Materials For Bone Tissue Engineering Research Articles

Surface-immobilized plant-derived osteopontin as an effective platform to promote osteoblast adhesion and differentiation.

In this report, recombinant human osteopontin synthesized in tobacco plants (p-rhOPN) is introduced as a potential bioactive molecule that can promote osteoblast adhesion and differentiation. A glass substrate (SiO2/Si-OH) grafted with poly(acrylic acid) (SiO2/Si-PAA) was prepared by surface-initiated reversible addition-fragmentation chain transfer polymerization and used as a carboxyl-rich platform for the chemical conjugation of p-rhOPN. The PAA grafting and subsequent p-rhOPN immobilization were confirmed by water contact angle, Fourier transform-infrared spectroscopy, X-ray photoelectron spectroscopy and atomic force microscopy analyses. Indirect ELISA quantification revealed that the p-rhOPN immobilization efficiency was above 95% and the surface coverage was a function of the p-rhOPN concentration. MC-3T3-E1 cells cultured on the SiO2/Si-PAA substrate immobilized with various concentrations (0.6-30 ng/mL) of p-rhOPN (SiO2/Si-p-rhOPN) exhibited superior cell spreading compared to those cultured on SiO2/Si-OH or gelatin-modified glass substrate (SiO2/Si-gelatin). Polymerase chain reaction analysis indicated that the SiO2/Si-p-rhOPN substrates with high level of immobilized p-rhOPN promoted MC-3T3-E1 cell differentiation, as demonstrated by the higher transcript expression levels of the osteogenic differentiation regulatory gene, Runt-related transcription factor 2, compared to cells cultured on SiO2/Si-OH or SiO2/Si-gelatin. Given that p-rhOPN can be more economically produced than the commercially available OPN derived from human or mammalian sources, then, together with its well-preserved biological function in spite of being chemically conjugated to the substrates, it is likely that p-rhOPN could be more broadly applied for the development of materials for bone tissue engineering with a promising medical and commercial value.

Antibacterial Surface Coating for Bone Scaffolds Based on the Dark Catalytic Effect of Titanium Dioxide.

Biomaterials which promote tissue integration and resist microbial colonisation are required in bone tissue engineering to prevent biomaterial-associated infections. Surface modification of established materials for bone tissue engineering, such as TiO2, have emerged as promising anti-infective strategies. Interestingly, the antibacterial activity of TiO2 in the form of particles can be enhanced by combining it with H2O2, even in the absence of irradiation. However, it remains unknown whether TiO2 surfaces elicit a similar effect. In this study, the antibacterial effect of porous TiO2 scaffolds generated by the catalytic decomposition of H2O2 in the absence of light (dark catalysis) was investigated. Porous ceramic foams were fabricated and sol-gel coated for high catalytic activity. Degradation of methylene blue in the presence of 3% H2O2 increased by 80% for the sol-gel-coated surfaces. The degradation kinetics indicate that intermediate free radicals that form at the liquid-TiO2 interface are responsible for the oxidative behavior of the surface. TiO2 surfaces were further pretreated with 30% H2O2 for prolonged oxidative behavior. The biological response toward such surfaces was assessed in vitro. S. epidermidis biofilms formed on modified surfaces showed reduced viability compared to nonmodified surfaces. Further, the same surface modification showed no cytotoxic effects on MC3T3 preosteoblasts. However, the results from the conducted genotoxicity assay were inconclusive, and further studies are needed to exclude ROS-mediated DNA damage. To conclude, this study provides evidence that a simple surface modification based on the dark catalytic effect of TiO2 can be used to create antibacterial surface properties for ceramic bone scaffolds.

Trends in Development of Bioresorbable Calcium Phosphate Ceramic Materials for Bone Tissue Engineering

The review of results obtained by the authors in the design of bioresorbable calcium phosphate ceramic materials intended for bond tissue defect regeneration is presented. The development of composite materials and new data in the fabrication of octacalcium phosphate bioresorbable ceramic are elucidated as well.

