Osteogenic Properties of Novel Methylsulfonylmethane-Coated Hydroxyapatite Scaffold

Despite numerous advantages of using porous hydroxyapatite (HAp) scaffolds in bone regeneration, the material is limited in terms of osteoinduction. In this study, the porous scaffold made from nanosized HAp was coated with different concentrations of osteoinductive aqueous methylsulfonylmethane (MSM) solution (2.5, 5, 10, and 20%) and the corresponding MH scaffolds were referred to as MH2.5, MH5, MH10, and MH20, respectively. The results showed that all MH scaffolds resulted in burst release of MSM for up to 7 d. Cellular experiments were conducted using MC3T3-E1 preosteoblast cells, which showed no significant difference between the MH2.5 scaffold and the control with respect to the rate of cell proliferation (p > 0.05). There was no significant difference between each group at day 4 for alkaline phosphatase (ALP) activity, though the MH2.5 group showed higher level of activity than other groups at day 10. Calcium deposition, using alizarin red staining, showed that cell mineralization was significantly higher in the MH2.5 scaffold than that in the HAp scaffold (p < 0.0001). This study indicated that the MH2.5 scaffold has potential for both osteoinduction and osteoconduction in bone regeneration.     

Effect of SiO2 in situ cross-linked CS/PVA on SrFe12O19 scaffolds prepared by 3D gel printing for targeting

The design of magnetic composite scaffolds with superior properties has the potential to construct a targeted delivery platform with hyperthermia. In this study, strontium hexaferrite (SrFe₁₂O₁₉, SrM) magnetic nanoparticles (MNPs) were obtained by the chemical precipitation method. Non-toxic cross-linked biogels were prepared for adhesive ceramic scaffolds, and chitosan/polyvinylalcohol (CS/PVA)-bonded SrM magnetic nanoscaffolds were successfully prepared by 3D gel printing (3DGP) method. The effects of PVA physical cross-linking and in situ formed SiO₂ on the properties of CS-bonded scaffolds were evaluated, and the compressive strengths were increased from 6.13 ± 2.45 MPa to 8.80 ± 2.02 MPa and 17.18 ± 2.15 MPa, respectively. The results showed that the saturation magnetization of SiO₂/CS/PVA/SrM composite scaffolds was 59.96 emu/g. In vitro immersion experiments showed that the degradation rates of SiO₂/CS/PVA/SrM scaffolds were 4.90% after 28 days, and the in situ SiO₂ improved the deposition of calcium salts on the scaffolds. The experiments showed that the SrM magnetic scaffolds could not only concentrate magnetic fields to improve the efficiency of targeting deposition but also achieve a weak targeting process without external magnetic field assistance. In vitro cell proliferation test showed that MC3T3-E1 cells had good adhesion and proliferation on the surface of SiO₂/CS/PVA/SrM scaffolds, which indicated that the scaffolds may be used for bone repairing.     

Effect of Morphological Characteristics and Biomineralization of 3D-Printed Gelatin/Hyaluronic Acid/Hydroxyapatite Composite Scaffolds on Bone Tissue Regeneration

The use of porous three-dimensional (3D) composite scaffolds has attracted great attention in bone tissue engineering applications because they closely simulate the major features of the natural extracellular matrix (ECM) of bone. This study aimed to prepare biomimetic composite scaffolds via a simple 3D printing of gelatin/hyaluronic acid (HA)/hydroxyapatite (HAp) and subsequent biomineralization for improved bone tissue regeneration. The resulting scaffolds exhibited uniform structure and homogeneous pore distribution. In addition, the microstructures of the composite scaffolds showed an ECM-mimetic structure with a wrinkled internal surface and a porous hierarchical architecture. The results of bioactivity assays proved that the morphological characteristics and biomineralization of the composite scaffolds influenced cell proliferation and osteogenic differentiation. In particular, the biomineralized gelatin/HA/HAp composite scaffolds with double-layer staggered orthogonal (GEHA20-ZZS) and double-layer alternative structure (GEHA20-45S) showed higher bioactivity than other scaffolds. According to these results, biomineralization has a great influence on the biological activity of cells. Hence, the biomineralized composite scaffolds can be used as new bone scaffolds in bone regeneration.     

