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Better histology and biochemistry for osteoblasts cultured on titanium-doped bioactive glass: bioglass 45S5 compared with iron-, titanium-, fluorine- and boron-containing bioactive glasses.

In the present study we used an established cell culture model to compare Bioglass 45S5 with four other bioactive glasses. Small substitutions or additions of certain ions like iron, titanium, fluorine or boron modified the basic 45S5 glass network. We used several histological and biochemical parameters to interpret the results found in terms of the used model. Regarding 45S5 as a reference, we found that osteoblasts cultured on iron-doped bioactive glass showed a more flattened morphology, and both lower proliferation rate and osteoblast expression. Osteoblasts cultured on titanium-doped glasses also showed a flattened morphology, but higher proliferation and remarkably higher osteoblast expression. On fluorine- and boron-containing glasses the osteoblasts showed a rather compact morphology, a normal proliferation but only moderate osteoblast expression. With microprobe analysis it was shown that the formation of calcium and phosphorus on titanium-doped glass was relatively lower and the release of sodium slower when compared with 45S5. Osteoblasts cultured on titanium-doped bioactive glasses demonstrated superior histological and biochemical parameters when compared with the other glass types. Further research into the physico-chemical properties and the in vivo behaviour of doped bioactive glasses is recommended.

Alkaline Phosphatase↗

Effect of polyacrylic acid on the apatite formation of a bioactive ceramic in a simulated body fluid: fundamental examination of the possibility of obtaining bioactive glass-ionomer cements for orthopaedic use.

Glass-ionomer cements, which consist of CaO-Al2O3-SiO2-CaF2 glass powders and a polyalkenoic acid solution, such as polyacrylic acid (PAA), have been widely used in dentistry. They set rapidly without any shrinkage, the lack of temperature increase on reaction, and develop high mechanical strength. Therefore, if bioactive glass-ionomer cements can be obtained, such cements are expected to be useful as cements for fixing orthopaedic implants to the surrounding bone. In the present study, to examine the possibility of obtaining bioactive glass-ionomer cements, the effect of PAA on the apatite formation on bioactive ceramics in a simulated body fluid was investigated. It was revealed that presence of even a small quantity of PAA inhibits the apatite formation in the body environment. It is speculated that when glass-ionomer cements are implanted into the body, PAA can be released from the glass-ionomer cements and inhibits the apatite formation on their surfaces. It is reasonable to suppose that this will occur with any glass-ionomer cement that contains PAA. Therefore, it might be considered difficult to obtain bioactive glass-ionomer cements.

Acrylic Resins↗

Bioactivity modulation of bioactive materials in view of their application in osteoporotic patients.

The application of bioactive ceramic coatings to prostheses confers strength to a material (ceramic or biological glass) that exerts beneficial effects on bone-tissue growth but that itself lacks the toughness and stability required of an implant device. The rate of bioactivity is related to the chemical reactivity of the material and causes interface dissolution, precipitation and ion-exchange reactions. Ceramics may differ in sintering temperature and thus exhibit differences in their in vitro dissolution features and in vivo performance. To test these effects, in vitro and in vivo studies were carried out on two biocompatible biological glasses and a ceramic of proven bioactivity in view of their potential utilization as covering materials. In addition, a modified chitosan was adsorbed on the surface of a series of hydroxyapatite (HA) samples. Human fibroblasts and/or osteoblasts were used for the in vitro tests, and normal (INT) and osteoporotic (OVX) rats, normal rabbits and sheep for the in vivo studies. Similar chemical changes were observed in both glasses, suggesting that these materials underwent modifications directly dependent on their biological environment. The in vivo tests point to the possibility of improving the bioactivity of ceramic substrates with chitosan. However, the different behaviour of the materials in vitro and in vivo suggests that these tests should be conducted in parallel.

Journal Article↗

Histological and histomorphometrical analysis of bioactive glass and fibre reinforced bioactive glass dental root implants.