Encapsulation of Rat Bone Marrow Derived Mesenchymal Stem Cells in Alginate Dialdehyde/Gelatin Microbeads with and without Nanoscaled Bioactive Glass for In Vivo Bone Tissue Engineering.

Alginate dialdehyde (ADA), gelatin, and nano-scaled bioactive glass (nBG) particles are being currently investigated for their potential use as three-dimensional scaffolding materials for bone tissue engineering. ADA and gelatin provide a three-dimensional scaffold with properties supporting cell adhesion and proliferation. Combined with nanocristalline BG, this composition closely mimics the mineral phase of bone. In the present study, rat bone marrow derived mesenchymal stem cells (MSCs), commonly used as an osteogenic cell source, were evaluated after encapsulation into ADA-gelatin hydrogel with and without nBG. High cell survival was found in vitro for up to 28 days with or without addition of nBG assessed by calcein staining, proving the cell-friendly encapsulation process. After subcutaneous implantation into rats, survival was assessed by DAPI/TUNEL fluorescence staining. Hematoxylin-eosin staining and immunohistochemical staining for the macrophage marker ED1 (CD68) and the endothelial cell marker lectin were used to evaluate immune reaction and vascularization. After in vivo implantation, high cell survival was found after 1 week, with a notable decrease after 4 weeks. Immune reaction was very mild, proving the biocompatibility of the material. Angiogenesis in implanted constructs was significantly improved by cell encapsulation, compared to cell-free beads, as the implanted MSCs were able to attract endothelial cells. Constructs with nBG showed higher numbers of vital MSCs and lectin positive endothelial cells, thus showing a higher degree of angiogenesis, although this difference was not significant. These results support the use of ADA/gelatin/nBG as a scaffold and of MSCs as a source of osteogenic cells for bone tissue engineering. Future studies should however improve long term cell survival and focus on differentiation potential of encapsulated cells in vivo.

Industrial-scale fabrication of an osteogenic and antibacterial PLA/silver-loaded calcium phosphate composite with significantly reduced cytotoxicity.

In this study, we report an industrial-scale fabrication method of a multifunctional polymer composite as a scaffold material for bone tissue engineering. This study successfully demonstrated the potential of applying industrial polymer processing technologies to produce specially functionalized tissue engineering scaffolds. With the inclusion of a newly synthesized multifunctional additive, silver-doped-calcium phosphate (silver-CaP), the composite material exhibited excellent osteogenic inducibility of human adipose-derived stem cells (hASC) and satisfactory antibacterial efficacy against Escherichia coli and Staphylococcus aureus. Also, relative to previously reported methods of direct loading silver particles into polymeric materials, our composite exhibited significantly reduced silver associated cytotoxicity. The enhanced biocompatibility could be a significant advantage for materials to be used for regenerative medicine applications where clinical safety is a major consideration. The impact of different silver loading methodologies on hASC' osteogenic differentiation was also studied. Overall, the results of this study indicate a promising alternative approach to produce multifunctional scaffolds at industrial-scale with higher throughput, lower cost, and enhanced reproducibility. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 900-910, 2019.

Preparation and osteogenic properties of poly ( L-lactic acid)/lecithin porous scaffolds with open pore structure

