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Isolation and characterization of plasmin-generated bioactive fragments of IGFBP-3.

Insulin-like growth factor-binding protein-3 (IGFBP-3) was digested with plasmin, and the proteolytic fragments were isolated by HPLC and tested for bioactivity as measured by stimulation of glucose uptake in microvessel endothelial cells. Two of the pooled fractions of the digest stimulated glucose uptake. The major bioactive pool, at an estimated protein concentration <50 ng/ml, stimulated glucose uptake to 150% of control with greater stimulation and 220% of control at approximately 250 ng/ml. Two fragments were present in the bioactive fraction, the dominant one migrating at approximately 20,000 and the other at approximately 8,000. Both fragments bound 125I-labeled insulin-like growth factor and [3H]heparin. NH2-terminal amino acid analysis of the bioactive peak yielded two sequences. One, representing the majority of the material, had an NH2-terminal sequence identical to IGFBP-3; the second fragment began at amino acid 202 of IGFBP-3. In contrast to the bioactive fragments, intact IGFBP-3, at concentrations up to 130 microgram/ml, had no bioactivity. These findings demonstrate that IGFBP-3 can be degraded into fragments that have potent bioactivities that are not present in the intact IGFBP-3 molecule.

Amino Acid Sequence↗

Antiequine chorionic gonadotropin (eCG) antibodies generated in goats treated with eCG for the induction of ovulation modulate the luteinizing hormone and follicle-stimulating hormone bioactivities of eCG differently.

In dairy goats, treatments associating a progestogen and the equine chorionic gonadotropin (eCG) are the easiest way to induce and synchronize estrus and ovulation and to permit artificial insemination (AI) and/or out of season breeding. From the first treatment, the injection of eCG induces, in some females, the production of anti-eCG antibodies (Abs) that will interfere with the effectiveness of subsequent treatments. These anti-eCG Abs delay the preovulatory LH surge and the ovulation time, leading to poor fertility of the treated females. In this study, by in vitro bioassays, we show that anti-eCG Abs can positively or negatively modulate the LH and/or FSH bioactivities of eCG. Moreover, the modulation level of eCG bioactivity does not depend on the anti-eCG Ab affinity for eCG, as shown by surface plasmon resonance technology. The specificity of anti-eCG Abs tested by competitive ELISA highlighted the importance of a glycan environment in the recognition mechanism, especially the sialic acids specific to eCG. The different effects of anti-eCG Abs on eCG bioactivities could be explained by two hypotheses. First, steric hindrance preventing the interaction of eCG with its receptors would explain the inhibitory effect of some anti-eCG Abs; second, a conformational change in eCG by anti-eCG Abs could induce inhibition or potentiation of eCG bioactivities. It is significant that these modulations of eCG bioactivities by anti-eCG Abs impact mainly on the FSH bioactivity of eCG, which is essential for ovarian stimulation and subsequent fertility after treatment and AI, and to a lesser extent on LH bioactivity.

Animals↗

Mode of secretion of bioactive luteinizing hormone in man.

The episodic nature of gonadotropin secretion was originally defined by RIA of circulating LH concentrations. We analyzed the pulsatile release of biologically active LH by measuring plasma LH concentrations in the rat interstitial cell testosterone bioassay. A computer algorithm to discriminate true biological signals (LH pulses) from background variation was applied to serially sampled LH data from seven men and seven postmenopausal women. Our results indicate the following. 1) In all subjects, mean bioactive LH values were considerably higher than immunoactive levels, (41.4 +/- 15.1 and 450 +/- 243 mIU/ml vs. 10.2 +/- 2.3 and 83 +/- 35 for men and postmenopausal women, respectively). There was a corresponding 4-fold increase in the total area under the bioactive LH secretion profile compared with that defined for the immunoactive hormone. 2) The absolute amplitude of the bioactive LH peaks was 0.5- to 11-fold higher than the immunoactive values. 3) Although the majority of the LH peaks were coincident by bioassay and RIA, significant dissociation occurred in 20% and 28% of the total LH peaks (rat interstitial cell testosterone bioassay and RIA) in men and postmenopausal women, respectively. 4) Significant increases in the bioactive to immunoactive ratio over interpulse bioactive to immunoactive levels occurred in 98% of the pulses in the men and 83% of those in postmenopausal women. Also, in two men, peaks of LH bioactivity exceeding 100 mIU were followed by major increases in serum testosterone concentrations. These findings demonstrate the value of bioactive LH determinations and indicate that LH is secreted in pulses of high biological activity. The in vitro LH bioassay provides a sensitive and appropriate estimate of functionally active LH in the circulation.