Bioactive glass has the ability to bond with bone, but it cannot be used as a load bearing device due to its limited mechanical properties. By reinforcing bioactive glass with a ductile second phase, a structurally reliable material is obtained. The aim of the present study was to evaluate histologically and morphometrically the interfacial behaviour of submerged composite dental root implants. Therefore, bulk and composite implants were subgingivally installed in the partially edentulous jaws of Beagle dogs and harvested after 4 and 16 months. Histologically, the connection between the implants and bone tissue could be clearly demonstrated. This bone connection is mainly located at the cortical bone level. In the vicinity of the infraalveolar nerve a fibrous tissue contact was found. It is shown that surgical trauma, motion at the glass to tissue interface, and gross ion dissolution from the material adversely affect the interfacial osteogenesis. If these factors are controlled, bone bonding is found over a larger area than the initial area of contact between the implant and bone tissue. This means that bone grows out along the implant surface, starting from the initial contact area. No difference was observed between the interfacial behaviour of bulk bioactive glass and intact fibre reinforced bioactive glass implants.

Animals↗

Effects of ceramic component on cephalexin release from bioactive bone cement consisting of Bis-GMA/TEGDMA resin and bioactive glass ceramics.

The purpose of this study was to elucidate the effect of amount of ceramic cement powder on drug release from bioactive bone cement. The associated bone-bonding strength was also investigated. The bioactive bone cement under investigation consisted of bisphenol-alpha-glycidyl methacrylate (Bis-GMA), triethylene-glycol dimethacrylate (TEGDMA) resin and a combination of apatite- and wollastonite-containing glass-ceramic (A-W GC) powder. A-W GC powder (50%, 70% and 80% w/w) containing 5% cephalexin (CEX) powder hardened within 5 min after mixing with Bis-GMA/TEGDMA resin. The compressive strength of the cement with or without drug increased with increasing the amount of ceramic powder. The compressive strength of the 80% ceramic cement without the incorporation of cephalexin was 194 MPa. This compressive strength was about 3 times higher than that for polymethylmethacrylate cement. After the cement was implanted in the proximal metaphysis of the tibiae of male rabbits, the failure load for the cement was found to increase with increasing of the amount of ceramic powder. This finding suggested that the cement formed a bonding with bone. In vitro CEX release from bioactive bone cement pellets in a simulated body fluid at pH 7.25 and 37 degrees C continued for more than 2 weeks. Drug release profile followed the Higuchi equation initially, but not at later stages. The drug release rate increased with increasing amount of ceramic powder in the mixture. Since the pore volume of the cement increased with increasing of amount of ceramic powder, the drug diffused in the pores between the ceramics particle and polymer matrix. As hydroxyapatite precipitated on the cement surface, the drug release rate decreased, as observed at the later release stage. These results suggest that varying the amount of ceramic powder in the cement system could control the drug release rate from bioactive bone cement.

Absorbable Implants↗

Long-term in vivo bioactivity and degradability of bulk sol-gel bioactive glasses.

Melt-derived bioactive glasses have been used with success in various clinical applications for over 10 years. Recently, particles of sol-gel-derived bioactive glasses with an initial high specific surface area have been shown to exhibit excellent osteoconductive properties as well as significant degradability. In this work, we explored the long-term in vivo bioactivity and degradability of bulk sol-gel-derived glasses in a rabbit model. Two sol-gel compositions (58S and 77S Bioglass) were used. Bulk 45S5 Bioglass was used as a control. Both sol-gel-derived glasses demonstrated osteoconductive properties similar to 45S5 Bioglass. In addition, absorbability was observed for both sol-gel-derived glasses starting after 12 weeks of implantation. Total absorption reached 40% after 52 weeks. No degradation could be measured in the case of bulk 45S5 melt-derived Bioglass within 1 year of implantation. The degradation process was highly time dependent, as demonstrated by regression analysis. New bone formation was found to fill in areas that had been resorbed, similar to bone remodeling. This absorbability can be assumed to be at least partially related to an osteoclastic resorption as viable osteoclasts-like cells were found to be in direct contact with the glass surfaces.

