Papers by Keyword: Biomimetic Process

Abstract: A surface-mediated gene transfer system using DNA-calcium phosphate (CaP) composite layers (D-CaP layers) would be useful in tissue engineeing. In previous studies, D-CaP layers were fabricated in supersaturated CaP solutions prepared using chemical reagents. In this study, a so-called RKM solution prepared using clinically approved infusion fluids was employed as a supersaturated CaP solution. A D-CaP layer consisting of submicron spherical particles was successfully fabricated on a polystyrene substrate by immersing the substrate in the RKM solution for 24 h. When the immersion period was prolonged from 24 to 72 h, amount of CaP and DNA on the substrate increased. However, the gene transfer capability of the D-CaP layer for the CHO-K1 cells was kept unchanged irrespective of the immersion period. In the RKM solution process, immersion period of 24 h was found to be long enough for gene transfer application of the D-CaP layer. More importantly, the D-CaP layer fabricated by the RKM solution process exhibited a significantly higher gene transfer capability than our previous D-CaP layer fabricated in the conventional CaP solution with the same DNA concentration. The RKM solution process for the fabrication of D-CaP layers was found to be advantageous to the previous process in terms of not only safety but the layers gene transfer capability.

Abstract: Calcium phosphate (CaP) coating is an effective technique for surface functionalization of biomaterials. The objective of our research is to prepare calcium phosphate (CaP) coatings on a hydroxyapatite/collagen (HAp/Col) nanocomposite and subsequently provide it with gene delivery function through the immobilization of DNA in the coating. We have specifically selected the HAp/Col nanocomposite since it has the high potential as bone substitutes due to its similar composition, nanostructure, and biological properties to those of human bone. CaP coatings consisting of different sized particles were prepared on the HAp/Col nanocomposite membrane by immersing it in supersaturaterd CaP solutions (so-called RKM solutions) with the varied Ca and P concentration levels. We immobilized DNA in the CaP coatings together with lipid and fibronectin by supplementing DNA, lipid, and fibronectin to the RKM solutions (DLF-RKM solutions). Gene transfer capability of the resulting HAp/Col nanocomposite membrane was improved with decreasing concentration level of the DLF-RKM solution. It was confirmed that the present CaP coating technique was effective in providing the HAp/Col nanocomposite membrane with gene transfer capability and that the Ca and P concentration level of the DLF-RKM solution was a controlling factor affecting the gene transfer efficiency.

Abstract: Organic polymer–hydroxyapatite (HA) hybrid was expected as a promising bone substitute. In this work, poly(vinyl alcohol) (PVA) was used as a polymer substrate and the PVA-HA hybrid was prepared by a biomimetic process. The resultant PVA-HA composites were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis and scanning electron microscopy. It was found that HA could deposite on PVA surface when PVA soaked in water for a little swelling and then was carried out CaP treatment previously. The formed HA showed bone-like structure and had a good interaction with PVA substrate. The resultant hybrid might exhibit good osteoconductivity and biocompatibility, and could be expected as a useful scaffold for bone tissue regeneration.

Abstract: Development of the organic-inorganic hybrids composed of apatite crystals and organic polymer is expected to be an attractive material that has mechanical properties similar to natural bone as well as bone-bonding ability, i.e. bioactivity. It is reported that the carboxyl groups (-COOH) on the surfaces of the organic substrates act as a catalyst for induction of heterogeneous nucleation of apatite. The present authors previously showed that the apatite was successfully deposited on the polyglutamic acid gels containing abundant carboxyl groups through the biomimetic process, when they were priorly treated with calcium chloride solution. In this study, we fabricated the polyglutamic acid gels with different degree of cross-linking. Effect of the cross-linking on their ability of the apatite formation was examined in simulated body fluid (SBF). It was suggested that the apatite deposition on the polyglutamic acid gels is governed not only by the amount of –COOH that induces the heterogeneous nucleation of the apatite, but also by swelling property that controls local increase in degree of supersaturation with respect to the apatite.

