Nano-scaled hydroxyapatite/polymer composite I. Coating of sintered hydroxyapatite particles on poly(gamma-methacryloxypropyl trimethoxysilane)grafted silk fibroin fibers through chemical bonding.

The inorganic-organic composite consisting of nano-scaled hydroxyapatite (HAp) and silk fibroin (SF) fibers was prepared through covalent linkage to develop a novel biomaterial for a soft-tissue-compatible material. The preparation of the composite was conducted through the three-step procedure consisting of chemical modification using 2-methacryloxyethyl isocyanate (MOI) monomer to introduce vinyl groups on SF, poly(gamma-methacryloxypropyl trimethoxysilane) (MPTS) graft-polymerization on SF, and coupling process between the surface of polyMPTS-grafted SF and HAp nano-particles. The amount of the graft-polymerization of polyMPTS through vinyl groups was well controlled by the reaction time. The nano-crystals were subsequently coated on the grafted fibers by heating at 120 degrees C for 2 h in a vacuum. The crystalline structure of the SF substrate did not change in the procedure. In the SEM observation of the composite surface, it was found that the bonded nano-crystals were separated and partially aggregated with several crystals attached on the SF fiber surface. The HAp particles adhered more strongly on the SF surface with separation or aggregation of several crystals than on the surface of the original SF after ultrasonic treatment.

Preparation of porous composite implant materials by in situ polymerization of porous apatite containing epsilon-caprolactone or methyl methacrylate.

Biodegradable and biostable composite foams were formed from porous apatite cement infiltrated with epsilon-caprolactone (CL) or methylmethacrylate (MMA) using a high over vacuum. For CL composites in situ polymerization was induced using trace water as an initiator and heating at 120 degrees C for up to 10 days or at 80 degrees C for 60 days. MMA composites were polymerized using AIBN initiator at 70 degrees C for 8 h. CL preparations gave composites with a polycaprolactone (PCL) number average of molecular weight (Mn) up to the maximum of 7.1 x 10(3) g/mol after 10 days and 16.8 x 10(3) g/mol after 60 days. The PCL and PMMA contents were close to 50 and 40 wt%, respectively, polymer was present as a thin coating on the apatite crystal plates and was evenly distributed throughout the samples. Re-evacuation of apatite saturated with monomer during preparation ensured that the upwards of 200 nm microchannels within the apatite cement were largely free of polymer, and the overall macroporous structure of the apatite foams was partly retained. Maximum compressive strengths increased from 9 MPa to 37 and 64 MPa for PCL and PMMA composites, respectively. The water drop contact angle of the PCL composite was 64 degrees, and therefore suitable for cell attachment. PMMA composite surfaces were more hydrophobic. Composites were subjected to corona discharge to induce suitable moderate hydrophilicity at the surface.

A hydroxyapatite coating covalently linked onto a silicone implant material.

A novel composite consisting of hydroxyapatite (HAp) microparticles covalently coupled onto a silicone sheet was developed. Initially, an acrylic acid (AAc) -grafted silicone sheet with a 16.7 microg/cm(2) surface graft density was prepared by corona-discharge treatment. The surface of sintered, spherical, carbonated HAp particles with an average diameter of 2.0 microm was subsequently modified with amino groups. The amino group surface density of the HAp particles was calculated to be approximately one amino molecule per 1.0 nm(2) of particle surface area. These samples were characterized with Fourier transform infrared spectrometry and X-ray photoelectron spectroscopy. After the formation of ammonium ionic bonds between both samples under aqueous conditions, they were reacted at 180 degrees C for 6 h in vacuo to form covalent bonds through a solid-phase condensation. The HAp particles were coupled to the AAc-grafted silicone surface by a covalent linkage. Further improvements in the adhesive and bioactive properties of the HAp-coated silicone material are expected.

Preparation and characterization of apatite deposited on silk fabric using an alternate soaking process.

Apatite-deposited silk fabric composite materials were developed using a new alternate soaking process. The characteristics of deposited apatite were studied using scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared spectrophotometry (FTIR), and X-ray photoelectron spectroscopy (XPS). Apatite weight increased with alternating soaking in a calcium solution [200 mM aqueous calcium chloride solution buffered with tris(hydroxymethyl) aminomethane and HCl (pH 7.4)] and a phosphate solution (120 mM aqueous disodium hydrogenphosphate) changed every hour. SEM showed that apatite deposited after 21 or more repeated soakings was over 20 microm thick. XRD showed that with alternate soakings, the apatite crystals deposited on silk fabric elongated along the c axis. FTIR and XPS indicated the existence of carbonate, HPO(4)(2-), and Na(+) ions in addition to constituent ions of hydroxyapatite. A loss of HPO(4)(2-) and Na(+) ions in the deposit upon further soaking might be associated with an increasing apatite crystallinity. Apatite deposited on silk by the alternate soaking process was a deficient apatite containing carbonate, HPO(4)(2-), and Na(+) ions as in a natural bone tissue. Thus, this apatite-silk composite material might be potentially bioactive.