To investigate the preparation and osteogenic properties of poly ( L-lactic acid)(PLLA)/lecithin porous scaffolds with open pore structure. PLLA/lecithin porous scaffolds with different lecithin contents (0, 5%, 10%, 20%, 30%, 40%, 50%) were prepared by thermally induced phase separation (groups A, B, C, D, E, F, and G, respectively). Scanning electron microscopy (SEM) was used to observe the surface morphology of the scaffolds. Wide-angle X-ray diffraction (XRD) and differential scanning calorimetry (DSC) were used to detect the crystallinity of the scaffolds. The water uptake ability of the scaffolds was measured. The cell growth and viability of bone marrow mesenchymal stem cells (BMSCs) of mouse on each scaffold was assessed by cell counting kit 8 (CCK-8) method. The osteogenic differentiation ability of BMSCs on each scaffold was evaluated by alkaline phosphatase (ALP) activity. Finally, a critical-size rat calvarial bone defect model was used to evaluate the osteogenesis of the scaffolds in vivo. Micro-CT was used to reconstruct the three-dimensional model of the defect area, and the bone volume and bone mineral density were quantitatively analyzed. SEM results showed that the lecithin could slightly reduce the pore size; when lecithin content was 50%, platelet-like structure could be observed on the scaffolds. Wide angle XRD and DSC showed that the crystallinity of scaffolds gradually decreased with the increase of lecithin content. The water uptake ability test showed that the hydrophilicity of scaffolds increased with the increase of lecithin content. CCK-8 assay showed that cell activity gradually increased with the increase of culture time. After 7 days of culture, the absorbance ( A) value of groups C, D, E, and F were significantly higher than that of groups A, B, and G ( P<0.05), but no significant difference was found among groups C, D, E, and F ( P>0.05). After 14 days of osteogenic induction, with the increase of lecithin content, there was a significant difference in ALP activity of each group. The ALP activity in groups D, E, F, and G were significantly higher than that in groups A, B, and C ( P<0.05). In vivo, the results of Micro-CT examination and bone volume and bone mineral density showed that the scaffolds with 30% lecithin had the best repairing effect. Prepared by thermally induced phase separation, the cytocompatibility, osteogenic differentiation, and bone repair ability of the PLLA/lecithin porous scaffold is obviously better than that of pure PLLA scaffold. PLLA/lecithin porous scaffold with suitable lecithin content is a promising scaffold material for bone tissue engineering.

Aligned porous chitosan/graphene oxide scaffold for bone tissue engineering

Chitosan/graphene oxide composites (CGs) were developed using directional freezing to create aligned porous three-dimensional (3D) scaffolds resembling bone lamellae. The contact provided by the anisotropic scaffolds played a critical role in guiding the alignment of MC3T3-E1 cells along the longitudinal direction. The higher mechanical strength, shape-memory, cell alignment guiding capabilities, and protein adsorption ability, render these composites potentially useful as scaffold materials in bone tissue engineering.

A Review on Properties of Natural and Synthetic Based Electrospun Fibrous Materials for Bone Tissue Engineering.

Bone tissue engineering is an interdisciplinary field where the principles of engineering are applied on bone-related biochemical reactions. Scaffolds, cells, growth factors, and their interrelation in microenvironment are the major concerns in bone tissue engineering. Among many alternatives, electrospinning is a promising and versatile technique that is used to fabricate polymer fibrous scaffolds for bone tissue engineering applications. Copolymerization and polymer blending is a promising strategic way in purpose of getting synergistic and additive effect achieved from either polymer. In this review, we summarize the basic chemistry of bone, principle of electrospinning, and polymers that are used in bone tissue engineering. Particular attention will be given on biomechanical properties and biological activities of these electrospun fibers. This review will cover the fundamental basis of cell adhesion, differentiation, and proliferation of the electrospun fibers in bone tissue scaffolds. In the last section, we offer the current development and future perspectives on the use of electrospun mats in bone tissue engineering.

Osteogenic extracellular matrix sheet for bone tissue regeneration.

The application of extracellular matrix (ECM) sheets without a scaffold is not extensively reported in bone regenerative medicine. The aim of the present study was to demonstrate that an osteogenic ECM sheet (OECMS) can retain ECM integrity and growth factors to enhance bone formation in a rat non-union model. OECMS was produced from osteogenic cell sheets (OCS). Collagen and growth factor [bone morphogenetic protein 2 (BMP-2), vascular endothelial growth factors (VFGFs), basic fibroblast growth factor (bFGF) and transforming growth factor β1 (TGF-β1)] concentrations in the OECMS were quantified by enzyme-linked immunosorbent assay (ELISA). Next, hydroxyapatite (HA) constructs combined with OECMSs were implanted subcutaneously into the rats' backs to evaluate their osteoinductive capacity by histological evaluation. In addition, OECMSs were implanted in a rat femoral non-union model. 18 male Fischer 344 inbred rats were divided into OECMS and control groups. Fracture healing was evaluated by radiological and histological analyses at 2, 5 and 8 weeks and biological analysis at 8 weeks. Collagen I and growth factors were retained in the OECMSs. Osteoid formation was identified in the HA combined with OECMS at 4 weeks. Enhanced bone regeneration at the non-union of the OECMS group was confirmed at 5 and 8 weeks. Biomechanical testing revealed a significantly higher maximum bending load in the OECMS group as compared to the control group at 8 weeks. The results demonstrated that OECMS retained BMP-2 and TGF-β1 and high osteoinductive and osteoconductive capacity. As such, OECMS represents a potential new scaffold-free material for bone tissue engineering.