Abstract: Thick coatings of bone morphogenetic protein (BMP)-calcium phosphate nano-composite were prepared by a biomimetic process, in which substrates were immersed in modified simulated body fluid containing rh-BMP2 at room temperature and pressure. In vitro analyses showed that the calcium phosphate and BMP formed stable and uniform coating of composite on surface of substrate, and that the composite derive significant improvement in tissue formation, suggesting an osteoinductive bioactive surface.

Abstract: Surgical implant coatings and grafts for tissue replacement have been made by porous surface materials to improve the implant to bone attachment. In this work, porous titanium samples were produced via powder metallurgy techniques and submitted to the biomimetic process in order to enhance its osteoconductivity. This process allows a nucleation and growth of a calcium phosphate film which makes a chemical bond with titanium. Therefore, it avoids the looseness of this film from substrate. The samples were chemically treated, heat treated at different temperatures and soaked into a modified body fluid solution (mSBF) during periods of 2 and 7 days. Samples with and without pretreatments and not soaked in mSBF were used as controls. SEM and EDX analyses detected a calcium phosphate phase on the sample surfaces treated at 400°C and 600°C and soaked in mSBF for 2 and 7 days. The results demonstrated the potential of the methodology applied for obtaining a bonelike apatite film on porous titanium samples processed by powder metallurgy.

Abstract: Natural bone has excellent mechanical properties such as high fracture toughness and high flexibility. These properties are achieved by specific microstructure of natural bone that is composed of the organic collagen and inorganic apatite. On the basis of these findings, apatite-polymer hybrids are expected as novel bone substitutes having excellent mehcanical performances and high bone-bonding ability, i.e. bioactivity. In this study, we attempted preparation of apatite-polyglutamic acid hybrids through biomimetic process that mimics the principle of biomineralization. Simple chemical modification of the polyglutamic acid gel with 1 M (= mol/L) calcium chloride solution provided the gel with apatite-forming ability in simulated body fluid (SBF, Kokubo solution). This type of hybrid is also useful for designing bioactive bone substitutes with injectability, since viscosity of the polyglutamic acid gel can be easily controlled according to degree of cross-linking.

Abstract: Organic-inorganic hybrids composed of organic polymer and apatite is quite attractive as novel bone-repairing materials since it has mechanical performance analogous to those of natural bone as well as bone-bonding ability, i.e. bioactivity. To fabricate such an apatite-polymer hybrid, biomimetic process has been recently paid much attention. In this process, bone-like apatite is deposited on the surfaces of organic substrates in simulated body fluid (SBF, Kokubo solution) having ion concentrations analogous to those of human extracellular fluid or more concentrated solutions. Previous studies showed that the apatite deposition is triggered by a catalytic effect of carboxyl groups (COOH) on the surfaces of the organic substrates. In this study, we examined apatite deposition on natural polypeptides derived from crops in a biomimetic solution. We selected gluten derived from wheat and zein derived from corn. Both of gluten and zein formed bone-like apatite on their surfaces in a solution that has inorganic ion concentrations 1.5 times those of simulated body fluid, when they were treated with 1 mol/L calcium chloride solution. High content of acidic amino acids such as glutamic acid and aspartic acid in gluten and zein would give large amount of carboxyl groups effective for the apatite nucleation.

Abstract: Apatite micropattern was fabricated by a combination of biomimetic process and transcription of resist pattern. We optimized some fabrication conditions such as the height of resist pattern, temperature, concentrations and pH of simulated body fluid(SBF). Consequently, we successfully obtained apatite micropattern widely and homogeneously on a substrate in a short fabrication period.

Abstract: Our previous study presented that sericin, a kind of silk protein, had high ability of apatite formation under a condition mimicking body environment when the sericin contains high content of β sheet structure. To confirm the effectiveness of β sheet structure on apatite nucleation, we attempted to synthesize of polypeptide containing β sheet structure and investigate apatite formation in 1.5SBF that has 1.5 times the inorganic ion concentrations of human blood plasma. Poly(FEFEFEFG) was synthesized as the polypeptide consisting mainly of β sheet structure. Formation of hydroxyapatite was observed on the film of the poly(FEFEFEFG) after soaking in 1.5SBF within 7 days. We could confirm that β sheet structure was effective even in the synthetic polypeptide.