Adult↗

Bioactive composite materials for tissue engineering scaffolds.

Synthetic bioactive and bioresorbable composite materials are becoming increasingly important as scaffolds for tissue engineering. Next-generation biomaterials should combine bioactive and bioresorbable properties to activate in vivo mechanisms of tissue regeneration, stimulating the body to heal itself and leading to replacement of the scaffold by the regenerating tissue. Certain bioactive ceramics such as tricalcium phosphate and hydroxyapatite as well as bioactive glasses, such as 45S5 Bioglass, react with physiologic fluids to form tenacious bonds with hard (and in some cases soft) tissue. However, these bioactive materials are relatively stiff, brittle and difficult to form into complex shapes. Conversely, synthetic bioresorbable polymers are easily fabricated into complex structures, yet they are too weak to meet the demands of surgery and the in vivo physiologic environment. Composites of tailored physical, biologic and mechanical properties as well as predictable degradation behavior can be produced combining bioresorbable polymers and bioactive inorganic phases. This review covers recent international research presenting the state-of-the-art development of these composite systems in terms of material constituents, fabrication technologies, structural and bioactive properties, as well as in vitro and in vivo characteristics for applications in tissue engineering and tissue regeneration. These materials may represent the effective optimal solution for tailored tissue engineering scaffolds, making tissue engineering a realistic clinical alternative in the near future.

Biocompatible Materials↗

Aldose reductase inhibition increases CNTF-like bioactivity and protein in sciatic nerves from galactose-fed and normal rats.

The impact of exaggerated polyol pathway flux on ciliary neurotrophic factor (CNTF)-like bioactivity and expression of CNTF in rat sciatic nerve was examined after 2 months of galactose intoxication. Polyol content was elevated (P < 0.001) and motor nerve conduction velocity reduced (P < 0.05) in galactose-fed rats compared with control animals or control and galactose-fed rats treated with the aldose reductase inhibitor (ARI) Ponalrestat. CNTF-like bioactivity in the galactose-fed group was reduced to 30% of that assayed in the control group (P < 0.001). ARI treatment significantly increased CNTF-like bioactivity by 60% compared with the untreated galactose group (P < 0.05) but did not restore it to control levels. Unexpectedly, bioactivity in ARI-treated control animals was increased by nearly 250% compared with untreated controls (P < 0.005). In addition to the deficit in CNTF bioactivity in untreated galactose rats, the expression of protein, but not of mRNA, was reduced (P < 0.05). In ARI-treated control and galactose-fed rats, the expression of CNTF peptide was significantly enhanced above control levels (both P < 0.05). Concomitant with the reduction in CNTF levels, there was a shift in the axonal size-frequency distribution of myelinated fibers toward smaller axons in galactose-fed rats that was prevented by ARI treatment. Since galactose feeding has little impact on levels of CNTF mRNA, these observations suggest that deficits in CNTF-like bioactivity may result from a posttranscriptional modification of neurotrophic protein expression or turnover. Unlike other functional and structural disorders in galactose neuropathy, factors other than polyol accumulation may contribute to the deficit in CNTF-like bioactivity.

Aldehyde Reductase↗

Serum androgen bioactivity during 5alpha-dihydrotestosterone treatment in elderly men.

The androgens used in the treatment of age-related androgen decline have different bioactivities that cannot be evaluated with conventional detection methods for serum steroids. We have recently developed a recombinant cell bioassay for the determination of androgen bioactivity in human serum that is based on androgen-specific interaction between the ligand-binding domain (LBD) and the N-terminal region of the androgen receptor (AR). In this work, we examined the effect of topically applied 5alpha-dihydrotestosterone (DHT; 7.5-10 g of 2.5% DHT gel daily for 6 months) on circulating androgen bioactivity in 14 men (age range, 51-63 years) with symptoms of andropause and pretreatment serum testosterone less than 15 nM, or serum sex hormone-binding globulin concentration greater than 30 nM, or both. The mean (+/-SEM) pretreatment androgen bioactivity was 3.3 +/- 0.3 nM testosterone equivalents, and the levels correlated with serum testosterone concentration (r =.55, P <.05). DHT gel treatment induced a sixfold increase (from 1.5 +/- 0.1 nM to 9.0 +/- 0.7 nM) in mean serum DHT level, whereas endogenous testosterone and estradiol levels measured with radioimmunoassays were suppressed by approximately 70% and approximately 50%, respectively (P <.0001). Concomitantly, serum androgen bioactivity increased by sevenfold (from 3.3 +/- 0.3 to 23.6 +/- 2.8 nM testosterone equivalents; P <.0001). We conclude that DHT gel therapy in elderly men significantly increases their circulating androgen bioactivity as measured with a mammalian cell bioassay. An androgen-specific bioassay such as ours may enable investigation of other androgens with different bioactivities, such as selective AR modulators.

