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The infection of implanted biomaterial scaffolds presents a major challenge. Existing therapeutic solutions, such as antibiotic treatment and silver nanoparticle-containing scaffolds are becoming increasingly impractical because of the growth of antibiotic resistance and the toxicity of silver nanoparticles. We present here a novel concept to overcome these limitations, an electrospun polycaprolactone (PCL) scaffold functionalised with zinc oxide nanowires (ZnO NWs). This study assessed the antibacterial capabilities and biocompatibility of PCL/ZnO scaffolds. The fabricated scaffolds were characterised by SEM and EDX, which showed that the ZnO NWs were successfully incorporated and distributed in the electrospun PCL scaffolds. The antibacterial properties were investigated by co-culturing PCL/ZnO scaffolds with Staphylococcus aureus. Bacterial colonisation was reduced to 51.3% compared to a PCL-only scaffold. The biocompatibility of the PCL/ZnO scaffolds was assessed by culturing them with HaCaT cells. The PCL scaffolds exhibited no changes in cell metabolic activity with the addition of the ZnO nanowires. The antibacterial and biocompatibility properties make PCL/ZnO a good choice for implanted scaffolds, and this work lays a foundation for ZnO NWs-infused PCL scaffolds in the potential clinical application of tissue engineering. [ABSTRACT FROM AUTHOR]

Barrier membranes that are used for guided tissue regeneration (GTR) therapy usually lack bioactivity and the capability to promote new bone tissue formation. However, the incorporation of an osteogenic agent into polymeric membranes seems to be the most assertive strategy to enhance their regenerative potential. Here, the manufacturing of composite electrospun membranes made of poly ("-caprolactone) (PCL) and particles of a novel bioactive glass composition (F18) is described. The membranes were mechanically and biologically tested with tensile strength tests and tissue culture with MG-63 osteoblast-like cell line, respectively. The PCL-F18 composite membranes demonstrated no increased cytotoxicity and an enhanced osteogenic potential when compared to pure PCL membranes. Moreover, the addition of the bioactive phase increased the membrane tensile strength. These preliminary results suggested that these new membranes can be a strong candidate for small bone injuries treatment by GTR technique. [ABSTRACT FROM AUTHOR]

Despite the phenomenal success of implants particularly in the realms of dentistry and orthopaedics, there are still challenges to overcome. The failure of implants resulting from infection, prosthetic loosening, and non-union continue to be the most notorious examples. The cascade of fracture healing and bone repair, especially with the presence of an implant, is complex because it involves a multifaceted immune response alongside the intricate process of bone formation and remodelling. Bone loss is a serious clinical problem that is frequently accompanied by chronic inflammation, illustrating that there is a convoluted relationship between inflammation and bone erosion. The effects of pro-inflammatory factors play a significant role in initiating and maintaining osteoclastogenesis that results in bone resorption by osteoclasts. This is because there is a disruption of the relative ratio between Receptor Activator of Nuclear Factor κB-Ligand (RANKL) and osteoprotegerin (OPG), which is central to modulating bone repair and remodelling. This review aims to provide a background to the bone remodelling process, the bone repair cascade post-implantation, and the associated complications. Furthermore, current clinical solutions that can influence bone formation via either internal or extrinsic mechanisms will be described. These efficacious treatments for osteolysis via targeting the RANKL/OPG ratio may be crucial to reducing the incidence of related implant failures in the future. [ABSTRACT FROM AUTHOR]

Hydroxyapatite and fluorhydroxyapatite (F)HA nanoparticles were synthesised in the presence of branched poly(acrylic acid)s (PAA) synthesised via reversible addition–fragmentation chain transfer polymerisation and compared to those synthesised in the presence of linear PAA. Analysis of the resulting nanoparticles using Fourier transform infrared spectroscopy, powder X-ray diffraction and transition electron microscopy found that the polymer was included within the nanoparticle samples and affected their morphology with nanoparticles synthesised in the presence of branched PAA being more acicular and smaller overall. [ABSTRACT FROM AUTHOR]

