The future of bioactive ceramics.

Two important worldwide needs must be satisfied in the future; (1) treatment of the deteriorating health of an aging population and, (2) decreasing healthcare costs to meet the needs of an increased population. The ethical and economic dilemma is how to achieve equality in quality of care while at the same time decreasing cost of care for an ever-expanding number of people. The limited lifetime of prosthetic devices made from first-generation nearly inert biomaterials requires new approaches to meet these two large needs. This paper advises an expanded emphasis on: (1) regeneration of tissues and (2) prevention of tissue deterioration to meet this growing need. Innovative use of bioactive ceramics with genetic control of in situ tissue responses offers the potential to achieve both tissue regeneration and prevention. Clinical success of use of bioactive glass for bone regeneration is evidence that this concept works. Likewise the use of micron sized bioactive glass powders in a dentifrice for re-mineralization of teeth provides evidence that prevention of tissue deterioration is also possible. This opinion paper outlines clinical needs that could be met by innovative use of bioactive glasses and ceramics in the near future; including: regeneration of skeletal tissues that is patient specific and genetic based, load-bearing bioactive glass-ceramics for skeletal and ligament and tendon repair, repair and regeneration of soft tissues, and rapid low-cost analysis of human cell-biomaterial interactions leading to patient specific diagnoses and treatments using molecularly tailored bioceramics.

Preparation of poly(L-lactic acid)-polysiloxane-calcium carbonate hybrid membranes for guided bone regeneration.

A novel poly(L-lactic acid) (PLLA)/calcium carbonates hybrid membrane containing polysiloxane was prepared using aminopropyltriethoxysilane (APTES) for biodegradable bone-guided regeneration. Carboxy groups in the PLLA made a chemical bond with amino groups in APTES, resulting in the formation of the hybrid membrane. The polysiloxane-hybridized PLLA was an amorphous phase. The membrane formed hydroxycarbonate apatite (HCA) on its surface after 3d of soaking in simulated body fluid (SBF). X-ray energy-dispersive spectroscopy showed that the HCA layer includes Si with Ca and P. After soaking the membrane in SBF, almost no Si was present in SBF. The membrane is expected to be a satisfactory substrate for the formation of the silicon-containing HCA layer using the SBF-soaking method. A result of osteoblast-like cellular proliferation on the membrane and the membrane coated with silicon-containing HCA showed no cell toxicity. The membrane coated with silicon-containing HCA had much higher cell-proliferation ability than the membrane.

Optimising bioactive glass scaffolds for bone tissue engineering.

A 3D scaffold has been developed that has the potential to fulfil the criteria for an ideal scaffold for bone tissue engineering. Sol-gel derived bioactive glasses of the 70S30C (70 mol% SiO2, 30 mol% CaO) composition have been foamed to produce 3D bioactive scaffolds with hierarchical interconnected pore morphologies similar to trabecular bone. The scaffolds consist of a hierarchical pore network with macropores in excess of 500 microm connected by pore windows with diameters in excess of 100 microm, which is thought to be the minimum pore diameter required for tissue ingrowth and vasularisation in the human body. The scaffolds also have textural porosity in the mesopore range (10-20 nm). The scaffolds were sintered at 600, 700, 800 and 1000 degrees C. As sintering temperature was increased to 800 degrees C the compressive strength increased from 0.34 to 2.26 MPa due to a thickening of the pore walls and a reduction in the textural porosity. The compressive strength is in the range of that of trabecular bone (2-12 MPa). Importantly, the modal interconnected pore diameter (98 microm) was still suitable for tissue engineering applications and bioactivity is maintained. Bioactive glass foam scaffolds sintered at 800 degrees C for 2 h fulfill the criteria for an ideal scaffold for tissue engineering applications.

Dose- and time-dependent effect of bioactive gel-glass ionic-dissolution products on human fetal osteoblast-specific gene expression.