Aging↗

Topochemical design of bioactive peptides and peptidomimetics.

For the studies of bioactive peptides, our laboratories have been employed an integrated approach including synthesis, bioassays, and conformational analysis. To obtain highly potent, selective and metabolically stable analogs, peptidomimetics such as peptide backbone modifications (retro-inverso structures), constrained amino acids, and cyclic structures have been incorporated into many bioactive peptide sequences. The conformational studies of the resulting analogs have led to topochemical models for the bioactivities of those peptides. This lecture will be focused on the results of such studies on opioids and somatostatin. We have synthesized numerous opioid analogs with various peptidomimetics based on three classes: enkephalins, dermorphin-deltorphins, and morphiceptins. Many of these analogs exhibit high potency, selectivity, and metabolic stability. Conformational studies of these analogs have enabled us to define the structural characteristics necessary for bioactivities of morphiceptins, dermorphins, enkephalins, and deltorphins. From these results, we can propose conformational models responsible for bioactivities at the mu- and delta-receptors. Our studies of cyclic somatostatin analogs are based on the highly active Merck analog c(-Pro6-Phe7-D-Trp8-Lys9-Thr10-Phe11-) (where the superscripts denote position in native somatostatin). To investigate the topochemical preference of backbone and side chains, unusual amino acids, including beta-methylphenylalanine7 or 11, beta-methyltryptophan8, as well as backbone modifications such as retro-inverso structures have been incorporated. The bioactivity profiles of these peptidomimetic molecules provide much information on the effects of backbone and side chain constraints on bioactivity.

Amino Acid Sequence↗

Compositional dependence of bioactivity of glasses in the system Na2O-K2O-MgO-CaO-B2O3-P2O5-SiO2.

The bioactivity, i.e., bone-bonding ability, of 26 glasses in the system Na2O-K2O-MgO-CaO-B2O3-P2O5-SiO2 was studied in vivo. This investigation of bioactivity was performed to establish the compositional dependence of bioactivity, and enabled a model to be developed that describes the relation between reactions in vivo and glass composition. Reactions in vivo were investigated by inserting glass implants into rabbit tibia for 8 weeks. The glasses and the surrounding tissue were examined using scanning electron microscopy (SEM), light microscopy, and energy-dispersive X-ray analysis (EDXA). For most of the glasses containing < 59 mol % SiO2, SEM and EDXA showed two distinct layers at the glass surface after implantation, one silica-rich and another containing calcium phosphate. The build-up of these layers in vivo was taken as a sign of bioactivity. The in vivo experiments showed that glasses in the investigated system are bioactive when they contain 14-30 mol % alkali oxides, 14-30 mol % alkaline earth oxides, and < 59 mol % SiO2. Glasses containing potassium and magnesium bonded to bone in a similar way as bioactive glasses developed so far.

Animals↗

Surface reaction layer formation in vitro on a bioactive glass fiber/polymeric composite.

In order to provide a fixation vehicle between a polymeric composite femoral hip prosthesis and bone tissue, we fabricated bioactive glass fibers. The glass fibers had a tensile strength of 596 MPa, 14 times that of bulk bioactive glass. After immersion in protein-free simulated body fluid for 10 days, we observed the development of a calcium phosphate layer (specifically, partially crystallized, calcium-deficient carbonated hydroxyapatite) on the surface of the glass fibers. The stages of the surface reaction layer formation were similar to those of 45S5 bioactive glass although the kinetics of the reaction layer formation were slower. We combined the bioactive glass fibers with a polymeric matrix to form a fiber-reinforced composite material and observed the formation of a calcium phosphate layer on the surface of the glass fibers within the composite material after immersion in both protein-free and protein-containing simulated body fluids. The rate of reaction layer formation was reduced in the presence of proteins. In both protein-free and protein-containing solutions, a "halo" of bioactivity reactions was observed on the surface of the polymer in regions surrounding the glass fibers. Our results suggest these glass fibers and glass fiber composites will exhibit bioactivity reactions in vivo.

