Preparation and in vitro characterization of novel bioactive glass ceramic nanoparticles

SiO(2)-CaO-P(2)O(5) ternary bioactive glass ceramic (BGC) nanoparticles with different compositions were prepared via a three-step sol-gel method. Polyethylene glycol was selected to be used as the surfactant to improve the dispersion of the nanoparticles. The morphology and composition of these BGC nanoparticles were observed by ESEM and EDX. All the BGC particles obtained in this method were about 20 nm in diameter. XRD analysis demonstrated that the different compositions can result in very different crystallinities for the BGC nanoparticles. Bioactivity tests in simulated body fluid solution (SBF), and degradability in phosphate buffer solution (PBS), were performed in vitro. SEM, EDX, and XRD were employed to monitor the surface variation of neat poly(L-lactic acid), PLLA, foam and PLLA/BGC porous scaffolds during incubation. The BGC nanoparticles with lower phosphorous and relative higher silicon content exhibited enhanced mineralization capability in SBF and a higher solubility in PBS medium. Such novel nanoparticles may have potential to be used in different biomedical applications, including tissue engineering or the orthopedic field.     

Formation of bone-like apatite on poly(L-lactic acid) fibers by a biomimetic process.

Bone-like apatite coating on poly(L-lactic acid) (PLLA) fibers was formed by immersing the fibers in a modified simulated body fluid (SBF) at 37 degrees C and pH 7.3 after hydrolysis of the fibers in water. The ion concentrations in SBF were nearly 1.5 times of those in the human blood plasma. The apatite was characterized by scanning electron microscopy (SEM) with energy dispersive X-ray spectroscopy (EDX), X-ray photoelectron spectroscopy (XPS), thin-film X-ray diffraction, and Fourier transform infrared spectroscopy. After 15 days of incubation in SBF, an apatite layer with about 5-6 microm thickness was formed on the surface of the fibers. This apatite had a Ca/P ratio similar to that of natural bone. The mass of apatite coated PLLA fibers increased with extending the incubation time. After 20 days incubation, the fibers increased their mass by 25.8 +/- 2.1%. The apatite coating had no significant effect on the tensile properties of PLLA fibers. In this article, the bone-like apatite coating on three-dimensional PLLA braids was also studied. The motivation for this apatite coating was that it might demonstrate enhanced osteoconductivity in the future studies when they serve as biodegradable scaffolds in tissue engineering.     

A study on the in vitro degradation properties of poly(l-lactic acid)/β-tricalcuim phosphate(PLLA/β-TCP) scaffold under dynamic loading

The objective of this study was to investigate the effects of dynamic loading on the in vitro degradation properties of poly(l-lactic acid)/β-tricalcuim phosphate(PLLA/β-TCP) composite scaffolds. These scaffolds were prepared by a technique, namely solvent self-proliferating/model compressing/particulate leaching, and they were incubated in a customized simulated body fluid (SBF) flow system under dynamic loading conditions for 6 weeks. The bioactivity and the degradation behaviors of the composite scaffolds were systematically investigated through the formation of apatite, the mass and porosity changes, the molecular weight changes of PLLA, the compressive strength changes, etc. Results show a high level of apatite deposition on the scaffolds, suggesting their good bioactivity in the SBF. Changes in mass, porosity, molecular weight and compressive strength of the scaffolds happened more under dynamic loading conditions than that under flow only SBF conditions. Dynamic loading with the investigated frequency promoted the degradation of the scaffolds, but did not markedly deteriorate the mechanical properties of the scaffolds. All the results suggest that the composite scaffolds have great potential to be applied in bone replacements or repairs under the in vivo load-bearing conditions.     