State of the Art Biofabrication Technologies and Materials for Bone Tissue Engineering

Bone tissue engineering is a field whose relevance is paramount, especially for the treatment of musculoskeletal-related disabilities. Failure of conventional methods to create physiologically relevant bone materials has prompted exploring several 3D-printing and additive manufacturing processes, including bioprinting, selective laser sintering, electrospinning and stereolithography. These technologies emerged in conjunction with new materials such as Hyperelastic Bone™, graphene and thermoplastics coupled with cell-laden hydrogels. This work will review these current state-of-the-art materials and technologies, their impact on advancements in bone tissue engineering and will highlight future considerations for the field.

Migration critically meditates osteoblastic differentiation of bone mesenchymal stem cells through activating canonical Wnt signal pathway.

Basic cellular events, such as focal adhesion and cytoskeleton organization, have been reported to be actively involved in fate decision process of stem cells, besides chemical and physical cues. Stem cell migration is critical in regulating various stem cell functions, but its influence on MSC differentiation into specific lineages has been rarely exploited. In this study, we used RGD-modified substrates to regulate cell motility though different RGD concentrations and systematically analyzed the correlation between osteoblastic differentiation and cell migration, as well as the role of Wnt signaling pathway. High motility correlated well with the significantly enhanced potential of the MSCs to differentiate into the osteoblastic lineage, as suggested by the significant up-regulations of Runx2, ALP, OCN expressions. The results also suggested that enhanced MSC migration efficiently activated the canonical Wnt-β-catenin pathway and stimulated transcription activities leading to osteoblastic differentiation, likely through internal forces generated dynamically during migration. Blockage of the Wnt-β-catenin pathway through artificial down-regulation of LRP5/6 expression significantly suppressed the osteoblastic differentiation for samples with high MSC motilities, further corroborating the critical involvement of Wnt/β-catenin pathway in the cell migration induced mechanotransduction and MSC differentiation into osteoblastic lineage. Our findings provide important insight for understanding the complicate mechanisms involved in MSC fate selection process and bone regeneration, and would have significant implications in the optimal design of bone tissue engineering materials through regulating cell motility.

Chitosan-hydroxyapatite nanocomposites: Effect of interfacial layer on mechanical and dielectric properties

The aim of this work is to investigate properties of interfacial layer in chitosan-hydroxyapatite nanoparticles (CS-nHAp) films. Simultaneous mechanical and dielectric percolation phenomena in films are observed for 30–40% of nHAp loadings. Structure, electrical and mechanical properties of composites and their dependencies on the hydroxyapatite concentrations are investigated by SEM, thermogravimetry, FTIR spectroscopy, nanoindentation and dielectric spectroscopy measurements. Hardness, reduced elastic modulus, conductivity and dielectric constant exhibit a strong dependence on nanoparticle concentration such that maximum values of referred properties are obtained at 30–40 wt % of nHAp. A three-phase model is used to include chitosan matrix, nanoparticles of hydroxyapatite and interfacial layer with dielectric constant higher than that of chitosan and hydroxyapatite. This layer between nanoparticles and matrix is due to strong interactions between chitosan's side groups with PO43− of hydroxyapatite. By performing simulations of spectroscopy measurements in the Hz-GHz frequency range we show that a maximum in all properties is traceable to the existence of overlapping and decreasing of thickness of interfacial layer. Our study proposes a systematic approach to design smart materials for bone tissue engineering.

