Surface-active biomaterials.

Since the discovery in 1969 of a man-made surface-active material that would bond to bone, a range of materials with the same ability has been developed. These include glass, glass-ceramic, and ceramic materials which have a range of reaction rates and from which it should be possible to select a surface-active material for a specific application. The available materials and their similarities, differences, and current clinical applications are reviewed.

Enhanced differentiation and mineralization of human fetal osteoblasts on PDLLA containing Bioglass composite films in the absence of osteogenic supplements.

This study investigates the cellular response of fetal osteoblasts to bioactive resorbable composite films consisting of a poly-D,L-lactide (PDLLA) matrix and bioactive glass 45S5 Bioglass (BG) particles at three different concentrations (0% (PDLLA), 5% (P/BG5), and 40% (P/BG40)). Using scanning electron microscopy (SEM) we observed that cells were less spread and elongated on PDLLA and P/BG5, whereas cells on P/BG40 were elongated but with multiple protrusions spreading over the BG particles. Vinculin immunostaining revealed similar distribution of focal adhesion contacts on all cells independent of substratum, indicating that all materials permitted cell adhesion. However, when differentiation and maturation of fetal osteoblasts was examined, incorporation of 45S5 BG within the PDLLA matrix was found to significantly (p < 0.05) enhance alkaline phosphatase enzymatic activity and osteocalcin protein synthesis compared to tissue culture polystyrene controls and PDLLA alone. Alizarin red staining indicated extracellular matrix mineralization on both P/BG5 and P/BG40, with significantly more bone nodules formed than on PDLLA. Real time RT-PCR revealed that expression of bone sialoprotein was also affected by the BG containing films compared to controls, whereas expression of Collagen Type I was not influenced. By performing these investigations in the absence of osteogenic factors it appears that the incorporation of BG stimulates osteoblast differentiation and mineralization of the extracellular matrix, demonstrating the osteoinductive capacity of the composite.

Biomaterials: a forecast for the future.

The survivability half-life of prostheses made with current bio-inert materials is approximately 15 years, depending upon clinical applications. Bioactive materials improve device lifetime but have mechanical limitations. This paper proposes that biomaterials research needs to focus on regeneration of tissues instead of replacement. Alternatives are: use hierarchical bioactive scaffolds to engineer in vitro living cellular constructs for transplantation, or use resorbable bioactive particulates or porous networks to activate in vivo the mechanisms of tissue regeneration.

The bonding of Bioglass to a cobalt-chromium surgical implant alloy.

A biocompatible composite implant system was developed by coating Bioglass onto cobalt-chromium alloy substrates. Strong bonding between glass and metal was obtained by immersion of preoxidized implants into molten Bioglass under controlled conditions. The thin, adherent Bioglass coating provides the capability of bonding directly to bone, while the underlying metal substrate gives the composite implants sufficient strength to be used in load bearing applications.

Bioglass composites: a potential material for dental application.

Bioglass, a promising material for dental applications, can be reinforced with ductile stainless steel fibres. Three aspects of the fibre-reinforced bioglass composites are discussed. They are the interface between the glass and the metal fibres, the mechanical properties of the composites and their in vivo bonding behaviour. The importance of a good interfacial bond between the glass and the metal fibres is outlined. The improvement in strength and toughness, due to the fibres, is explained. The in vivo bonding behaviour of the bioglass composite is checked under statically loaded conditions.

Development and in vitro characterisation of novel bioresorbable and bioactive composite materials based on polylactide foams and Bioglass for tissue engineering applications.

Bioactive and bioresorbable composite materials were fabricated using macroporous poly(DL-lactide) (PDLLA) foams coated with and impregnated by bioactive glass (Bioglass) particles. Stable and homogeneous Bioglass coatings on the surface of PDLLA foams as well as infiltration of Bioglass particles throughout the porous network were achieved using a slurry-dipping technique in conjunction with pre-treatment of the foams in ethanol. The quality of the bioactive glass coatings was reproducible in terms of thickness and microstructure. Additionally, electrophoretic deposition was investigated as an alternative method for the fabrication of PDLLA foam/Bioglass composite materials. In vitro studies in simulated body fluid (SBF) were performed to study the formation of hydroxyapatite (HA) on the surface of PDLLA/Bioglass composites. SEM analysis showed that the HA layer thickness rapidly increased with increasing time in SBF. The high bioactivity of the PDLLA foam/Bioglass composites indicates the potential of the materials for use as bioactive, resorbable scaffolds in bone tissue engineering.