Design and characterization of a novel chitosan/nanocrystalline calcium phosphate composite scaffold for bone regeneration

To meet the challenge of regenerating bone lost to disease or trauma, biodegradable scaffolds are being investigated as a way to regenerate bone without the need for an auto- or allograft. Here, we have developed a novel microsphere-based chitosan/nanocrystalline calcium phosphate (CaP) composite scaffold and investigated its potential compared to plain chitosan scaffolds to be used as a bone graft substitute. Composite and chitosan scaffolds were prepared by fusing microspheres of 500-900 microm in diameter, and porosity, degradation, compressive strength, and cell growth were examined. Both scaffolds had porosities of 33-35% and pore sizes between 100 and 800 . However, composite scaffolds were much rougher and, as a result, had 20 times more surface area/unit mass than chitosan scaffolds. The compressive modulus of hydrated composite scaffolds was significantly higher than chitosan scaffolds (9.29 +/- 0.8 MPa vs. 3.26 +/- 2.5 MPa), and composite scaffolds were tougher and more flexible than what has been reported for other chitosan-CaP composites or CaP scaffolds alone. Using X-ray diffraction, scaffolds were shown to contain partially crystalline hydroxyapatite with a crystallinity of 16.7% +/- 6.8% and crystallite size of 128 +/- 55 nm. Fibronection adsorption was increased on composite scaffolds, and cell attachment was higher on composite scaffolds after 30 min, although attachment rates were similar after 1 h. Osteoblast proliferation (based on dsDNA measurements) was significantly increased after 1 week of culture. These studies have demonstrated that composite scaffolds have mechanical properties and porosity sufficient to support ingrowth of new bone tissue, and cell attachment and proliferation data indicate composite scaffolds are promising for bone regeneration.     

Homogeneous chitosan/poly(L-lactide) composite scaffolds prepared by emulsion freeze-drying

The combination between chitosan (CS)-based hydrophilic extracellular matrix polysaccharide and polylactide (PLA)-based hydrophobic biodegradable aliphatic polyester is a challenge in the biomaterials field. This study investigated the formation of homogeneous chitosan/poly(L-lactide) (CS/PLLA) porous composite scaffold using a novel emulsion freeze-drying technique. An oil-in-water (O/W) emulsification system was used in the presence of surfactant Tween-80, in which CS solution was used as the water phase and PLLA solution was used as the oil phase. The composite scaffolds showed well interconnected pore structures and homogenous distribution of CS and PLLA when the PLLA volume fraction was not higher than 50%. Once the PLLA content increased to 75%, SEM micrographs demonstrated that the two components present phase separation region. FT-IR analysis revealed that there are strong hydrogen bond interactions between CS and PLLA components. The porosity of the CS/PLLA composites was in the range of 85-90% and showed a slight decrease with increasing PLLA dose. The mechanical properties of the composites lay between that of the pure CS and the PLLA scaffold. The compressive strength increased from 0.17 to 0.21 MPa, while the compressive modulus increased from 2.37 to 3.38 MPa as the PLLA contents increased from 25 to 75%. In vitro cytotoxicity was evaluated by MTT assay. The results indicated that MC3T3-E1 cell viability and proliferation in the CS/PLLA scaffold were comparable to that in the CS scaffold, and much higher than that in the PLLA scaffold. The successful hydrophilic polysaccharide and hydrophobic polyester system offers a new delivery method of growth factors and a novel scaffold design for tissue engineering.     

Rapid Mineralization of Electrospun Scaffolds for Bone Tissue Engineering

We investigated different techniques to enhance calcium phosphate mineral precipitation onto electrospun poly(_L-lactide) (PLLA) scaffolds when incubated in concentrated simulated body fluid (SBF), 10×SBF. The techniques included the use of vacuum, pre-treatment with 0.1 M NaOH and electrospinning gelatin/PLLA blends as means to increase overall mineral precipitation and distribution throughout the scaffolds. Mineral precipitation was evaluated using environmental scanning electron microscopy, energy dispersive spectroscopy mapping and the determination of the mineral weight percents. In addition we evaluated the effect of the techniques on mechanical properties, cellular attachment and cellular proliferation on scaffolds. Two treatments, pre-treatment with NaOH and incorporation of 10% gelatin into PLLA solution, both in combination with vacuum, resulted in significantly higher degrees of mineralization (16.55 and 15.14%, respectively) and better mineral distribution on surfaces and through the cross-sections after 2 h of exposure to 10×SBF. While both scaffold groups supported cell attachment and proliferation, 10% gelatin/PLLA scaffolds had significantly higher yield stress (1.73 vs 0.56 MPa) and elastic modulus (107 vs 44 MPa) than NaOH-pre-treated scaffolds.     

