Fabrication and characterization of poly(ε-caprolactone)/gelatin nanofibrous scaffolds for retinal tissue engineering

Different ratios of poly(ε-caprolactone) (PCL) and gelatinwere used to fabricate scaffolds for regeneration of retinal pigment epithelium (RPE) layer. Physical and chemical characterizations were performed and the behavior of human RPE cells on the scaffolds was evaluated subsequently. An increase in gelatin content in the scaffold enhanced hydrophilicity, RPE cell attachment, proliferation, and spreading over PCL scaffolds. Granular and cytoplasmic expressions of RPE65 and Cytokeratin 8/18 markers confirmed the presence of RPE cells. It was believed that PCL/gelatin scaffolds could be used as substrates to replace RPE extracellular matrix to facilitate regeneration of RPE layer in retinal diseases.     

Characterization of the morphology and hydrophilicity of chitosan/caffeic acid hybrid scaffolds

The morphology and affinity of a scaffold influence the attachment of cells to its surfaces. In this study, the morphology and hydrophilicity of chitosan/caffeic acid hybrid scaffolds were investigated. Grafting caffeic acid onto chitosan hybrid scaffolds by using high levels of potassium persulfate produced scaffolds with looser morphology and higher porosity, as indicated by scanning electron microscopy (SEM) and porosity analysis. SEM analysis showed that the prepared scaffolds had a macroporous morphology with interconnected pores. Differential scanning calorimetry (DSC) revealed that the scaffolds’ hydrophilicity decreased after caffeic acid grafting. The scaffolds were cultured with human osteosarcoma UMR-106 cells, but SEM analysis showed that cell attachment was poor. However, calcification of the scaffolds promoted the attachment of UMR-106 cells onto the scaffold. This study shows that calcified chitosan/caffeic acid hybrid scaffolds could be suitable for use in hard-tissue engineering.     

Design,Fabrication, and Characterization of Novel Porous Conductive Scaffolds for Nerve Tissue Engineering

Highly conductive polypyrrole/graphene (PYG) nanocomposite was synthesized with chemical oxidation process via emulsion polymerization and used for the preparation of novel porous conductive gelatin/chitosan-based scaffolds. The effect of PYG loading on various properties of scaffolds was investigated. The obtained results indicated that by introducing PYG into the polymeric matrix, the porosity and swelling capacity decreased while electrical conductivity and Young's modulus demonstrated increasing trend. The in vitro biodegradation test revealed that pure gelatin/chitosan matrix lost 80% of its weight after six weeks in the presence of lysozyme whilst the biodegradation rate was significantly lower for the conductive scaffolds. Furthermore, Schwann cell attachment and proliferation were evaluated by MTT assay and SEM image and the results revealed significant cell biocompatibility of the conductive scaffold with low amount of PYG. The results confirmed the potential of gelatin/chitosan/PYG compounding as a suitable biomaterial for using in nerve tissue engineering applications in which electrical stimulation plays a vital role.     

Fabrication and Properties of Gelatin/Chitosan Composite Hydrogel

Gelatin/chitosan hydrogels were prepared by using glutaraldehyde as crosslinker. The porous structure was confirmed by scanning electron microscope (SEM). Swelling ratios of the hydrogels with various ratio of gelatin to chitosan and crosslinker reagent dosage were studied in phosphate-buffered saline (PBS). In addition, in-vitro cytotoxicity was assessed via MTT assay with fibroblastic cell cultured in hydrogel extractions. It was found that by increasing glutaraldehyde dosage and chitosan content, the swelling ratio of the hydrogels decreased in buffer solutions. The MTT test showed that the gelatin/chitosan hydrogel clearly presented adequate cell viability, non-toxicity, and suitable properties. Therefore, these developed blends, based on gelatin and chitosan has broadened the number of choices of biomaterials to be potentially used in biomedical applications such as biomaterial, drug delivery vehicles and skin tissue engineering.     

