Molecular biologic comparison of new bone formation and resorption on microrough and smooth bioactive glass microspheres

In a recent in vitro study, chemical microroughening of a bioactive glass surface was shown to enhance attachment of MG-63 osteoblastic cells to glass. The current study was designed to delineate the effects of microroughening on the gene expression patterns of bone markers during osteogenesis and new bone remodeling on bioactive glass surface in vivo. With the use of a rat model of paired comparison, a portion of the medullary canal in the proximal tibia was evacuated through cortical windows and filled with microroughened or smooth bioactive glass microspheres. The primary bone-healing response and subsequent remodeling were analyzed at 1, 2, and 8 weeks, respectively, by radiography, pQCT, histomorphometry, BEI-SEM, and molecular biologic analyses. The expression of various genes for bone matrix components (type I collagen, osteocalcin, osteopontin, osteonectin) and proteolytic enzymes (cathepsin K, MMP-9) were determined by Northern analysis of the respective mRNAs. Paired comparison showed significant differences in the mRNAs levels for specific bone matrix components at 2 weeks: osteopontin was significantly higher (p =.01) and osteonectin significantly lower (p =.05) in bones filled with microroughened microspheres than in those filled with smooth microspheres. Bones filled with microrough microspheres also showed significantly increased ratios of cathepsin K and MMP-9 (both markers of osteoclastic resorption) to type I collagen (p =.02 and p =.02, respectively) at 2 weeks and a significantly increased expression of MMP-9 at 8 weeks (p =.05). The pQCT, histomorphometric, and BEI-SEM analyses revealed no significant differences in the pattern of bone-healing response. Based on these results, microroughening of a bioactive glass surface could trigger temporal changes in the expression of specific genes especially by promoting the resorption part of new bone-remodeling processes. Future studies are needed to evaluate if the observed changes of gene expression are directly related to the microrough surface of any biomaterial or are biomaterial specific.     

Characterization of microrough bioactive glass surface: surface reactions and osteoblast responses in vitro

The current study characterized the in vitro surface reactions of microroughened bioactive glasses and compared osteoblast cell responses between smooth and microrough surfaces. Three different bioactive glass compositions were used and surface microroughening was obtained using a novel chemical etching method. Porous bioactive glass specimens made of sintered microspheres were immersed in simulated body fluid (SBF) or Tris solutions for 1, 6, 24, 48, or 72 h, and the formation of reaction layers was studied by means of a scanning electron microscope/energy dispersive X-ray analysis (SEM/EDXA). Cell culture studies were performed on bioactive glass disks to examine the influence of surface microroughness on the attachment and proliferation of human osteoblast-like cells (MG-63). Cell attachment was evaluated by means of microscopic counting of in situ stained cells. Cell proliferation was analyzed with a nonradioactive cell proliferation assay combined with in situ staining and laser confocal microscopy. The microroughening of the bioactive glass surface increased the rate of the silica gel layer formation during the first hours of the immersion. The formation of calcium phosphate layer was equal between control and microroughened glass surfaces. In cell cultures on bioactive glass, the microrough surface enhanced the attachment of osteoblast-like cells but did not have an effect on the proliferation rate or morphology of the cells as compared with smooth glass surface. In conclusion, microroughening significantly accelerated the early formation of surface reactions on three bioactive glasses and had a positive effect on initial cell attachment.     

Creation of microrough surface on sintered bioactive glass microspheres

Bioactive glasses are surface-active, generally silica-based, synthetic materials that form a firm chemical bond to bone. The aim of this study was to further enhance the bioactivity of glasses by creating a microroughness on their surface. Microroughness increases potential surface area for cell attachment and biomaterial-cell interactions. Three bioactive glasses of different composition were studied. Each material was flame-sprayed into microspheres, and a selected fraction of the spheres (250-300 microm) was sintered to form porous bioactive glass specimens. To create microrough surfaces, different acid etching techniques were tested. Atomic force microscopy (AFM) and back-scattered electron imaging of scanning electron microscopy (BEI-SEM) were used to characterize surface roughness. The degree of roughness was measured by AFM. A novel chemical-etching method, developed through intensive screening of different options, was found consistently to create the desired microroughness, with an average roughness value (R(a)) of 0.35-0.52 microm and a root mean-square roughness value (R(rms)) of 0.42-0.64 microm. Microroughening of the glass surface was obtained even in the internal parts of the porous glass matrices. Measured by BEI-SEM, the etching of a bioactive glass surface did not interfere with the formation of the characteristic surface reactions of bioactive glasses. This was confirmed by immersing the etched and control glass bodies in a simulated body fluid and tris(hydroxymethyl) aminomethane/HCl. The etching process did not significantly affect the mechanical strength of the sintered bioactive glass structures. Based on these experiments, it seems possible to create a reproducible microroughness of appropriate size on the surface of porous bioactive glass. The biologic benefits of such a surface treatment need to be validated with in vivo experiments.     