In vitro evaluation of niobia added soda lime borosilicate bioactive glasses

In the present study, systematic and detailed investigations have been reported, in terms of bioactivity, cytocompatibility and mechanical hardness of a synthetic material that is proposed as a bone implant material in in-vitro conditions. The objective of the present study is to explore the effect of niobium addition on bioactivity and cytocompatibility of soda lime borosilicate bioglasses. Synthesis of bioglasses is carried out by conventional melt-quenching method. Simulated body fluid is prepared by Tas method. The bioactivity study was done for twenty-one days of immersion in simulated body fluid. XRD, FTIR, and SEM-EDS are showed formation of hydroxyapatite layer on the niobium added soda lime borosilicate glass even at a higher concentration (10 mol %) of niobium content. Degradation test was performed as per ISO 10993. The results obtained from degradation studies have demonstrated that weight loss of niobia mixed soda lime borosilicate glasses decreased with the increase of niobia. Cytocompatibility study has been studied by MTT Assay with osteoblast cells (MG-63). Cell viability is not affected by the addition of niobium content. Niobium added soda lime borosilicate glasses are both bioactive and cytocompatible. Hence it can be concluded that niobia mixed borosilicate glasses may be used as potential material in bone tissue engineering.

Tissue engineered bone mimetics to study bone disorders ex vivo: Role of bioinspired materials

Recent advances in materials development and tissue engineering has resulted in a substantial number of bioinspired materials that recapitulate cardinal features of bone extracellular matrix (ECM) such as dynamic inorganic and organic environment(s), hierarchical organization, and topographical features. Bone mimicking materials, as defined by its self-explanatory term, are developed based on the current understandings of the natural bone ECM during development, remodeling, and fracture repair. Compared to conventional plastic cultures, biomaterials that resemble some aspects of the native environment could elicit a more natural molecular and cellular response relevant to the bone tissue. Although current bioinspired materials are mainly developed to assist tissue repair or engineer bone tissues, such materials could nevertheless be applied to model various skeletal diseases in vitro. This review summarizes the use of bioinspired materials for bone tissue engineering, and their potential to model diseases of bone development and remodeling ex vivo. We largely focus on biomaterials, designed to re-create different aspects of the chemical and physical cues of native bone ECM. Employing these bone-inspired materials and tissue engineered bone surrogates to study bone diseases has tremendous potential and will provide a closer portrayal of disease progression and maintenance, both at the cellular and tissue level. We also briefly touch upon the application of patient-derived stem cells and introduce emerging technologies such as organ-on-chip in disease modeling. Faithful recapitulation of disease pathologies will not only offer novel insights into diseases, but also lead to enabling technologies for drug discovery and new approaches for cell-based therapies.

Graphene-Oxide-Decorated Microporous Polyetheretherketone with Superior Antibacterial Capability and In Vitro Osteogenesis for Orthopedic Implant.

Due to its similar elastic modulus of human bones, polyetheretherketone (PEEK) has been considered as an excellent cytocompatible material. However, the bioinertness, poor osteoconduction, and weak antibacterial activity of PEEK limit its wide applications in clinics. In this study, a facile strategy is developed to prepare graphene oxide (GO) modified sulfonated polyetheretherketone (SPEEK) (GO-SPEEK) through a simple dip-coating method. After detailed characterization, it is found that the GO closely deposits on the surface of PEEK, which is attributed to the π-π stacking interaction between PEEK and GO. Antibacterial tests reveal that the GO-SPEEK exhibits excellent suppression toward Escherichia coli. In vitro cell attachment, growth, differentiation, alkaline phosphatase activity, quantitative real-time polymerase chain reaction analyses, and calcium mineral deposition all illustrate that the GO-SPEEK substrate can significantly accelerate the proliferation and osteogenic differentiation of osteoblast-like MG-63 cells compared with those on PEEK and SPEEK groups. These results suggest that the GO-SPEEK has an improved antibacterial activity and cytocompatibility in vitro, showing that the developed GO-SPEEK has a great potential as the bioactive implant material in bone tissue engineering.

