不同分子量壳聚糖膜性质的研究

分别以分子量为130，000、220，000、300，000、550，000道尔顿的壳聚糖制备壳聚糖膜，并研究了各膜的表面结构、结晶性、力学特性、渗透性、透光透气性、吸附性、生物降解性等。结晶表明壳聚糖膜的各种特性和壳聚糖的分子量相关，高分子量的壳聚糖膜表面较为光滑，透光性较好，透气性、渗透性和生物降解性较差；低分子量的壳聚糖膜表面较为粗糙，透气性、渗透性和生物降解性较好，但透光性较差。经分析认为膜的结晶性和超微结构决定了不同分子量壳聚糖膜具有不同的性质。     

植入式葡萄糖传感器膜材料壳聚糖与Nafion的组织相容性比较

背景：壳聚糖是天然高分子多糖,可单独或者与其他材料复合制作敷料、药物、基因载体、生物涂层、组织工程支架、传感器膜材料等。 目的：了解壳聚糖作为植入式葡萄糖传感器膜材料的组织相容性,并与Nafion膜进行对比。 方法：制备壳聚糖膜并对其理化性质进行表征,比较壳聚糖膜皮下植入与肌肉植入、Nafion膜肌肉植入的生物相容性。 结果与结论：壳聚糖膜的厚度、溶胀率、表观密度等理化参数可以通过浓度、铸膜液体积来控制;壳聚糖膜能生物降解,63 d皮下植入的降解率为(17.0±9.9)%,说明壳聚糖的体内降解速度较慢。壳聚糖膜皮下植入引起的炎症反应较肌肉植入重,63 d后形成的纤维包膜比肌肉植入要厚(P < 0.05);肌肉植入Nafion与壳聚糖膜引起材料周围纤维包膜厚度差异无显著性意义(P > 0.05),两者均在15 d以后趋于稳定。证明壳聚糖膜能生物降解,与Nafion膜均有较好的组织相容性。     

壳聚糖膜降解动物实验研究

本研究首次采用新兴天然生物材料壳聚糖做膜材料 ,研制了可降解吸收性壳聚糖膜 ,并通过动物体内植入实验研究其生物降解性 ,经组织学光镜观察考察其组织相容性。结果表明 ,壳聚糖膜是一种性能优良的生物降解性材料 ,具有较好的组织相容性 ,可用于壳聚糖植入膜的研制     

壳聚糖膜的降解性研究

用不同性能地壳聚糖为膜材料制备壳聚糖膜，通过体外酸解，酶解试验及动物体内植入试验研究其降解。试验结果表明，壳聚糖是一种可生物降解性膜材料，壳聚糖膜的降解性与其脱乙酰度，介质酸性强弱及溶菌酶等因素有关。壳聚糖膜在动物体内降解比较缓慢，２０天约降解８．２％。     

生物降解性防术后粘连膜的实验研究

本文首次采用新型医用天然高分子材料壳聚糖作膜材料，制备了可降解吸收防术后粘连膜，并通过动物实验研究其生物降解性和生物相容性．初步研究结果表明，壳聚糖具有很好的成膜性。壳聚糖膜在小鼠体内可以缓慢降解，并具有较好的生物相容性，是一种很有发展前景的天然防术后粘连膜材料。     

壳聚糖—明胶混合膜的制备及其生物降解性研究

将壳聚糖与明胶按一定比例混合制膜，通过体外降解及动物体内实验研究了其降解性和生物相容性，结果表明壳聚糖－明胶混合膜在小鼠体现人降解速度较快，并具有较好的生物相容性，溶菌酶对混合膜的生物降解有促进作用。     

壳聚糖/PLGA乳化膜预防鸡趾屈肌腱粘连的实验研究

目的研究壳聚糖／PLGA乳化膜预防鸡趾鞘管区屈肌腱粘连的效果。方法雌性成年种禽45只，随机分成3组．每组15只。A组为对照组，B组为PLGA膜组，C组为壳聚糖／PLGA乳化膜组。将左足三、四趾趾浅屈肌腱切除，横断趾深屈肌腱，作改良Kessler法缝合．B、C两组分别局部包裹PLGA膜及壳聚糖／PLGA乳化膜。术后2、4、6周取材，分别行大体观察、组织学检查和生物力学测定。结果组织学检查显示：3组肌腱愈合进程无明显差异，B组局部白细胞浸润较其他两组明显。两实验组缝合处粘连评分及将肌腱牵出鞘管所需最大力量与对照组相比差异有显著统计学意义（P〈0．05）。结论局部应用PLGA膜及壳聚糖／PLGA乳化膜均能减轻肌腱术后粘连，前者局部炎症反应较明显，而后者无明显炎症反应发生．因而壳聚糖／PLGA乳化膜是一种较理想的预防肌腱粘连的可降解材料。     