Bioactive glass microspheres as osteopromotive inlays in macrotextured surfaces of Ti and CoCr alloy bone implants: trapezoidal surface grooves without inlay most efficient in resisting torsional forces

We have tested the efficacy of porous bioactive glass (BG) inlays in enhancement of implant osseointegration. A total of 24 sheep underwent bilateral surgical implantation of three parallel implants on the anteromedial cortical surface of each tibia. The disc-shaped implants made of Ti6Al4V or cobalt chromium (CoCr) alloys had two parallel surface grooves (trapezoidal space with bottom widening) filled with sintered 100% bioactive glass microspheres or a selected mixture of bioactive and biocompatible glass microspheres. The surface of uncoated control implants was smooth, grit-blasted or had unfilled grooves. A subgroup of control smooth CoCr implants was coated with two or three BG layers. Implant incorporation with bone was evaluated using torque testing to failure, scanning electron microscopy and morphometry at 12 and 25 weeks. A total of 144 in vivo implants and 16 ex vivo cemented control implants were analyzed. Control Ti6Al4V implants with unfilled trapezoidal grooves showed highest torsional failure loads with excellent ingrowth of new bone and remodeling of ingrown bone into lamellar bone. Implants with BG inlays and microroughened control Ti6Al4V implants showed significantly lower torsional failure loads than control Ti6Al4V implants with unfilled grooves. In conclusion, BG inlays failed to enhance biological implant fixation. Macrotextured surface was more effective than grit-blasting in promotion of mechanical incorporation.     

Surface characteristics and primary bone marrow stromal cell response of a nanostructured strontium-containing oxide layer produced on a microrough titanium surface

Strontium (Sr) has been successfully used for the treatment of osteoporotic bone, increasing new bone formation while reducing bone resorption by stimulating proliferation and differentiation of osteoblastic cells and inhibiting osteoclast function. In this study, Sr-incorporated Ti oxide layer was produced on clinically relevant osteoconductive implant surface, that is, a grit-blasted microrough Ti surface, by a simple hydrothermal treatment with the expectation of utilizing the osteoblast response enhancement effect of Sr for the future applications as a more osteoconductive surface of the permanent load-bearing endosseous implants, without altering the original microrough surface features of grit-blasted Ti at the micron-scale. This surface exhibits a hierarchical structure (i.e., a nanoscale surface architecture of the Sr-incorporated Ti oxide layer (SrTiO(3)) imposed on micron-scale rough Ti structure) and Sr ion release into physiological solution. In vitro experiments using primary mouse bone marrow stromal cells (BMSCs) revealed that the hydrothermally produced SrTiO(3) coating promotes both the early and late cell response of BMSCs grown on a microrough Ti surface, with notably enhanced attachment, spreading, focal adhesion, alkaline phosphatase activity, and expression of critical integrins and osteoblastic phenotype genes. These results indicate that a hydrothermally produced SrTiO(3) coating improves the osteoconductivity of the microrough Ti surface by enhancing both the early and late cell response of BMSCs.     

Effects of microrough and hierarchical hybrid micro/nanorough surface implants on osseointegration in ovariectomized rats: A longitudinal in vivo microcomputed tomography evaluation

