A study on the fabrication of porous chitosan/gelatin network scaffold for tissue engineering

A novel porous chitosan/gelatin scaffold for tissue engineering was prepared via polyelectrolyte complex formation, freeze drying and post‐crosslinking with glutaraldehyde. The porosity and mean pore diameters could be controlled within 30∼100 µm by varying the original water content and the freezing conditions. Dipping the scaffolds in poly(lactic acid) provided good mechanical properties making it a promising candidate towards tissue engineering. © 2000 Society of Chemical Industry     

Preparation and characterization of chitosan/PEG/gelatin composites for tissue engineering

Chitosan scaffolds have gained much attention in tissue engineering. However, brittleness and low biodegradability limit scaffolds application, especially in use as guided tissue regeneration membranes (GTRm) in surgical operations. The first objective of this work is to improve the brittleness of the chitosan membrane, which is not desired for use via adding polyethylene glycol (PEG) to chitosan, and the second objective is to accelerate the degradation rate by blending gelatin with the binary chitosan‐PEG mixture. The addition of PEG softened the blend membrane in vision and in touch. The tensile compliant increased from 7.87 × 10^?3 (MPa^?1) for chitosan membrane to 3.63 × 10^?1 (MPa^?1) for chitosan‐PEG‐gelatin (CPG) membrane. Degradation results in vitro indicated that CPG membrane degraded faster and weight loss increased more significantly than chitosan membrane and the lowest tensile strength of CPG membrane could meet the requirement of the application. CPG membrane showed significant improvement in degradation and flexibility in comparison with the chitosan membrane. Cell adhesion, viability, and proliferation onto the external surface of CPG membrane with C2C12 cell had been evaluated in vitro and quantified by a methyl thiazolyl tetrazolium (MTT) reduction assay. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Preparation and characterization of novel chitosan/nanodiopside/nanohydroxyapatite composite scaffolds for tissue engineering applications

Chitosan/nanodiopside/nanohydroxyapatite (CS/nDP/nHAp) composite scaffolds were prepared from the mixture of chitosan, nDP, and nHAp in different inorganic/organic weight ratios by using the freeze-drying method. The prepared nHAp and composite scaffolds were investigated using BET, TG, FT-IR, SEM, EDS, and XRD techniques. The composite scaffolds had 50–85% porosities with interlinked porous networks. Moreover, investigation of the cell proliferation, adhesion, and viability using MTT test, and mouse preosteoblast cell proved the cytocompatibile nature of the composite scaffolds with improved cell attachment and proliferation. All these results essentially illustrated that this composite could be a potential for bone tissue engineering applications.     

Characterization of chitosan/citrate and chitosan/acetate films and applications for wound healing

In this work, we aimed to develop a scaffold of chitosan (CS) with a porous sponge structure for an artificial skin. The scaffolds were prepared from both CS/citric and CS/acetic solutions. In addition, the cast films were also prepared from the same solutions to compare some of their properties. They were characterized using WAXD, FTIR, DSC, tensile measurements, and SEM observation. It was found that CS/acetate had low crystallinity but CS/citrate was in an amorphous state, resulting in a large ductility with rubbery softness. Despite the different morphologies of CS/citrate and CS/acetate scaffolds, both scaffolds exhibited the wound healing effect available for tissue engineering. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Preparation and characterization of novel chitosan/zeolite scaffolds for bone tissue engineering applications

In this study, chitosan-based novel scaffolds containing zeolite A were fabricated by freeze-drying technique. The nanocomposite scaffolds were prepared from chitosan and zeolite A nanocrystals with different amounts (0.5, 1.0, and 2.0%) in aqueous media. The zeolite A nanocrystals and nanocomposite scaffolds were characterized by using FTIR, X-ray powder diffraction, scanning electron microscope, and thermogravimetric analysis. The scaffolds were seeded with bone marrow-derived human mesenchymal stem cell line (UE7T-13), and cell attachment, viability, and cytotoxicity assays were performed. In vitro cytotoxicity of scaffolds toward human mesenchymal stem cell line was evaluated through the evaluation of cell viability and cell attachment assays.     