Preparation and characterization of Chitosan and PLGA‐based scaffolds for tissue engineering applications

Three dimensional (3D) biodegradable porous scaffolds play a crucial role in bone tissue repair. In this study, four types of 3D polymer/hydroxyapatite (HAp) composite scaffolds were prepared by freeze drying technique in order to mimic the organic/inorganic nature of the bone. Chitosan (CH) and poly(lactic acid‐co‐glycolic acid) (PLGA) were used as the polymeric part and HAp as the inorganic component. Properties of the resultant scaffolds, such as morphology, porosity, degradation, water uptake, mechanical and thermal stabilities were examined. 3D scaffolds having interconnected macroporous structure and 77–89% porosity were produced. The pore diameters were in the range of 6 and 200 µm. PLGA and HAp containing scaffolds had the highest compressive modulus. PLGA maintained the strength by decreasing water uptake but increased the degradation rate. Scaffolds seeded with SaOs‐2 osteoblast cells showed that all scaffolds were capable of encouraging cell adhesion and proliferation. The presence of HAp particles caused an increase in cell number on CH‐HAp scaffolds compared to CH scaffolds, while cell number decreased when PLGA was incorporated in the structure. CH‐PLGA scaffolds showed highest cell number on days 7 and 14 compared to others. Based on the properties such as interconnected porosity, high mechanical strength, and in vitro cell proliferation, blend scaffolds have the potential to be applied in hard tissue treatments. POLYM. COMPOS., 36:1917–1930, 2015. © 2014 Society of Plastics Engineers     

3D gel printing of magnetic hydrogel scaffolds assisted by in-situ gelation in a water level-controlled crosslinker bath

Polyvinylalcohol/chitosan (PVA/CS) is an excellent dual-network hydrogel material, but some significant challenges remain in fabricating composites with specific structures. In this study, 3D gel printing (3DGP) combined with a water-level controlled crosslinker bath was proposed for the rapid in-situ prototyping of PVA/CS/Fe₃O₄ magnetic hydrogel scaffolds. Specifically, the PVA/CS/Fe₃O₄ hydrogels were extruded into the crosslinker water to achieve rapid in-situ gelation, improving the printability of hydrogel scaffolds. The effect of the PVA/CS ratio on the rheological and mechanical properties of dual-network magnetic hydrogels was evaluated. The printing parameters were systematically optimized to facilitate the coordination between the crosslinking water bath and printer. The different crosslinking water baths were investigated to improve the printability of PVA/CS/Fe₃O₄ hydrogels. The results showed that the printability of the sodium hydroxide (NaOH) crosslinker was significantly better than that of sodium tripolyphosphate (TPP). The magnetic hydrogels (PVA: CS= 1: 1) crosslinked by NaOH had better compressive strength, swelling rate, and saturation magnetization of 1.17 MPa, 92.43%, and 22.19 emu/g, respectively. The MC3T3-E1 cell culture results showed that the PVA/CS/Fe₃O₄ scaffolds promoted cell adhesion and proliferation, and the scaffolds crosslinked by NaOH had superior cytocompatibility. 3DGP combined with a water-level controlled crosslinker bath offers a promising approach to preparing magnetic hydrogel materials.     