This study aimed to evaluate the effects of microrough and hierarchical hybrid micro/nanorough surface implants on osseointegration in ovariectomized rats at different time points. Implants with machined, microrough and hierarchical hybrid micro/nanorough surfaces were inserted into the distal femurs of ovariectomized Sprague-Dawley female rats. At weeks 0, 4, and 12 following implantation, in vivo microcomputed tomography (micro-CT) scanning was used to assess bone microarchitectural changes. After 12 weeks, all the rats were sacrificed, and the femurs with implants were harvested for histological analysis and pull-out test. For the Micro-CT analysis, the trabecular number and the bone volume ratio increased significantly in the microrough group (p < 0.01) and micro/nanorough group (p < 0.01) compared with the machined group. The trabecular separation decreased significantly in the micro/nanorough group (p < 0.01) compared with the other two groups. For the maximum pull-out forces and the bone-implant-contact analyses, significant statistical differences were found among the three groups, with the following sequence: micro/nanorough group > microrough group > machined group. The results indicate that the microrough and the hierarchical hybrid micro/nanorough surfaces of the implant can be beneficial to osseointegration under osteoporotic conditions, and the hierarchical hybrid micro/nanorough surface is more efficient.     

Coating of VEGF-releasing scaffolds with bioactive glass for angiogenesis and bone regeneration

Bioactive glasses are potentially useful as bone defect fillers, and vascular endothelial growth factor (VEGF) has demonstrated benefit in bone regeneration as well. We hypothesized that the specific combination of prolonged localized VEGF presentation from a matrix coated with a bioactive glass may enhance bone regeneration. To test this hypothesis, the capacity of VEGF-releasing polymeric scaffolds with a bioactive glass coating was examined in vitro and in vivo using a rat critical-sized defect model. In the presence of a bioactive glass coating, we did not detect pronounced differences in the differentiation of human mesenchymal stem cells in vitro. However, we observed significantly enhanced mitogenic stimulation of endothelial cells in the presence of the bioactive glass coating, with an additive effect with VEGF release. This trend was maintained in vivo, where coated VEGF-releasing scaffolds demonstrated significant improvements in blood vessel density at 2 weeks versus coated control scaffolds. At 12 weeks, bone mineral density was significantly increased in coated VEGF-releasing scaffolds versus coated controls, while only a slight increase in bone volume fraction was observed. The results of this study suggest that a bioactive glass coating on a polymeric substrate participates in bone healing through indirect processes which enhance angiogenesis and bone maturation and not directly on osteoprogenitor differentiation and bone formation. The mass of bioactive glass used in this study provides a comparable and potentially additive, response to localized VEGF delivery over early time points. These studies demonstrate a materials approach to achieve an angiogenic response formerly limited to the delivery of inductive growth factors.     

Peri- and intra-implant bone response to microporous Ti coatings with surface modification

Bone growth on and into implants exhibiting substantial surface porosity is a promising strategy in order to improve the long-term stable fixation of bone implants. However, the reliability in clinical applications remains a point of discussion. Most attention has been dedicated to the role of macroporosity, leading to the general consensus of a minimal pore size of 50–100 μm in order to allow bone ingrowth. In this in vivo study, we assessed the feasibility of early bone ingrowth into a predominantly microporous Ti coating with an average thickness of 150 μm and the hypothesis of improving the bone response through surface modification of the porous coating. Implants were placed in the cortical bone of rabbit tibiae for periods of 2 and 4 weeks and evaluated histologically and histomorphometrically using light microscopy and scanning electron microscopy. Bone with osteocytes encased in the mineralized matrix was found throughout the porous Ti coating up to the coating/substrate interface, highlighting that osseointegration of microporosities (<10 μm) was achievable. The bone trabeculae interweaved with the pore struts, establishing a large contact area which might enable an improved load transfer and stronger implant/bone interface. Furthermore, there was a clear interconnection with the surrounding cortical bone, suggesting that mechanical interlocking of the coating in the host bone in the long term is possible. When surface modifications inside the porous structure further reduced the interconnective pore size to the submicrometer level, bone ingrowth was impaired. On the other hand, application of a sol–gel-derived bioactive glass–ceramic coating without altering the pore characteristics was found to significantly improve bone regeneration around the coating, while still supporting bone ingrowth.     