Absorbable Implants↗

Effects of ions in aqueous media on hydroxyapatite induction by silica gel and its relevance to bioactivity of bioactive glasses and glass-ceramics.

Hydroxyapatite induction by a synthesized pure silica hydrogel was examined in various simulated body fluids (SBFs) having different magnesium, calcium, and phosphate ion concentrations as well as pH values. The silica hydrogel generated biologically active apatite on its surface by taking up calcium and phosphorous ionic groups from a surrounding SBF that was prepared to emulate the human plasma in inorganic composition. The induction period for apatite nucleation on the surface of the silica was largely decreased with the addition of a small amount of the calcium or phosphate ions to the SBF and with an increase in pH, but increased with the addition of magnesium ion. Bioactivity of bioactive materials like Bioglass and glass-ceramic A-W was well interpreted in terms of the rate of apatite formation reflected in these results. Moreover, the results provide the basic knowledge for designing new bioactive materials.

Biocompatible Materials↗

The bone response of oxidized bioactive and non-bioactive titanium implants.

A number of experimental and clinical data on so-called oxidized implants have reported promising outcomes. However, little is investigated on the role of the surface oxide properties and osseointegration mechanism of the oxidized implant. Sul [On the Bone Response to Oxidized Titanium Implants: The role of microporous structure and chemical composition of the surface oxide in enhanced osseointegration (thesis). Göteborg: Department of Biomaterials/Handicap Research, University of Göteborg, Sweden; 2002; Biomaterials 2003; 24: 3893-3907] recently proposed two action mechanisms of osseointegration of oxidized implants, i.e. mechanical interlocking through bone growth in pores/other surface irregularities (1) and biochemical bonding (2). The aim of the present study is two-fold: (i) investigating the role of the implant surface chemistry on bone responses; (ii) investigating the validity of the biochemical bonding theory of the oxidized, bioactive bone implants with specific implant surface chemistry. Two groups of oxidized implants were prepared using micro arc oxidation process and were then inserted in rabbit bone. One group consisted of magnesium ion incorporated implants (MgTiO implant), the other consisted of TiO2 stoichiometry implants (TiO implant). Surface oxide properties of the implants were characterized with various surface analytic techniques. After 6 weeks of follow up, the mean peak values of removal torque of Mg implants dominated significantly over TiO implants (p < or = 0.0001). Bonding failure generally occurred in the bone away from the bone to implant interface for the MgTiO implant and mainly occurred at the bone to implant interface for the TiO implant that consisted mainly of TiO2 chemistry and significantly rougher surface as compared to the MgTiO implant. Between bone and the Mg- incorporated implant surface, ionic movements and ion concentrations gradient were detected. The current in vivo experimental data may provide positive evidence for the surface chemistry-mediated biochemical bonding theory of oxidized bioactive implants. However, the present study does not rule out potential synergy effects of the oxide thickness, micro-porous structure, crystal structure and surface roughness on improvements of bone responses to oxidized bioactive implants.

Absorbable Implants↗

Well-ordered mesoporous bioactive glasses (MBG): a promising bioactive drug delivery system.

The local drug release system is considered to be an alternative to treat the bone infection. In this paper, well-ordered mesoporous bioactive glasses (MBG) with high specific surface area have been synthesized in aqueous solution by a two-step acid-catalyzed self-assembly process combined with hydrothermal treatment. Gentamicin was encapsulated into the MBG by adsorption method and in vitro release of gentamicin from MBG was performed in distilled water and modified simulated body fluid (SBF), respectively. The results showed that the amount of drug loading of MBG was three times more than that of conventional sol-gel 58S. The outcomes of drug release in distilled water and in SBF showed that M58S effectively decreased the initial burst. During the release period, gentamicin was released from the M58S at a much lower release rate as compared to that from 58S after soaking in distilled water and SBF. Furthermore, the drug release was sensitive to the pH and ionic concentration of the release medium suggesting possible controls of the release rate. In addition, in contrast to conventional sol-gel 58S, M58S had higher ability to induce hydroxyapatite (HAp) formation. Therefore, well-ordered mesoporous bioactive glasses might be used as a bioactive drug release system for preparation of bone implant materials.

