Fabrication and characterization of chitosan/kefiran electrospun nanofibers for tissue engineering applications

Chitosan (CS)-based nanofibrous scaffolds are very promising in tissue engineering applications. However, electrospinning of CS is not possible unless using toxic solvents such as trifluoroacetic acid or by blending with other polymers. In the present study, we investigated CS-based nanofibers' fabrication by blending it with kefiran as a natural polysaccharide. A series of solutions with various CS to kefiran ratios were prepared and underwent electrospinning. The effects of main process parameters, including applied voltage and needle tip-to-collector distance on nanofibers' diameter and morphology, were also studied. Nanofibers containing 80% CS and 20% Kefiran with an average diameter of 81 ± 17 nm were successfully electrospun. Thermogravimetric analysis indicated the presence of both polymers in blend nanofibers. The diameter of CS/kefiran nanofibers increased with enhanced applied voltage, while needle tip-to-collector distance did not significantly affect the mean diameters. Appropriate viability of l929 cells on the obtained scaffolds was demonstrated utilizing Alamar blue assay. Also, cell attachment onto the fiber surface was confirmed by scanning electron microscopy. Results indicated that CS/kefiran nanofibrous scaffolds would be promising for tissue engineering applications.     

Fabrication and characterization of chitosan/PVA with hydroxyapatite biocomposite nanoscaffolds

Nanofibrous biocomposite scaffolds of chitosan (CS), PVA, and hydroxyapatite (HA) were prepared by electrospinning. The scaffolds were characterized by FTIR, SEM, TEM, and XRD techniques. Tensile testing was used for the characterization of mechanical properties. Mouse fibroblasts (L929) attachment and proliferation on the nanofibrous scaffold were investigated by MTT assay and SEM observation. FTIR, TEM, and XRD results showed the presence of nanoHA in the scaffolds. The scaffolds have porous nanofibrous morphology with random fibers in the range of 100–700 nm diameters. The CS/PVA (90/10) fibrous matrix (without HA) showed a tensile strength of 3.1 ± 0.2 MPa and a tensile modulus 10 ± 1 MPa with a strain at failure of 21.1 ± 0.6%. Increase the content of HA up to 2% increased the ultimate tensile strength and tensile modulus, but further increase HA up to 5–10% caused the decrease of tensile strength and tensile modulus. The attachment and growth of mouse fibroblast was on the surface of nanofibrous structure, and cells' morphology characteristics and viability were unaffected. A combination of nanofibrous CS/PVA and HA that mimics the nanoscale features of the extra cellular matrix could be promising for application as scaffolds for tissue regeneration, especially in low or nonload bearing areas. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

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.     

Glutaraldehyde‐crosslinked chitosan/hydroxyapatite bone repair scaffold and its application as drug carrier for icariin

Chitosan/hydroxyapatite (CS/HA) bone repair scaffolds crosslinked by glutaraldehyde (GA) were prepared. Characterization of morphology, structure, mechanical property, and porosity of scaffolds were evaluated. The influences of CS viscosity, HA content, and crosslinking degree on properties of scaffolds were discussed. SEM images showed that CS/HA scaffolds were porous with short rod‐like HA particles dispersing evenly in CS substrate. When [η]_CS = 5.75 × 10^?4, HA content = 65%, and crosslinking degree = 10%, the resulting CS/HA scaffolds had a flexural strength of 20 MPa and porosity of 60%, which could meet the requirements of bone repair materials. The scaffolds were used as drug carriers for icariin, and the impacts of loading time and crosslinking degree of scaffolds on drug‐loading dose were discussed. The suitable loading time was 24 h and it would be better to keep crosslinking degree no more than 10%. The drug release behavior demonstrated that the icariin‐loading CS/HA scaffolds could achieve basic drug sustained release effect. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 1539–1547, 2013     

3D mesoporous bioactive glass/silk/chitosan scaffolds and their compatibility with human adipose-derived stromal cells