Additive manufacturing technologies enable the creation of very precise and well-defined structures that can mimic hierarchical features of natural tissues. In this article, we describe the development of a manufacturing technology platform to produce innovative biodegradable membranes that are enhanced with controlled microenvironments produced via a combination of selective laser melting techniques and conventional electrospinning. This work underpins the manufacture of a new generation of biomaterial devices that have significant potential for use as both basic research tools and components of therapeutic implants. The membranes were successfully manufactured and a total of three microenvironment designs (niches) were chosen for thorough characterisation. Scanning electron microscopy analysis demonstrated differences in fibre diameters within different areas of the niche structures as well as differences in fibre density. We also showed the potential of using the microfabricated membranes for supporting mesenchymal stromal cell culture and proliferation. We demonstrated that mesenchymal stromal cells grow and populate the membranes penetrating within the niche-like structures. These findings demonstrate the creation of a very versatile tool that can be used in a variety of tissue regeneration applications including bone healing. [ABSTRACT FROM AUTHOR]

Barrier membranes used for the treatment of bone tissue defects caused by periodontitis lack the ability to promote new bone tissue regeneration. However, the addition of an osteogenic component to membranes may enhance their regenerative potential. Here the manufacturing of composite membranes made of poly(caprolactone) and strontium-substituted bioactive glass is described using the solution-electrospinning technique, with particles located both inside and on the surface of the fibers. All membranes are characterized using scanning electron microscopy and energy dispersive X-ray spectroscopy, and glass dissolution from within the fibers is investigated in water. In vitro material cytotoxicity is determined using a rat osteosarcoma cell line. Electrospun fibers exhibit porous surfaces and regions of increased diameter where the particles are accumulated. The glass dissolves after immersion in water, releasing dissolution products that are associated with increased pH. Further evidence suggests accelerated polymer degradation due to interactions between both components, which may provide the additional benefit of reducing the pH changes associated with glass dissolution. All compositions are biocompatible in vitro, with the exception of membranes with >50 μg of glass on their surface. In conclusion, these membranes show great potential for bone healing applications, including guided bone regeneration and scaffolds for musculoskeletal tissue engineering. [ABSTRACT FROM AUTHOR]

Hydroxyapatite nanoscale particles (nHA) were prepared by wet chemical precipitation using four different synthesis methods. Differences in physico-chemical properties including morphology, particle-size, and crystallinity were investigated following alteration of critical processing parameters. The nanoparticles were also studied using X-ray diffraction (XRD), Fourier Transform infrared spectroscopy in attenuated total reflectance mode (FTIR-ATR), and transmission electron microscopy (TEM) with energy dispersive X-ray (EDS) spectrometry. The results showed that the particles obtained were composed of nHA, with different morphologies and aspect ratios (1.5 to 4) and degrees of crystallinity (40% to 70% following calcination) depending on the different process parameters of the synthesis method used, such as temperature, ripening time and pH. This study demonstrated that relatively small adjustments to processing conditions of different wet chemical preparation methods significantly affect the morphological and chemical characteristics of nHA. For the predicable preparation of biomimetic nHA for specific applications, the selection of both production method and careful control of processing conditions are paramount. [ABSTRACT FROM AUTHOR]

Poly(2-ethyl-(2-pyrrolidone)methacrylate)–hyaluronic acid hydrogels based on the free radical polymerization of 2-ethyl-(2-pyrrolidone)methacrylate combined with hyaluronic acid, using N,N′-methylenebisacrylamide or triethylene glycol dimethacrylate, as cross-linking agents, were considered for tissue engineering applications. Bovine articular chondrocytes were seeded onto the poly(2-ethyl-(2-pyrrolidone)methacrylate)–hyaluronic acid hydrogels, under orbital agitation, for a total of 40 days. The engineered cell-constructs were characterized according to cell proliferation, morphology and distribution as well as the biochemical composition of the tissue formed. The chondrocytes were found to be attached and presented a typical spherical morphology. Cells were able to proliferate and synthesize a hyaline-like matrix rich in glycosaminoglycans and collagen type II which were mainly located on the superficial area. Increased content of individual components poly(2-ethyl-(2-pyrrolidone)methacrylate) and hyaluronic acid, in triethylene glycol dimethacrylate–cross-linked networks led to enhanced cell distribution and total glycosaminoglycans content, supporting their potential application for the repair of cartilaginous tissues. [ABSTRACT FROM AUTHOR]