Nanohydroxyapatite/Graphene Nanoribbons Nanocomposites Induce in Vitro Osteogenesis and Promote in Vivo Bone Neoformation.

Nanomaterials based on graphene oxide nanoribbons (GNR) and nanohydroxyapatite (nHAp) serve as attractive materials for bone tissue engineering. Herein, we evaluated the potential of nHAp/GNR toward in vitro analysis of specific genes related to osteogenesis and in vivo bone regeneration using animal model. Three different concentrations of nHAp/GNR composites were analyzed in vitro using a cytotoxicity assay, and osteogenic potential was determined by ALP, OPN, OCN, COL1, and RUNX2 genes and alkaline phosphatase assays. In vivo bone neoformation using a well-established in vivo rat tibia defect model was used to confirm the efficiency of the optimized composite. The scaffolds were nontoxic, and the osteogenesis process was dose-dependent (at 200 μg mL-1 of nHAp/GNR) compared to controls. The in vivo results showed higher bone neoformation after 15 days of nHAp/GNR implantation compared to all groups. After 21 days, both nHAp/GNR composites showed better lamellar bone formation compared to control. We attributed this enhanced bone neoformation to the high bioactivity and surface area presented by nHAp/GNR composites, which was systematically evaluated in previous studies. These new in vivo results suggest that nHAp/GNR composites can be exploited for a range of strategies for the improved development of novel dental and orthopedic bone grafts to accelerate bone regeneration.

Cuttlebone as a Marine-Derived Material for Preparing Bone Grafts

The use of synthetic materials for biomedical applications still presents issues owing to the potential for unfavourable safety characteristics. Currently, there is increasing interest in using natural, marine-derived raw materials for bone tissue engineering. In our study, the endoskeleton of the mollusc Sepia, i.e. cuttlebone (CB), was used with regenerated cellulose (RC) to prepare three-dimensional composite bone grafts. CB microparticles were mechanically immobilised within a cellulose gel, resulting in a macroporous structure upon lyophilisation. The interconnected porous structure of the regenerated cellulose/cuttlebone (RC/CB) composite was evaluated by micro-computed tomography. The porosity of the composite was 80%, and the pore size predominantly ranged from 200 to 500μm. The addition of CB microparticles increased the specific scaffold surface by almost threefold and was found to be approximately 40mm-1. The modulus of elasticity and compressive strength of the RC/CB composite were 4.0 ± 0.6 and 22.0 ± 0.9MPa, respectively. The biocompatibility of the prepared RC/CB composite with rat hepatocytes and extensor digitorum longus muscle tissue was evaluated. The obtained data demonstrated that both the composite and cellulose matrix samples were non-cytotoxic and had no damaging effects. These results indicate that this RC/CB composite is a novel material suitable for bone tissue-engineering applications.

Repair of bone defects in rat radii with a composite of allogeneic adipose-derived stem cells and heterogeneous deproteinized bone

BackgroundIn the bone tissue engineering domain, seed cells, scaffold and cell-scaffold composites are three focuses. In this study, the feasibility of using allogeneic adipose-derived stem cells(ADSCs) combined with heterogeneous deproteinized bone (HDB) to repair segmental radial defects was investigated by observing the repair of the defect area.MethodsADSCs were cultured in vitro, purified, antigen-detected and osteogenic differentiation potency-measured; then, the ADSCs of the third generation were seeded into HDB to prepare an ADSCs-HDB composite partly with osteogenesis induced cells. Sixty Wistar rats were randomly divided into four groups with 15 in each group. A bone defect (4 mm in length) was created at the left radius in each rat. Two kinds of ADSCs-HDB composites were implanted in the ADSCs osteogenesis group or ADSCs group; HDB was implanted in the negative control group; nothing was filled in the blank control group. The bone defect repair was evaluated by gross observation, molybdenum target X-ray examination and histological analyses after surgery.ResultsGross observation: the bone defect area was completely filled and difficult to recognize in the ADSCs osteogenesis group. The connection of the ADSCs group was strong, but the implants were clearly identifiable. The joints of the negative control group were slightly thick but the connection was unstable. In the blank control group, kermesinus tissue was between the two ends and bones were not connected after 8 weeks. Molybdenum target X-ray examinations: In the ADSCs osteogenesis group, evident bridges in the graft were observed in the defects in the fourth week; the defects were filled with new bone completely and a marrow cavity appeared at 8 weeks. In the ADSCs group, there were some callus formations, but the radial defect was still obvious at 8 weeks. In the negative control group, fracture lines were clear. In the blank control group, no osseous bridges were observed, which resulted in bone nonunion eventually in 8 weeks. There were significant differences in the callus density between experimental groups and the blank control group at 4 and 8 weeks (P < 0.01). Histological measures showed that the rate and quality of the new bone formation and remodelling was significantly different between the experimental and control groups.ConclusionsA composite of ADSCs-HDB has a strong osteogenic ability. It can repair segmental bone defects well and is promising to serve as grafting material in bone tissue engineering.