Human adipose-derived stem/stromal cells (hASCs) have been popularly studied as cell-based therapy in the field of regenerative medicine due to their ability to differentiate into several cell types. In this study, in order to improve the mechanical strength and bioactivity of scaffolds for bone tissue engineering, three types of mesoporous bioactive glasses with different shapes and compositions were dispersed in the silk fibroin/chitosan (SF/CS)-based scaffolds, which were fabricated with a combination of freezing and lyophilization. The characteristic and physical properties of these composite scaffolds were evaluated. The biocompatibility was also assessed through hASCs in vitro tests. Both Alamar Blue® and Live/Dead assay® revealed that the spherical mesoporous bioactive glass doped scaffolds enhanced cell viability and proliferation. Furthermore, the addition of spherical mesoporous bioactive glass into SF/CS scaffolds encouraged hASC osteogenic differentiation as well. These results suggested that this composite scaffold can be applicable material for bone regeneration.     

Characterization of PLGA/Chitosan Electrospun Nano-Biocomposite Fabricated by Two Different Methods

Nano-biocomposites composed of poly (lactide-co-glycolide) (PLGA) and chitosan (CS) were electrospun using two fabrication methods. In the single nozzle method, the CS nano-powders dispersed in PLGA solutions were electrospun through a single nozzle but in the double-nozzle method, PLGA and CS were simultaneously electrospun from two syringes and the electrospun PLGA nanofiber and electrosprayed CS nanoparticles were mixed and collected on the rotating drum (randomly oriented [A] and aligned [B]) to prepare the nano-biocomposite membrane. The PLGA/CS scaffolds were prepared at the different ratios. The single-nozzle method was associated with decreasing fiber diameter when the CS content was increased and exhibited improve mechanical and hydrophilic properties.     

Macroporous chitosan/carboxymethylcellulose blend membranes and their application for lysozyme adsorption

The adsorption of lysozyme was investigated with novel macroporous chitosan (CS)/carboxymethylcellulose (CMC) blend membranes. The CS/CMG blend membranes were prepared by a simple solution‐blending method with glutaraldehyde as a crosslinking agent for CS and with silica particles as porogens. The CS/CMC blend membranes were insoluble in aqueous media when the CMC concentration in the membranes did not exceed 30 mol %. The protein adsorption on these membranes from aqueous solutions containing different amounts of lysozyme at different pHs was investigated in batch systems. The results showed that the lysozyme adsorption capacity had a maximum at pH 9.2, and this indicated that the CS/CMC blend membranes could act as cation‐exchange membranes. Moreover, the blend membranes showed the best adsorption properties for lysozyme when the CMC concentration was 20 mol %. In addition, the lysozyme adsorption capacity of the blend membranes increased with an increase in the initial lysozyme concentration and the adsorption temperature. The maximum adsorption capacity of the macroporous CS/CMC blend membranes was as high as 240 mg/g (170 mg/mL), and more than 95% of the adsorbed lysozyme was desorbed in a pH buffer at 11.8. The blend membranes also demonstrated good reusability after several adsorption–desorption cycles. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 96: 1267–1274, 2005     

Enhancing the mechanical and barrier properties of chitosan/graphene oxide composite films using trisodium citrate and sodium tripolyphosphate crosslinkers

This study investigated the effect of ionic crosslinking on the mechanical, barrier, and optical properties of chitosan (CS) and CS/graphene oxide (CSGO) composite films using trisodium citrate (CIT) and sodium tripolyphosphate (TPP) solutions of different concentrations (0.5, 1.0. 2.0, and 3.0% w/v). Successful crosslinking was confirmed by Fourier-transform infrared spectroscopy. The hydrophilicity and light transmittance decreased significantly (p < 0.05) with the increase in concentration of both crosslinking agents. The CS films crosslinked with 3.0% w/v TPP exhibited significant (p < 0.05) improvements in barrier properties, achieving a 51% decrease of water vapor permeability and 59% decrease in oxygen permeability, in comparison to neat CS film. In addition, TPP-crosslinked CSGO films experienced an 82% and 42% improvement in tensile strength and elongation at break, respectively. Overall, crosslinked CS and CSGO films possess significantly improved properties and have great potential to be further studied as food packaging materials.     