Poly(lactic-co-glycolic) acid (PLGA) has attracted considerable interest as a base material for biomedical applications due to its: (i) biocompatibility; (ii) tailored biodegradation rate (depending on the molecular weight and copolymer ratio); (iii) approval for clinical use in humans by the U.S. Food and Drug Administration (FDA); (iv) potential to modify surface properties to provide better interaction with biological materials; and (v) suitability for export to countries and cultures where implantation of animal-derived products is unpopular. This paper critically reviews the scientific challenge of manufacturing PLGA-based materials with suitable properties and shapes for specific biomedical applications, with special emphasis on bone tissue engineering. The analysis of the state of the art in the field reveals the presence of current innovative techniques for scaffolds and material manufacturing that are currently opening the way to prepare biomimetic PLGA substrates able to modulate cell interaction for improved substitution, restoration, or enhancement of bone tissue function. [ABSTRACT FROM AUTHOR]

Poly[2-ethyl(2-pyrrolidone) methacrylate] and hyaluronic acid hydrogels were synthesized via free-radical polymerization of 2-ethyl(2-pyrrolidone) methacrylate, hyaluronic acid and different crosslinkers. The ability of these hydrogels to induce apatite formation by incubating in simulated body fluid was investigated. The effect of hyaluronic acid content, crosslinkers and immersion time on mineralization behaviour and interface properties as well as the metabolic activity of different cultured cells were also determined. The bioactivity of the poly[2-ethyl(2-pyrrolidone) methacrylate] and hyaluronic acid hydrogels along with cell viability data indicated their potential application in bone tissue engineering. [ABSTRACT FROM AUTHOR]

Potassium fluorrichterite (KNaCaMgSiOF) glass-ceramics were modified by either increasing the concentration of calcium in the glass (GC5), or by the addition of PO to produce potassium fluorrichterite-fluorapatite (GP2). The solubility of the stoichiometric composition (GST), GC5 and GP2 were measured using the standard test described in ISO 6872:1995 (Dental Ceramics). Ion release profiles were determined for Si, Ca, Mg, Na, K and P using inductively coupled plasma mass spectrometry and fluoride ion (F) concentration was measured using an ion-selective electrode. The cytotoxicity of all compositions was assessed using cultured rat osteosarcoma cells (ROS, 17/2.8). Cell response was qualitatively assessed using scanning electron microscopy (SEM) and quantitatively using the Alamar blue assay. GST was the least soluble and also released the lowest concentration of ions following immersion in water. Of the modified compositions, GC5 demonstrated intermediate solubility but the greatest ion release while GP2 exhibited the highest solubility. This was most likely due to GC5 having the greatest proportion of residual glass following crystallisation. The mass loss exhibited by GP2 may have been due in part to the partial disintegration of the surface of specimens during solubility testing. SEM demonstrated that all compositions supported the growth of healthy ROS cells on their surfaces, and this data was further supported by the quantitative Alamar blue assay. [ABSTRACT FROM AUTHOR]

Novel chitosan/polybutylene succinate fibre-based scaffolds (C-PBS) were seeded with bovine articular chondrocytes in order to assess their suitability for cartilage tissue engineering. Chondrocytes were seeded onto C-PBS scaffolds using spinner flasks under dynamic conditions, and cultured under orbital rotation for a total of 6 weeks. Non-woven polyglycolic acid (PGA) felts were used as reference materials. Tissue-engineered constructs were characterized by scanning electron microscopy (SEM), hematoxylin-eosin (H&E), toluidine blue and alcian blue staining, immunolocalization of collagen types I and II, and dimethylmethylene blue (DMB) assay for glycosaminoglycans (GAG) quantification at different time points. SEM showed the chondrocytes' typical morphology, with colonization at the surface and within the pores of the C-PBS scaffolds. These observations were supported by routine histology. Toluidine blue and alcian blue stains, as well as immunohistochemistry for collagen types I and II, provided qualitative information on the composition of the engineered extracellular matrix. More pronounced staining was observed for collagen type II than collagen type I. Similar results were observed with constructs engineered on PGA scaffolds. These also exhibited higher amounts of matrix glycosaminoglycans and presented a central region which contained fewer cells and little matrix, a feature that was not detected with C-PBS constructs. [ABSTRACT FROM AUTHOR]