Osteoblast attachment and mineralized nodule formation on rough and smooth 45S5 bioactive glass monoliths.

Human primary osteoblast responses to smooth and roughened bioactive glass of 45S5 (Bioglass trade mark ) composition (46.1% SiO(2), 26.9% CaO, 2.6% P(2)O(5), 24.4% Na(2)O) were analysed in vitro. The smooth and rough surfaces had R(a) values and peak to valley distances of 0.04, 4.397, 2.027, and 21.328 microm, respectively. Cell attachment and morphology was observed using phalloidin staining of the actin cytoskeleton and revealed significant differences between smooth and rough surfaces. Cells that were spiky in appearance on the rough compared to the smooth surface formed an organized actin matrix much later on the rough surface. Scanning electron microscopy revealed many cell filipodia extending from more rounded cell bodies on the rough surface. A significantly greater number of nodules on the rough surface was observed, and these were shown to mineralize when supplemented with beta-glycerophosphate and dexamethasone. Raman spectroscopy confirmed the presence of hydroxyapatite in the mineralized cultures showing a definite peak at 964 cm(-1). FTIR analysis showed hydroxyapatite formation occurred more rapidly on the rough surface. This study demonstrates that although initial cell morphology was less advanced on the roughened surface, the cells were able to form mineralized nodules in greater numbers. This may have implications to bone tissue engineering using bioactive glasses.

Osteoblast responses to tape-cast and sintered bioactive glass ceramics.

The advantage of tape-cast bioactive glasses lies in the manufacturing procedure, which allows the build-up of layers and, therefore, the production of complex shapes. This, therefore, has applications to tissue engineering, where specific shapes are required such as repair of craniofacial defects. The bioactivity of tape-cast discs sintered at temperatures ranging from 800 degrees C to 1000 degrees C and for 3 or 6 h was analyzed by FTIR. Tape-cast discs were used to culture primary human osteoblasts, and cell attachment, cell death, collagen production, nodule formation, and mineralization were studied. These responses were dependent upon Si and Na release profiles of the tape-cast discs, and development of the hydroxyapatite layer.

In vitro release kinetics of proteins from bioactive foams.

This study describes an approach to obtaining 3-D scaffolds for tissue engineering that allows the incorporation and release of biologically active proteins to stimulate cell function. Laminin was adsorbed on the textured surfaces of binary 70S30C (70 mol % SiO(2), 30 mol % CaO) and ternary 58S (60 mol % SiO(2), 36 mol % CaO, 4 mol % P(2)O(5)) foams. The covalent bonds between the binding sites of the proteins and the ligands on the scaffolds' surfaces did not denaturate the proteins. In vitro studies show that the foams modified with chemical groups and coated with laminin were bioactive, as demonstrated by the formation of a crystalline hydroxy carbonate apatite (HCA) layer formed on the surfaces of the foams upon exposure to simulated body fluid (SBF). The release of proteins from the foams also was investigated. Sustained and controlled release from the scaffolds over a 30-day period was achieved. Laminin release from the bioactive foams followed the dissolution rate of the material network. These results suggest that bioactive foams have the potential to act as scaffolds for soft-tissue engineering with a controlled release of proteins that can induce tissue formation or regeneration.

In vitro bioactivity of S520 glass fibers and initial assessment of osteoblast attachment.

