chitosan and its derivatives, carboxymethyl chitosan (CMC) and N-methylene phosphonic chitosan (NPC), were used to modify the surface of PLLA films by an entrapment method. The surface properties of original and modified PLLA films were measured by using water contact angle measurement and X-ray photoelectron spectroscopy (XPS). Subsequently, the cytocompatibility of these PLLA films was investigated by testing osteoblasts-like cytocompatibility, cell attachment, cell proliferation, alkaline phosphatase activity, and cell cycle. Experimental results indicated that the hydrophilicity of the modified films was improved and the surface of the modified PLLA films became enriched with chitosan and its derivatives. Moreover, the surface modification with chitosan and its derivatives significantly promoted osteoblasts-like compatibility of PLLA films. This surface modification, combining the individual advantages of PLLA with good mechanical property and chitosan as well as its derivatives with good cytocompatibility, is a promising method to prepare desirable biomaterials.     

壳聚糖改性丝素合金膜的制备及其相关性质研究

以无毒氧化葡萄糖醛作交联剂 ,采用溶液共混交联法制备壳聚糖改性丝素合金膜。用 FTIR、DSC表征其结构 ,测定其等电点、力学性能、不同 p H条件下的溶胀率和对模型药物 5 - Fu的渗透性。结果表明 :改性丝素合金膜中丝素和壳聚糖分子间存在着强烈的氢键相互作用及良好的相容性。改性膜的等电点对应的 p H值是 5 .35 ,而丝素膜的等电点是 4 .5。改性膜的力学性能优于单组分膜 ,当壳聚糖含量为 4 0 %～ 6 0 %时 ,具有最大的抗张强度和拉伸率 ,分别为 71.4～ 72 .7MPa和 2 .96～ 3.82 %。改性丝素合金膜对 5 - Fu的渗透量与壳聚糖的含量和时间成正相关关系 ,渗透系数随 p H值增大 (5→ 9)先逐渐减小然后略有增大 ,在 p H=7时最小。     

可降解性壳聚糖复合膜的研制与生物学评价

用冷冻干燥法研制了壳聚糖—羧甲基壳聚糖复合膜(以下简称复合膜)，通过体外及动物实验对复合膜进行了生物学评价。结果表明，该复合膜不仅具有良好的表面相容性，而且具有良好的结构相容性。可以满足引导组织再生(GTR)术用膜的基本要求。