Biocompatible Materials↗

Effect of surface reaction stage on fibronectin-mediated adhesion of osteoblast-like cells to bioactive glass.

Bioactive glasses and ceramics enhance bone formation and bond directly to bone, and have emerged as promising substrates for bone tissue engineering applications. Bone bioactivity involves physicochemical surface reactions and cellular events, including cell attachment to adsorbed extracellular matrix proteins. The effects of fibronectin (Fn) adsorption and glass surface reaction stage on the attachment of osteoblast-like cells (ROS 17/2.8) to bioactive glass were analyzed. Bioactive glass disks were pretreated in a simulated physiologic solution to produce three reaction layers: unreacted glass (BG0), amorphous calcium phosphate (BG1d), and carbonated hydroxyapatite (BG7d). Synthetic hydroxyapatite (sHA) and nonreactive borosilicate glass (CG) were used as controls. A spinning disk device which applied a linear range of forces to attached cells while maintaining uniform chemical conditions at the interface was used to quantify cell adhesion. The number of adherent cells decreased in a sigmoidal fashion with applied force, and the resulting detachment profile provided measurements of adhesion strength. For the same amount of adsorbed Fn, cell adhesion was higher on surface-reacted bioactive glasses (BG1d and BG7d) than on BG0, CG, and sHA. For all substrates, cell attachment was primarily mediated by the RGD binding site of Fn, as demonstrated by blocking experiments with antibodies and RGD peptides. Cell adhesion strength increased linearly with adsorbed Fn surface density. Analysis of this fundamental relationship revealed that improved adhesion to reacted bioactive glasses resulted from enhanced cell receptor-Fn interactions, suggesting substrate-dependent conformational changes in the adsorbed Fn.

Biocompatible Materials↗

Prodrug bioactivation and oncolysis of diffuse liver metastases by a herpes simplex virus 1 mutant that expresses the CYP2B1 transgene.

BACKGROUND: Herpes simplex virus 1 (HSV-1) infection of cancer cells results in viral replication with cell destruction and liberation of progeny virion that infect adjacent tumor cells. rRp450 is a novel replication-conditional HSV-1 mutant that expresses both the endogenous herpes viral thymidine kinase gene and the rat p450 CYP2B1 transgene; p450 bioactivates such cancer prodrugs as cyclophosphamide. METHODS: Viral cytotoxicity and replication assays were performed in colon carcinoma cells as well as primary human hepatocytes. For in vivo studies, diffuse liver metastases were generated by inoculating MC26 cells into the portal system of BALB/c mice. Mice were treated with control media, rRp450, or rRp450 plus cyclophosphamide. RESULTS: Cytopathic effects induced by rRp450 replication in colon carcinoma cells were equivalent to those induced by wild type HSV-1 in vitro. Assays developed to separate cytotoxicity mediated by viral replication from cytotoxicity mediated by chemotherapy confirmed that HSV-1 thymidine kinase bioactivates ganciclovir and CYP2B1 bioactivates cyclophosphamide in rRp450-infected cells. rRp450 mediated cytotoxicity in the presence of cyclophosphamide was increased by 21% to 30% above that achieved by viral replication alone. Cyclophosphamide bioactivation produced bystander killing of colon carcinoma cells but not hepatocytes. In contrast to these effects of cyclophosphamide, rRp450 mediated cytotoxicity was reduced in the presence of ganciclovir. These findings are explained by further experiments showing that bioactivation of cyclophosphamide only minimally affected HSV-1 replication in colon carcinoma cells, whereas bioactivation of ganciclovir markedly attenuated HSV-1 replication. In vivo studies revealed a substantial decrease in hepatic tumor burden in all rRp450-treated animals compared to controls. The addition of cyclophosphamide augmented rRp450's in vivo anti-neoplastic effect. CONCLUSIONS: The rRp450 mutant HSV-1 is highly oncolytic against colon carcinoma cells both in vitro and in vivo. rRp450 displays preferential replication in colon carcinoma cells compared to normal hepatocytes. Activation of cyclophosphamide by the p450 transgene augmented the anti-neoplastic effects of rRp450 without simultaneously decreasing viral replication. Oncolysis induced by HSV-1 replication combined with cyclophosphamide prodrug activation warrants further investigation as a potential therapy for colon carcinoma liver metastases.