Microarchitectural and mechanical characterization of chitosan/hydroxyapatite/demineralized bone matrix composite scaffold

Porous degradable scaffolds are used extensively in bone tissue engineering. As well as material type, the architectural and mechanical characterizations of scaffolds are important to facilitate cell and tissue growth. Matrices composed of hydroxyapatite (HA), chitosan (CS) and demineralized bone matrix (DBM) may create an appropriate environment for the regeneration of bones. In this study, CS/HA/DBM scaffolds with sufficient structural integrity and high interconnected porosity were produced using different combinations of CS, HA and DBM. Both mechanical and biological properties of porous scaffolds were determined by local microarchitecture whose parameters were quantified based on micro computed tomography (Micro-CT) analysis. Within porosity range of 48–65%, the ranges of average compressive modulus and ultimate strength of the scaffolds were 3 ± 1–6 ± 1 kPa and 11 ± 2–24 ± 2 kPa, respectively. With the increase of HA concentration at the equal weight of DBM, the average trabecular thickness and trabecular separation increased and bone surface/volume ratio decreased, resulting in higher volume fraction and lower total porosity. In vitro, MC3T3-E1 preosteoblast cells were used to investigate cell attachment, spreading and proliferation on the scaffolds via hematoxyline and eosin (HE), scanning electron microscopy (SEM) and MTS assay. The results showed that MC3T3-E1 cells adhered to the surface of composite scaffolds, cell number increased with culture time. Cell viability increased with the HA particles decreased, changed little with the DBM increased. Consideration of the microarchitectural and mechanical characterization and biocompatibility of the scaffolds, 3:3:1.5 and 3:5:1.5 groups were believed to be the best in our study.     

Novel hydroxyapatite/tussah silk fibroin/chitosan bone-like nanocomposites

The nanocomposite particles of hydroxyapatite–tussah silk fibroin (HA–TSF) and HA–chitosan (HA–CS) were developed by biomimetic synthesis using Ca(NO₃)₂ and Na₃PO₄ as the precursors of inorganic phase in the presence of TSF and CS. Both nanocomposite particles were carbonate-substituted HA with low crystallinity. TSF and CS induced preferential alignment of HA crystallites along the direction of c-axis, and the induction effect of TSF was more than of CS. HA–TSF and HA–CS nanocomposite particles were found to be needle-like in the shape with a typical size of 100–200 nm in length and about 20 nm in width, and 115–250 nm in length and about 25 nm in width, respectively, as the result of the preferential arrangement of HA crystallites along c-axis intensified by TSF and CS. Based on both nanocomposite particles, the bone-like nanocomposites of HA–TSF/CS and HA–CS/TSF with the same compositions were prepared by isostatic pressing using CS and TSF concentrated solutions as adhesive composition, respectively. However, the two bone-like nanocomposites exhibited significantly different mechanical properties. The compressive strength, compressive modulus, and bending strength of HA–TSF/CS composite were significantly higher than of HA–CS/TSF composite. The fracture mechanism of both composites was analyzed by SEM observation. The study result indicates that HA–TSF/CS nanocomposite is an ideal candidate for bone substitute materials.     

Shape memory effect of chitosan/glycerol composite film in mixed water/ethanol solution

In this paper, a chitosan (CS)/glycerol (GL) composite film was prepared, and the solution-responsive feature of the composite film in a mixed water/ethanol solution was investigated. The composite film began to show obvious shape recovery behavior when the water content of the mixed solution was more than 50% v/v. In mixed solutions, when the molar ratio of water molecules and ethanol molecules was more than 2:1, apart from the combination with the ethanol molecules, the residual water molecules could interact with the CS molecules, promoting the formation of hydrogen bonds. The stress stored in CS molecules was released, and the shape memory effect of the composite film was triggered. In addition, the effect of water content in the mixed solutions on the structural, crystal, thermal stability and mechanical properties of the composite film was characterized. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47037.     