生物降解性防术后粘加膜的实验研究

本文首先采用新型医用天然高分子材料壳聚糖作膜材料，制备了可降解吸收的防术后粘连膜，并通过动物实验研究其生物降解性和生物相容性。初步研究结果表明，壳聚糖具有很好的成膜性。壳莫大 半导体环境可以缓慢降解，并具有较好的生物相容性，是一种很有发展前景的天然防术后宫膜材料。     

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。     

改性壳聚糖防粘连膜修复坐骨神经损伤

背景:研究表明壳聚糖与动物及人体具有较好的生物相容性、可降解性,可支持许旺细胞在壳聚糖膜上生长,而且能够明显抑制成纤维细胞生长。目的:观察改性壳聚糖防粘连膜对大鼠坐骨神经再生修复的影响。方法:切断60只SD大鼠双侧坐骨神经,缝合外膜,随机在一侧坐骨神经缝合处包裹改性壳聚糖防粘连膜,以另一侧为对照。术后20,30,40d进行电生理及组织学检测,观察改性壳聚糖防粘连膜对大鼠坐骨神经损伤修复的影响。结果与结论:早期改性壳聚糖防粘连膜治疗侧神经断端的炎性反应较对照侧明显,随着膜的自行降解,炎症反应逐渐减轻,神经缝合口纤维组织增生减少。与对照侧相比,大鼠坐骨神经损伤30d后,改性壳聚糖防粘连膜治疗侧神经传导速度恢复快,比目鱼肌记录到的神经-肌肉电潜伏期缩短(P0.05)。30d后各时点与对照侧比较,改性壳聚糖防粘连膜治疗侧坐骨神经再生轴索密度大于对照侧(P0.05)。说明改性壳聚糖防粘连膜虽早期会加重周围神经损伤的炎性反应,但随着膜的降解,炎症反应逐渐减轻,可减轻神经缝合口纤维组织增生,防止粘连,因而有利于神经传导速度恢复及轴索生长。     

载盐酸四环素壳聚糖缓释膜的制备

背景：壳聚糖具有良好的生物相容性、生物可降解性及较好的抗菌活性。 目的：使用流延法制备载有不同盐酸四环素的壳聚糖载药纳米纤维膜,观察其缓释性能和抑菌性能。 方法：采用流延法制备厚度为0.03 mm的载有不同含量(0,3%,5%,10%,20%)盐酸四环素的壳聚糖载药缓释膜,测定载药率,绘制盐酸四环素缓释曲线。分别用液体培养和固体培养检测载药缓释膜的体外抑菌性能,用磷酸盐缓冲液观察载药缓释膜的降解性能。 结果与结论：随盐酸四环素含量的增加,缓释膜载药率降低,突释量增大。载药壳聚糖膜可有效抑制金黄色葡萄球菌的生长,并随盐酸四环素含量的增加,抑菌效果提高,当盐酸四环素含量超过10%时,载药壳聚糖膜抑菌率的变化不明显。盐酸四环素的加入加快了壳聚糖膜降解,并随着盐酸四环素含量的增加,降解速率增大,当盐酸四环素载药量超过10%时,降解可在8 d内完成。相比较得出,盐酸四环素含量在10%时,在疗效和性价比上是较好的选择。     

Fabrication,Characterization of Chitosan/Nanosilver Film and Its Potential Antibacterial Application

In this work chitosan/silver nanoparticle films were synthesized by a simple photochemical method of reduction of silver ions in an acidic solution of AgNO₃ and chitosan. This is a novel, cheap, easy, quick and in situ approach to prepare chitosan films loaded with silver nanoparticles (AgNPs). Chitosan used here is a natural polymer and acts as a very good chelating and stabilizing agent; thus, this approach of formation of chitosan/silver nanoparticle films is proved to be an excellent 'green approach' for the synthesis of metal nanoparticle composites. The presence of silver nanoparticles was confirmed from the transmission electron microscopy (TEM), X-ray diffraction (XRD) and thermogravimetric analysis (TGA) of the film. The surface plasmon resonance (SPR) obtained at 400 nm also confirmed the presence of nanosilver in the chitosan film. The developed chitosan–nanosilver films demonstrated excellent antibacterial action against model bacteria, Escherichia coli and Bacillus. This approach can be easily used in the large-scale production of such silver-nanoparticles-loaded chitosan films. These films can be used as antimicrobial packaging materials, as wound dressings and can also be grafted onto various implants.     