Bioactive glasses dissolve upon immersion in culture medium, and release their constitutive ions into solution. There has been some evidence suggesting that these ionic-dissolution products influence osteoblast-specific processes. Here, the effect of 58S sol-gel-derived bioactive glass (60% SiO(2), 36% CaO, 4% P(2)O(5), in molar percentage) on primary osteoblasts derived from human fetal long bone explant cultures is investigated, and it is hypothesized that critical concentrations of sol-gel-dissolution products (consisting of a combination of simple inorganic ions) can enhance osteoblast phenotype in vitro by affecting the expression of a number of genes associated with the differentiation and extracellular matrix deposition processes. Cells were exposed to a range of 58S dosages continuously for a period of 4-14 days in monolayer cultures. Quantitative real-time RT-PCR analysis of a panel of osteoblast-specific markers showed a varied gene expression pattern in response to the material. The highest concentration of Ca and Si tested (96 and 50 ppm, respectively) promoted upregulation of gene expression for most markers (including alkaline phosphatase, osteocalcin, and osteopontin) at the latest time point, compared to non-58S-treated control, although this observation was not statistically significant. The same 58S concentration produced higher ALP activity levels and increased proliferation throughout the culture period, compared to lower dosages tested; however, the results generated were again not statistically significant. The data overall suggest that no significant effect can be ascribed to the ionic products of 58S bioactive gel-glass dissolution tested here and their ability to stimulate osteoblastic marker gene expression.

Bioactive glasses beyond bone and teeth: emerging applications in contact with soft tissues.

The applications of bioactive glasses (BGs) have to a great extent been related to the replacement, regeneration and repair of hard tissues, such as bone and teeth, and there is an extensive bibliography documenting the role of BGs as bone replacement materials and in bone tissue engineering applications. Interestingly, many of the biochemical reactions arising from the contact of BGs with bodily fluids, in particular the local increase in concentration of various ions at the glass-tissue interface, are also relevant to mechanisms involved in soft tissue regeneration. An increasing number of studies report on the application of BGs in contact with soft tissues, aiming at exploiting the well-known bioactive properties of BGs in soft tissue regeneration and wound healing. This review focuses on research, sometimes involving preliminary in vitro studies but also in vivo evidence, that demonstrates the suitability of BGs in contact with tissues outside the skeletal system, which includes studies investigating vascularization, wound healing and cardiac, lung, nerve, gastrointestinal, urinary tract and laryngeal tissue repair using BGs in various forms of particulates, fibers and nanoparticles with and without polymer components. Potentially active mechanisms of interaction of BGs and soft tissues based on the surface bioreactivity of BGs and on biomechanical stimuli affecting the soft tissue-BG collagenous bonding are discussed based on results in the literature.

Nodule formation and mineralisation of human primary osteoblasts cultured on a porous bioactive glass scaffold.

The aim of this study was to analyse human osteoblast responses to a porous bioactive glass scaffold. It was hypothesised that osteoblasts would attach, proliferate and form mineralised nodules in response to culture on the bioactive glass. As dissolution products are a key feature of bioactive glasses, this was measured by inductively coupled plasma optical emission spectroscopy to determine effects of both the glass surface and ion release. Osteoblasts attached and proliferated on the foams as demonstrated by scanning electron microscopy. Nodule formation was also observed in the pores of the glass and also in conditioned medium containing dissolution products at certain concentrations and these nodules were shown to be mineralised by alizarin red staining. Undiluted dissolution products from the foams however caused significant apoptosis suggesting an ion concentration dependent response.

PDLLA/Bioglass composites for soft-tissue and hard-tissue engineering: an in vitro cell biology assessment.

The aim of this study was to examine the effect of increased content of 45S5 Bioglass (0-40 wt%) in poly(dl-lactic acid) (PDLLA) porous foams on the behaviour of MG-63 (human osteosarcoma cell line) and A549 cells (human lung carcinoma cell line). The ability of these cell lines to grow on bioactive composites was quantitatively investigated in order to assess the potentiality for their use in hard and soft-tissue engineering. Two hours after cell seeding, an increase of cell adhesion according to the increased content of Bioglass((R)) present in the foams for both cell types was observed. Cell proliferation studies performed over a period of 4 weeks showed a better aptitude of the A549 cells to proliferate on PDLLA foams containing 5 wt% Bioglass when compared to the proliferation on foams with 40 wt% Bioglass. A lower proliferation rate was obtained for cells on pure PDLLA. Scanning electron microscopy analysis showed for both cell types the presence of cells inside the porous structure of the foams. These results confirmed the biocompatibility of PDLLA/Bioglass composite foams and the positive effect of Bioglass on MG-63 cell behaviour and also showed for the first time the possibility for human lung epithelial type II cells to adhere and proliferate on these porous scaffolds. In addition, we describe a positive effect of 45S5 Bioglass on A549 cell behaviour in a dose-dependent manner, indicating the potential of using PDLLA/Bioglass composites with an optimal concentration of 45S5 Bioglass not only in bone tissue engineering but also in lung tissue engineering.

