壳聚糖支架在组织工程中的应用

综述了壳聚糖作为细胞生长载体在软骨组织工程，骨组织工程和皮肤组织工程等方面的应用进展，表明壳聚糖有望成为优异的组织工程支架材料。     

Preparation and characterization of macroporous chitosan/wollastonite composite scaffolds for tissue engineering

Chitosan/wollastonite composite scaffolds were prepared by a thermally induced phase separation method. The microstructure, mechanical performance and in vitro bioactivity of the composite scaffolds were investigated. The composite scaffolds were macroporous and wollastonite particles were dispersed uniformly on the surface of the pore walls. Scanning electron microscope images of the composite scaffolds demonstrated that the scaffolds had interconnected pores with diameters from 60 to 200 microm. Both the pore size and structure were affected by freezing temperature. The mechanical performance of the composite scaffolds was improved compared to that of pure chitosan scaffolds. The in vitro bioactivity of the scaffolds was evaluated by soaking samples in simulated body fluid and the apatite layer was observed on the surface of the pore walls of the composite scaffolds. Our results suggest that the incorporation of wollastonite into chitosan could enhance both the mechanical strength and the in vitro bioactivity of the resultant composite. The macroporous chitosan/wollastonite scaffolds may be a potential candidate for application in tissue engineering.     

Effects of different cross-linking conditions on the properties of genipin-cross-linked chitosan/collagen scaffolds for cartilage tissue engineering

A cross-linking reagent is required to improve mechanical strength and degradation properties of biopolymers for tissue engineering. To find the optimal preparative method, we prepared diverse genipin-cross-linked chitosan/collagen scaffolds using different genipin concentrations and various cross-linking temperatures and cross-linking times. The compressive strength increased with the increasing of genipin concentration from 0.1 to 1.0%, but when concentration exceeded 1.0%, the compressive strength decreased. Similarly, the compressive strength increased with the increasing of temperature from 4 to 20°C, but when temperature reached 37°C, the compressive strength decreased. Showing a different trend from the above two factors, the effect of cross-linking time on the compressive strength had a single increasing tendency. The other results also demonstrated that the pore size, degradation rate and swelling ratio changed significantly with different cross-linking conditions. Based on our study, 1.0% genipin concentration, 20°C cross-linking temperature and longer cross-linking time are recommended.     

Fabrication and characterization of PCL/gelatin/chitosan ternary nanofibrous composite scaffold for tissue engineering applications

In the present study, we have fabricated a ternary composite nanofibrous scaffold from PCL/gelatin/chitosan, by electrospinning technique, using a solvent system—chloroform/methanol for polycaprolactone (PCL) and acetic acid for gelatin and chitosan, for tissue engineering applications. Field emission scanning electron microscopy (FE-SEM) was used to investigate the fiber morphology of the scaffold and it was found that the fiber morphology was influenced by the concentrations of PCL, gelatin, and chitosan in polymer solution during electrospinning. X-ray diffraction, Fourier transform infrared, and thermogravimetric (TG) analysis results showed some interactions among the molecules of PCL, gelatin, and chitosan within the scaffold. In-vitro cell culture studies were done by seeding L929 mouse fibroblasts on fabricated composite scaffold, which confirmed the cell viability, high cell proliferation rate, and cell adhesion on composite scaffold as indicated by MTT assay, DNA quantification, and FE-SEM analysis of cell-scaffold construct. Thus, the ternary composite scaffold made from the combination of PCL (synthetic polymer), gelatin, and chitosan (natural polymer) may find potential application in tissue engineering.     