Potassium fluorrichterite (KNaCaMgSiOF) glass-ceramics were modified by either increasing the concentration of calcium (GC5) or by the addition of PO (GP2). The stoichiometric composition (GST), GC5 and GP2 were soaked in simulated body fluid (SBF) along with 45S5-type bioglass as a control. After immersion, surface analyses were performed using thin-film X-ray diffraction (TF-XRD), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS) and Fourier-transform infrared (reflection) spectroscopy (FT-IR). All compositions showed the formation of a calcium phosphate rich surface layer in SBF; GST, GP2 and the bioglass control within 7 days of immersion and GC5 after 14 days. It was concluded that all compositions were likely to be osteoconductive in vivo , with GP2 providing the best performance in terms of the combination of rapid formation of the surface layer and superior mechanical properties. This glass-ceramic system has potential as a load bearing bioceramic for fabrication of medical devices intended for skeletal tissue repair. [ABSTRACT FROM AUTHOR]

Advances in nanotechnology have been exploited to develop new biomaterials including nanocrystalline hydroxyapatite (nHA) with physical properties close to those of natural bone mineral. While clinical data are encouraging, relatively little is understood regarding bone cells' interactions with synthetic graft substitutes based on this technology. The aim of this research was therefore to investigate the in vitro response of both osteoblast cell lines and primary osteoblasts to an nHA paste. Cellular metabolic activity was assessed using the cell viability reagent PrestoBlue and quantitative, real-time PCR was used to determine gene expression related to osteogenic differentiation. A potential role of calcium-sensing receptor (CaSR) in the response of osteoblastic cells to nHA was also investigated. Indirect contact of the nHA paste with human osteoblastic cells (Saos-2, MG63, primary osteoblasts) and human bone marrow-derived mesenchymal stem cells enhanced the cell metabolic activity. The nHA paste also stimulated gene expression of runt-related transcription factor 2, collagen 1, alkaline phosphatase, and osteocalcin, thereby indicating an osteogenic response. CaSR was not involved in nHA paste-induced increases in cellular metabolic activity. This investigation demonstrated that the nHA paste has osteogenic properties that contribute to clinical efficacy when employed as an injectable bone graft substitute. [ABSTRACT FROM AUTHOR]

Chain silicate glass ceramics based on K-fluorrichterite (KNaCaMg5Si8O22F2, KFR) have potential for use in restorative dentistry and biomedical applications because they may be cast to shape and, when cerammed, have high biaxial flexural strength and fracture toughness. Excess CaO in canasite-based chain silicate compositions is known to enhance their bioactivity. Therefore, two new glass ceramic compositions have been fabricated with 5 mol% CaF2 (Glass A) and 5 mol% CaO (Glass B) substituted for MgO in the KFR formula unit. The phase evolution of Glasses A and B was studied in detail using X-ray diffraction and transmission electron microscopy. In addition, their mechanical properties were assessed. For Glass A, the fracture toughness (FT=2.66±0.02 MPa·m1/2) and biaxial flexural strength (BFS=227.3±24.5 MPa) were optimized for samples heat treated at 900°C for 4 h. In Glass B, however, the best FT (2.08±0.02 MPa·m1/2) and BFS (217.4±4.4 MPa) were obtained at 950°C. In view of their excellent mechanical properties and castability, Glasses A and B are considered potential candidates for fabrication of custom medical devices in restorative dentistry and moderate load-bearing reconstructive bone surgery. [ABSTRACT FROM AUTHOR]