Fabrication of macroporous chitosan/poly(l-lactide) hybrid scaffolds by sodium acetate particulate-leaching method

For tissue engineering applications, 3-D macroporous chitosan/poly(l-lactide) (PLLA) scaffolds were prepared by the particulate-leaching method using sodium acetate as the porogen in an acidic water/dioxane solution. The stability and dispersity of chitosan on the chitosan/PLLA hybrid scaffolds were determined by measuring water contact angles, establishing crystallinity using X-ray diffraction, and using eosin staining to observe the chitosan under a light microscope. The porous structure of the particulate-leached chitosan/PLLA scaffolds was investigated in terms of pore morphology, interconnectivity, etc. by using scanning electron microscopy. Chitosan/PLLA scaffolds produced by particulate-leaching showed a highly porous structure and improved stability and dispersity of chitosan as compared to pure PLLA and chitosan-coated PLLA scaffolds. The highly porous structure that resulted from a high concentration of chitosan improved the efficiency of cell adhesion after culturing cells for 4?h. After 48?h, the cultured cells showed increased cell proliferation on the hybrid scaffolds. Thus, particulate-leached chitosan/PLLA scaffolds can be applied to tissue engineering of various types, including the industrial membrane field.     

壳聚糖/N-乙烯基吡咯烷酮接枝共聚物的制备及其性能

以壳聚糖（CS）和N-乙烯基吡咯烷酮（NVP）为原料,本文采用过氧化氢和抗坏血酸共引发剂引发,制备壳聚糖/N-乙烯基吡咯烷酮的接枝共聚物（CS-g-NVP）,研究了引发剂配比、反应温度和反应时间等因素对接枝共聚反应的影响。通过红外光谱、核磁共振和热重分析等对产物进行表征,并考察其吸湿性和亲水性能。结果表明,接枝共聚反应的最佳条件为：引发剂抗坏血酸/过氧化氢的摩尔比为1∶1、反应温度60℃、反应时间12h。接枝共聚产物的吸湿性优于壳聚糖,在饱和氯化钙和饱和硫酸铵环境下产物的吸湿率分别为3.68%和23.1%,高于原料壳聚糖的0.41%和9.77%,产物具有良好的亲水性和水溶性,能溶解于酸性和碱性水溶液中。     

壳聚糖/生物活性玻璃多孔复合支架的制备及界面增强作用

采用卵磷脂作为表面活性剂对溶胶-凝胶生物玻璃进行表面改性,并采用冷冻干燥法制备用于骨和软骨组织工程的壳聚糖/生物活性玻璃复合多孔支架(chitosan/bioglass porous composite scaffolds,CS/BGS),观察CS/BGS的显微形貌并测定抗压强度,探讨生物玻璃的表面改性对CS/BGS显微结构及力学强度的影响。研究表明:采用冷冻干燥法可以制备具有一定强度的三维连通的CS/BGS,且孔隙率达到90%以上。通过对生物玻璃表面改性可以在一定程度上提高CS/BGS的抗压强度。     

Preparation and characterization of magnetite/hydroxyapatite/chitosan nanocomposite by in situ compositing method

Chitosan is an important kind of biomaterial that is widely used in medical applications. One of the key concerns about its use is the preparation of composites used for bone engineering. Aim of this study concerns the preparation of three‐dimensional nanocomposites having potential use in bone repair and regeneration. The magnetite/hydroxyapatite/chitosan nanocomposites were prepared via in situ compositing method by preparing precursor solutions and molds with chitosan membrane. These nanocomposites were characterized by chemical, spectroscopic, magnetic, and morphological methods. X‐ray diffraction analysis results demonstrate the formation of magnetite and hydroxyapatite in the chitosan matrix. FTIR analysis indicates that inorganic nanoparticles were chemically bound to the amino and hydroxyl groups in CS molecules. From TG/DTA data, it can be concluded that during preparation raw materials were almost perfectly incorporated into the nanocomposites, and the decrease in decomposition temperatures indicates the formation of chemical bonds between inorganic nanoparticles and chitosan molecules. TEM results show that the maximum size of inorganic particles in the magnetite/hydroxyapatite/chitosan nanocomposites was under 50 nm, and these particles were dispersed homogeneously in the chitosan matrix. From the magnetic measurement, it could be concluded that the nanocomposites were superparamagnetic, which is also the peculiarity of nanomagnetites. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Preparation,characterization, and salicylic acid release behavior of chitosan/poly(vinyl alcohol) blend microspheres