Drug Delivery Systems↗

Bioactive polyacetylenes in food plants of the Apiaceae family: occurrence, bioactivity and analysis.

Many bioactive compounds with known effects on human physiology and disease have been identified through studies of plants used in traditional medicine. Some of these substances occur also in common food plants, and hence could play a significant role in relation to human health. Food plants of the Apiaceae plant family such as carrots, celery and parsley, contain a group of bioactive aliphatic C17-polyacetylenes. These polyacetylenes have shown to be highly toxic towards fungi, bacteria, and mammalian cells, and to display neurotoxic, anti-inflammatory and anti-platelet-aggregatory effects and to be responsible for allergic skin reactions. The effect of these polyacetylenes towards human cancer cells, their human bioavailability and their ability to reduce tumour formation in a mammalian in vivo model indicates that they may also provide benefits for health. The present state of knowledge on the occurrence of polyacetylenes in Apiaceae food plants, their biochemistry and bioactivity is presented in this review as well as relatively new methods for the isolation and quantification of these compounds from plants, plant products and biological fluids.

Acetylene↗

Interactions of bioactive glasses with osteoblasts in vitro: effects of 45S5 Bioglass, and 58S and 77S bioactive glasses on metabolism, intracellular ion concentrations and cell viability.

In a cell culture model of murine osteoblasts three particulate bioactive glasses were evaluated and compared to glass (either borosilicate or soda-lime-silica) particles with respect to their effect on metabolic activity, cell viability, changes in intracellular ion concentrations, proliferation and differentiation. 45S5 Bioglass caused extra- and intracellular alkalinization, a rise in [Ca2+]i and [K+]i, a small plasma membrane hyperpolarization, and an increase in lactate production. Glycolytic activity was also stimulated when cells were not in direct contact with 45S5 Bioglass particles but communicated with them only through the medium. Similarly, raising the pH of culture medium enhanced lactate synthesis. 45S5 Bioglass had no effect on osteoblast viability and, under most conditions, did not affect either proliferation or differentiation. Bioactive glasses 58S and 77S altered neither the ion levels nor enhanced metabolic activity. It is concluded that: (1) some bioactive glasses exhibit well-defined effects in osteoblasts in culture which are accessible to experimentation; (2) 45S5 Bioglass causes marked external and internal alkalinization which is, most likely, responsible for enhanced glycolysis and, hence, cellular ATP production; (3) changes in [H+] could contribute to alternations in concentrations of other intracellular ions; and (4) the rise in [Ca2+]i may influence activities of a number of intracellular enzymes and pathways. It is postulated that the beneficial effect of 45S5 on in vivo bone growth and repair may be due to some extent to alkalinization, which in turn increases collagen synthesis and crosslinking, and hydroxyapatite formation.

Animals↗

Induction of bioactivity of a non-bioactive glass-ceramic by a chemical treatment.

Glass-ceramic A-W(Al), which was prepared by heat treatment of a MgO-CaO-SiO2-P2O5-Al2O3 glass to precipitate crystalline apatite and wollastonite, shows a higher mechanical strength than glass-ceramic A-W, which was prepared by heat treatment of a MgO-CaO-SiO2-P2O5 glass to precipitate the same types of crystalline phases. The former, however, does not show bone-bonding ability, i.e. bioactivity, whereas the latter shows it. In the present study, in order to induce bioactivity of glass-ceramic A-W(Al), it was treated with HCl or NaOH solutions with different concentrations, and its bioactivity was evaluated by examining the apatite formation on its surface in a simulated body fluid (SBF) with ion concentrations nearly equal to those of human blood plasma. When the glass-ceramic A-W(Al) was pretreated with HCl aqueous solutions with concentrations over 0.1 M, it formed the bone-like apatite on its surface in SBF. This was attributed to the formation of a hydrated silica on its surface by the HCl treatment.