Determining relevance of a weight-bearing ovine model for bone ingrowth assessment

A weight-bearing ovine model was used to quantify cancellous bone ingrowth and remodeling in porous-coated implants over 6, 12, and 24 weeks in situ. The null hypothesis for the investigation was that there would be no significant difference between the amount of cancellous bone ingrowth and rate of remodeling in this ovine model compared to a reported human bilateral implant model. Bone ingrowth progressed from 20.1 +/- 8.2% at 6 weeks in situ to 23.8 +/- 7.9% at 12 weeks, and 30 +/- 5.1% at 24 weeks. Fluorochrome analysis demonstrated a mineral apposition rate of 1.07 +/- 0.28 microm/day for bone at the porous-coating interface, whereas host bone remodeling at 0.89 +/- 0.23 microm/day. Histological analysis showed no adverse tissue or inflammatory response. The null hypothesis was supported in that regression analysis demonstrated that the amount of cancellous bone ingrowth over time (p = 0.545) and mineral apposition rate over time (p = 0.089) in this ovine model was not significantly different than reported human bilateral knee data. The results of this study appear to validate the ovine model for use in understanding skeletal attachment of porous-coated implants to cancellous bone in humans.     

Extracellular matrix formation and mineralization on a phosphate-free porous bioactive glass scaffold using primary human osteoblast (HOB) cells

Sol-gel derived bioactive glasses of the 70S30C (70mol% SiO2, 30mol% CaO) composition have been foamed to produce 3D bioactive scaffolds with hierarchical interconnected pore morphologies similar to trabecular bone. The aim of this study was to investigate primary human osteoblast response to porous bioactive glass scaffolds. The scaffolds supported osteoblast growth and induced differentiation, within the 3-week culture period, as depicted by enhanced ALPase enzymatic activity, without the addition of supplementary factors such as ascorbic acid, beta-glycerophosphate and dexamethasone. This is the first time this has been observed on a bioactive glass that does not contain phosphate. Deposition of extracellular matrix was also confirmed by enhanced production of the extracellular matrix protein collagen type I. SEM showed indications of mineralized bone nodule formation without the addition of growth factors. The 70S30C bioactive glass scaffolds therefore fulfil many of the criteria for an ideal scaffold for bone tissue engineering applications.     

Bioactive polymethyl methacrylate-based bone cement: comparison of glass beads, apatite- and wollastonite-containing glass-ceramic, and hydroxyapatite fillers on mechanical and biological properties

A new bioactive bone cement (designated GBC) consisting of polymethyl methacrylate (PMMA) as an organic matrix and bioactive glass beads as an inorganic filler has been developed. The bioactive beads, consisting of MgO-CaO-SiO(2)-P(2)O(5)-CaF(2) glass, have been newly designed, and a novel PMMA powder was selected. The purpose of the present study was to compare this new bone cement GBC's mechanical properties in vitro and its osteoconductivity in vivo with cements consisting of the same matrix as GBC and either apatite- and wollastonite-containing glass-ceramic (AW-GC) powder (designated AWC) or sintered hydroxyapatite (HA) powder (HAC). Each filler added to the cements amounted to 70 wt %. The bending strength of GBC was significantly higher than that of AWC and HAC (p < 0.0001). Cements were packed into intramedullar canals of rat tibiae in order to evaluate osteoconductivity as determined by an affinity index. Rats were sacrificed at 2, 4, and 8 weeks after operation. An affinity index, which equaled the length of bone in direct contact with the cement expressed as a percentage of the total length of the cement surface, was calculated for each cement. At each time interval studied, GBC showed a significantly higher affinity index than AWC or HAC up to 8 weeks after implantation (p < 0.03). The value for GBC increased significantly with time up to 8 weeks (p < 0.006). The handling property of GBC was comparable with that of PMMA bone cement. Our study revealed that the higher osteoconductivity of GBC was due to the higher bioactivity of the bioactive glass beads at the cement surface and the lower solubility of the new PMMA powder to MMA monomer. In addition, it was found that the smaller spherical shape and glassy phase of the glass beads gave GBC strong enough mechanical properties to be useful under weight-bearing conditions. GBC shows promise as an alternative with improved properties to the conventionally used PMMA bone cement.     