Homogeneous Chitosan/Poly(L-Lactide) Composite Scaffolds Prepared by Emulsion Freeze-Drying

The combination between chitosan (CS)-based hydrophilic extracellular matrix polysaccharide and polylactide (PLA)-based hydrophobic biodegradable aliphatic polyester is a challenge in the biomaterials field. This study investigated the formation of homogeneous chitosan/poly(L-lactide) (CS/PLLA) porous composite scaffold using a novel emulsion freeze-drying technique. An oil-in-water (O/W) emulsification system was used in the presence of surfactant Tween-80, in which CS solution was used as the water phase and PLLA solution was used as the oil phase. The composite scaffolds showed well interconnected pore structures and homogenous distribution of CS and PLLA when the PLLA volume fraction was not higher than 50%. Once the PLLA content increased to 75%, SEM micrographs demonstrated that the two components present phase separation region. FT-IR analysis revealed that there are strong hydrogen bond interactions between CS and PLLA components. The porosity of the CS/PLLA composites was in the range of 85–90% and showed a slight decrease with increasing PLLA dose. The mechanical properties of the composites lay between that of the pure CS and the PLLA scaffold. The compressive strength increased from 0.17 to 0.21 MPa, while the compressive modulus increased from 2.37 to 3.38 MPa as the PLLA contents increased from 25 to 75%. In vitro cytotoxicity was evaluated by MTT assay. The results indicated that MC3T3-E1 cell viability and proliferation in the CS/PLLA scaffold were comparable to that in the CS scaffold, and much higher than that in the PLLA scaffold. The successful hydrophilic polysaccharide and hydrophobic polyester system offers a new delivery method of growth factors and a novel scaffold design for tissue engineering.     

Effects of bioactive glass nanoparticles on the mechanical and biological behavior of composite coated scaffolds

Biphasic calcium phosphates (BCP) scaffolds are widely used for bone tissue regeneration. However, brittleness, low mechanical properties and compromised bioactivities are, at present, their major disadvantages. In this study we coated the struts of a BCP scaffold with a nanocomposite layer consisting of bioactive glass nanoparticles (nBG) and polycaprolactone (PCL) (BCP/PCL-nBG) to enhance its mechanical and biological behavior. The effect of various nBG concentrations (1-90 wt.%) on the mechanical properties and in vitro behavior of the scaffolds was comprehensively examined and compared with that for a BCP scaffold coated with PCL and hydroxyapatite nanoparticles (nHA) (BCP/PCL-nHA) and a BCP scaffold coated with only a PCL layer (BCP/PCL). Introduction of 1-90 wt.% nBG resulted in scaffolds with compressive strengths in the range 0.2-1.45 MPa and moduli in the range 19.3-49.4 MPa. This trend was also observed for BCP/PCL-nHA scaffolds, however, nBG induced even better bioactivity and a faster degradation rate. The maximum compressive strength (increased ～14 times) and modulus (increased ～3 times) were achieved when 30 wt.% nBG was added, compared with BCP scaffolds. Moreover, BCP/PCL-nBG scaffolds induced the differentiation of primary human bone-derived cells (HOBs), with significant up-regulation of osteogenic gene expression for Runx2, osteopontin and bone sialoprotein, compared with the other groups.     