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     

Size and morphology-controlled synthesis of mesoporous hydroxyapatite nanocrystals by microwave-assisted hydrothermal method

We report the rapid microwave-assisted hydrothermal synthesis of mesoporous hydroxyapatite (HAp) nanocrystals with controlled size, morphology, and surface area using various organic modifiers as regulators. The products were analyzed for their crystalline nature, phase purity, morphology, particle size and pore size distribution. Results indicated that ascorbic acid, cetyltrimethyl ammonium bromide (CTAB) and polyvinylpyrrolidone (PVP) play an important role to obtain needle like, rod like and fiber like mesoporous HAp nanocrystals with different specific surface area by controlling growth habit of HAp along c-axis. In addition, the prepared samples were B-type carbonated HAp similar to bone minerals. Therefore, the present approach can be a promising way to obtain precursor for making tissue engineering scaffolds, drug/protein delivery carriers and bone fillers with tunable characteristics.     

Influences of doping approach on conductivity of composite alkaline solid polymer electrolyte PVA–HA–KOH–H2O

Hydroxyapatite (HA) was selected as dopant and synthesized by a sonochemical method in this study. PVA–HA–KOH–H₂O composite alkaline solid polymer electrolyte (ASPE) was prepared by solution casting method. Two different doping routes, i.e., the physical mixing (PM) and in situ synthesis (ISS) were considered. The influences of the ASPE composition such as HA content, KOH content and water content on the conductivity were investigated. The ASPE prepared by ISS route has a porous structure which is important for ion transfer. Accordingly, the conductivity of the ASPE prepared by ISS is higher than that of by PM. A highest conductivity of about 0.1 S/cm is reached for ASPE prepared by ISS route at 70% water content for m(PVA):m(KOH):m(HA) = 10:14:10 system, higher than that of by PM route, 0.08 S/cm. The nickel/zinc cell shows better electrochemical properties assembled with the ASPE prepared by ISS approach than by PM approach.     

Three-dimensional silk impregnated HAp/PHBV nanofibrous scaffolds for bone regeneration

Three-dimensional silk fibroin impregnated poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) nanofibrous scaffolds with or without hydroxyapatite (HAp) were prepared by wet-electrospinning method followed by freeze-drying. Scaffolds with cotton wool-like structure have the average fiber diameter of 450–850?nm with 80–85% porosity. In-vitro cell culture tests using MG-63 osteosarcoma human cells revealed improved cell viability, alkaline phosphatase (ALP) activity and total cellular protein amount on the silk impregnated scaffolds compared to PHBV and HAp/PHBV scaffolds after 10 days of cell culture. Immunohistochemical analyses on the silk impregnated scaffolds showed that HAp triggered cell penetration and type I collagen production. Besides, HAp mineralization tendency increased with a decrease in percent crystallinity of the scaffolds comprising HAp and silk after 4 weeks of incubation in simulated body fluid. Consequently, cotton wool-like HAp/PHBV-SF scaffold would be a promising candidate as a bone-filling material for tissue regeneration.     

Effects of pore size on the mechanical and biological properties of stereolithographic 3D printed HAp bioceramic scaffold

In this study, hydroxyapatite (HAp) scaffolds with the pore size of 400, 500, and 600 μm were prepared by stereolithographic 3D printing (SL-3DP). The effects of pore size on mechanical and biological properties of the HAp scaffolds were investigated. Firstly, the macro- and microstructure of the HAp scaffolds were observed. Then, the compressive strength of the HAp scaffolds were tested. Finally, the biological properties of the HAp scaffolds were further characterized in vitro by the synthetic body fluid (SBF) solution immersion testing, as well as by using the cell proliferation and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. From this study, it was found that the HAp scaffold with a pore size of 600 μm had the most promising application prospect.     