Assessment of polyglycolic acid mesh and bioactive glass for soft-tissue engineering scaffolds.

Sufficient neovascularization of neotissue is currently a limiting factor for the engineering of large tissue constructs. 45S5 Bioglass has been investigated extensively in bone tissue engineering but there has been relatively little previous research on its application to soft-tissue engineering. The objectives of this study were to investigate the use of 45S5 Bioglass in soft-tissue engineering scaffolds using in vitro and in vivo models. A fibroblast cell line (208F) was used for in vitro evaluation of surfaces coated with 45S5 Bioglass. Increased proliferation of fibroblasts was observed after growth on polystyrene surfaces coated with low concentrations (0.01-0.2%wt/vol) of 45S5 Bioglass for 24 h in vitro, determined as a change in total cell number by measuring lactate dehydrogenase. At higher concentrations of 45S5 Bioglass and longer periods of incubation (48 and 72 h) on coated surfaces, cell proliferation was reduced. Light microscopy revealed that the morphology of fibroblasts grown on 45S5 Bioglass-coated surfaces was not altered at low concentrations, but at higher concentrations fibroblasts became vacuolated. Enzyme-linked immunosorbent assay of conditioned culture medium collected from fibroblasts grown for 24 h on surfaces coated with low concentrations of 45S5 Bioglass (0.01%wt/vol) was found to contain significantly higher concentrations of vascular endothelial growth factor. Histological examination of polyglycolic acid (PGA)/45S5 Bioglass composite scaffolds that had been implanted subcutaneously into rats revealed that 45S5 Bioglass-coated meshes were well tolerated. Light microscopy revealed that neovascularization into 45S5 Bioglass-coated meshes was significantly increased at 28 and 42 days. Electron microscopy revealed fibroblasts adhering closely to the PGA mesh but not to 45S5 Bioglass particles. The apparent ability of 45S5 Bioglass incorporated into scaffolds to increase neovascularization would be extremely beneficial during the engineering of larger soft-tissue constructs.

Time- and concentration-dependent effects of dissolution products of 58S sol-gel bioactive glass on proliferation and differentiation of murine and human osteoblasts.

Bone loss is a significant clinical problem, and treatments utilizing donated graft material are limited. To meet future demands in the healthcare industry, there has been a shift of outlook toward the use of bioactive materials for tissue regeneration. A number of in vivo and in vitro studies have highlighted the potential of the bioactive glass ceramic 45S5 Bioglass as a synthetic regenerative scaffold. The application of sol-gel processing techniques has led to the synthesis of mesoporous bioactive glasses with greater textural and compositional variety. In this study, we evaluated the effects of supplemented tissue culture medium containing up to 203 ppm silica prepared by static soaking of particles of 58S sol-gel bioactive glass (58% SiO(2), 33% CaO, 9% P(2)O(5)) on the in vitro proliferation and differentiation of murine and human primary osteoblasts. These extracts had a higher silica content than those used previously in studies of 45S5 Bioglass, because of the faster rates of ion exchange permitted by the higher surface area-to-volume ratio of mesoporous glass. We found that osteoblasts from both species increased their proliferation in response to the glass-conditioned medium. In addition, the extent to which supplemented medium could alter cell differentiation varied with time in culture. Proliferation induced by supplemented medium paralleled effects induced by treatment with basic fibroblast growth factor, a known mitogenic growth factor for osteoblasts. Bone nodule formation was also increased by exposure to the glass-conditioned medium and this effect was positively correlated with the dose of glass used to prepare the medium. Apoptosis was stimulated by glass-conditioned medium in murine osteoblasts, but inhibited in human osteoblasts. These data demonstrate the bioactive effects of dissolution products derived from sol-gel materials on primary osteoblasts and complements in vivo studies that indicate the suitability of this material as a bone graft substitute.

