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Biomedical subjects

Masao Yoshinari

Publications and source records attributed to Masao Yoshinari.

At least 19 recordsLinked to original sources

Adhesion of mouse fibroblasts on hexamethyldisiloxane surfaces with wide range of wettability.

Surface wettability is an important physicochemical property of biomaterials, and it would be more helpful for understanding this property if a wide range of wettability are employed. This study focused on the effect of surface wettability on fibroblast adhesion over a wide range of wettability using a single material without changing surface topography. Plasma polymerization with hexamethyldisiloxane followed by oxygen (O2)-plasma treatment was employed to modify the surfaces. The water contact angle of sample surfaces varied from 106 degrees (hydrophobicity) to almost 0 degrees (super-hydrophilicity). O2 functional groups were introduced on polymer surfaces during O2-plasma treatment. The cell attachment study confirmed that the more hydrophilic the surface, the more fibroblasts adhered in the initial stage that includes super-hydrophilic surfaces. Cells spread much more widely on the hydrophilic surfaces than on the hydrophobic surfaces. There was no significant difference in fibroblast proliferation, but cell spreading was much greater on the hydrophilic surfaces. The fibronectin adsorbed much more on a hydrophilic surface while albumin dominated on a hydrophobic surface in a competing mode. These findings suggest the importance of the surface wettability of biomaterials on initial cell attachment and spreading. The degree of wettability should be taken into account when a new biomaterial is to be employed.

Animals↗

Fibronectin immobilization using water-soluble carbodiimide on poly-L-lactic acid for enhancing initial fibroblast attachment.

The aim of this study is to evaluate the influence of fibronectin immobilization on poly-L-lactic acid (PLA) films on the initial attachment of human gingival fibroblasts. Carboxylic acid groups are chemically introduced on the PLA films' surface by surface hydrolysis with 0.5 M NaOH. The contact angle of PLA surface with respect to double-distilled water decreases significantly after NaOH hydrolysis. X-ray photoelectron spectroscopy (XPS) also reveals significantly higher intensities of C(C=O)/C(C-O) after NaOH hydrolysis. Fibronectin is immobilized onto the hydrolyzed PLA surface through a condensation reaction between the carboxylic acid groups on the hydrolyzed PLA surface and the amino groups of fibronectin using water-soluble carbodiimide. XPS analysis shows that the fibronectin-immobilized PLA surface is enriched with nitrogen atoms. The immobilization of fibronectin significantly enhances the number of initially attached human gingival fibroblasts on the PLA surface. No obvious differences in morphology are noted between fibroblasts cultured on native PLA and on fibronectin-immobilized PLA. Fibronectin can be immobilized onto the PLA surface after NaOH hydrolysis and this is effective in enhancing the initial attachment of human gingival fibroblasts.

Adsorption↗

Adsorption behavior of antimicrobial peptide histatin 5 on PMMA.

Adsorption of antimicrobial peptide histatin 5 on a poly(methyl methacrylate) denture base may serve to prevent biofilm formation, leading to a reduction of denture-induced stomatitis. This study focused on adsorption behavior of histatin 5 onto PMMA surfaces modified using a cold plasma technique and the effectiveness of histatin 5 adsorption for reducing Candida albicans biofilm formation by the quartz crystal microbalance-dissipation (QCM-D) technique. PMMA spin-coated specimens were treated with oxygen (O(2)) plasma using a plasma surface modification apparatus. The amount of histatin 5 adsorbed onto the PMMA treated with O(2) plasma is more than six times greater than that adsorbed onto untreated PMMA. The degree of histatin 5 adsorption had a negative correlation with the contact angle, whereas that of zeta-potential showed no significant correlation. XPS analysis revealed that the introduction of the carboxyl and O(2) functional groups were observable on the O(2) plasma-treated PMMA. Increased surface hydrophilicity and the formation of the carboxyl could be responsible for histatin 5 adsorption on plasma-treated PMMA. There is no significant difference between histatin-adsorbed PMMA and control PMMA for C. albicans initially attached. On the contrary, the amount of C. albicans colonization on histatin-adsorbed PMMA was significantly less than the control.

Adsorption↗

Control of bisphosphonate release using hydroxyapatite granules.