Previous work by Miller et al. (2000–2004) has demonstrated that canasite-based glass ceramics have potential for use as biocompatible glass ceramics in hard-tissue augmentation. Several compositional modifications with respect to the stoichiometric formula (K2Na4Ca5Si12O30F4) were studied and biocompatibility in simulated body fluid was reported. However, the mechanism(s) of crystallization were not investigated in detail. The purpose of this study was to examine the early stages of nucleation and growth in four glass compositions using X-ray diffraction and transmission electron microscopy. In stoichiometric compositions (CAN1), laths of predominantly frankamenite homogeneously nucleate throughout the glass at ∼700°C without the presence of a nucleating phase. However, in Na2O-deficient compositions (CAN2), CaF2 particles (650°C) act as nucleating sites for canasite laths (700°C). In CaO-rich compositions (CAN3), CaF2 particles (650°C) once again act as nucleating sites but for xonotlite (700°C) rather than canasite laths. Instead, frankamenite and canasite crystallize to become the dominant phases at >700°C. In P2O5-modified compositions (CAN4), CaF2 and fluorapatite, present on cooling, act as nucleating agents for canasite (750°C). [ABSTRACT FROM AUTHOR]

Stoichiometric K-fluorrichterite (Glass A) and the same composition with 2 mol% P2O5 added (Glass B) were prepared and then heat-treated isothermally from 550°-1000°C with 50°C intervals. Samples were characterized using X-ray diffraction (XRD) and transmission electron microscopy (TEM). The biaxial flexural strength and indentation fracture toughness of heat-treated glass specimens were also determined for both materials. XRD traces and TEM images showed similar phase evolution and fine microstructures for both systems at ≤950°C, with mica and diopside reacting with residual glass to form K-fluorrichterite as the temperature was increased from 650°C. However, in Glass B, fluorapatite was also present at >800°C. In contrast, coarser microstructures were observed at 1000°C, with larger K-fluorrichterite (20 µm) and enstatite (10 µm) crystals in Glasses A and B, respectively. The highest fracture toughness (2.69 ± 0.01 MPa·m½) and biaxial strength (242.6 ± 3.6 MPa) were recorded for Glass B heat-treated at 1000°C. This was attributed to the presence of enstatite coupled with an interlocked lath-like crystalline microstructure. [ABSTRACT FROM AUTHOR]

Guided bone regeneration (GBR) is employed to encourage the formation of new bone in osseous defects by restricting the infiltration of soft tissues. While a variety of membranes have been evaluated for this surgical procedure, the nonresorbable material of choice is currently expanded polytetrafluoroethylene (e-PTFE). A new alkali-cellulose membrane produced by a biotechnological process has been developed as an alternative to e-PTFE for GBR. In this study, the biocompatibility of this novel alkali-cellulose membrane and e-PTFE was compared using tissue culture and an in vivo GBR model. In vitro both materials supported the attachment, migration and differentiation of osteoblast-like cells in culture for up to 3 weeks. The in vivo model was based upon full-thickness transcortical bone defects in the mandibular rami of Sprague-Dawley rats. The right rami were used as controls, contralateral defects being covered bucally and lingually with either e-PTFE or alkali-cellulose membranes. Pathological and histomorphometric analysis was undertaken at 4 and 10 weeks postimplantation. Bone regeneration associated with alkali-cellulose membranes was predominantly endochondral in type in contrast to e-PTFE which induced direct bone formation (intramembranous ossification). The amount of new bone formed in defects was similar for both types of membrane, but alkali-cellulose membranes induced significantly greater inflammatory response; characterized by lymphocytes, macrophages and multinucleated giant cells. Degradation and possible exposure of individual cellulose fibres may account for the poor performance of alkali-cellulose membranes in vivo. This animal and in vitro study indicates that when choosing a nonresorbable membrane for GBR, e-PTFE membranes are likely to perform better than those produced from alkali-cellulose. [ABSTRACT FROM AUTHOR]

Injectable nanoscale hydroxyapatite (nHA) systems are highly promising biomaterials to address clinical needs in bone tissue regeneration, due to their excellent biocompatibility, bioinspired nature, and ability to be delivered in a minimally invasive manner. Bulk strontium-substituted hydroxyapatite (SrHA) is reported to encourage bone tissue growth by stimulating bone deposition and reducing bone resorption, but there are no detailed reports describing the preparation of a systematic substitution up to 100% at the nanoscale. The aim of this work was therefore to fabricate systematic series (0–100 atomic% Sr) of SrHA pastes and gels using two different rapid-mixing methodological approaches, wet precipitation and sol-gel. The full range of nanoscale SrHA materials were successfully prepared using both methods, with a measured substitution very close to the calculated amounts. As anticipated, the SrHA samples showed increased radiopacity, a beneficial property to aid in vivo or clinical monitoring of the material in situ over time. For indirect methods, the greatest cell viabilities were observed for the 100% substituted SrHA paste and gel, while direct viability results were most likely influenced by material disaggregation in the tissue culture media. It was concluded that nanoscale SrHAs were superior biomaterials for applications in bone surgery, due to increased radiopacity and improved biocompatibility. [ABSTRACT FROM AUTHOR]