Preparation and characterization of collagen/PLA,chitosan/PLA,and collagen/chitosan/PLA hybrid scaffolds for cartilage tissue engineering

In this study, three-dimensional (3D) porous scaffolds were developed for the repair of articular cartilage defects. Novel collagen/polylactide (PLA), chitosan/PLA, and collagen/chitosan/PLA hybrid scaffolds were fabricated by combining freeze-dried natural components and synthetic PLA mesh, where the 3D PLA mesh gives mechanical strength, and the natural polymers, collagen and/or chitosan, mimic the natural cartilage tissue environment of chondrocytes. In total, eight scaffold types were studied: four hybrid structures containing collagen and/or chitosan with PLA, and four parallel plain scaffolds with only collagen and/or chitosan. The potential of these types of scaffolds for cartilage tissue engineering applications were determined by the analysis of the microstructure, water uptake, mechanical strength, and the viability and attachment of adult bovine chondrocytes to the scaffolds. The manufacturing method used was found to be applicable for the manufacturing of hybrid scaffolds with highly porous 3D structures. All the hybrid scaffolds showed a highly porous structure with open pores throughout the scaffold. Collagen was found to bind water inside the structure in all collagen-containing scaffolds better than the chitosan-containing scaffolds, and the plain collagen scaffolds had the highest water absorption. The stiffness of the scaffold was improved by the hybrid structure compared to plain scaffolds. The cell viability and attachment was good in all scaffolds, however, the collagen hybrid scaffolds showed the best penetration of cells into the scaffold. Our results show that from the studied scaffolds the collagen/PLA hybrids are the most promising scaffolds from this group for cartilage tissue engineering.     

Development of polycaprolactone/chitosan blend porous scaffolds

Polycaprolactone (PCL) and chitosan were blended to fabricate porous scaffolds for tissue-engineering applications by employing a concentrated acetic acid solution as solvent and salt particles as porogen. These scaffolds showed well-controlled and interconnected porous structures. The pore size and porosity of the scaffolds could be effectively modulated by selecting appropriate amounts and sizes of porogen. The results obtained from compressive mechanical measurements indicated that PCL/chitosan could basically retain their strength in their dry state compared to individual components. In a hydrated state, their compressive stress and modulus could be still well maintained even though the weight ratio of chitosan reached around 50 wt%.     

不同条件对SA/GT/CS三元复合水凝胶微球溶胀性能的影响

将海藻酸钠/明胶共混液滴入舍有CaCl2的壳聚糖醋酸溶液中,制备出SA/CS/GT三元复合水凝胶微球.改变sA/GT的质量配比、盐离子浓度、温度,考察不同条件下对复合微球溶胀度的影响.结果表明:当SA : GT质量比为10:0.6、10:0.8,CaCl2浓度为3%～4 %,所成微球的溶胀性能较好;随温度的升高,溶胀度不断增大.     

Fabrication and characterization of gold nanoparticle-doped electrospun PCL/chitosan nanofibrous scaffolds for nerve tissue engineering

In the field of nerve tissue engineering, nanofibrous scaffolds could be a promising candidate when they are incorporated with electrical cues. Unique physico-chemical properties of gold nanoparticles (AuNPs) make them an appropriate component for increasing the conductivity of scaffolds to enhance the electrical signal transfer between neural cells. The aim of this study was fabrication of AuNPs-doped nanofibrous scaffolds for peripheral nerve tissue engineering. Polycaprolactone (PCL)/chitosan mixtures with different concentrations of chitosan (0.5, 1 and 1.5) were electrospun to obtain nanofibrous scaffolds. AuNPs were synthesized by the reduction of HAuCl₄ using chitosan as a reducing/stabilizing agent. A uniform distribution of AuNPs with spherical shape was achieved throughout the PCL/chitosan matrix. The UV–Vis spectrum revealed that the amount of gold ions absorbed by nanofibrous scaffolds is in direct relationship with their chitosan content. Evaluation of electrical property showed that inclusion of AuNPs significantly enhanced the conductivity of scaffolds. Finally, after 5 days of culture, biological response of Schwann cells on the AuNPs-doped scaffolds was superior to that on as-prepared scaffolds in terms of improved cell attachment and higher proliferation. It can be concluded that the prepared AuNPs-doped scaffolds can be used to promote peripheral nerve regeneration.

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Preparation and cytocompatibility of silk fibroin / chitosan scaffolds

One challenge in soft tissue engineering is to find an applicable scaffold, not only having suitable mechanical properties, porous structures, and biodegradable properties, but also being abundant in active groups and having good biocompatibility. In this study, a three-dimensional silk fibroin/chitosan (SFCS) scaffold was successfully prepared with interconnected porous structure, excellent hydrophilicity, and proper mechanical properties. Compared with polylactic glycolic acid (PLGA) scaffold, the SFCS scaffold further facilitated the growth of HepG2 cells (human hepatoma cell line). Keeping the good cytocompatibility and combining the advantages of both fibroin and chitosan, the SFCS scaffold should be a prominent candidate for soft tissue engineering, for example, liver.     