The efficacy of hydroxyapatite (HAp) as a carrier was investigated to establish a method of local administration of bisphosphonates (Bps), which has currently been administered systemically. HAp granules (300-500 microm in size) with different physicochemical features were prepared by altering the sintering temperature. To ascertain the physicochemical properties of the HAp granules, their crystallinity was assessed using X-ray diffraction, the surface morphology was examined under scanning electron microscopy, and the specific surface area and calcium dissolution were evaluated. Different Bps-HAp composites were subsequently prepared and the concentration of Bps released from these composites was measured. The influence of Bps-HAp composites on the rate of osteoclast survival was also evaluated. The results revealed that (1) HAp solubility depends on the sintering temperature; (2) The concentration of released Bps could be controlled by regulating the sintering temperature of HAp as a carrier; and (3) Bps released from Bps-HAp composites reduced the number of osteoclasts. These findings indicated that Bps-HAp composites could be locally administered as a drug delivery system to areas with bone resorption.

Animals↗

Oxygen plasma surface modification enhances immobilization of simvastatin acid.

Simvastatin acid (SVA) has been reported to stimulate bone formation with increased expression of BMP-2. Therefore, immobilization of SVA onto dental implants is expected to promote osteogenesis at the bone tissue/implant interface. The aim of this study was to evaluate the immobilization behavior of SVA onto titanium (Ti), O(2)-plasma treated titanium (Ti + O(2)), thin-film coatings of hexamethyldisiloxane (HMDSO), and O(2)-plasma treated HMDSO (HMDSO + O(2)) by using the quartz crystal microbalance-dissipation (QCM-D) technique. HMDSO surfaces were activated by the introduction of an OH group and/or O(2)-functional groups by O(2)-plasma treatment. In contrast, titanium surfaces showed no appreciable compositional changes by O(2)-plasma treatment. The QCM-D technique enabled evaluation even at the adsorption behavior of a substance with a low molecular weight such as simvastatin. The largest amount of SVA was adsorbed on O(2)-plasma treated HMDSO surfaces compared to untreated titanium, HMDSO-coated titanium, and O(2)-plasma treated titanium. These findings suggested that the adsorption of SVA was enhanced on more hydrophilic surfaces concomitant with the presence of an OH group and/or O(2)-functional group resulting from the O(2)-plasma treatment, and that an organic film of HMDSO followed by O(2)-plasma treatment is a promising method for the adsorption of SVA in dental implant systems.

Adsorption↗

The bisphosphonate pamidronate on the surface of titanium stimulates bone formation around tibial implants in rats.

Many materials with differing surfaces have been developed for clinical implant therapy in dentistry and orthopedics. We analyzed the quantity of new bone formed in vivo around calcium-immobilized titanium implants with surfaces modified using pamidronate (PAM), a nitrogen-containing bisphosphonate (N-BP), implants of pure titanium, and titanium implants immobilized with calcium ions. New bone formation was visualized using fluorescent labeling (calcein blue and alizarin complexone) with intravenous injection at 1 and 3 weeks after implantation. After 4 weeks, undecalcified sections were prepared, and new bone formation around the implants was examined by morphometry using confocal laser scanning microscopy images. After 1 week, more new bone formed around the PAM-immobilized implant than around the calcium-immobilized and pure titanium implants. This was also seen with the new bone formation after 3 weeks. After 4 weeks, significantly more new bones were formed around the BP-immobilized implant than around the calcium ion-implanted and pure titanium implants. The new N-BP-modified titanium surface stimulates new bone formation around the implant, which might contribute to the success of implant therapy.

Animals↗

Corrosion behavior and surface characterization of titanium in solution containing fluoride and albumin.

The objective of this study was to demonstrate the role of albumin on the corrosion behavior of titanium in a solution containing 2.0 g/l fluoride and either 0.1 or 1.0 g/l albumin. The corrosion behavior and surface characterization of passive films on titanium immersed in such a solution were examined. In addition, the change in pH and the concentration of dissolved titanium in the solution were examined. The results showed that the corrosion of titanium in a solution containing fluoride was distinct, and that adding albumin to the solution containing fluoride suppressed corrosion. Fluorine was detected on the titanium surface immersed in the solution containing fluoride, and dissolution of the titanium was confirmed. The titanium immersed in a solution containing both fluoride and albumin had an albumin film regardless of the albumin concentration. In addition, the amount of dissolved titanium from the titanium immersed in the solution was less than when the solution contained no albumin. It was suggested that the formation of adsorbed albumin films on or in the passive film acted to not only protect the titanium from attack by the fluoride but also suppressed dissolution of the titanium-fluoride compounds.