Bioactivity of gel-glass powders in the CaO-SiO2 system: a comparison with ternary (CaO-P2O5-SiO2) and quaternary glasses (SiO2-CaO-P2O5-Na2O).

Bioactive glasses react chemically with body fluids in a manner that is compatible with the repair processes of the tissues. This results in the formation of an interfacial bond between the glasses and living tissue. Bioactive glasses also stimulate bone-cell proliferation. This behavior is dependent on the chemical composition as well as the surface texture of the glasses. It has been recently reported that gel-derived monolith specimens in the binary SiO2 - CaO are bioactive over a similar molar range of SiO2 content as the previously studied ternary CaO-P2O5-SiO2 system. In this report, the preparation and bioactivity of the binary gel-glass powder with 70 mol % SiO2 is discussed and its bioactivity is compared with the melt-derived 45S5 (quaternary) Bioglass and sol-gel-derived 58S (ternary) bioactive gel-glass compositions. Dissolution kinetic parameters K(1) and K(2) were also computed based on the silicon release for all glass powders. It was shown that the simple two-component SiO2-CaO gel-glass powder is bioactive with comparable dissolution rates as the clinically used melt-derived 45S5 Bioglass powder and extensively studied sol-gel-derived 58S gel-glass powder.

Structural studies of bioactivity in sol-gel-derived glasses by X-ray spectroscopy.

Extended X-ray absorption fine structure spectroscopy and X-ray absorption near edge structure, X-ray fluorescence spectroscopy, and X-ray powder diffraction have been used to study the local calcium environment in four sol-gel-derived bioactive calcium silicate glasses of the general formula (CaO)(x)(SiO(2))(1-x). The formation of a hydroxyapatite layer on the composition with the highest bioactivity (x = 0.3) with time has been studied, in an in vitro environment, by immersion in simulated body fluid (SBF) at 37 degrees C. The calcium oxygen environment in the four compositions has been shown to be six-coordinate in character. Both the extended X-ray absorption fine structure spectroscopy and X-ray absorption near edge structure show a gradual increase in coordination number and Ca--O bond distance with longer exposure to SBF. X-ray fluorescence show that calcium is quickly lost from the samples on exposure to SBF and the calcium concentration then recovers with time. There is clear evidence that the recovery of calcium content is due to the formation of a CaO-P(2)O(5)-rich layer. Annealing of samples at 650 degrees C shows the presence of what, on the length scales probed by X-ray diffraction, appears to be noncrystalline calcium phosphate after 1 h of exposure to an SBF solution, which becomes more crystalline on longer exposure.

Factors affecting the structure and properties of bioactive foam scaffolds for tissue engineering.

Resorbable 3D macroporous bioactive scaffolds have been produced for tissue-engineering applications by foaming sol-gel-derived bioactive glasses of the 58S (60 mol% SiO2, 36 mol% CaO, 4 mol% P2O5) composition with the aid of a surfactant. Bioactive glasses are known to have the ability to regenerate bone, and to release ionic biological stimuli that promote bone-cell proliferation by gene activation. The foams exhibit a hierarchical structure, with interconnected macropores (10-500 microm), which provide the potential for tissue ingrowth and mesopores (2-50 nm), which enhance bioactivity and release of ionic products. Many factors in the sol-gel and foaming processes can be used to control these pore sizes and distributions. This work concentrates on the effect of the processing temperature, gelling agent concentration, and the amount of water used for the foam generation on the structure, pore morphology, and the properties of the foam scaffold. The simplest and most reproducible method for controlling the modal pore diameter was by the amount of water added during the foaming process. The in vitro dissolution and bioactivity of the bioactive foams were compared to that of unfoamed monoliths and powders (< 20 microm in diameter) of the same composition.

Binary CaO-SiO(2) gel-glasses for biomedical applications.