Blend microspheres of chitosan (CS) with poly(vinyl alcohol) (PVA) were prepared as candidates for oral delivery system. CS/PVA microspheres containing salicylic acid (SA), as a model drug, were obtained using the coacervation‐phase separation method, induced by addition of a nonsolvent (sodium hydroxide solution) and then crosslinked with glutaraldehyde (GA) as a crosslinking agent. The microspheres were characterized by Fourier transform infrared spectroscopy, differential scanning calorimetry (DSC), and scanning electron microscopy. Percentage entrapment efficiency, particle size, and equilibrium swelling degree of the microsphere formulations were determined. The results indicated that these parameters were changed by preparation conditions of the microspheres. Effects of variables such as CS/PVA ratio, pH, crosslinker concentration, and drug/polymer (d/p) ratio on the release of SA were studied at three different pH values (1.2, 6.8, and 7.4) at 37°C. It was observed that SA release from the microspheres increased with decreasing CS/PVA ratio and d/p ratio whereas it decreased with the increase in the extent of crosslinking. It may also be noted that drug release was much higher at pH 1.2 than that of at pH 6.8 and 7.4. The highest SA release percentage was obtained as 100% for the microspheres prepared with PVA/CS ratio of 1/2, d/p ratio of 1/2, exposure time to GA of 5 min, and concentration of GA 1.5% at the end of 6 h. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Characterization of gamma irradiated concentrated aqueous solutions of chitosan/sodium alginate blends and their drug uptake‐release characters

Blends films based on different ratios of concentrated aqueous solutions of chitosan (CS) and sodium alginate (AG) in the presence of 1% of glutaraldehyde, as a cross‐linking agent for chitosan, were prepared by solution casting and then exposed to gamma irradiation. The formed blends were characterized by IR spectroscopic analysis, differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). The uptake‐release properties of CS/AG blends, taking ketoprofen as an example for drug, were also investigated. DSC thermograms of CS/AG blends revealed good miscibility was sustained between CS and AG. The water uptake and gel content of CS/AG blends was found to decrease by increasing the ratio of AG in the initial solution. The IR spectra indicated the formation of cross‐linking and hydrogen bonding, while the TGA study showed that the CS/AG blends displayed higher thermal stability than pure CS polymer. Based on Fick's law, it was demonstrated that the main parameters affecting the release of ketoprofen drug from the CS/AG blend hydrogels were composition and pH. © 2011 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Fabrication of a porous wall and higher interconnectivity scaffold comprising gelatin/chitosan via combination of salt-leaching and lyophilization methods

A novel gelatin/chitosan scaffold with higher porosity and interconnectivity was designed through salt-leaching/lyophilization (SLL) method. The properties of the fabricated scaffolds were compared with conventional scaffolds, which are obtained by thermally induced phase-separation (TIPS) method. The scaffolds made by phase-separation method have high tensile strength, but suffer from less channel interconnectivity, pore uniformity and also low surface porosity. The microstructure, porosity, phosphate-buffered saline (PBS) solution absorption and tensile strength of the prepared scaffolds by SLL method were studied. In this work, SLL as a two-step technique is introduced for creating porosity to improve both channel interconnectivity and pore uniformity for water-soluble polymers in comparison with the TIPS method. The SLL technique includes two mechanisms: the first, leaching of mixed sodium chloride crystals and particles created during recrystallization of the dissolved NaCl and the second, phase separation during lyophilization at the pore walls. These two steps in porosity formation lead to special pore morphology, which is more suitable for cell culturing because of higher interconnectivity and rich surface porosity in comparison with the phase-separated scaffolds. The prepared scaffolds, using this technique with different salt/polymer ratios and salt crystal size, have 91?C97% porosity and 94?C190???m mean pore size with tensile strength of 72?C215?kPa and PBS solution absorption between 12.4 and 19 times dry weight. The pore size of scaffolds prepared using the SLL method could be adjusted independently of polymer solution concentration. These scaffolds have a great potential in skin tissue engineering application.     