Natural origin hydroxyapatite scaffold as potential bone tissue engineering substitute

Fish scales derived natural hydroxyapatite (FS-HAp) scaffolds were prepared through solvent casting technique, which could mimic the structure of cortical and cancellous bone tissues of body system. The hydroxyapatite (HAp) biomaterial was synthesized by thermal decomposition of chemically treated fish scales. Fabricated scaffolds were characterized through morphological analysis, volumetric shrinkage, mechanical tests, and in vitro, in vivo biological studies. The projected scaffolds successfully mimic the cancellous/cortical bone system in terms of structure, porosity, mechanical strength, and exhibit excellent bioactive behavior. The FS-HAp scaffolds manifest good mechanical behaviors with Vickers Hardness (HV) of ~0.78 GPa, 0.52 GPa compressive stress, 190 MPa tensile stress and ~35% porosity on sintering at 1200 °C. In vitro and in vivo studies suggest these nontoxic HAp scaffolds graft with osteoconductive support, facilitating new cell growth on the developed scaffold surface. The graded grafts have a great potential for application as traumatized tissue augmentation substitute, and ideal for load-bearing bone applications.     

Nanohydroxyapatite-coated hydroxyethyl cellulose/poly (vinyl) alcohol electrospun scaffolds and their cellular response

The present study focused on the preparation of nanohydroxyapatite (nHA)-coated hydroxyethyl cellulose/polyvinyl alcohol (HEC/PVA) nanofibrous scaffolds for bone tissue engineering application. The electrospun HEC/PVA scaffolds were mineralized via alternate soaking process. FESEM revealed that the nHA was formed uniformly over the nanofibers. The nHA mineralization enhanced the tensile strength and reduced the elongation at breakage of scaffolds. The wettability of the nanofibrous scaffolds was significantly improved. The in vitro biocompatibility of scaffolds was evaluated with human osteosarcoma cells. nHA-coated scaffolds had a favorable effect on the proliferation and differentiation of osteosarcoma cell and could be a potential candidate for bone regeneration.     

In situ synthesis and characterization of nano-size hydroxyapatite in poly(vinyl alcohol) matrix

Hydroxyapatite (HAp) crystals were prepared via an in situ biomimetic process in the presence of poly(vinyl alcohol) (PVA). The effect of polymer amount and its molecular weight on the physical properties of the HAp crystals were investigated. X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FT-IR) analysis, confirmed the formation of the crystalline HAp at room temperature. Microstructural features such as size and morphology of the resulting HAp samples were characterized using BET, scanning and transmission electron microscopy. The results indicate that the development (size and shape) of the HAp nanocrystals precipitated in an aqueous solution of PVA was influenced by the molecular weight of the polymer in such a way that smallest crystallite size was observed in the case of PVA with the highest molecular weight. It is believed that the HAp formation was initiated through the interaction of Ca²⁺ ions with the negative side groups on the polymer surface. The larger number of reaction sites in the PVA polymer with higher molecular weight led to a higher number of HAp nuclei and therefore smaller crystallite size.     

Electrospun poly(L‐lactic acid)/hydroxyapatite composite fibrous scaffolds for bone tissue engineering

Poly(L ‐lactic acid) (PLLA) is one of the most studied synthetic biodegradable polymeric materials as a bone graft substitute. Taking into account the osteoconductive property of hydroxyapatite (HAp), we prepared fibrous matrices of PLLA without and with HAp particles in amounts of 0.25 or 0.50% (w/v, based on the volume of the base 15% w/v PLLA solution in 70:30 v/v dichloromethane/tetrahydrofuran). These fibrous matrices were assessed for their potential as substrates for bone cell culture. The presence of HAp in the composite fibre mats was confirmed using energy dispersive X‐ray spectroscopy mapping. The average diameters of both neat PLLA and PLLA/HAp fibres, as determined using scanning electron microscopy, ranged between 2.3 and 3.5 µm, with the average spacing between adjacent fibres ranging between 5.7 and 8.5 µm. The porosity of these fibrous membranes was high (ca 97–98%). A direct cytotoxicity evaluation with L929 mouse fibroblasts indicated that the neat PLLA fibre mats released no substance at a level that was toxic to the cells. The presence of HAp particles at 0.50% w/v in the PLLA fibrous scaffolds not only promoted the attachment and the proliferation of MC3T3‐E1 mouse pre‐osteoblastic cells, but also increased the expression of osteocalcin mRNA and the extent of mineralization after the cells had been cultured on the scaffolds for 14 and 21 days, respectively. The results obtained suggested that the PLLA/HAp fibre mats could be materials of choice for bone tissue engineering. Copyright © 2009 Society of Chemical Industry     