Biomimetic porous collagen/hydroxyapatite scaffold for bone tissue engineering

Bone defect and osteochondral injury frequently occur due to diseases or traumatism and bring a crucial challenge in orthopedics. The hybrid scaffold has shown promise as a potential strategy for the treatment of such defects. In this study, a novel biomimetic porous collagen (Col)/hydroxyapatite (HA) scaffold was fabricated through assembling layers of Col containing gradual amount of HA under the assistance of “iterative layering” freeze‐drying process. The scaffold presents a double gradient of highly interconnected porosity and HA content from top to bottom, mimicking the inherent physiological structure of bone. Owing to the biomimetic structure and component, significant increase of cell proliferation, alkaline phosphatase activity, and osteogenic differentiation in vitro was observed, illustrating potential application of the excellent Col/HA scaffold as a promising strategy for bone tissue engineering. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 45271.     

纳米纤维应用于组织工程的研究进展

组织工程支架的关键作用是起到引导细胞繁殖、生长,促进组织修复的一个过程。纳米纤维支架由于具有特殊的纳米效应,而更有利于细胞的黏附、增殖、功能化,因而被广泛应用于组织工程。本文分别介绍了纳米纤维支架在各类组织工程中应用,包括皮肤和创伤敷料组织工程、血管组织工程、神经组织工程、骨组织工程、软骨组织工程中应用的研究进展,同时介绍了纳米纤维支架药物控制释放中的应用。指出目前纳米技术还不成熟,需要从制备工艺的优化、基因工程的引入及纳米材料安全性能的科学评价等几方面解决纳米纤维支架在组织修复工程中面临的问题。     

Preparation and properties of high performance gelatin-based hydrogels with chitosan or hydroxyethyl cellulose for tissue engineering applications

High performance gelatin-based biocompatible hybrid hydrogels are developed using functionalized polyethylene glycol as a cross-linker in presence of chitosan or hydroxyethyl cellulose. Tensile test shows robust and tunable mechanical properties and reveals non-linear and J-shaped stress-strain curves similar to those found for native extracellular matrix. Degradation study demonstrates that the mass loss and change in mechanical properties are dependent on hydrogel composition and cross-linking density. Structural features of the hydrogels are confirmed by infrared spectroscopy. A preliminary biological evaluation is carried out using rat myoblasts and human fibroblasts cell lines. The results show that all hydrogels allow cell adhesion and proliferation during four days culture, hence, they might have a great potential for use in the biomedical applications.     

Thin natural gelatin/chitosan nanofibrous scaffolds for retinal pigment epithelium cells

Damage of the retinal pigmented epithelial cells causes various diseases such as age-related macular degeneration in retinal tissue. Nowadays, scientists are attempting to replace lost retinal cells with healthy and efficient cells that provide better conditions for recovering and preventing blindness. In this study, gelatin/chitosan nanofibrous scaffolds with mean diameters of 180?nm were fabricated for subretinal space through electrospinning. Thickness and morphology of the gelatin–chitosan scaffolds were analyzed by scanning electron microscopy (SEM). The results showed that the high rate of degradation, i.e., 90% damage was obtained after 1 month. The cell viability of gelatin/chitosan nanofibers were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The SEM results of cultured RPE on gelatin/chitosan scaffolds showed the appropriate adhesion of cells on the substrate. The results of the identity of RPE cells cultured on the scaffolds indicated that a large number of cells strongly expressed RPE65 and cytokeratin 8/18.     

组织工程支架材料及制备方法研究现状

组织工程支架已经广泛应用于皮肤、软骨、心血管、心脏等各种组织的修复中.组织工程材料分为天然材料和合成材料两大类,均需要具备良好的生物相容性.组织工程支架的制备方法主要有相分离、冷冻干燥、发泡、颗粒浸出、静电纺丝、3D打印等.现对组织工程支架所用到的材料、制备方法以及组织工程支架的应用进行综述.     

Preparation and characterization of hydroxyapatite/chitosan–gelatin network composite

A novel composite composed of hydroxyapatite (HA) and a network formed via cocrosslinking of chitosan and gelatin with glutaraldehyde was developed. Two preparation methods are described in detail. A porous material, with similar organic–inorganic constituents to that of natural bone, was made by a unique sol–gel method. The formation of the network in the presence of HA was characterized using IR analysis. The morphology of the composites was also examined using SEM. In addition, XRD was applied to estimate the change in the component crystal. The results indicate that the presence of HA does not retard the formation of the chitosan/gelatin network. On the other hand, the polymer matrix has hardly any influence on the high crystallinity of HA. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 77: 2929–2938, 2000     

Preparation and characterization of novel sodium alginate/chitosan two ply composite membranes