Preparation and characterization of biomimetic silk fibroin/chitosan composite nanofibers by electrospinning for osteoblasts culture

ABSTRACT: In this study, we have successfully fabricated electrospun bead-free silk fibroin [SF]/chitosan [CS] composite nanofibers [NFs] covering the whole range of CS content (0%, 25%, 50%, 75%, and 100%). SF/CS spinning solutions were prepared in a mixed solvent system of trifluoroacetic acid [TFA] and dichloromethane. The morphology of the NFs was observed by scanning electron microscope, and the average fiber diameter ranges from 215 to 478 nm. Confocal laser scanning microscopy confirms the uniform distribution of SF and CS within the composite NFs. To increase biocompatibility and preserve nanostructure when seeded with cells in culture medium, NFs were treated with an ethanol/ammonia aqueous solution to remove residual TFA and to change SF protein conformation. After the chemical treatment, SF/CS NFs could maintain the original structure for up to 54 days in culture medium. Properties of pristine and chemically treated SF/CS NFs were investigated by Fourier transform infrared spectroscopy [FT-IR], X-ray diffraction [XRD], and thermogravimetry/differential scanning calorimetry [TG/DSC]. Shift of absorption peaks in FT-IR spectra confirms the conformation change of SF from random coil to β-sheet by the action of ethanol, which is also consistent with the SF crystalline diffraction patterns measured by XRD. From TG/DSC analysis, the decomposition temperature peaks due to salt formation from TFA and protonated amines disappeared after chemical treatment, indicating complete removal of TFA by binding with ammonium ions during the treatment. This was also confirmed with the disappearance of F1s peak in X-ray photoelectron spectroscopy spectra and disappearance of TFA salt peaks in FT-IR spectra. The composite NFs could support the growth and osteogenic differentiation of human fetal osteoblastic [hFOB] cells, but each component in the composite NF shows distinct effect on cell behavior. SF promotes hFOB proliferation while CS enhances hFOB differentiation. The composite SF/CS NFs will be suitable for bone tissue engineering applications by choosing a suitable blend composition.PACS: 87.85.jf; 87.85.Rs; 68.37.Hk.     

交联壳聚糖/聚乙烯醇/蜗牛黏液复合膜的制备及性能研究

采用延流法制备了香兰素（V）交联的壳聚糖/聚乙烯醇/蜗牛黏液（CS/PVA/SM）复合膜,并通过热重分析仪（TG）、扫描电子显微镜（SEM）和万能材料试验机等研究了不同CS/SM配比对复合膜光学性能、水蒸气和氧气阻隔能力、力学性能、热力学性能及生物降解性能等的影响。结果表明,CS/PVA/SM复合膜为可降解的亲水性薄膜,当CS溶液/SM溶液体积比为5/3时,复合膜性能优良,其抗氧化活性为87.51 %,其水蒸气透过率比纯CS膜降低了75.16 %,不透明度降低了87.74 %,拉伸强度提高了16.04 %,断裂伸长率提高了28.26倍;随着SM含量的增加,复合膜的热稳定性有所降低;CS溶液/SM溶液体积比为5/1、5/2和5/3时,复合膜表现出良好的相容性;SM的添加使复合膜具有很好的延展性和柔韧性,V的添加提高了复合膜的拉伸强度和抗氧化能力;所制备的CS/PVA/SM复合膜在食品包装领域中有潜在的应用前景。