Bioactive glass fibers are attractive materials for use as tissue-engineering scaffolds and as the reinforcing phase for resorbable bioactive composites. The bioactivity of S520 glass fibers (52.0 mol % SiO(2), 20.9 Na(2)O, 7.1 K(2)O, 18.0 CaO, and 2.0 P(2)O(5)) was evaluated in two media, simulated body fluid (SBF) and Dulbecco's modified Eagle's medium (DMEM), for up to 20 days at 37 degrees C. Hydroxyapatite formation was observed on S520 fiber surfaces after 5 h in SBF. After a 20-day immersion, a continuous hydroxyapatite layer was present on the surface of samples immersed in SBF as well as on those samples immersed in DMEM [fiber surface area to solution volume ratio (SA:V) of 0.10 cm(2)/mL]. Backscattered electron imaging and EDS analysis revealed that the hydroxyapatite layer formation was more extensive for samples immersed in SBF. Decreasing the SA:V ratio to 0.05 cm(2)/mL decreased the time required to form a continuous hydroxyapatite surface layer. ICP was used to reveal Si, Ca, and P release profiles in DMEM after the 1st h (15.1, 83.8, and 29.7 ppm, respectively) were similar to those concentrations previously determined to stimulate gene expression in osteoblasts in vitro (16.5, 83.3, and 30.4 ppm, respectively). The tensile strength of the 20-microm diameter fibers was 925 +/- 424 MPa. Primary human osteoblast attachment to the fiber surface was studied by using SEM, and mineralization was studied by using alizarin red staining. Osteoblast dorsal ruffles, cell projections, and lamellipodia were observed, and by 7 days, cells had proliferated to form monolayer areas as shown by SEM. At 14 days, nodule formation was observed, and these nodules stained positive for alizarin red, demonstrating Ca deposition and, therefore mineralization.

Comparison of ossicular replacement materials in a mouse ear model.

Four biomaterials, UF45S5 Bioglass, Silastic, Plasti-Pore, and Proplast, were used to replace the incus in a mouse ear model. Bioglass, a bioactive glass ceramic, compared favorably with the other test materials in maintaining surgical positioning between malleus and stapes and remaining stable to a blast of nitrogen gas and to pick manipulation. In a short-term animal study, Bioglass showed histocompatibility comparable to that of these other implant materials now used in ossicular replacement surgery in humans.

Using 45S5 bioglass cones as endosseous ridge maintenance implants to prevent alveolar ridge resorption: a 5-year evaluation.

Bioglass cones acting as space fillers after removal of tooth roots delay the resorption of alveolar ridges. In 1987, 242 implants in 29 patients with a mean postimplantation interval of 19.9 months were reported by the authors. Bioglass cones had been fitted snugly at least 2 mm below the alveolar crest, and dentures were placed no sooner than 6 weeks following tooth removal; 2.9% had been lost and 3.7% developed dehiscences. The present report on 168 implants in 20 recalled patients (mean postimplantation interval of 63.2 months) revealed a loss of 14.3% of the implants and 7.7% of the implants requiring recontouring. Literature indicates highest survival rates for implants in the anterior mandible; however, the present data demonstrate a statistically significant retention rate in the anterior maxilla. With this high rate of Bioglass cone retention (85.7%) after 5 years, their placement into fresh sockets to maintain the alveolar ridge is recommended.

Mechanical properties of bioactive glasses, glass-ceramics and composites.

The application of bioactive glass and glass-ceramics has been widely documented over the past twenty years but the high modulus and low fracture toughness has made them less applicable for clinical, load bearing, applications. The development of non-resorbable polyethylene and polysulphone matrices for these materials has improved the mechanical properties. However, the primary concern of whether the bioactivity of the composites is reduced is still unresolved. The more recent development of resorbable carrier systems, dextran and collagen, for bioactive glasses does not introduce such problems, hence making this form of composite suitable for novel soft tissue applications. The development of a simple quality index has enabled some of the materials described within this paper to be ranked by their ability to replace bone, thus enabling possible new research directions to be emphasized.

Tissue response to Bioglass endosseous ridge maintenance implants.