Albumins↗

Changes in the distribution of laminin-5 during peri-implant epithelium formation after immediate titanium implantation in rats.

Laminin-5 (Ln-5), a component of the basement membrane (BM), regulates epithelial cell migration and adhesion. This study used anti-Ln-5 (gamma2chain) antibody to investigate the distribution of Ln-5 during the formation of peri-implant epithelium (PIE) in rats, and compared it to the distribution of Ln-5 during oral mucosa formation after tooth extraction. One day after extraction, the junctional epithelium (JE) had disappeared. After 3 days, new epithelium formed from the oral sulcular epithelium (OSE) and extended horizontally over the wound with Ln-5-positive cells at the leading edge. After 5 days, the epithelium extending from the OSE on each side of the wound joined and formed additional new epithelium. The new epithelium expressed Ln-5 in the BM. After 1-2 weeks, the oral epithelium (OE) extending from the sides of the wound joined in the center. Thereafter, OSE and new epithelium disappeared, and only OE remained covering the wound. Three days after implantation (titanium), no JE remained. New epithelium formed from the keratinized OSE extending apically with Ln-5-positive cells. After 1-2 weeks, the new epithelium became the PIE and spread further apically facing the implant surface. Ln-5 was expressed at the PIE-connective tissue interface, but not at the implant-PIE interface. Finally, after 4 weeks, Ln-5 was expressed at the implant-PIE interface, and the PIE was non-keratinized epithelium. These findings suggest that Ln-5 induces cell migration during PIE formation, and that PIE originates from OSE. Furthermore, they support the hypothesis that Ln-5 contributes to the attachment of PIE to titanium, regardless of the delay in the synthesis and deposition of Ln-5 at the titanium-PIE interface.

Animals↗

Ultrastructural localization of laminin-5 (gamma2 chain) in the rat peri-implant oral mucosa around a titanium-dental implant by immuno-electron microscopy.

Laminin-5 (Ln-5) is an important molecule associated with epithelial cell adhesion and migration. In the gingiva around the tooth, Ln-5 localizes within basement membranes between the junctional epithelium (JE) and the tooth or connective tissue. Recently, we reported that in the oral mucosa around a dental implant, Ln-5 is expressed within the basement membranes at the implant-peri-implant epithelium (PIE) interface, and at the PIE-connective tissue interface. However, the ultrastructural localization of Ln-5 within or along the PIE has not yet been reported. Therefore, peri-implant oral mucosa was treated with anti-Ln-5 (gamma2 chain) antibody and examined using immuno-electron microscopy. Ln-5 was localized in the cells of the innermost-third layer and basal layer of the PIE. A 100-nm-wide Ln-5-positive internal basal lamina (basement membrane) and hemidesmosomes as adhesion structures were formed at the apical portion of the implant-PIE interface. However, at the upper-middle portion of the interface, these adhesion structures were not observed. Furthermore, at the PIE-connective tissue interface, the Ln-5-positive external basal lamina (basement membrane) and hemidesmosomes were partially deficient. Judging from these findings, we concluded that Ln-5 contributes to the attachment of the PIE to the titanium surface, and that PIE attached to titanium at the apical portion of the dental implant-PIE interface.

Animals↗

Influence of fabrication techniques on retention force of fiber-reinforced composite posts.

The purpose of this study was to compare the retention force of FRC posts which were built up using direct and direct-indirect fabrication techniques with two fiber-reinforced core build-up systems (FibreKor and i-TFC). Posts were cemented in endodontically treated bovine single roots with resin cement using either direct or direct-indirect technique. Following which, the retention force of post-and-cores and fracture sites were examined. It was found that both the retention force and fracture site depended on the fabrication technique and resin cements. Post-and-cores built up with direct-indirect technique had greater retention force than those fabricated using direct technique. Fracture modes most frequently observed were adhesive failure at Resin/Dentin interface, a mixture of adhesive failures at Post/Resin and Resin/Dentin interfaces as well as cohesive failure of resin cements. Based on the results of this study, we concluded that when it comes to post-and-core build-up for endodontically treated tooth, the direct-indirect technique is more effective than the direct technique on the retention force of FRC posts.

Analysis of Variance↗

Characterization and protein-adsorption behavior of deposited organic thin film onto titanium by plasma polymerization with hexamethyldisiloxane.