In situ grown fibrous composites of poly(dl-lactide) and hydroxyapatite as potential tissue engineering scaffolds

In situ grown hydroxyapatite (HA) within electrospun poly(dl-lactide) (PDLLA) fibers were initially investigated as potential tissue engineering scaffolds with respect to the mechanical performances, biomineralization capability, degradation behaviors, cell growth and differentiation profiles. The tensile strength and Young’s moduli of in situ grown composites (IGC) were 8.2 ± 1.1 and 63.5 ± 5.6 MPa, respectively, which were significantly higher than those of blend electrospun composites (BEC) with 25.2% of HA inoculation. The interactions between HA and matrix polymers were approved by the red-shifts of CO stretching and OH- stretching modes and the increases in glass transition temperatures of fibrous composites. The localization of apatite phase on the fiber surface improved the biomineralization capability and enhanced the morphological stability of the fibers and fibrous mats even when the degradation of matrix polymers was detected. The cell viability and alkaline phosphatase levels were significantly higher for composites IGC, indicating favorable scaffolds for cell proliferation and osteogenic differentiation.     

Counterions present in syntheses induce the precipitation of two different populations of Sr-containing hydroxyapatite crystals

The ions found in biological environment during biomineralization as well as the counterions present in syntheses, i.e., the reagents employed, play an important role in the precipitation and stabilization of apatites. In this work, precipitation of strontium-containing hydroxyapatites at different concentrations (0, 20, 40, 60, 80 and 100% Sr²⁺) was performed in the presence of two counterions typically present in biological environments: Na⁺ and Cl⁻. It was demonstrated that the presence of Na⁺/Cl⁻ in the synthesis environment led to the precipitation of a biphasic hydroxyapatite (HAp) system formed by two non-miscible phases: Ca-rich HAp and Sr-rich HAp. The biphasic system was observed for intermediate Sr²⁺ concentrations (20, 40 and 60%) and exhibited greater lattice microstrain compared with the single-phase systems of Ca HAp (0%) and Sr HAp (100%). Although Na⁺/Cl⁻ were inserted into both Ca- and Sr-rich HAp phases, Cl⁻ ions were preferentially accommodated into the enlarged Sr-rich HAp structure. The presence of Cl⁻ ions in the Sr-rich HAp phase decreased microstrain compared to the Ca-rich phase. After calcination, the biphasic system was transformed into a completely miscible Ca/Sr HAp phase without the formation of other phosphates or oxides.     

Development of bone scaffold using Puntius conchonius fish scale derived hydroxyapatite: Physico-mechanical and bioactivity evaluations

Naturally derived Hydroxyapatite (HAp) from fish scale is finding wide applications in the development of bone scaffold to promote bone regeneration. But porous HAp scaffold is fragile in nature making it unsuitable for bone repair or replacement applications. Thus, it is essential to improve the mechanical property of HAp scaffolds while retaining the interconnected porous structure for tissue ingrowth in vivo. In this study solvent casting particulate leaching technique is used to develop novel Puntius conchonius fish scale derived HAp bone scaffold by varying the wt.% of the HAp from 60 to 80% in PMMA matrix. Physico-chemical, mechanical, structural and bioactive properties of the developed scaffolds are investigated. The obtained results indicate that HAp-PMMA scaffold at 70?wt % HAp loading shows optimal properties with 7.26?±?0.45?MPa compressive strength, 75?±?0.8% porosity, 8.0?±?0.68% degradation and 190?±?11% water absorption. The obtained results of the scaffold can meet the physiological demands to guide bone regeneration. Moreover, in vitro bioactivity analysis also confirms the formation of bone like apatite in the scaffold surface after 28 days of SBF immersion. Thus, the developed scaffold has the potential to be effectively used in bone tissue engineering applications.     