Surface modification of bioactive glass nanoparticles and the mechanical and biological properties of poly(L-lactide) composites

Novel bioactive glass (BG) nanoparticles/poly(L-lactide) (PLLA) composites were prepared as promising bone-repairing materials. The BG nanoparticles (Si:P:Ca=29:13:58 weight ratio) of about 40nm diameter were prepared via the sol-gel method. In order to improve the phase compatibility between the polymer and the inorganic phase, PLLA (M(n)=9700Da) was linked to the surface of the BG particles by diisocyanate. The grafting ratio of PLLA was in the vicinity of 20 wt.%. The grafting modification could improve the tensile strength, tensile modulus and impact energy of the composites by increasing the phase compatibility. When the filler loading reached around 4 wt.%, the tensile strength of the composite increased from 56.7 to 69.2MPa for the pure PLLA, and the impact strength energy increased from 15.8 to 18.0 kJ m(-2). The morphology of the tensile fracture surface of the composite showed surface-grafted bioactive glass particles (g-BG) to be dispersed homogeneously in the PLLA matrix. An in vitro bioactivity test showed that, compared to pure PLLA scaffold, the BG/PLLA nanocomposite demonstrated a greater capability to induce the formation of an apatite layer on the scaffold surface. The results of marrow stromal cell culture revealed that the composites containing either BG or g-BG particles have much better biocompatibility compared to pure PLLA material.     

Bioactive glass/polymer composite scaffolds mimicking bone tissue

The aim of this work was the preparation and characterization of scaffolds with mechanical and functional properties able to regenerate bone. Porous scaffolds made of chitosan/gelatin (POL) blends containing different amounts of a bioactive glass (CEL2), as inorganic material stimulating biomineralization, were fabricated by freeze-drying. Foams with different compositions (CEL2/POL 0/100; 40/60; 70/30 wt %/wt) were prepared. Samples were crosslinked using genipin (GP) to improve mechanical strength and thermal stability. The scaffolds were characterized in terms of their stability in water, chemical structure, morphology, bioactivity, and mechanical behavior. Moreover, MG63 osteoblast-like cells and periosteal-derived stem cells were used to assess their biocompatibility. CEL2/POL samples showed interconnected pores having an average diameter ranging from 179 ± 5 μm for CEL2/POL 0/100 to 136 ± 5 μm for CEL2/POL 70/30. GP-crosslinking and the increase of CEL2 amount stabilized the composites to water solution (shown by swelling tests). In addition, the SBF soaking experiment showed a good bioactivity of the scaffold with 30 and 70 wt % CEL2. The compressive modulus increased by increasing CEL2 amount up to 2.1 ± 0.1 MPa for CEL2/POL 70/30. Dynamical mechanical analysis has evidenced that composite scaffolds at low frequencies showed an increase of storage and loss modulus with increasing frequency; furthermore, a drop of E' and E″ at 1 Hz was observed, and for higher frequencies both moduli increased again. Cells displayed a good ability to interact with the different tested scaffolds which did not modify cell metabolic activity at the analyzed points. MTT test proved only a slight difference between the two cytotypes analyzed. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A 100A:2654-2667, 2012.     

Composite coating of bonelike apatite particles and collagen fibers on poly L-lactic acid formed through an accelerated biomimetic coprecipitation process

Collagen and apatite were coprecipitated as a composite coating on poly L-lactic acid (PLLA) in an accelerated biomimetic process. The incubation solution contained collagen (1 g/L) and simulated body fluid with 5 times inorganic ionic concentrations as human blood plasma. The coating formed on PLLA films and scaffolds after a 24-h incubation was characterized by using energy-dispersive X-ray spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy (SEM). It was shown that the coating contained carbonated bonelike apatite and collagen, which was similar in composition to natural bone. SEM showed a complex composite coating of submicron bonelike apatite particulates combined with collagen fibrils. It is expected that such biocomposite coating may better facilitate cell interaction and osteoconductivity. This work provided an efficient process to obtain bonelike apatite/collagen composite coating, which is potentially useful in bone tissue engineering.     