Biological assessment of the bone-screw interface after insertion of uncoated and hydroxyapatite-coated pedicular screws in the osteopenic sheep

The sheep seems to be a promising model of osteoporosis and biomaterial osteointegration in osteopenic bone. The long-term ovariectomized sheep model was used for the biological investigation of bone healing around uncoated and hydroxyapatite (HA)-coated pedicle screws in osteopenic bone. Four sheep were ovariectomized and four sheep were sham-operated. Twenty-four months after surgery, the animals were implanted with uncoated and HA-coated stainless steel screws in the lumbar vertebral pedicles. Four months later, bone-to-implant contact, bone ingrowth, and bone hardness were measured around screws. Uncoated stainless steel presented significantly (p < 0.0005) lower bone-to-implant contact in healthy and osteopenic bone compared with HA-coated stainless steel. HA significantly improved bone ingrowth in healthy bone (p < 0.05) compared with uncoated stainless steel. Osteopenia significantly (p < 0.05) reduced the area of bone ingrowth around the screw threads for both types of implants. In the inner thread area, bone microhardness significantly increased (p < 0.05) in HA-coated surface versus uncoated for healthy and osteopenic bone. HA coating significantly enhances bone-to-implant contact also in osteopenic bone in comparison with uncoated stainless steel surfaces. Bone ingrowth and mineralization are ameliorated by the osteoconductive HA coating. However, osteopenia seems to greatly influence bone ingrowth processes around the implanted screws regardless of the characteristics of the material surface.     

The behavior of novel hydrophilic composite bone cements in simulated body fluids

Composite bone cements were formulated with bioactive glass (MgO--SiO(2)--3CaO. P(2)O(5)) as the filler and hydrophilic matrix. The matrix was composed of a starch/cellulose acetate blend (SCA) as the solid component and a mixture of methylmethacrylate/acrylic acid (MMA/AA) as the liquid component. The curing parameters, mechanical properties, and bioactive behavior of these composite cements were determined. The addition of up to 30 wt % of glass improved both compressive modulus and yield strength and kept the maximum curing temperature at the same value presented by a typical acrylic-based commercial formulation. The lack of a strongly bonded interface (because no coupling agent was used) had important effects on the swelling and mechanical properties of the novel bone cements. However, bone cements containing AA did not show a bioactive behavior, because of the deleterious effect of this monomer on the calcium phosphate precipitation on the polymeric surfaces. Formulations without AA were prepared with MMA or 2-hydroxyethyl methacrylate (HEMA) as the liquid component. Only these formulations could form an apatite-like layer on their surface. These systems, therefore, are very promising: They are bioactive, hydrophilic, partially degradable, and present interesting mechanical properties. This combination of properties could facilitate the release of bioactive agents from the cement, allow bone ingrowth in the cement, and induce a press-fitting effect, improving the interfaces with both the prosthesis and the bone.     

The effect of mesoporous bioactive glass on the physiochemical,biological and drug-release properties of poly(dl-lactide-co-glycolide) films

Poly(lactide-co-glycolide) (PLGA) has been widely used for bone tissue regeneration. However, it lacks hydrophilicity, bioactivity and sufficient mechanical strength and its acidic degradation by-products can lead to pH decrease in the vicinity of the implants. Mesoporous bioactive glass (MBG) with highly ordered structure (pore size 2–50 nm) possesses higher bioactivity than non-mesoporous bioactive glass (BG). The aim of this study is to investigate the effect of MBG on the mechanical strength, in vitro degradation, bioactivity, cellular response and drug release of PLGA films and optimize their physicochemical, biological and drug-delivery properties for bone tissue engineering application. The surface and inner microstructure, mechanical strength and surface hydrophilicity of MBG/PLGA and BG/PLGA films were tested. Results indicated that MBG or BG was uniformly dispersed in the PLGA films. The incorporation of MBG into PLGA films significantly improved their tensile strength, modulus and surface hydrophilicity. MBG/PLGA resulted in an enhanced mechanical strength, in vitro degradation (water absorbance, weight loss and ions release), apatite-formation ability and pH stability in simulated body fluids (SBF), compared to BG/PLGA. MBG/PLGA and BG/PLGA films enhanced human osteoblastic-like cells (HOBs) attachment, spreading and proliferation compared to PLGA. HOBs differentiation was significantly upregulated when cells were cultured on 30 MBG/PLGA for 14 days, compared to 30 BG/PLGA. MBG/PLGA enhanced the accumulative release of dexamethazone (DEX) at early stages (0–200 h) compared to BG/PLGA, however, after 200 h, DEX-release rates for MBG/PLGA was slower than that of BG/PLGA. The contents of MBG in PLGA films can control the amount of DEX released. Taken together, MBG/PLGA films possessed excellent physicochemical, biological and drug-release properties, indicating their potential application for bone tissue engineering by designing 3D scaffolds according to their corresponding compositions.     