Bioactive monetite‐containing whisker‐like fibers reinforced chitosan scaffolds

The aim of this work was to develop bioactive chitosan scaffolds reinforced with monetite‐containing whisker‐like fibers. The fibers synthesized by homogeneous precipitation were characterized as monetite/hydroxyapatite short fibers (MAFs), using XRD, FTIR and SEM. The pure chitosan and MAFs/chitosan composite scaffolds were produced by freeze‐drying, and characterized with respect to porosity, pore size, swelling behavior, compressive strength and modulus, and in vitro bioactivity. The incorporation of MAFs in chitosan matrices led to increase the pore size, according to the evaluation by FE‐SEM, and decrease the porosity of composite scaffolds. The swelling ratio decreased as MAFs content of scaffolds increased. The compressive strength and modulus of scaffolds were improved by an increase in MAFs content. The noncross‐linked scaffolds with a chitosan: MAFs weight ratio of 1:1 (CW3) showed a porosity of 75.5%, and the strength and modulus of 259 kPa and 2.8 MPa in dry state, respectively. The crosslinking by glutaraldehyde resulted in improved mechanical properties. The strength and modulus of cross‐linked CW3 scaffolds in wet state reached to 345 kPa and 1.8 MPa, respectively. The in vitro bioactivity of the reinforced scaffolds, evaluated by FE‐SEM/EDS, XRD, and ATR‐FTIR, was confirmed by the formation of a carbonated apatite layer on their surfaces when they soaked in simulated body fluid (SBF). The results of this initial study indicate that the monetite‐containing whisker‐like fibers may be an appropriate reinforcement of chitosan scaffolds.     

The Role of Bloom Index of Gelatin on the Interaction with Retinal Pigment Epithelial Cells

Biocompatible materials are of considerable interest in the development of cell/drug delivery carriers for therapeutic applications. This paper investigates the effects of the Bloom index of gelatin on its interaction with retinal pigment epithelial (RPE) cells. Following two days of culture of ARPE-19 cells with gelatin samples G75-100, G175, and G300, the in vitro biocompatibility was determined by cell proliferation and viability assays, and glutamate uptake measurements, as well as cytokine expression analyses. The mitochondrial dehydrogenase activity in the G300 groups was significantly lower than that of G75-100 and G175 groups. The Live/Dead assays also showed that the gelatin samples G300 induced mild cytotoxicity. In comparison with the treatment of gelatins with low Bloom index, the exposure to high Bloom strength gelatins markedly reduced the glutamate uptake capacity of ARPE-19 cells. One possible explanation for these observations is that the presence of gelatin samples G300 with high viscosity in the medium may affect the nutrient availability to cultured cells. The analyses of pro-inflammatory cytokine IL-6 expression at both mRNA and protein levels showed that the gelatins with low Bloom index caused less cellular inflammatory reaction and had more acceptable biocompatibility than their high Bloom strength counterparts. These findings suggest that the Bloom index gives influence on cellular responses to gelatin materials.     

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.     

Preparation of enhanced three-dimensional porous chitosan scaffolds by acetylation and aqueous extraction

We have devised a method to prepare a 3-dimensional (3D) porous acetylated chitosan scaffold for use as a cell adhesion matrix in tissue engineering applications. The scaffold was prepared by molding a mixture of chitosan and gelatin (as porogen), then removing the uncomplexed porogen by aqueous extraction from the freeze-dried material prior to acetylation. The extent of chitosan acetylation according to the reaction time was observed by X-ray diffraction (XRD) analysis. The differences between the aqueous-extracted and control phase-separated chitosan scaffolds in terms of pore morphology and interconnectivity were examined by scanning electron microscopy (SEM), enzymatic degradation, and surface roughness tests. The fibroblast cell line NIH-3T3 was used to test relative cell affinities for the acetylated versus untreated (control) chitosan scaffolds. The acetylated 3D porous scaffolds showed high interconnectivity and improved biocompatibility properties. Thus, these scaffolds may be very useful for a variety of tissue engineering applications.     