Sodium alginate/chitosan (SA/CS) two ply composite membranes were prepared by casting and solvent evaporation technique. NaHCO₃ was used as a porogen additive to form pores in the interior of the composite membranes and glycerol was introduced as a plasticizer. The water uptake capacity, mechanical strength, oxygen permeation property, and in vitro cytotoxicity were evaluated to test the feasibility to utilize the composite membranes for wound dressing. The average pore size, water uptake capacity, and oxygen permeation property of the composite membranes could be adjusted by the ratio of NaHCO₃ in the SA solution. The SA/CS two ply composite membranes showed high water uptake capacity, suitable mechanical strength, excellent oxygen permeability, and good biocompatible. It indicates that the SA/CS two ply composite membranes are suitable for wound dressing application. It provides a simple but promising platform to fabricate wound dressing using natural polymers. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 2007     

Hierarchical bioceramic scaffold for tissue engineering: A review

Bioceramic scaffolds have a promising application in bone-tissue engineering field. However, bioceramic scaffolds exhibit low fracture toughness; hence, to overcome this problem, hierarchical bioceramic scaffold or bioceramic scaffolds coated with polymer are produced. Starting with the fundamental requirements for bioceramic scaffold, this article provides detailed information on recent developments of method to produce porous bioceramics scaffold and hierarchical bioceramic scaffold. Chemical modifications to enhance interfacial adhesion and formation of interpenetrating network structures between the bioceramic scaffold and the natural polymer layer are discussed in this article. Areas of future research are highlighted at the end of this review.     

Nanofibrous scaffold prepared by electrospinning of poly(vinyl alcohol)/gelatin aqueous solutions

A series of nanofibrous scaffolds were prepared by electrospinning of poly(vinyl alcohol) (PVA)/gelatin aqueous solution. PVA and gelatin was dissolved in pure water and blended in full range, then being electrospun to prepared nanofibers, followed by being crosslinked with glutaraldehyde vapor and heat treatment to form nanofibrous scaffold. Field emission scanning electron microscope (FESEM) images of the nanofibers manifested that the fiber average diameters decreased from 290 to 90 nm with the increasing of gelatin. In vitro degradation rates of the nanofibers were also correlated with the composition and physical properties of electrospinning solutions. Cytocompatibility of the scaffolds was evaluated by cells morphology and MTT assay. The FESEM images revealed that NIH 3T3 fibroblasts spread and elongated actively on the scaffolds with spindle‐like and star‐type shape. The results of cell attachment and proliferation on the nanofibrous scaffolds suggested that the cytotoxicity of all samples are grade 1 or grade 0, indicating that the material had sound biosafety as biomaterials. Compared with pure PVA and gelatin scaffolds, the hybrid ones possess improved biocompatibility and controllability. These results indicate that the PVA/gelatin nanofibrous have potential as skin scaffolds or wound dressing. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Preparation of biomimetic three-dimensional gelatin/montmorillonite–chitosan scaffold for tissue engineering

A novel gelatin/montmorillonite–chitosan (Gel/MMT–CS) nanocomposite scaffold was prepared via the intercalation process and the freeze-drying technique, using the ice particulates as the porogen materials. Properties including pore structure, water adsorption content, in vitro degradation and tensile strength were investigated. It was demonstrated that the introduced intercalation structure endowed the Gel/MMT–CS scaffold with good mechanical properties and a controllable degradation rate. Scanning of the electron microscope images revealed that the scaffold obtained was highly porous and suitable for the implanted cells to adhere and grow. The mitochondrial activity assay provided good evidences of cells viability on the Gel/MMT–CS membranes, giving an indication of possible application as a matrix for tissue engineering.     

Properties of polyelectrolyte complex films of chitosan and gelatin

A series of chitosan–gelatin complexes was prepared by varying the ratio of constituents. Differential scanning calorimetry was used to determine the amount of the different states of water. The interaction between chitosan and gelatin was checked by IR and X-ray analysis and was related to mechanical strength. The results indicate that the water take-up of a chitosan–gelatin complex is depressed by strong interactions within networks. Chitosan can improve the tensile strength of complex films, and even with high water content these can keep appropriate tensile strength and higher elongation. © 1999 Society of Chemical Industry     

Synthesis,characterization and in-vitro behavior of natural chitosan-hydroxyapatite-diopside nanocomposite scaffold for bone tissue engineering