Tailoring properties of porous Poly (vinylidene fluoride) scaffold through nano-sized 58s bioactive glass

The biological properties of porous poly (vinylidene fluoride) (PVDF) scaffolds fabricated by selective laser sintering were tailored through nano-sized 58s bioactive glass. The results showed that 58s bioactive glass distributed evenly in the PVDF matrix. There were some exposed particles on the surface which provided attachment sites for biological response. It was confirmed that the scaffolds had highly bioactivity by the formation of bone-like apatite in simulated body fluid. And the bone-like apatite became dense with the increase in 58s bioactive glass and culture time. Moreover, the scaffolds were suitable for cell adhesion and proliferation compared with the PVDF scaffolds without 58s bioactive glass. The research showed that the PVDF/58s bioactive glass scaffolds had latent application in bone tissue engineering.     

Combined effect of BMP-2 gene transfer and bioactive glass microspheres on enhancement of new bone formation

Adenovirus-mediated recombinant human BMP-2 (RAdBMP-2) gene transfer has been found to have significant osteoinductive properties. The hypothesis of the current study was that bioactive glass surface could provide favorable osteoconductive conditions for cellular action of osteoinductive RAdBMP-2 gene transfer. In the rat proximal tibia, a portion of the medullary cavity was evacuated and filled with bioactive glass microspheres and injected with adenovirus carrying the human BMP-2 gene (BG/RAdBMP-2). Control defects filled with BG microspheres were injected with adenovirus carrying the LacZ reporter gene (BG/RAdLacZ) or saline (BG). Empty control defects were also used. Bone healing response was analyzed at 4 days, and at 2 and 8 weeks by radiography, peripheral quantitative computed tomography (pQCT), histomorphometry, and backscattered electron imaging of scanning electron microscopy (BEI-SEM) equipped with energy dispersive X-ray analysis (EDXA). In empty controls, the amount of intramedullary new bone peaked at 2 weeks, whereas defects filled with bioactive glass with and without RAdBMP-2 gene transfer showed a constant time-related increase of intramedullary new bone. At 8 weeks, there was significantly more new bone in defects treated with BG and RAdBMP-2 than in defects left to heal without filling (p < 0.001). Compared with the other controls (BG only or BG/RAdLacZ), the difference was not significant. In the current model, the osteopromotive effect of bioactive glass microspheres appears synergistic with the osteoinductive action of BMP-2 gene transfer, or one overshadows the other, as no additive effect was observed.     

Peripheral quantitative computed tomography in evaluation of bioactive glass incorporation with bone

This laboratory study examined the feasibility of non-invasive, in vivo peripheral quantitative computed tomography (pQCT) method in evaluation of bioactive glass incorporation with bone. An intramedullary defect model of the rat tibia was applied. The defect was filled with bioactive glass microspheres (diameter of 250-315 microm) or was left to heal without filling (empty controls). The results of the pQCT analysis were compared with those of histomorphometry. In the control defects, there was a good correlation (r2 = 0.776, p < 0.001) between the pQCT density of the intramedullary space and the amount of new bone measured by histomorphometry. In the defects filled with bioactive glass, the use of thresholding techniques of the applied pQCT system (Stratec XCT Research M) failed in separation of new bone formation and bioactive glass particles. However, detailed analysis of the pQCT attenuation profiles showed time-related changes which well matched with the histomorphometric results of new bone formation both in control and bioactive glass filled defects. The biphasic pQCT attenuation profiles of bioactive glass filled defects could be separated into two distinct peaks. In statistical analysis of various variables, the center (i.e. the value of attenuation) of the major attenuation peak was found to be the most significant indicator of the incorporation process. The center of the peak initially decreased (during the first 4 weeks of healing) and thereafter increased. These two phases probably reflect the primary resorption and reactivity of the bioactive glass microspheres in vivo followed by secondary new bone formation on their surfaces. Based on these results, pQCT-method seems to be suitable for in vivo follow-up of the bioactive glass incorporation processes. Although the imaging technique is not able to discriminate the individual microspheres from invading new bone unambiguously, the attenuation profiling seems to give adequate information about the state of the incorporation process. This information may help to establish non-invasive imaging techniques of synthetic bone substitutes for preclinical and clinical testing of their efficacy.     