仿生型壳聚糖-明胶/溶胶凝胶生物玻璃复合支架的制备及其矿化性能研究

利用冷冻干燥法制备出用于骨和软骨组织工程的壳聚糖-明胶/溶胶凝胶生物玻璃(CS-Gel/SGBG)仿生型复合多孔支架,并进行了孔隙率的测定和显微形貌的观察;探讨了各组分不同用量对CS-Gel/SGBG复合支架显微结构的影响以及复合支架在模拟生理体液中的仿生矿化性能。研究表明,通过调节各组分的不同用量,可以制备出三维连通的复合多孔支架,且孔隙率达到90%以上;在模拟生理体液中浸泡后发现CS-Gel/SGBG支架表面有大量结晶态类骨碳酸羟基磷灰石生成,表明复合支架有良好的生物矿化性能。     

视网膜色素上皮细胞培养及其超微结构的观察

目的 体外培养视网膜色索上皮(retinal pigment epithelium RPE)细胞并对其进行超微结构观察,为RPE细胞在细胞和分子水平研究奠定基础。方法 采用酶消化法对RPE细胞进行取材和培养,利用光镜和电镜进行观察及分析。结果 培养的细胞贴壁生长,融合后呈单层、镶嵌式排列;电镜下可见到细胞表面绒毛、细胞之间复合体、细胞间质少及细胞排列紧密等上皮组织的特征性结构,细胞内可直接观察到大量黑色素颗粒。结论 为各种损伤致RPE细胞形态学改变的研究奠定了基础。     

Study on the properties of the electrospun silk fibroin/gelatin blend nanofibers for scaffolds

Silk fibroin (SF)/gelatin blend nanofibers membranes as scaffolds were fabricated successfully via electrospinning with different composition ratios in formic acid. The formation of intermolecular hydrogen bonds and the conformational transition of SF provided scaffolds with excellent mechanical properties. FTIR and DTA analysis showed the SF/gelatin nanofibers had more β‐sheet structures than the pure SF nanofibers. The former's breaking tenacity increased from 0.95 up to 1.60 MPa, strain at break was 7.6%, average fiber diameter was 89.2 nm, porosity was 87%, and pore diameter was 142 nm. MTT, H&E stain, and SEM results showed that the adhesion, spreading, and proliferation of human umbilic vein endothelium cells (HUVECs) and mouse fibroblasts on the SF/gelatin nanofibers scaffolds were definitely better than that on the SF nanofibers scaffolds. The scaffolds could replace the natural ECM proteins, support long‐term cell growth, form three‐dimensional networks of the nanofibrous structure, and grow in the direction of fiber orientation. Our results prove that the addition of gelatin improved the mechanical and biological properties of the pure SF nanofibers, these SF/gelatin blend nanofiber membranes are desirable for the scaffolds and may be a good candidate for blood vessel engineering scaffolds. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Mechanical and structural properties of polylactide/chitosan scaffolds reinforced with nano-calcium phosphate

In this study, chitosan/polylactide scaffolds reinforced with nano-calcium phosphate (average crystallite size of 16.5?nm) (CP) were fabricated to create a material with excellent properties for bone tissue engineering applications via freeze-casting method. The structural and mechanical properties of nanocomposite scaffolds were studied by increasing amount of chitosan/poly lactide ratio and nano-CP content in both dry and hydrate states, which reflected the exact status of scaffolds in a real biomechanical environment. The morphologies of the nanocomposite scaffolds were viewed using scanning electron microscopy (SEM) and all the scaffolds exhibited a high porosity (up to 92?%) with open pores of 38?C387???m average diameters, which decreased with increased chitosan/polylactide ratio and nano-CP content. Also, SEM photograph of the cross-sectional area of the scaffold showed nano-CP was dispersed all over the polymer matrix thoroughly. The results of mechanical tests showed that the compressive modulus (E) and compressive stress (??) enhanced, when chitosan and nano-CP increased. X-ray diffraction analysis indicated typical chitosan, polylactic acid and nano-CP peaks and showed that the increase in nano-CP weight percentage increased its peak intensities. In addition, the effect of pore-size distributions of the scaffolds with the same composition was studied in relation to mechanical properties. The results showed substantial differences in the pore-size distributions of scaffolds with the same composition prepared, which have no effect on their dry states.     