Apatites↗

Antibiotic delivery system using bioactive bone cement consisting of Bis-GMA/TEGDMA resin and bioactive glass ceramics.

A novel drug delivery system containing cephalexin (CEX) as a model drug using a new bioactive bone cement consisting of 15% bisphenol-alpha-glycidyl methacrylate (Bis-GMA), 15% triethylene-glycol dimethacrylate (TEGDMA) resin and 70% apatite- and wollastonite-containing glass-ceramic (A-W GC) powder was investigated. A-W GC powder containing CEX powder hardened within 5 min after mixing with Bis-GMA/TEGDMA resin, and furthermore its compressive strength was expected to be higher than that of polymethylmethacrylate cement. In vitro CEX release from bioactive bone cement pellets in a simulated body fluid at pH 7.25 and 37 degrees C continued for more than 2 weeks. The drug release rate increased with increasing amount of CEX in the mixture. All of the drug release profiles followed the Higuchi equation at the initial stage, but not at later stages. As hydroxyapatite was precipitated out on the cement surface, the drug release rate decreased. These results suggest that the CEX release rate from bioactive bone cement could be controlled by varying the amount of drug in the cement system.

Biomechanical Phenomena↗

Thiazolidinedione bioactivation: a comparison of the bioactivation potentials of troglitazone, rosiglitazone, and pioglitazone using stable isotope-labeled analogues and liquid chromatography tandem mass spectrometry.

Troglitazone, a thiazolidinedione (TZD) type insulin sensitizer for the treatment of diabetes, was withdrawn from the U.S. market after several fatal cases of hepatotoxicity. Although the mechanism(s) of these idiosyncratic adverse reactions are not completely understood, circumstantial evidence suggests at least a partial contribution of reactive metabolite formation. Despite isolated case reports of hepatotoxicity, the other TZD derivatives pioglitazone and rosiglitazone are comparatively safe. Herein, we report on the bioactivation potential of these drugs and their TZD ring isotope-labeled 2-(15)N-3,4,5-(13)C(3) analogues in rat and human liver microsomes supplemented with glutathione (GSH). Screening for GSH adducts as surrogate markers for reactive intermediate formation was performed by liquid chromatography tandem mass spectrometry. Chemical characterization of the GSH conjugates was conducted by acquisition of their respective product ion spectra and the comparison between unlabeled and stable isotope-labeled TZD derivatives. The data suggest that all drugs undergo bioactivation processes via a common metabolic activation on the TZD ring, yielding disulfide type GSH conjugates as evidenced by the loss of labeled positions in the TZD moiety. Additional bioactivation processes leading to GSH adducts not involving TZD ring scission were evident for troglitazone. In human liver microsomes at low substrate concentrations, only troglitazone yielded a predominant GSH adduct not involving TZD ring scission. This property may contribute, together with other factors such as the relatively high dose administered as well as its potential to induce hepatic cholestasis and oxidative stress, to the hepatotoxicity of this drug.

Animals↗

Effects of water-soluble component content on cephalexin release from bioactive bone cement consisting of bis-GMA/TEGDMA resin and bioactive glass ceramics.