Bioactivity and osteoblast responses of novel biomedical nanocomposites of bioactive glass nanofiber filled poly(lactic acid)

Biomedical nanocomposites constituted of bioactive ceramic and resorbable polymer have shown promise for the successful regeneration of bone tissues. We developed herein a novel nanocomposite made up of a bioactive glass in a nanofibrous form and a degradable synthetic polymer, poly(lactic acid) (PLA). The glass nanofiber with a bioactive composition was generated via an electrospinning process with an average diameter of approximately 320 nm. The nanofiber was homogenized with PLA solution at various concentrations (up to 35% nanofiber), followed by drying and thermal pressing to produce dense nanocomposites. The nanocomposites showed an internal morphology of uniformly dispersed nanofibers within the PLA matrix. The nanocomposites induced rapid formation of a hydroxycarbonate apatite layer on the surface under a simulated physiological medium. As the amount of bioactive nanofiber increased (from 5 to 25%), the in vitro bioactivity of the nanocomposite was improved. The osteoblast responses to the nanocomposites (compositions with 5 and 25% nanofiber) were assessed in terms of cell proliferation, differentiation, and mineralization. Osteoblasts attached and grew well on the nanocomposites and secreted collagen protein at initial culturing periods. The differentiation of cells, as assessed by the expression of alkaline phosphatase, was significantly improved on the nanocomposites as compared to those on pure PLA. Moreover, the mineralized product by the cells was observed to be significantly higher on the nanocomposites with respect to pure PLA. The newly developed nanocomposite constituted of bioactive nanofiber and degradable polymer is considered as a promising bone regeneration matrix with its excellent bioactivity and osteoblast responses.     

The influence of novel bioactive glasses on in vitro osteoblast behavior

Implant success requires a direct bond between bone and implant surface. Bioinert implants, such as titanium alloys, are commonly plasma-spray-coated with a bone-bonding, bioactive material such as hydroxyapatite. Such coatings tend to be chemically and topographically inhomogeneous without reproducible properties. A family of bioactive glasses that can be enameled and reliably adheres to titanium alloy has been developed. In this study the cytocompatibility of two of these glass compositions was tested in the as-cast condition. The effects of these glasses on the early and late events of osseous tissue formation in vitro were determined with MC3T3-E1.4 mouse osteoblast-like cells. MC3T3-E1.4 cells were cultured on glasses containing 55 and 50 wt % SiO(2), with titanium alloy (Ti6Al4V) and tissue culture polystyrene as controls. Cellular adhesion and proliferation, and alkaline phosphatase activity were studied over 5 to 15 days in culture. Qualitative and quantitative assays of mineralization were conducted. The osteoblast-like cells showed increased proliferation when grown on a bioactive glass containing 50 wt % silica. However, the adhesion, differentiation and mineralization behavior were similar on both glass compositions used in this study. These bioactive glasses proved to be cytocompatible substrata for osteoblast-like cell culture, and yielded higher cellular proliferation than titanium alloy.     

Preconditioned 70S30C bioactive glass foams promote osteogenesis in vivo

Bioactive glass scaffolds (70S30C; 70% SiO₂ and 30% CaO) produced by a sol–gel foaming process are thought to be suitable matrices for bone tissue regeneration. Previous in vitro data showed bone matrix production and active remodelling in the presence of osteogenic cells. Here we report their ability to act as scaffolds for in vivo bone regeneration in a rat tibial defect model, but only when preconditioned. Pretreatment methods (dry, pre-wetted or preconditioned without blood) for the 70S30C scaffolds were compared against commercial synthetic bone grafts (NovaBone® and Actifuse®). Poor bone ingrowth was found for both dry and wetted sol–gel foams, associated with rapid increase in pH within the scaffolds. Bone ingrowth was quantified through histology and novel micro-CT image analysis. The percentage bone ingrowth into dry, wetted and preconditioned 70S30C scaffolds at 11 weeks were 10 ± 1%, 21 ± 2% and 39 ± 4%, respectively. Only the preconditioned sample showed above 60% material–bone contact, which was similar to that in NovaBone and Actifuse. Unlike the commercial products, preconditioned 70S30C scaffolds degraded and were replaced with new bone. The results suggest that bioactive glass compositions should be redesigned if sol–gel scaffolds are to be used without preconditioning to avoid excess calcium release.