Bioactive materials are routinely used in dental and orthopaedic applications. The concept was first introduced in 1971, with the discovery of 45S5 Bioglass, which is known to develop an interfacial bond between the implant and the host tissue. This glass is composed of SiO(2), CaO, P(2)O(5) and Na(2)O. Since then numerous glasses and glass ceramics with similar compositions have been extensively studied for clinical applications. Until 1990 it was accepted that P(2)O(5) and Na(2)O were necessary components for the glass composition to be bioactive. However, calcium silicate glasses with high SiO(2) content are impossible to produce using the traditional melt-quench method. This is due to the liquid-liquid immiscibility region that is present between 0.02 and 0.3 mole fraction of CaO and in terms of bioactivity, high CaO compositions were inferior to those quaternary bioactive glass compositions already in existence. In the last few years several studies have been reported regarding the production of CaO-SiO(2) glasses via the sol-gel processing technique. This report summarises the findings of the past and the present and also outlines potential of these calcium silicate gel-glasses in the field of biomaterials.

Compositional and microstructural design of highly bioactive P2O5-Na2O-CaO-SiO2 glass-ceramics.

Bioactive glasses having chemical compositions between 1Na(2)O-2CaO-3SiO(2) (1N2C3S) and 1.5Na(2)O-1.5CaO-3SiO(2) (1N1C2S) containing 0, 4 and 6 wt.% P(2)O(5) were crystallized through two stage thermal treatments. By carefully controlling these treatments we separately studied the effects on the mechanical properties of two important microstructural features not studied before, crystallized volume fraction and crystal size. Fracture strength, elastic modulus and indentation fracture toughness were measured as a function of crystallized volume fraction for a constant crystal size. Glass-ceramics with a crystalline volume fraction between 34% and 60% exhibited a three-fold improvement in fracture strength and an increase of 40% in indentation fracture toughness compared with the parent glass. For the optimal crystalline concentration (34% and 60%) these mechanical properties were then measured for different grain sizes, from 5 to 21 µm. The glass-ceramic with the highest fracture strength and indentation fracture toughness was that with 34% crystallized volume fracture and 13 µm crystals. Compared with the parent glass, the average fracture strength of this glass-ceramic was increased from 80 to 210 MPa, and the fracture toughness from 0.60 to 0.95 MPa.m(1/2). The increase in indentation fracture toughness was analyzed using different theoretical models, which demonstrated that it is due to crack deflection. Fortunately, the elastic modulus E increased only slightly; from 60 to 70 GPa (the elastic modulus of biomaterials should be as close as possible to that of cortical bone). In summary, the flexural strength of our best material (215 MPa) is significantly greater than that of cortical bone and comparable with that of apatite-wollastonite (A/W) bioglass ceramics, with the advantage that it shows a much lower elastic modulus. These results thus provide a relevant guide for the design of bioactive glass-ceramics with improved microstructure.

Twenty-first century challenges for biomaterials.

During the 1960s and 1970s, a first generation of materials was specially developed for use inside the human body. These developments became the basis for the field of biomaterials. The devices made from biomaterials are called prostheses. Professor Bill Bonfield was one of the first to recognize the importance of understanding the mechanical properties of tissues, especially bone, in order to achieve reliable skeletal prostheses. His research was one of the pioneering efforts to understand the interaction of biomaterials with living tissues. The goal of all early biomaterials was to 'achieve a suitable combination of physical properties to match those of the replaced tissue with a minimal toxic response in the host'. By 1980, there were more than 50 implanted prostheses in clinical use made from 40 different materials. At that time, more than three million prosthetic parts were being implanted in patients worldwide each year. A common feature of most of the 40 materials was biological 'inertness'. Almost all materials used in the body were single-phase materials. Most implant materials were adaptations of already existing commercial materials with higher levels of purity to eliminate release of toxic by-products and minimize corrosion. This article is a tribute to Bill Bonfield's pioneering efforts in the field of bone biomechanics, biomaterials and interdisciplinary research. It is also a brief summary of the evolution of bioactive materials and the opportunities for tailoring the composition, texture and surface chemistry of them to meet five important challenges for the twenty-first century.

Fabricating sol-gel glass monoliths with controlled nanoporosity.