Antineoplastic Agents, Alkylating↗

An investigation of bioactive glass powders by sol-gel processing.

Bioactive glass powders, with a composition of SiO 2-CaO-P 2O 5, have been successfully synthesized via a sol-gel process at considerably lower temperatures than required for conventional melting methods. Bioactive glass powders made via conventional methods form an interfacial bond with bone when they are implanted. Bonding is correlated with the formation of a surface hydroxyapatite layer. This study examined the formation of a hydroxyapatite layer in Tris-buffered solution as a function of SiO 2 content of sol-gel derived powders. A FT-IRRS technique was used to monitor the formation of the hydroxyapatite on the surface of the powders. X-ray diffraction analysis and BET were also used to characterize the chemical and physical properties of the sol-gel derived bioactive powders. It was discovered that: (a) the rate of hydroxyapatite formation decreased with increasing SiO 2 content for powders whose SiO 2 content was less than 90 mol%; (b) a hydroxyapatite film does not form for the powders whose SiO 2 content is more than 90 mol%; (c) the SiO 2 limit, beyond which the powders lost their bioactivity, was much higher for bioactive glass powders made through sol-gel process (90%) than those made by conventional melting methods (60%). These results indicate that it is possible to significantly expand the bioactive composition range through microstructural control made possible by sol-gel processing techniques.

Glass↗

Interfacial tensile strength between polymethylmethacrylate-based bioactive bone cements and bone.

We have developed two types of polymethylmethacrylate (PMMA)-based bioactive bone cements containing bioactive glass beads (designated GBC) or apatite-wollastonite containing glass-ceramic powder (designated AWC) as the filler. A new method was used to evaluate the bone-cement interfacial strength of these bioactive bone cements. Two types of bioactive bone cements (GBC and AWC) and PMMA cement (CMW-1) were put in a frame attached to the smooth tibial metaphyseal cortex of the rabbit and polymerized in situ. The load required to detach the cement from the bone was measured at 4, 8, and 16 weeks after implantation. The interfacial tensile strength of GBC and AWC showed significantly higher values than PMMA cement from 4 weeks, and increased with time. For GBC, strength reached a maximum value of 12.39 +/- 1.79 kgf 16 weeks after implantation. Histological examination of rabbit tibiae up to 16 weeks demonstrated no intervening layer between the bioactive bone cements and the bone, whereas fibrous tissue was observed at the interface between the PMMA cement and the bone. From this study, we conclude that PMMA-based bioactive bone cements have a relatively higher adhesiveness at the interface than the conventionally used PMMA cement, showing potential as a promising alternative.

Animals↗

Effect of wollastonite ceramics and bioactive glass on the formation of a bonelike apatite layer on a cobalt base alloy.

A biomimetic method was used to promote a bioactive surface on a cobalt base alloy (ASTM F-75). The metallic substrates were alkali treated and some of the samples were subsequently heat treated. The treated samples were immersed in simulated body fluid (SBF) on granular particles of either bioactive glass or wollastonite. For comparative purposes, no bioactive system was used in some tests. Three different methods were used for the immersion of the samples in SBF: 1) 21 days in SBF, 2) 21 days in 1.5 SBF, and 3) 7 days in SBF followed by 14 days in 1.5 SBF (re-immersion method). A bonelike apatite layer was formed on all the samples placed on wollastonite and bioactive glass particles. The morphology of the apatite layer formed by using the re-immersion method and wollastonite closely resembled the existing bioactive systems. No apatite layer was observed on the samples treated without bioactive material and soaked for 21 days in SBF or 1.5 SBF, apart from the substrates treated by using the re-immersion method. The heat treatment delayed the apatite formation in all the cases studied.

Alloys↗

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.

Biocompatible Materials↗

Synthesis of bioactive PMMA bone cement via modification with methacryloxypropyltri-methoxysilane and calcium acetate.