Preparation and cytocompatibility of silk fibroin/chitosan scaffolds

One challenge in soft tissue engineering is to find an applicable scaffold, not only having suitable mechanical properties, porous structures, and biodegradable properties, but also being abundant in active groups and having good biocompatibility. In this study, a three-dimensional silk fibroin/chitosan (SFCS) scaffold was successfully prepared with interconnected porous structure, excellent hydrophilicity, and proper mechanical properties. Compared with polylactic glycolic acid (PLGA) scaffold, the SFCS scaffold further facilitated the growth of HepG2 cells (human hepatoma cell line). Keeping the good cytocompatibility and combining the advantages of both fibroin and chitosan, the SFCS scaffold should be a prominent candidate for soft tissue engineering, for example, liver.     

Fabrication and characterization of chitosan microspheres agglomerated scaffolds for bone tissue engineering

In the presented paper authors describe a method for bone scaffolds fabrication. The technique is based on the agglomeration of chitosan microspheres. The fabrication process is complex and consists of a few steps: chitosan spheres extrusion, scaffold formation by compression followed by the spheres agglomeration and bonding with cross-linking agent (STPP, sodium tripolyphosphate). The described method allows manufacturing of porous materials with controllable shape, pore size distribution and their interconnectivity. In this technique 3D scaffold porosity can be regulated by altering spheres diameter. Authors studied influence of cross-linker concentrations and time of cross-linking process on the scaffold morphology, mechanical properties, enzymatic degradation rate (in the presence of lysozyme) and human osteoblasts response. Surface morphology and topography were evaluated by SEM. Porosity and pore interconnectivity were observed via μCT scanning. Mechanical tests showed that chitosan scaffolds perform compression characteristic (Young Modulus) similar to natural bone. Cytotoxicity established by XTT assay confirmed that most of the developed composite materials do not show toxic properties. Osteoblast adhesion and morphology were analyzed by SEM and optical microscopy.     

Comparative study on the role of gelatin,chitosan and their combination as tissue engineered scaffolds on healing and regeneration of critical sized bone defects: an in vivo study

Gelatin and chitosan are natural polymers that have extensively been used in tissue engineering applications. The present study aimed to evaluate the effectiveness of chitosan and gelatin or combination of the two biopolymers (chitosan–gelatin) as bone scaffold on bone regeneration process in an experimentally induced critical sized radial bone defect model in rats. Fifty radial bone defects were bilaterally created in 25 Wistar rats. The defects were randomly filled with chitosan, gelatin and chitosan–gelatin and autograft or left empty without any treatment (n?=?10 in each group). The animals were examined by radiology and clinical evaluation before euthanasia. After 8?weeks, the rats were euthanized and their harvested healing bone samples were evaluated by radiology, CT-scan, biomechanical testing, gross pathology, histopathology, histomorphometry and scanning electron microscopy. Gelatin was biocompatible and biodegradable in vivo and showed superior biodegradation and biocompatibility when compared with chitosan and chitosan–gelatin scaffolds. Implantation of both the gelatin and chitosan–gelatin scaffolds in bone defects significantly increased new bone formation and mechanical properties compared with the untreated defects (P?<?0.05). Combination of the gelatin and chitosan considerably increased structural and functional properties of the healing bones when compared to chitosan scaffold (P?<?0.05). However, no significant differences were observed between the gelatin and gelatin–chitosan groups in these regards (P?>?0.05). In conclusion, application of the gelatin alone or its combination with chitosan had beneficial effects on bone regeneration and could be considered as good options for bone tissue engineering strategies. However, chitosan alone was not able to promote considerable new bone formation in the experimentally induced critical-size radial bone defects.     