Preparation and characterization of porous scaffold composite films by blending chitosan and gelatin solutions for skin tissue engineering

Biodegradable polymers have significant potential in biotechnology and bioengineering. However, for some applications, they are limited by their inferior mechanical properties and unsatisfactory compatibility with cells and tissues. In the present investigation blends of chitosan and gelatin with various compositions were produced as candidate materials for biomedical applications. Fourier transform infrared spectral analysis showed good compatibility between these two biodegradable polymers. The composite films showed improved tensile properties, highly porous structure, antimicrobial activities, low water dissolution, low water uptake and high buffer uptake compared to pure chitosan or gelatin films. These enhanced properties could be explained by the introduction of free ? OH, ? NH₂ and ? NHOCOCH₃ groups of the amorphous chitosan in the blends and a network structure through electrostatic interactions between the ammonium ions (? NH³⁺) of the chitosan and the carboxylate ions (? COO^?) of the gelatin. Scanning electron microscopy images of the blend composite films showed homogeneous and smooth surfaces which indicate good miscibility between gelatin and chitosan. The leafy morphologies of the scaffolds indicate a large and homogeneous porous structure, which would cause increased ion diffusion into the gel that could lead to an increase in stability in aqueous solution, buffer and temperature compared to the gelatin/chitosan system. In vivo testing was done in a Wistar rat (Rattus norvegicus) model and the healing efficiencies of the scaffolds containing various compositions of chitosan were measured. The healing efficiencies in Wistar rat of composites with gelatin to chitosan ratios of 10:3 and 10:4 were compared with that of a commercially available scaffold (Eco‐plast). It was observed that, after 5 days of application, the scaffold with a gelatin to chitosan ratio of 10:3 showed 100% healing in the Wistar rat; however, the commercial Eco‐plast showed only a little above 40% healing of the dissected rat wound. Copyright © 2012 Society of Chemical Industry     

Surface treatment with amino acids of porous collagen based scaffolds to improve cell adhesion and proliferation

The purpose of this study was to improve the biocompatibility of glutaraldehyde (GA) cross‐linked chitosan coated collagen scaffold for cartilage tissue regeneration. In order to prevent the potential toxicity of GA, we treated the designed scaffold with either glutamic acid or glycine. Amino acid treated scaffolds were characterized by scanning electron microscopy (SEM) techniques. Afterward, chondrocyte interaction with the composite scaffold was investigated assessing cell adhesion and proliferation using Hoechst staining and MTT cell proliferation assay, respectively. The SEM analyses of the scaffolds’ surface and cross‐section confirmed the adhesion of amino acids on the surface of the scaffolds. We also observed that scaffolds’ porosity was reduced due to the coverage of the pores by chitosan and amino acids, leading to low porosity. The use of amino acid improved the chondrocyte adhesion and proliferation inside the scaffolds’ pores when cells were cultured onto the chitosan‐coated collagen scaffolds. Overall, our in vitro results suggest the use of amino acid to improve the biocompatibility of natural polymer composite scaffold being crosslinked with glutaraldehyde. Such scaffold has improved mechanical properties; biocompatibility thus may be useful for tissue regeneration such as cartilage.