Bone cement consisting of polymethylmethacrylate (PMMA) powder and methylmethacrylate (MMA) liquid is clinically used for fixation of implants such as artificial hip joints. However, it does not show bone-bonding ability, i.e., bioactivity. The lack of bioactivity would be one of factors which cause loosening between the cement and the implant. The present authors recently showed the potential of bioactive PMMA-based bone cement through modification with gamma-methacryloxypropyltrimethoxysilane (MPS) and calcium acetate. In this study, the effects of the kinds of PMMA powder on setting time, apatite formation and compressive strength were investigated in a simulated body fluid (Kokubo solution). The cement modified with calcium acetate calcined at 220 degrees C could set within 15 min when the PMMA powder had an average molecular weight of 100,000 or less. The addition of calcium acetate calcined at 120 degrees C in the PMMA powder required a much longer period for setting. The modified cements formed an apatite layer after soaking in the Kokubo solution within 1 day for cement starting from PMMA powder with a molecular weight of 100,000 or less. Compressive strengths of the modified cements were more than 70 MPa for cements starting from 100,000 and 56,000 in molecular weight. After soaking in Kokubo solution for 7 days, the modified cement consisting of PMMA powder of 100,000 in molecular weight showed a smaller decrease in compressive strength than that consisting of 56,000 in molecular weight. These results indicate that bioactive PMMA cement can be produced with appropriate setting time and mechanical strength when PMMA powders with a suitable molecular weight are used. Such a type of design of bioactive PMMA bone cement leads to a novel development of bioactive material for bone substitutes.

Acetates↗

New chemical treatment for bioactive titanium alloy with high corrosion resistance.

It was recently claimed that titanium metal and its alloys can bond to the living bone, without being coated by apatite (VPS coatings), but by being chemically and heat-treated. The bioactivity of treated titanium is of interest because of the opportunity to obtain orthopaedic or dental implants presenting, at the same time, high toughness, strength and fatigue resistance as well as bone-bonding ability. The bioactive behaviour of the treated implants is due to the presence of a modified surface, which, during soaking in body fluid, promotes the precipitation of apatite. The apatite formed is strongly bonded to the substrate and promotes living bone bonding. In this work were characterised samples of Ti-6Al-7Nb alloy with surfaces presenting a different chemical and mechanical state. The aim of the research was twofold. The first objective was to characterise chemically and heat-treated samples with different surface topography, in order to define the best conditions for osteogenic integration. The second aim was to assess the corrosion behaviour of the bioactive implants, because they expose a microporous and quite thin modified surface layer. No-treated and passivated samples, with a surface state closed to that nowadays used on implants, were used as reference. The surface structure, morphology, electrochemical behaviour and bioactivity of the different samples were assessed by means of XRD, SEM-EDS, anodic polarizations, open circuit measurements and in-vitro tests. Results evidence that it is possible to modify the surface of the Ti-6Al-7Nb alloy in order to obtain the formation of a bioactive layer and that the substrate roughness influences the characteristics of the surface layer formed. It was also evidenced that the as treated surfaces present inadequate corrosion behaviour, so a new two-step chemical treatment has been developed in order to obtain a bioactive material with good corrosion resistance.

Biocompatible Materials↗

Bioactivity of CaSiO3/poly-lactic acid (PLA) composites prepared by various surface loading methods of CaSiO3 powder.

Mixing bioactive ceramic powders with polymers is an effective method for generating bioactivity to the polymer-matrix composites but it is necessary to incorporate up to 40 vol% of bioactive ceramic powder. However, such a high mixing ratio offsets the advantages of the flexibility and formability of polymer matrix and it would be highly advantageous to lower the mixing ratio. Since surface loading of ceramic powders in the polymer is thought to be an effective way of reducing the mixing ratio of the ceramic powder while maintaining bioactive activity, CaSiO(3)/poly-lactic acid (PLA) composites were prepared by three methods; (1) casting, (2) spin coating and (3) hot pressing. In methods (1) and (2), a suspension was prepared by dissolving PLA in chloroform and dispersing CaSiO(3) powder in it. The suspension was cast and dried to form a film in the case of method (1) while it was spin-coated on a PLA substrate in method (2). In method (3), CaSiO(3) powder was surface loaded on to a PLA substrate by hot pressing. The bioactivity of these samples was investigated in vitro using simulated body fluid (SBF). Apatite formation was not observed in the samples prepared by method (1) but some apatite formation was achieved by mixing polyethylene glycol (PEG) with the PLA, producing a porous polymer matrix. In method (2), apatite was clearly observed after soaking for 7 days. Enhanced apatite formation was observed in method (3), the thickness of the resulting apatite layers becoming about 20 microm after soaking for 14 days. Since the amount of CaSiO(3) powder used in these samples was only <or=0.4 vol%, it is concluded that this preparation method is very effective in generating bioactivity in polymer-matrix composites by loading with only very small amounts of ceramic powder.

Biocompatible Materials↗