改性天然胶乳/壳聚糖/明胶三元复合材料制备研究

采用正交试验方法,考察了原料的配比、交联剂的用量及交联温度等对复合材料性能的影响。较佳的制备工艺条件分别为改性天然胶乳/壳聚糖/明胶配比60/20/20,交联剂用量0.004%,交联温度60℃。随改性天然胶乳的加入,复合膜柔韧性增强,力学性能下降;壳聚糖对复合膜的力学性能贡献大于明胶。     

Fabrication of chitosan microparticles loaded in chitosan and poly(vinyl alcohol) scaffolds for tissue engineering application

In recent decades, the use of microparticle-mediated drug delivery is widely applied in the field of biomedical application. Here, we report the new dressing material with ciprofloxacin-loaded chitosan microparticle (CMP) impregnated in chitosan (CH) and poly(vinyl alcohol) (PVA) scaffold for effective delivery of drug in a sustained manner to the wound site. Moreover, the peculiar physiochemical and structural properties of the CH–CMP scaffold has proved better tensile strength and excellent swelling to achieve 82% of drug release. In vitro biocompatibility was done for both scaffold using NIH 3T3 fibroblasts and human keratinocytes (HaCaT) cell lines. In vitro fluorescent activity showed distinct biocompatibility with good cell adhesion and proliferation. However, the CH–CMP scaffold showed best result to act as promising biomaterial in effective drug delivery in tissue engineering.     

甲醇、戊二醛交联剂对丝素蛋白/明胶多孔支架的性能影响

为了对比甲醇、戊二醛两种交联剂对丝素蛋白/明胶复合多孔支架的性能影响,采用冷冻干燥法等比例制备该支架,分两组分别用甲醇和戊二醛进行交联。通过观察支架的微观形貌,测量支架的孔隙率、吸水率、溶胀率,测试热稳定性及力学性能,比较经两种交联剂处理后支架结构和性能的变化。结果表明,经戊二醛交联后的支架孔隙分布更加规则、均匀,孔隙率、吸水率、溶胀率更高,力学性能更强。采用戊二醛交联丝素蛋白/明胶复合多孔支架,能够使支架性能更加优良。     

原位水化法制备羟基磷灰石/壳聚糖复合支架材料

以含Ca2+和PO34-的溶液为无机相,壳聚糖(chitosan,CS)溶液为高分子相,采用原位水化法制备羟基磷灰石(hydroxyapatite,HAP)/CS复合多孔支架材料。XRD和IR的表征和分析表明水化24h后,复合支架中的钙磷盐从磷酸氢钙(dicalciumphos phate dehydrate,DCPD)转化为HAP。SEM和EDS显示15μm左右的棒状HAP颗粒均匀地分散在多孔支架的孔壁上,压缩强度的测试结果表明这种结构显著提高复合支架的力学性能。     

含纳米银的明胶/壳聚糖纳米纤维的制备及其抗菌性能研究

以浓度为88%的甲酸溶液作为纺丝溶剂,采用静电纺丝和紫外光照射还原的方法制备了含纳米银颗粒的明胶/壳聚糖纳米纤维。研究发现,壳聚糖的加入量低于明胶质量的3/16时可以得到纳米纤维,纤维平均直径随着硝酸银加入量的增大而减小,纤维表面纳米银的平均直径随着硝酸银加入量的增大而增大,在纺丝体系中硝酸银的加入量存在一个极限值。所制得含纳米银的明胶/壳聚糖纳米纤维对金黄色葡萄球菌和绿脓杆菌具有较好的抑菌性能,纺丝时加入1%硝酸银制得纳米纤维膜的抑菌率达到99%以上,这种抗菌型纳米纤维可以应用于医用敷料等领域。     

聚乙烯醇/壳聚糖/明胶水凝胶的合成及其对苯酚的吸附研究

以聚乙烯醇(PVA)、壳聚糖和明胶为原料、戊二醛为交联剂,在酸性溶液中通过共混交联反应合成了聚乙烯醇/壳聚糖/明胶水凝胶,单因素法讨论了影响水凝胶性能的因素如:反应物质量比、交联剂用量、反应温度、反应时间等聚乙烯醇/壳聚糖/明胶水凝胶,对不同浓度苯酚溶液的吸附研究.研究结果表明聚乙烯醇/壳聚糖/明胶水凝胶对苯酚有较好的吸附性能.