Nanotopography is known to affect cell response, but the mechanisms are unknown. It is therefore important to be able to produce biomaterial surfaces with controllable nanopore sizes and morphologies from materials that can be used to make templates (scaffolds) for tissue regeneration. A rapid method of fabrication of sol-gel monoliths with controlled nanopore size is described in this paper. Pores of 2 nm in diameter were achieved, termed micropores by IUPAC convention. Conventional sol-gel processing yields pores an order of magnitude larger. Gelling time was also reduced from many hours to a few minutes, without using a gelling agent, and large crack-free monoliths were synthesized within 1 week.

Hierarchical porous materials for tissue engineering.

Biological organisms have evolved to produce hierarchical three-dimensional structures with dimensions ranging from nanometres to metres. Replicating these complex living hierarchical structures for the purpose of repair or replacement of degenerating tissues is one of the great challenges of chemistry, physics, biology and materials science. This paper describes how the use of hierarchical porous materials in tissue engineering applications has the potential to shift treatments from tissue replacement to tissue regeneration. The criteria that a porous material must fulfil to be considered ideal for bone tissue engineering applications are listed. Bioactive glass foam scaffolds have the potential to fulfil all the criteria, as they have a hierarchical porous structure similar to that of trabecular bone, they can bond to bone and soft tissue and they release silicon and calcium ions that have been found to up-regulate seven families of genes in osteogenic cells. Their hierarchical structure can be tailored for the required rate of tissue bonding, resorption and delivery of dissolution products. This paper describes how the structure and properties of the scaffolds are being optimized with respect to cell response and that tissue culture techniques must be optimized to enable growth of new bone in vitro.

Controlling ion release from bioactive glass foam scaffolds with antibacterial properties.

Bioactive glass scaffolds have been produced, which meet many of the criteria for an ideal scaffold for bone tissue engineering applications, by foaming sol-gel derived bioactive glasses. The scaffolds have a hierarchical pore structure that is very similar to that of cancellous bone. The degradation products of bioactive glasses have been found to stimulate the genes in osteoblasts. This effect has been found to be dose dependent. The addition of silver ions to bioactive glasses has also been investigated to produce glasses with bactericidal properties. This paper discusses how changes in the hierarchical pore structure affect the dissolution of the glass and therefore its bioactivity and rate of ion delivery and demonstrates that silver containing bioactive glass foam scaffolds can be synthesised. It was found that the rate of release of Si and Ca ions was more rapid for pore structures with a larger modal pore diameter, although the effect of tailoring the textural porosity on the rate of ion release was more pronounced. Bioactive glass scaffolds, containing 2 mol% silver, released silver ions at a rate that was similar to that which has previously been found to be bactericidal but not high enough to be cytotoxic to bone cells.

Third-generation biomedical materials.

Whereas second-generation biomaterials were designed to be either resorbable or bioactive, the next generation of biomaterials is combining these two properties, with the aim of developing materials that, once implanted, will help the body heal itself.

Broad-spectrum bactericidal activity of Ag(2)O-doped bioactive glass.

Bioactive glass has found extensive application as an orthopedic and dental graft material and most recently also as a tissue engineering scaffold. Here we report an initial investigation of the in vitro antibacterial properties of AgBG, a novel bioactive glass composition doped with Ag(2)O. The bacteriostatic and bactericidal properties of this new material and of two other bioactive glass compositions, 45S5 Bioglass and BG, have been studied by using Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus as test microorganisms. Concentrations of AgBG in the range of 0.05 to 0.20 mg of AgBG per ml of culture medium were found to inhibit the growth of these bacteria. Not only was AgBG bacteriostatic, but it also elicited a rapid bactericidal action. A complete bactericidal effect was elicited within the first hours of incubation at AgBG concentrations of 10 mg ml(-1). 45S5 Bioglass and BG had no effect on bacterial growth or viability. The antibacterial action of AgBG is attributed exclusively to the leaching of Ag(+) ions from the glass matrix. Analytical measurements rule out any contribution to AgBG-mediated bacterial killing by changes in pH or ionic strength or the dissolution of other ionic species from the biomaterials. Our observations of the dissolution profiles of Ag(+) from AgBG in the presence and absence of bacteria are consistent with silver accumulation by the bacteria.