多孔聚乙烯醇/明胶软骨组织工程支架复合材料的制备及其生物相容性

背景：以明胶为基体制备的组织工程支架材料具有良好的生物相容性和生物降解性能,但存在力学性能低,降解速率难以控制的缺陷。 目的：制备一种软骨组织工程支架材料多孔聚乙烯醇/明胶复合物,并检测其理化性能和生物相容性。 方法：采用乳化发泡法制备聚乙烯醇/明胶多孔支架,并通过电镜分析、力学测试、皮下植入实验,检测材料孔径和孔隙率、IR光谱、力学性能和生物相容性。 结果与结论：多孔材料内部呈三维网状多孔结构,孔径均匀,有相似的孔隙率61.8%,含水率44.6%,抗拉强度为(5.01±0.03) MPa,抗压强度为(1.47±0.36) MPa,有较好的力学性能,IR光谱分析表明材料内部结构均匀。皮下植入后,炎症反应逐渐减轻,囊壁逐渐变薄,并趋于稳定,提示多孔聚乙烯醇/明胶支架材料具有较好的生物相容性和力学性能。     

Mineralization of hydroxyapatite in electrospun nanofibrous poly(L-lactic acid) scaffolds

A highly porous electrospun poly(L-lactic acid) (PLLA) nanofibrous scaffold was used as a matrix for mineralization of hydroxyapatite. The mineralization process could be initiated by immersing the electrospun scaffold in the simulated body fluids (SBF) at 37 degrees C for varying periods of time. Scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS), wide-angle X-ray diffraction (WAXD), Fourier transform infrared (FTIR), and Raman spectroscopy were used to characterize the composition and the structure of the deposited mineral on the nanofiber surface. Results indicated that the mineral phase was a carbonated apatite with thin flake-like nanostructures. The effects of functional groups on the scaffold surface and anionic additives in the incubation fluids on the nucleation and growth of the mineral were investigated. It was found that a minuscule amount of anionic additives (e.g., citric acid and poly-L-aspartic acid) in the SBF could effectively inhibit the mineral growth. Surface modification of the scaffold was carried out by hydrolysis of PLLA scaffold in NaOH aqueous solution, where carboxylic acid groups were produced without compromising the scaffold integrity. The mineralization process from modified PLLA electrospun scaffolds was significantly enhanced because the calcium ions can bind to the carboxylate groups on the fiber surface.     

Polylactic acid fibre-reinforced polycaprolactone scaffolds for bone tissue engineering

The employment of composite scaffolds with a well-organized architecture and multi-scale porosity certainly represents a valuable approach for achieving a tissue engineered construct to reproduce the middle and long-term behaviour of hierarchically complex tissues such as spongy bone. In this paper, fibre-reinforced composites scaffold for bone tissue engineering applications is described. These are composed of poly-L-lactide acid (PLLA) fibres embedded in a porous poly(epsilon-caprolactone) matrix, and were obtained by synergistic use of phase inversion/particulate leaching technique and filament winding technology. Porosity degree as high as 79.7% was achieved, the bimodal pore size distribution showing peaks at ca 10 and 200 microm diameter, respectively, accounting for 53.7% and 46.3% of the total porosity. In vitro degradation was carried out in PBS and SBF without significant degradation of the scaffold after 35 days, while in NaOH solution, a linear increase of weight lost was observed with preferential degradation of PLLA component. Subsequently, marrow stromal cells (MSC) and human osteoblasts (HOB) reached a plateau at 3 weeks, while at 5 weeks the number of cells was almost the same. Human marrow stromal cell and trabecular osteoblasts rapidly proliferate on the scaffold up to 3 weeks, promoting an oriented migration of bone cells along the fibre arrangement. Moreover, the role of seeded HOB and MSC on composite degradation mechanism was assessed by demonstrating a more relevant contribution to PLLA degradation of MSC when compared to HOB. The novel PCL/PLLA composite scaffolds thus showed promise whenever tuneable porosity, controlled degradability and guided cell-material interaction are simultaneously requested.     