Effect of ZnO reinforcement on the compressive properties,in vitro bioactivity,biodegradability and cytocompatibility of bone scaffold developed from bovine bone-derived HAp and PMMA

In this study, the effect of zinc oxide (ZnO) incorporation on the properties of Hydroxyapatite (HAp)/Poly(methyl methacrylate) (PMMA)/ZnO based composite bone scaffold is investigated. HAp is derived from calcination of bovine bone bio-waste and ZnO is synthesized by direct precipitation technique. Porous scaffolds are developed by gas foaming process using ammonium bicarbonate as the foaming agent and adding ZnO nanoparticles (NPs) at 2.5, 5, 7.5 and 10% (w/w) respectively. Incorporation of ZnO up to 5% (w/w) is found to significantly enhance the porosity, compressive strength, thermal stability and swelling properties of the developed scaffolds. In-vitro bioactivity and biodegradability assessment using simulated body fluid (SBF) show improved results of 5% ZnO loaded scaffolds. Furthermore, the composite scaffold show enhanced cytocompatibility during the in vitro cytotoxicity test performed using XTT assay. A comprehensive study on the scaffold properties shows that 5% ZnO composite scaffold exhibits the best-optimized properties suitable for bone tissue engineering applications.     

Fabrication of MgAl2O4 Spinel Scaffolds and Sonochemical Synthesis and Deposition of Hydroxyapatite Nanorods

Hydroxyapatite (HAp) is one of the most emerging biocompatible ceramic widely used as scaffolds in various biomedical applications such as orthopedics and dentistry. In spite of the superior properties for biomedical applications, they exhibit poor mechanical properties. This has lead to the concept of fabricating composite out of bioactive and bioinert material to derive bioactivity in combination with desirable mechanical properties. In this study, bioinert magnesium aluminate (MgAl₂O₄) spinel scaffolds are prepared through polymeric foam replication process followed by sintering. Sintered foams have exhibited good structural integrity and the stress–strain curves recorded during the uniaxial compression have shown a plateau indicating high‐energy absorption capability. Sonochemical process has been employed for the simultaneous synthesis and deposition of HAp formulation from the stoichiometric solution of precursors on the spinel scaffolds. Sonochemical process has resulted in the formation of phase pure HAp with unique morphology of nanorods coated throughout the three‐dimensional foam structures. Cytotoxicity evaluation of this new scaffold material has not shown any alteration in the viability, growth, and morphology of the cells. The new scaffold material thus developed is expected to have high potential for biomedical applications.     

Evaluation of mechanical and biocompatibility properties of hydroxyapatite/manganese dioxide nanocomposite scaffolds for bone tissue engineering application

The aim of this research was to evaluate the mechanical properties, biocompatibility, and degradation behavior of scaffolds made of pure hydroxyapatite (HA) and HA-modified by MnO₂ for bone tissue engineering applications. HA and MnO₂ were developed using sol-gel and precipitation methods, respectively. The scaffolds properties were characterized using X-ray diffraction (XRD), Fourier transform spectroscopy (FTIR), scanning electron microcopy (SEM), energy dispersive spectroscopy (EDS), and transmission electron microscopy (TEM). The interaction of scaffold with cells was assessed using in vitro cell proliferation and alkaline phosphatase (ALP) assays. The obtained results indicate that the HA/MnO₂ scaffolds possess higher compressive strength, toughness, hardness, and density when compared to the pure HA scaffolds. After immersing the scaffold in the SBF solution, more deposited apatite appeared on the HA/MnO₂, which results in the rougher surface on this scaffold compared to the pure HA scaffold. Finally, the in vitro biological analysis using human osteoblast cells reveals that scaffolds are biocompatible with adequate ALP activity.