Conical devices placed in the alveolar ridge after tooth extraction have been used clinically for several years to maintain the ridge morphology. In this way, the bone atrophy which occurs after extractions is minimized, and denture fit and function are enhanced. A system using such cones made from Bioglass (registered trademark of the University of Florida) and matching burs has been developed and tested clinically. Average four-year data show a retention rate of over 90%, which compares favorably with other systems using other materials (see Hench et al., 1991). Stanley et al. (to be published), in a review of the four-year clinical data, point out that a few of the cones, although firmly positioned within the alveolar ridge, have a radiolucent zone around the implant. In a clinical study, it is not possible to determine whether this radiolucent zone represents areas of fibrous capsule which are not attached to the implant and therefore compromise its long-term stability, or whether the soft tissue is adherent to the implant and thus contributes to its long-term stability. In a recent study, conical implants identical to those in the clinical trial were placed in the alveolar ridges of dogs and evaluated for up to two years. The adhesion of bone and soft tissue was measured and the development and stabilization of the reactive gel layer monitored. The findings in this animal study support the clinical observations and contribute to an explanation of the success of the Bioglass system in patients.

Bioactive glass-induced osteoblast differentiation: a noninvasive spectroscopic study.

Here, we report on a rapid, noninvasive biophotonics system using Raman spectroscopy to detect real-time biochemical changes in foetal osteoblasts (FOBs) following exposure to 45S5 Bioglass (BG)-conditioned media. Bio-Raman spectroscopy, combined with multivariate statistical analysis techniques (principal component analysis and least squares analysis), was able to noninvasively identify biochemical differences in FOBs cultured for different time periods and between FOBs exposed/or not to BG-conditioned media. Gene and protein expression studies were also performed for known markers of osteoblastic differentiation, namely, alkaline phosphatase, bone sialoprotein, and collagen type I. Quantitative RT-PCR confirmed upregulation of genes associated with osteoblast differentiation after exposure to BG-conditioned media. These results suggest that Raman spectroscopy can noninvasively detect biochemical changes in FOBs associated with differentiation. This technique could have important applications in the field of regenerative medicine by enabling rapid characterization of cell or organoid behavior on novel bioactive scaffolds without damage to either cell or biomaterial.

Preparation of bioactive glass-polyvinyl alcohol hybrid foams by the sol-gel method.

A new class of materials based on inorganic and organic species combined at a nanoscale level has received large attention recently. In this work the idea of producing hybrid materials with controllable properties is applied to obtain foams to be used as scaffolds for tissue engineering. Hybrids were synthesized by reacting poly(vinyl alcohol) in acidic solution with tetraethylorthosilicate. The inorganic phase was also modified by incorporating a calcium compound. Hydrated calcium chloride was used as precursor. A surfactant was added and a foam was produced by vigorous agitation, which was cast just before the gel point. Hydrofluoric acid solution was added in order to catalyze the gelation. The foamed hybrids were aged at 40 degrees C and vacuum dried at 40 degrees C. The hybrid foams were analyzed by Scanning Electron Microscopy, Mercury Porosimetry, Nitrogen Adsorption, X-ray Diffraction and Infra-red Spectroscopy. The mechanical behavior was evaluated by compression tests. The foams obtained had a high porosity varying from 60 to 90% and the macropore diameter ranged from 30 to 500 microm. The modal macropore diameter varied with the inorganic phase composition and with the polymer content in the hybrid. The surface area and mesopore volume decreased as polymer concentration increased in the hybrids. The strain at fracture of the hybrid foams was substantially greater than pure gel-glass foams.

Future developments and applications of biomaterials: an overview.

It is recommended that the emphasis of biomaterials research and development for the future should be to achieve improved reliability. Use of increasing numbers of implants per year coupled with decreasing long term (greater than 5 years) success rates are resulting in progressively larger numbers of reparative implant operations. This trend can be altered by emphasizing three areas of R&D: 1) Studies of composite biomaterial systems offering unique combinations of biological surface behavior and substrate mechanical performance; 2) Investigate mechanisms of interfacial reactions so that long term responses of the host-implant can be predicted; 3) Develop long term predictive relationships for biomaterials reliability based upon interfacial reactions, biomechanics, fracture mechanics, fatigue testing, and retrieval analysis. Brief examples of efforts to develop undrestanding in these three areas are described using bioglass coated metal and bioglass coated alumina implants.