Modification and cytocompatibility of biocomposited porous PLLA/HA-microspheres scaffolds

Poly(L-lactic acid) and hydroxyapatie (PLLA/HA) composite scaffolds have good properties and suit to use as bone tissue engineering. In this work, hollow HA microspheres (HAM) with poor crystallinity were fabricated by a flame-drying method. The HAM has the potential to be used to release drugs or proteins in addition to improve osteoconductivity. Different ratios of PLLA/HAM were used to prepare porous composite scaffolds using the thermally induced phase separation technique. The HAMs were randomly incorporated into the PLLA porous scaffolds. As the HAMs ratio was increased, the porous composite scaffolds changed from ladder-like into isotropic structure. In addition, the compressive strength of PLLA/HAMs composite scaffolds improved first and declined with the increasing of HAMs ratio in the scaffolds. In vitro experiment showed that PLLA/HAMs composite scaffolds improved the attachment, migration, and differentiation of osteoblastic cells. These results demonstrated that the PLLA/HAMs composite scaffolds were superior to plain PLLA scaffold for bone tissue engineering.     

Direct ink writing of highly porous and strong glass scaffolds for load-bearing bone defects repair and regeneration

The quest for synthetic materials to repair load-bearing bone lost because of trauma, cancer, or congenital bone defects requires the development of porous, high-performance scaffolds with exceptional mechanical strength. However, the low mechanical strength of porous bioactive ceramic and glass scaffolds, compared with that of human cortical bone, has limited their use for these applications. In the present work bioactive 6P53B glass scaffolds with superior mechanical strength were fabricated using a direct ink writing technique. The rheological properties of Pluronic® F-127 (referred to hereafter simply as F-127) hydrogel-based inks were optimized for the printing of features as fine as 30 μm and of three-dimensional scaffolds. The mechanical strength and in vitro degradation of the scaffolds were assessed in a simulated body fluid (SBF). The sintered glass scaffolds showed a compressive strength (136 ± 22 MPa) comparable with that of human cortical bone (100–150 MPa), while the porosity (60%) was in the range of that of trabecular bone (50–90%). The strength is ∼100-times that of polymer scaffolds and 4–5-times that of ceramic and glass scaffolds with comparable porosities. Despite the strength decrease resulting from weight loss during immersion in SBF, the value (77 MPa) is still far above that of trabecular bone after 3 weeks. The ability to create both porous and strong structures opens a new avenue for fabricating scaffolds for load-bearing bone defect repair and regeneration.     

Fabrication of three-dimensional polycaprolactone/hydroxyapatite tissue scaffolds and osteoblast-scaffold interactions in vitro

Computer-aided tissue-engineering approach was used to develop a novel precision extrusion deposition (PED) process to directly fabricate Polycaprolactone (PCL) and composite PCL/hydroxyapatite (PCL-HA) tissue scaffolds. The process optimization was carried out to fabricate both PCL and PCL-HA (25% concentration by weight of HA) with a controlled pore size and internal pore structure of the 0 degrees /90 degrees pattern. Two groups of scaffolds having 60% and 70% porosity and with pore sizes of 450 and 750 microm, respectively, were evaluated for their morphology and compressive properties using scanning electron microscopy (SEM) and mechanical testing. Our results suggested that inclusion of HA significantly increased the compressive modulus from 59 to 84 MPa for 60% porous scaffolds and from 30 to 76 MPa for 70% porous scaffolds. In vitro cell-scaffolds interaction study was carried out using primary fetal bovine osteoblasts to assess the feasibility of scaffolds for bone tissue-engineering application. The cell proliferation and differentiation were calculated by Alamar Blue assay and by determining alkaline phosphatase activity. The osteoblasts were able to migrate and proliferate over the cultured time for both PCL as well as PCL-HA scaffolds. Our study demonstrated the viability of the PED process to the fabricate PCL and PCL-HA composite scaffolds having necessary mechanical property, structural integrity, controlled pore size and pore interconnectivity desired for bone tissue engineering.     