     

Bone‐like apatite‐coated chitosan scaffolds: Characterization and osteoblastic activity

In this study, porous scaffolds were prepared from chitosan (2% w/v in acetic acid and deacetylation degree: DD > 85%) by freeze‐drying method, and freshly lyophilized scaffolds were stabilized with ethanol solutions. Bone‐like apatite formation on chitosan scaffolds was achieved by immersing the scaffolds into a novel concentrated simulated body fluid (10× SBF‐like solution) for different periods, i.e., 6 and 24 h. Scanning electron microscope views showed that the 6‐h treatment in 10× SBF‐like solution led to the formation of calcium phosphate nucleation sites on chitosan scaffolds, whereas the apatite particles showed characteristic cauliflower‐like morphology at the end of 24‐h treatment. X‐ray diffraction results supported the fact that mineral phase was made of hydroxyapatite. Osteogenic activities of untreated and SBF‐treated chitosan scaffolds were examined by preosteoblastic MC3T3 cell culture studies. The mitochondrial activity test showed that apatite‐coated scaffolds stimulated cell proliferation compared with uncoated scaffolds. Alkaline phosphatase and osteocalcine levels indicated that the differentiation of the cells on all scaffolds increased significantly from 15th day of culture to the 21th day of culture, especially for the cells on 24‐h SBF‐treated scaffolds. The results of this study indicated that 10× SBF‐like solution‐treated chitosan scaffolds may be evaluated for bone tissue engineering. POLYM. COMPOS., 31:1418–1426, 2010. © 2009 Society of Plastics Engineers     

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.     

In vitro response of retinal pigment epithelial cells exposed to chitosan materials prepared with different cross-linkers

The interaction between cells and biopolymers is the evaluation indicator of the biocompatibility of materials. The purpose of this work was to examine the responses of retinal pigment epithelial (RPE) cells to genipin (GP) or glutaraldehyde (GTA) cross-linked chitosan by means of cell viability assays, cytokine expression analyses, and apoptosis assays. Evaluations of non-cross-linked chitosan were conducted simultaneously for comparison. Both GP and GTA treated samples with the same extent of cross-linking (around 80%) were prepared by varying cross-linking time. Our results showed that GP cross-linking was carried out by either radical polymerization of the monomers or S(N)2 nucleophilic substitution reaction involving the replacement of the ester group on the monomer with a secondary amide linkage. On the other hand, GTA could react with free amino groups of chitosan, leading to the formation of either the Schiff bases or the Michael-type adducts with terminal aldehydes. The biocompatibility of non-cross-linked chitosan membranes was demonstrated by the absence of any signs of toxicity or inflammation reaction. The present study showed that the ARPE-19 cells exposed to GTA cross-linked chitosan membranes had significantly higher cytotoxicity, interleukin-6 levels, and number of TUNEL-positive nuclei than did those exposed to GP treated samples. In addition, the materials modified with GTA trigger apoptosis at an early stage and may induce toxicity in the RPE cells later. The findings suggest that while the chitosan molecules bridged by GP are satisfactorily cytocompatible, the counterparts treated by GTA do not seem to be tolerated. In terms of material safety, the GP cross-linked chitosan may be compatible with human RPE cells and may have a potential application as delivery carriers in the treatment of posterior segment diseases.     

Preparation of elastic chitosan/poly(<Emphasis Type="SmallCaps">l</Emphasis>-lactide-co-ε-caprolactone) macroporous scaffolds by acetylation and particulate leaching