Composite materials based on a combination of biodegradable polymers and bioactive ceramics, including chitosan and hydroxyapatite are discussed as suitable materials for scaffold fabrication. Diopside is a member of bioactive silicates; it is a good choice for hard tissue engineering because of its biocompatibility with host tissue and high mechanical strength. Chitosan and hydroxyapatite were extracted from shrimp shell and bovine bone, respectively and diopside nanoparticles were prepared by the sol-gel method. The present study reports on a chitosan composite which was reinforced by hydroxyapatite and diopside; the scaffolds were fabricated by the freeze-drying method. The so-produced chitosan-hydroxyapatite-diopside (CS-HA-DP) scaffolds were further cross-linked using tripolyphosphate (TPP) to achieve enhanced mechanical strength. The ratios of the ceramic components in composites were 5-58-37, 10-55-35, and 15-52-33 (diopside-hydroxyapatite-chitosan, w/w %). The physicochemical properties of scaffolds were investigated using Fourier-transform infrared spectrometry (FT-IR), X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS) techniques. The effect of scaffolds composition on bioactivity and biodegradability were studied well. To investigate mechanical properties of samples, compression test was done. Results showed that the composite scaffold with 5% DP has the highest mechanical strength. The porosity of composites dropped from 92% to 76% by increasing the amount of DP. Cytocompatibility of the scaffolds was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) activity, and cell attachment studies using human osteoblast cells. Results demonstrated no sign of toxicity and cells were found to be attached to the pore walls within the scaffolds; moreover, results illustrated that the developed composite scaffolds could be a potential candidate for tissue engineering.     

Preparation and characterization of a novel thermosensitive hydrogel based on chitosan and gelatin blends

In this study, a novel injectable in situ gelling thermosensitive hydrogel system based on chitosan and gelatin blends was designed and investigated. The addition of gelatin provides the correct buffering and other physicochemical conditions including control of hydrophobic interactions and hydrogen bonding, which are necessary to retain chitosan in solution at neutral pH near 4°C and furthermore to allow gel formation upon heating to body temperature. The chitosan/gelatin hydrogels were studied by FTIR, swelling, and rheological analysis. The rheological analysis evidenced the endothermic gelation of chitosan/gelatin solutions, which indicated their gelation temperatures and reflected the effect of gelatin concentration on the thermosensitive properties of gels. The morphology of this system was examined with laser scanning confocal microscopy and scanning electron microscopy. The images indicated that the gels were quite heterogeneous and porous. The investigation of these gels as vehicles for delivering bovine serum albumin as a model drug of protein showed that the system could sustain the release of the protein drug. These results show that chitosan/gelatin solutions can form gels rapidly at body temperature and have promising perspective for their use in local and sustained delivery of protein drug. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

聚己内酯/壳聚糖核壳结构纤维引导组织再生膜的制备及表征

高性能的引导组织再生膜是牙周引导组织再生术成功的关键,静电纺丝法因可仿生制备类细胞外基质结构,在引导组织再生膜研制方面显示出巨大潜力。本研究通过同轴静电纺丝法,以聚己内酯（PCL）为核层,壳聚糖（CS）为壳层,制备核壳结构的纳米纤维,并用香草醛对制备的纤维膜进行交联。利用扫描电子显微镜（SEM）、透射电子显微镜（TEM）、X 射线衍射（XRD）、傅里叶变换红外光谱（FTIR）、力学测试及细胞培养等手段对制备的纤维膜进行形貌、内部结构、化学组成、力学性能和细胞相容性表征。结构分析表明本研究成功制备了核壳结构的PCL-CS纤维膜。力学测试和亲疏水性测试结果表明交联后的纤维膜具有较好的耐水性和力学性能,断裂强度高出文献报道值近两倍;体外细胞培养结果显示MG-63细胞能在交联后的纤维膜上黏附和持续增殖,表明纤维膜具有较好的细胞相容性,在引导组织再生领域有较好的应用前景。     

Evaluation of PCL/chitosan electrospun nanofibers for liver tissue engineering

In this investigation, a nanofibrous scaffold was fabricated through electrospinning of polycaprolactone (PCL) and chitosan (CS) using a novel collector to make better orientation and pore size for cell infiltration. PCL/CS nanofibers with 90-rpm collector speed and 40° angle between collector wires of the new collector have fewer diameters with better pore, size and nanofibers orientation. Mechanical properties, roughness parameters, topology, structure, hydrophilicity, and cell growing were considered for liver tissue engineering. The cell culture was done using epithelial liver mouse cells. The developed electrospun PCL/CS scaffold using novel collector would be an excellent matrix for biomedical applications especially liver tissue engineering.