The effect of the amount of a water-soluble, lactose, on cephalexin (CEX) release from bioactive bone cement consisting of bisphenol-alpha-glycidyl methacrylate (bis-GMA), triethylene-glycol dimethacrylate (TEGDMA) resin and apatite- and wollastonite-containing glass-ceramic (A-W GC) powder was investigated. A-W GC powder containing 5% CEX and lactose powders hardened within 5 min after mixing with bis-GMA/TEGDMA resin, and furthermore its compressive strength was expected to be higher than that of polymethylmethacrylate cement. In vitro CEX release from bioactive bone cement pellets in a simulated body fluid at pH 7.25 and 37 degrees C continued for more than 2 wk. The drug-release rate increased with increasing amount of lactose powder in the mixture. CEX release profiles followed the Higuchi equation in the initial stage, but not in later stages. As hydroxyapatite was precipitated out on the cement surface, the CEX release rate decreased. The micropore distribution of the cements measured by mercury porosimetry also supported the variation in drug release due to cement porosity being mainly a result of the dissolution of lactose in the cements. These results suggest that the rate of CEX release from bioactive bone cement could be controlled by varying the amount of lactose in the cement system.

Journal Article↗

Bioactive evaluation of 45S5 bioactive glass fibres and preliminary study of human osteoblast attachment.

Bioactive glass fibres can be used as tissue engineering scaffolds. In this investigation, the bioactive response of 45S5 glass fibres was assessed in simulated body fluid (SBF). Preliminary attachment of osteoblasts to the fibre surface was assessed, as were the fibre tensile strength and fracture toughness. Fourier transform infrared spectroscopy (FTIR) analysis revealed that hydroxyapatite (HA) was formed on the fibres' surface after 2-4 days in SBF. Raman micro-spectroscopic analysis was used to monitor development of the HA layer during immersion. A correlation was found between increase in intensity of the PO4(3-) peak near 964cm(-1) and appearance of crystalline HA (P-O bending peaks) using FTIR. Such results are encouraging for in situ bioactivity monitoring, as Raman spectra are insensitive to the presence of water, unlike FTIR. Average tensile strength of 45S5 fibres (79 microm diameter) was 340+/-140 MPa. Fracture toughness, determined using fracture surface analysis, was 0.7+/-0.1 MPa m1/2. Confocal microscopy revealed osteoblasts attached and spread along the fibres after 15-90 min culture. Scanning electron microscopy analysis showed that cells with filopodia and dorsal ruffles remained attached after 14 days in culture. These results are encouraging, as cell adhesion is an important first step prior to proliferation and differentiation.

Biocompatible Materials↗

Characterization and in vitro bioactivity of zinc-containing bioactive glass and glass-ceramics.

Zinc-containing glass is prepared by the substitution of CaO in 58S bioactive glass with 0.5 and 4 wt% ZnO, and glass-ceramics are obtained by heat-treating the glass at 1,200 C. The bending strength and in vitro bioactivity of the glass and glass-ceramics are evaluated. The results indicate that Zn promotes the crystallization of SiO(2) and wollastonite in glass-ceramics, and proper crystallization can enhance the bending strength of the glass-ceramic. The in vitro results show that ZnO in glass retards the hydroxyapatite (HA) nucleation at the initial stage of simulated body fluid (SBF) soaking, but does not affect the growth of HA after long periods of soaking, and the ionic products of 58S4Z glass can stimulate the proliferation of osteoblast at certain concentrations. Osteoblasts attach well on both glass samples and glass-ceramic samples, but the high Si ion concentration released from glass samples restrains the proliferation of osteoblasts after 3 days of culture. In contrast, osteoblasts show good proliferation on glass-ceramic samples, and ZnO in glass-ceramics promotes the proliferation rate. The results in this study suggest that the glass and glass-ceramics with different ZnO content might be used as bioactive bone implant materials in different applications.

Animals↗

Porcine and human relaxin bioactivity: bioactivities of porcine relaxin and human relaxin do not differ in mice and rats.

This study compares the bioactivity of porcine relaxin-1 to that of recombinant human relaxin-2 in mice and rats. The effects of the two hormone preparations on elongation of the mouse interpubic ligament and both the wet weight and the extensibility of the rat cervix were compared. No difference in bioactivity was detected between porcine relaxin-1 and recombinant human relaxin-2 in either rodent. Therefore, decisions concerning which of the two available forms of relaxin to employ for in vivo experimentation in mice and rats can be made without concerns about relative bioactivity.

Amino Acid Sequence↗