Biphasic calcium phosphate nanocomposite porous scaffolds for load-bearing bone tissue engineering

A novel biodegradable nanocomposite porous scaffold comprising a beta-tricalcium phosphate (beta-TCP) matrix and hydroxyl apatite (HA) nanofibers was developed and studied for load-bearing bone tissue engineering. HA nanofibers were prepared with a biomimetic precipitation method. The composite scaffolds were fabricated by a method combining the gel casting and polymer sponge techniques. The role of HA nanofibers in enhancing the mechanical properties of the scaffold was investigated. Compression tests were performed to measure the compressive strength, modulus and toughness of the porous scaffolds. The identification and morphology of HA nanofibers were determined by X-ray diffraction and transmission electron microscopy, respectively. Scanning electron microscopy was used to examine the morphology of porous scaffolds and fracture surfaces to reveal the dominant toughening mechanisms. The results showed that the mechanical property of the scaffold was significantly enhanced by the inclusion of HA nanofibers. The porous composite scaffold attained a compressive strength of 9.8 +/- 0.3 MPa, comparable to the high-end value (2-10 MPa) of cancellous bone. The toughness of the scaffold increased from 1.00+/-0.04 to 1.72+/-0.02 kN/m, as the concentration of HA nanofibers increased from 0 to 5 wt %.     

Fabrication and characterization of gelatin-based biocompatible porous composite scaffold for bone tissue engineering

In this study, composite scaffolds were prepared with polyethylene oxide (PEO)-linked gelatin and tricalcium phosphate (TCP). Chitosan, a positively charged polysaccharide, was introduced into the scaffolds to improve the properties of the artificial bone matrix. The chemical and thermal properties of composite scaffolds were investigated by Fourier transform infrared spectroscopy, thermogravimetric analyzer, differential thermal analyzer. In vitro cytotoxicity of the composite scaffold was also evaluated and the sample showed no cytotoxic effect. The morphology was studied by SEM and light microscopy. It was observed that the prepared scaffold had an open interconnected porous structure with pore size of 230-354 μm, which is suitable for osteoblast cell proliferation. The mechanical properties were assessed and it was found that the composite had compressive modulus of 1200 MPa with a strength of 5.2 MPa and bending modulus of 250 MPa having strength of 12.3 MPa. The porosity and apparent density were calculated and it was found that the incorporation of TCP can reduce the porosity and water absorption. It was revealed from the study that the composite had a 3D porous microstructure and TCP particles were dispersed evenly among the crosslinked gelatin/chitosan scaffold. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 100A:3020-3028, 2012.     

Calcium phosphate deposition rate,structure and osteoconductivity on electrospun poly(l-lactic acid) matrix using electrodeposition or simulated body fluid incubation

Mineralized nanofibrous scaffolds have been proposed as promising scaffolds for bone regeneration due to their ability to mimic both nanoscale architecture and chemical composition of natural bone extracellular matrix. In this study, a novel electrodeposition method was compared with an extensively explored simulated body fluid (SBF) incubation method in terms of the deposition rate, chemical composition and morphology of calcium phosphate formed on electrospun fibrous thin matrices with a fiber diameter in the range ∼200–1400 nm prepared using 6, 8, 10 and 12 wt.% poly(l-lactic acid) (PLLA) solutions in a mixture of dichloromethane and acetone (2:1 in volume). The effects of the surface modification using the two mineralization techniques on osteoblastic cell (MC3T3-E1) proliferation and differentiation were also examined. It was found that electrodeposition was two to three orders of magnitude faster than the SBF method in mineralizing the fibrous matrices, reducing the mineralization time from ∼2 weeks to 1 h to achieve the same amounts of mineralization. The mineralization rate also varied with the fiber diameter but in opposite directions between the two mineralization methods. As a general trend, the increase of fiber diameter resulted in a faster mineralization rate for the electrodeposition method but a slower mineralization rate for the SBF incubation method. Using the electrodeposition method, one can control the chemical composition and morphology of the calcium phosphate by varying the electric deposition potential and electrolyte temperature to tune the mixture of dicalcium phosphate dihydrate and hydroxyapatite (HAp). Using the SBF method, one can only obtain a low crystallinity HAp. The mineralized electrospun PLLA fibrous matrices from either method similarly facilitate the proliferation and osteogenic differentiation of preosteoblastic MC3T3-E1 cells as compared to neat PLLA matrices. Therefore, the electrodeposition method can be utilized as a fast and versatile technique to fabricate mineralized nanofibrous scaffolds for bone tissue engineering.