壳聚糖/聚己内酯共混膜的制备及性能

背景：壳聚糖/聚己内酯共混材料在生物材料领域具有广泛的应用前景,但与蛋白、细胞反应机制尚不明确。 目的：观察壳聚糖/聚己内酯共混膜表面蛋白黏附和细胞活性。 方法：将不同配比的壳聚糖/聚己内酯混合溶液旋转涂膜法成膜。分别通过原子力显微镜、滴形分析仪、石英晶体天平和MTT比色法测量膜的表面形貌、亲疏水性、蛋白吸附和细胞增殖活性。 结果与结论：膜的表面形貌、亲疏水性、蛋白吸附和细胞增殖活性在很大程度上取决于壳聚糖和聚己内酯的质量配比。细胞在壳聚糖膜上具有较好的伸展形态,在聚己内酯膜上具有较高的增殖活性。     

Effects of the Surface Characteristics of Nano-Crystalline and Micro-Particle Calcium Phosphate/Chitosan Composite Films on the Behavior of Human Mesenchymal Stem Cells In Vitro

Calcium phosphate (CaP) compounds, the main inorganic constituent of mammalian bone tissues, are believed to support bone precursor cell growth and osteogenic differentiation. Chitosan, a deacetylated derivative of chitin, is a versatile biopolymer to offer broad possibilities for cell-based tissue engineering. In the present study, different scales of CaP crystals on chitosan membranes were prepared for culture of human mesenchymal stem cells (hMSCs) in vitro. A series of aqueous CaP suspensions with different concentrations were mixed with chitosan solution and chitosan/calcium phosphate (C/CaP) films were fabricated by the solvent-casting method. With different weight ratios of CaP in chitosan solution, the various surface characteristics of nano-amorphous (C/CaP 0.1), nano-crystalline (C/CaP 0.5) and micro-particle (C/CaP 2) CaP compounds were examined by scanning electron microscopy and electron dispersion spectroscopy. X-ray diffraction on micro-particles of CaP indicated the formation of crystalline hydroxyapatite. The behavior of hMSCs, including proliferation, cell spreading and osteogenic differentiation, was studied on the C/CaP films. In basal culture medium, the incorporation of CaP into chitosan films could promote the proliferation of hMSCs. The C/CaP 0.5 film with connected CaP nano-crystals had better cellular viability. The fluorescence microscope images at 14 days of culture revealed extensive networks of F-actin filaments of hMSCs on chitosan, C/CaP 0.1 and C/CaP 0.5 films. The cellular morphology on C/CaP 2 film with discrete CaP micro-particles was partly restrained. In osteogenic medium, the alkaline phosphatase (ALP) activity of hMSCs increased and showed the process of osteogenic differentiation. The ALP levels on C/CaP 2 film were higher than those on C/CaP 0.1 and C/CaP 0.5 films. These results demonstrated that the crystallinity and topography of CaP on chitosan membranes could modulate the behaviors of cultured hMSCs in vitro.     

Surface modification of poly (D,L-lactic acid) with chitosan and its effects on the culture of osteoblasts in vitro

Chitosan is a good biodegradable natural polymer, widely used in biomedical fields. In this study, chitosan was used to modify the surface of poly (D,L-lactic acid) (PDLLA) in order to enhance its cell affinity. The properties of a modified PDLLA surface and control were investigated by contact angle and electron spectroscopy for chemical analysis (ESCA), which indicated the changes in surface energy and chemical structure. Scanning electron microscopy (SEM) observation displayed differences in surface morphology between the chitosan-modified film and the control. These data reflected that PDLLA films could be modified with chitosan and in turn may affect the biocompatibility of the modified films. Therefore, adhesion and growth of osteoblasts on modified PDLLA films as well as control were studied. Cell morphologies on the films were examined by SEM and cell viability was evaluated using an MTT assay; the differentiated cell function was assessed by measuring alkaline phosphatase (ALP) activity. The ALP activity of modified PDLLA films was significantly higher than that found on the control (p < 0.01). The proliferation of osteoblasts on modified films was also found to be higher than that on the control (p < 0.05), suggesting that chitosan could be used to modify PDLLA and then enhance its cell biocompatibility.     