Chemical modification of silk fibroin with 2-methacryloyloxyethyl phosphorylcholine. II. Graft-polymerization onto fabric through 2-methacryloyloxyethyl isocyanate and interaction between fabric and platelets.

2-Methacryloyloxyethyl phosphorylcholine (MPC) was grafted onto silk fabric in a two-step heterogeneous system through the vinyl bonds of 2-methacryloyloxyethyl isocyanate (MOI) modified on the fabric. First, habutae silk fabric was modified with the MOI monomer in anhydrous dimethyl sulfoxide using di-n-butyltin (IV) dilaurate and hydroquinone at 35 degrees C. The saturated weight gain of modified MOI monomer on the fabric was 7.3 wt% versus the original silk. Second, graft polymerization with MPC onto the MOI modified silk was conducted using 2,2'-azo bis[2-(2-imidazolin-2-yl)propane dihydrochloride] (VA-044) as an azo polymerization initiator. The weight of the grafted MPC eventually gained was about 26.0 wt%. The MOI-modified and MPC-grafted silk fabrics were analyzed by Fourier transform infrared (FT-IR) spectroscopy. To confirm the improved biocompatibility of the silk fabric, platelet adhesion was preliminarily tested measuring lactate dehydrogenase. The number of platelets adhering to polyMPC-grafted silk fabric decreased by about one tenth compared to original and MOI-modified silk after 60 min of contact with human platelet-rich plasma (1.0 x 10(6) platelets cm(-2)).

Ca-adsorption and apatite deposition on silk fabrics modified with phosphate polymer chains.

Silk fabric was modified with polymethacryloyloxyethylphosphate (pMOEP) by graft copolymerization. Ca-adsorption onto pMOEP-grafted silk fabric was significantly enhanced compared to that onto original silk fabric. SEM observation indicated that some crystallites were deposited on the pMOEP-grafted silk fabric after 1 week of immersion in simulated body fluid, whereas no change occurred on the surface of the original silk fabric. X-ray diffraction showed that this crystallite contained hydroxyapatite. These results indicate that pMOEP-grafted silk fabric induce hydroxyapatite formation more effectively than the original silk fabric.

Novel functional polymers: poly(dimethylsiloxane)-polyamide multiblock copolymer. V. The interaction between biomolecules and the surface of aramid-silicone resins.

Multiblock copolymers consisting of aromatic polyamide(aramid) and poly(dimethylsiloxane) (PDMS) aramid-silicone resins (PASs) were synthesized by low temperature solution polycondensation, and PAS films were prepared by casting from an N,N'-dimethylacetamide solution. In this study, we investigated bovine serum albumin (BSA) adsorption, L929 cell adhesion, and tissue reaction on the surface of PAS in order to clarify the interaction between PAS and biomolecules. It was found that the amount of adsorbed biomolecules on PAS was extremely low in contrast with those on aramid and nylon films, and it was comparable to SILASTIC 500-1 film. This suppression of adsorption of biomolecules onto PAS seemed to be due to the low surface free energy of the outermost surface of PAS, where PDMS block was condensed.

[The diagnosis of a mutation of the insulin receptor by non-radioisotropical RT-PCR-SSCP analysis].

We evaluated a 24 years old diabetes woman with type A insulin resistant (patient "Yakushima") who had some typical symptoms as acanthosis nigricans, hirsutism and virilization. Insulin binding to the patient's erythrocytes was significantly decreased to about 30% of the normal control. In order to determine the mutation of the insulin receptor, we used for reverse transcript-polymerase chain reaction-single strand conformation polymorphism (RT-PCR-SSCP) analysis without using radiolabeled materials. We also analysed the nucleotide sequence with non-isotropical probe. Our results suggested that the mutation was heterozygous, and the patient had a new missense mutation substituted Asn461 for Thr461 in the alpha-subunit.

A novel biomaterial: poly(dimethylsiloxane)-polyamide multiblock copolymer I. Synthesis and evaluation of blood compatibility.

Aramid-silicone resins (PASs) consisting of aromatic polyamide (aramid) and poly(dimethyl-siloxane) (PDMS) segments were synthesized by low temperature solution polycondensation. For the evaluation of blood compatibility in vitro, two kinds of experiments were carried out. One was the thromboxane B2(TXB2) release test from platelets attaching to PAS and Biomer. The other was the observation of the platelet adhesion on the surfaces of PAS by scanning electron microscopy (SEM). The results indicated that PAS was bio-inert in vitro. The surface chemical composition of PAS films was investigated by means of electron probe micro analysis (EPMA), X-ray photoelectron spectroscopy (XPS), and dynamic contact angle measurements. The relationship between blood compatibility and surface composition of PAS is discussed.