For soft tissue engineering applications, 3-D macroporous acetylated chitosan/poly(l-lactideco-ε-caprolactone) (PLCL) scaffolds were prepared by acetylation and particulate leaching using sodium acetate in an acidic water/dioxane solution. Acetylated 5 wt% chitosan/PLCL scaffold of 90% porosity was determined and confirmed through various tests. The physiochemical properties of acetylated chitosan/PLCL hybrid scaffolds were examined by measuring water contact angles, pore morphology and interconnectivity using scanning electron microscopy (SEM), and dye release testing. In addition, mechanical properties such as tensile strength and bending stress recovery for determining the elasticity of scaffolds were measured. The fibroblast cell line NIH-3T3 was used to test relative cell affinities for the acetylated chitosan/PLCL vs. normal chitosan/PLCL films and porous scaffolds. The acetylated chitosan/PLCL films and scaffolds showed a high initial cell adhesion after 4 h of cell culture and increased cell proliferation compared to that of the control. The acetylated chitosan/PLCL scaffolds produced by particulate leaching showed a highly porous structure and improved the biocompatibility and stability of chitosan compared to that of chitosan-coated PLCL scaffolds. Thus, these scaffolds may be very useful for a variety of tissue engineering applications.     

Profound Re-Organization of Cell Surface Proteome in Equine Retinal Pigment Epithelial Cells in Response to In Vitro Culturing

The purpose of this study was to characterize the cell surface proteome of native compared to cultured equine retinal pigment epithelium (RPE) cells. The RPE plays an essential role in visual function and represents the outer blood-retinal barrier. We are investigating immunopathomechanisms of equine recurrent uveitis, an autoimmune inflammatory disease in horses leading to breakdown of the outer blood-retinal barrier and influx of autoreactive T-cells into affected horses’ vitrei. Cell surface proteins of native and cultured RPE cells from eye-healthy horses were captured by biotinylation, analyzed by high resolution mass spectrometry coupled to liquid chromatography (LC MS/MS), and the most interesting candidates were validated by PCR, immunoblotting and immunocytochemistry. A total of 112 proteins were identified, of which 84% were cell surface membrane proteins. Twenty-three of these proteins were concurrently expressed by both cell states, 28 proteins exclusively by native RPE cells. Among the latter were two RPE markers with highly specialized RPE functions: cellular retinaldehyde-binding protein (CRALBP) and retinal pigment epithelium-specific protein 65kDa (RPE65). Furthermore, 61 proteins were only expressed by cultured RPE cells and absent in native cells. As we believe that initiating events, leading to the breakdown of the outer blood-retinal barrier, take place at the cell surface of RPE cells as a particularly exposed barrier structure, this differential characterization of cell surface proteomes of native and cultured equine RPE cells is a prerequisite for future studies.     

Synthesis,Characterization and <Emphasis Type="Italic">in-vitro</Emphasis> Study of Chitosan/Gelatin/Calcium Phosphate Hybrid Scaffolds Fabricated Via Ion Diffusion Mechanism for Bone Tissue Engineering

In this study, biomimetic scaffolds were designed to investigate calcium phosphate formation via a double diffusion mechanism within a gelatin/chitosan hydrogel in biological pH and temperature. Three types of samples with initial percentages of chitosan (20, 30 and 40 wt. %) were prepared. Diffusion of calcium and phosphate ions through the hydrogel formed a precipitation layer. Samples were freeze dried to form porous scaffolds and soaked in glutaraldehyde to increase their mechanical properties. X-ray diffraction (XRD), Fourier transform infra-red (FTIR) spectroscopy and scanning electron microscopy (SEM) were employed to investigate the microstructure and to characterize the prepared scaffolds. Analysis of precipitation indicated the presence of brushite and hydroxyapatite. The amorphous calcium phosphate phase converted into crystalline hydroxyapatite after immersion in simulated body fluid which mimics the formation of hydroxyapatite in the human body. FTIR results suggested the presence of structural hydroxyl and phosphate bonds in the structure of the prepared scaffolds which could be due to the formation of hydroxyapatite. With increasing amount of chitosan in the composite scaffold, the water up-take ability was increased from 380 to 660 %, yield strength and Young’s modulus slightly decreased and the crystalinity of the precipitated phase increased. Mechanical properties obtained from the samples were in the range of cancellous bone. MTT assay results and alkaline phosphatase activity showed prepared scaffolds had proper biocompatibility.