Development of bFGF-Chitosan Matrices and Their Interactions with Human Dermal Fibroblast Cells

Chitosan (CH) is a naturally derived, biodegradable polymer of glucosamine with a variable frequency of N-acetyl-D-glucosamine units, and has been demonstrated to have numerous pharmacological and wound-healing properties. Biodegradable chitosan films were fabricated using a solvent casting technique and investigated for skin tissue-engineering applications. Basic fibroblast growth factor (bFGF) was incorporated into the CH matrices (1 μg/film) by 3 methods: adsorption, entrapment and covalent binding. Release rates and biological activity of the incorporated bFGF were monitored. Human dermal fibroblasts (HDF cells) were used as an in vitro model for cell response to CH and bFGF-CH films. Cell attachment, growth and acid-soluble collagen quantification were employed as an assessment of cell function. The fibroblasts were found to remain viable on the chitosan films and scaffolds. CH films without bFGF were compatible with HDF cells; however, the fibroblasts did not proliferate. The release profile of adsorbed and bound bFGF from CH films were similar (indicating that binding was not efficient) while entrapped bFGF was not released in the time frame studied. The concentration of bFGF released to the cell culture medium was not high enough to stimulate HDF proliferation. However, cell attachment was significantly increased in chitosan films with bFGF adsorbed onto the surface as compared to control surfaces. HDF cells grown on CH films produced significantly more collagen than those on control surfaces.     

Chitosan film enriched with an antioxidant agent, taurine, in fenestration defects

A natural polysaccharide, chitosan (poly-N-acetyl glucosaminoglycan), which is a nontoxic and bioabsorbable polymer, has been shown to have hemostatic and antibacterial effects. An amino acid, taurine, is considered to be beneficial for regulating the inflammation process. The purpose of this study was to investigate the synergistic effects of taurine and chitosan in the experimental defects at the vestibular bone of maxillary canine teeth in six dogs. Chitosan films were prepared as delivery system with or without taurine and placed in the randomly chosen defects. Biopsies were performed on the postoperative seventh day and routine histological procedures were performed for light and electron microscopic evaluations. For each group, 30 different microscopic areas were examined and the numbers of macrophages and neutrophils in these areas were counted. The mean numbers of both macrophages and neutrophils were found statistically different between the chitosan film incorporated with taurine and free chitosan groups (p < 0.0001 p > 0.05). In addition to the increase in cell counts in both groups, the cytological alterations were more obvious in the chitosan film group incorporated with taurine. Accordingly, taurine appears to enhance the acceleration effect of chitosan on wound healing at early periods. This effect could be considered beneficial in tissue repair in destructive diseases like periodontitis.     

Porous calcium phosphate coating over phosphorylated chitosan film by a biomimetic method

A porous calcium phosphate coating deposited on chitosan films was studied using scanning electron microscopy, energy-dispersive X-ray analysis, micro-Fourier transform infrared spectroscopy (micro-FTIR) and thin-film X-ray diffractometry (XRD). Chitosan films were first prepared by dissolving chitosan powder in dilute acetic acid and drying in a flat petri dish. The films were phosphorylated using urea and H3PO4 with the P content being 0.1-0.2 wt%. Phosphorylated films soaked in saturated Ca(OH)2 solution for 8 days led to the formation of a calcium phosphate precursor phase over the entire surface. This precursor phase stimulated the growth of a porous coating of calcium-deficient hydroxy apatite when immersed in 1.5 x SBF for more than 20 days. Phosphorylated films not treated with Ca(OH)2 did not show any calcium phosphate growth upon immersion in SBF solution. The precursor phase is thought to be octacalcium phosphate, which nucleates a HAP phase during SBF treatment. Initially, this treatment in SBF results in the formation of a single-layer calcium phosphate particles over the film surface. As immersion time in SBF increases, further nucleation and growth produce a porous HAP coating. The Ca/P ratio of the HAP coating is a function of SBF immersion time.     

Effects of the degree of deacetylation on the physicochemical properties and Schwann cell affinity of chitosan films

Chitosan is a potential material for the preparation of nerve repair conduits. In order to find a better chitosan for the application in peripheral nerve regeneration, the effects of the degree of deacetylation (DD) on the physicochemical properties and Schwann cell affinity of chitosan films have been evaluated. Six kinds of chitosan samples with similar molecular weight, but various DD in a range from 70.1 to 95.6% were prepared from one stock chitosan material and fabricated into films. X-ray diffraction analysis showed that there were more crystalline regions in the higher DD chitosan films. Swelling and mechanical property measurements revealed that the swelling index of chitosan films decreased and their elastic modulus and tensile strength increased with the increase in DD. The adsorption amount of fibronectin and laminin on chitosan films was measured by means of enzyme-linked immunosorbent assay (ELISA). Culture of adult rat Schwann cells on the films showed that the chitosan films with higher DD provided better substrata for Schwann cell spreading and proliferation. In conclusion, DD of chitosan plays an important role in their physicochemical properties and affinity with Schwann cells. The results suggest that chitosan with a DD higher than 90% is considered as a promising material for application in peripheral nerve regeneration.