Bone injuries that arise from trauma, cancer treatment, or infection are a major and growing global challenge. An increasingly ageing population plays a key role in this, since a growing number of fractures are due to diseases such as osteoporosis, which place a burden on healthcare systems. Current reparative strategies do not sufficiently consider cell-substrate interactions that are found in healthy tissues; therefore, the need for more complex models is clear. The creation of in vitro defined 3D microenvironments is an emerging topographically-orientated approach that provides opportunities to apply knowledge of cell migration and differentiation mechanisms to the creation of new cell substrates. Moreover, introducing biofunctional agents within in vitro models for bone regeneration has allowed, to a certain degree, the control of cell fate towards osteogenic pathways. In this research, we applied three methods for functionalizing spatially-confined electrospun artificial microenvironments that presented relevant components of the native bone stem cell niche. The biological and osteogenic behaviors of mesenchymal stromal cells (MSCs) were investigated on electrospun micro-fabricated scaffolds functionalized with extracellular matrix (ECM) proteins (collagen I), glycosaminoglycans (heparin), and ceramic-based materials (bioglass). Collagen, heparin, and bioglass (BG) were successfully included in the models without modifying the fibrous structures offered by the polycaprolactone (PCL) scaffolds. Mesenchymal stromal cells (MSCs) were successfully seeded in all the biofunctional scaffolds and they showed an increase in alkaline phosphatase production when exposed to PCL/BG composites. This research demonstrates the feasibility of manufacturing smart and hierarchical artificial microenvironments for studying stem cell behavior and ultimately the potential of incorporating these artificial microenvironments into multifunctional membranes for bone tissue regeneration [ABSTRACT FROM AUTHOR]

Preventing the development of osteomyelitis while enhancing bone regeneration is challenging, with relatively little progress to date in translating promising technologies to the clinic. Nanoscale hydroxyapatite (nHA) has been employed as a bone graft substitute, and recent work has shown that it may be modified with silver to introduce antimicrobial activity against known pathogens. The aim of this study was to incorporate silver-doped nHA into electrospun scaffolds for applications in bone repair. Silver-doped nHA was produced using a modified, rapid mixing, wet precipitation method at 2, 5, 10 mol.% silver. The silver-doped nHA was added at 20 wt.% to a polycaprolactone solution for electrospinning. Bacteria studies demonstrated reduced bacterial presence, with Escherichia coli and Staphylococcus aureus undetectable after 96 h of exposure. Mesenchymal stem cells (MSCs) were used to study both toxicity and osteogenicity of the scaffolds using PrestoBlue® and alkaline phosphatase (ALP) assays. Innovative silver nHA scaffolds significantly reduced E. coli and S. aureus bacterial populations while maintaining cytocompatibility with mammalian cells and enhancing the differentiation of MSCs into osteoblasts. It was concluded that silver-doped nHA containing scaffolds have the potential to act as an antimicrobial device while supporting bone tissue healing for applications in orthopedic and dental bone surgery. [ABSTRACT FROM AUTHOR]

Oral disease greatly affects quality of life, as the mouth is required for a wide range of activities including speech, food and liquid consumption. Treatment of oral disease is greatly limited by the dose forms that are currently available, which suffer from short contact times, poor site specificity, and sensitivity to mechanical stimulation. Mucoadhesive devices prepared using electrospinning offer the potential to address these challenges by allowing unidirectional site-specific drug delivery through intimate contact with the mucosa and with high surface areas to facilitate drug release. This review will discuss the range of electrospun mucoadhesive devices that have recently been reported to address oral inflammatory diseases, pain relief, and infections, as well as new treatments that are likely to be enabled by this technology in the future. [ABSTRACT FROM AUTHOR]