The Investigation of ZnO/Poly(vinylidene fluoride) Nanocomposites with Improved Mechanical, Piezoelectric, and Antimicrobial Properties for Orthopedic Applications.

Studies have shown that piezoelectric materials can be used as bioactively charged surfaces to induce tissue regeneration (like bone reconstruction). β-phase poly(vinylidene fluoride) (PVDF) and zinc oxide (ZnO) are regarded as potential bone tissue engineering materials because of their piezoelectricity and biocompatibility. Additionally, ZnO nanoparticles (NPs) are ideal materials to reduce infection. In this study, PVDF scaffolds doped with ZnO NPs (ZnO/PVDF) were prepared by electrospinning increasing ZnO concentrations and the ratio of the β-phase PVDF resulting in an enhanced the modulus, elongation to failure and load stress. In vitro osteoblast assays were also performed to determine material cytotoxicity and bone reconstruction potential. Results exhibited that the piezo-excited scaffolds had a 30% greater osteoblast density, compared to non-piezoelectric-excited groups after 3 days of culture. To evaluate the scaffold's antimicrobial properties, three common etiologic agents of orthopedic after-surgery infection, Staphylococcus aureus (SA), Methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli), were separately seeded and counted. Results of this in vitro study showed significantly reduced E. coli (45%), SA (48%) and MRSA (37%) density on the 1 mg/ml ZnO/PVDF scaffolds without piezo-excitation (compared to the control). Moreover, compared to the non-piezo-excited scaffolds, results showed for the first time that the density of the same bacteria on the piezo-excited scaffolds reduced by over 23%, 22%, and 33%, respectively. For the first time, the effects of novel ZnO/PVDF scaffolds on osteoblasts and bacteria were investigated here, and significantly decreased bacteria with increased osteoblast density on the piezo-excited scaffolds demonstrated that these scaffolds have a strong potential for numerous antibacterial orthopedic applications.

Hydroxyapatite/Polyvinyl Alcohol Composite Hydrogels for Bone and Cartilage Tissue Engineering

Composite hydrogels on the basis of hydroxyapatite (HAp) and polyvinyl alcohol (PVA) has been proposed as a promising materials for bone and cartilage tissue engineering. HAp/PVA composite hydrogels with phase ratio 50:50wt% and 70:30wt% were obtained via in situ wet chemical precipitation technique in combination with the freeze-thawing approach. The XRD studies of sintered products revealed that HAp/PVA composite hydrogels synthesized from PVA with degree of hydrolysis (DH) 98% and molecular weights (MW) 25 kDa and 78 kDa are more suitable for biomedical purposes due to the formation of stoichiometric HAp. Swelling studies indicated that HAp/PVA 50:50 (78 kDa, 88% and 98%) hydrogels after 24h of immersion swell ~4.25-6.5 times less than identical samples with phase composition of 70:30wt%, which is accounted to different number of intermolecular hydrogen bonds formed. After 16 subsequent freeze-thawing cycles (FTC), HAp/PVA 50:50 (78 kDa, 88% and 98%) hydrogels contain ~1.2 times higher content of crosslinked PVA than HAp/PVA 70:30 (78 kDa, 88% and 98%) hydrogel samples.