Plasma polymerized hexamethyldisiloxane (HMDSO) thin film was deposited onto titanium using a radio-frequency apparatus for the surface modification of titanium. A titanium disk was first polished using colloidal silica at pH=9.8. Plasma-polymerized HMDSO films were firmly attached to the titanium by heating the titanium to a temperature of approximately 250 degrees C. The thickness of the deposited film was 0.07-0.35mum after 10-60min of plasma polymerization. The contact angle with respect to double distilled water significantly increased after HMDSO coating. X-ray photoelectron spectroscopy revealed that the deposited thin film consisted of Si, C, and O atoms. No Ti peaks were observed on the deposited surface. The deposited HMDSO film was stable during 2-weeks immersion in phosphate buffer saline solution. Fourier transform reflection-absorption spectroscopy showed the formation of Si-H, Si-C, C-H, and Cz.dbnd6;O bonds in addition to Si-O-Si bonds. Quartz crystal microbalance-dissipation measurement demonstrated that the deposition of HMDSO thin films on titanium has a benefit for fibronectin adsorption at the early stage. In conclusion, plasma polymerization is a promising technique for the surface modification of titanium. HMDSO-coated titanium has potential application as a dental implant material.

Adsorption↗

The effect of surface pore size on the differentiation of rat bone marrow cells: morphological observations and expression of bone related protein mRNA.

The purpose of this study was to investigate the behavior of rat bone marrow cells (RBM) growing on surfaces with different pore sizes. RBM behavior on Millipore filters (MF-Millipore membrane filter) made from cellulose mixed esters with 5 different pore surfaces (0.45 microm, 1.2 microm, 3.0 microm, 5.0 microm and 8.0 microm) were compared in terms of morphological changes on the different pore sizes. Furthermore, the expressions of osteopontin and osteocalcin mRNAs were investigated. On the 1.2 microm and 3.0 microm pore surfaces, RBM attached to the substrate well, but cells on the 5.0 microm and 8.0 microm pore surfaces invaded deeply into the pores. Higher levels of both osteopontin and osteocalcin mRNA expression were always observed in cells cultured on the 1.2 microm filter. These results suggest that the 1.2 microm Millipore filter pore size is the most suitable for inducing RBM to differentiate into an osteoblastic phenotype among these surfaces and is probably related to production of the ECM but not to the phenomenon of cell spreading.

Animals↗

Fatigue strengths of particulate filler composites reinforced with fibers.

The aim of this study was to evaluate the dynamic fatigue strengths at 10(5) cycles and the strains of particulate filler composite resins with and without reinforcing fibers. An UHMWPE (Ribbond), a polyaromatic polyamide fiber (Fibreflex), and three glass fibers (GlasSpan, FibreKor, Vectris Frame) were used to reinforce the particulate filler composite resins. The fatigue properties were measured in three-point bending mode using a servohydraulic universal testing machine at a frequency of 5 Hz, until failure occurred or 10(5) cycles had been completed. The fatigue strengths at 10(5) cycles were determined by the staircase method. The fractured aspects of specimens were evaluated by an optical and scanning electron microscope. The fatigue strengths of particulate filler composite resins were 49-57 MPa, and those of fiber-reinforced were 90-209 MPa. Unidirectional glass fibers showed higher reinforcing effects on the fatigue strengths of composite resins. The strain of UHMWPE-reinforced composite was largest.

Analysis of Variance↗

Corrosion behavior and surface characterization of Ti-20Cr alloy in a solution containing fluoride.

The purpose of this study was to investigate the correlation between corrosion resistance and surface composition of an experimental Ti-20 mass% Cr casting alloy in a saline solution containing fluoride. The alloy had a greater resistance to corrosion in a fluoride-containing saline solution than did commercially pure titanium. However, with confirmed dissolution of titanium and chromium, it meant that the fluoride in the saline solution corroded the alloy slightly. X-ray photoelectron spectroscopy analysis revealed that the surface composition of the alloy consisted of titanium and chromium oxides containing hydroxide. The [Ti]/([Ti] + [Cr]) ratio in the surface oxide film decreased when immersed in fluoride-containing saline solution, that is, the surface oxide film became chromium-rich oxide. Therefore, the alloy obtained good corrosion resistance to fluoride due to formation of a chromium-rich oxide film.

Chromium Alloys↗

The initial attachment and subsequent behavior of osteoblastic cells and oral epithelial cells on titanium.

The interaction between implant materials and bone cells or oral epithelial (OE) cells contributes to the clinical success of dental implants. The functional activity of cells in contact with an implant is determined by its surface properties. Before cells attach, extracellular matrix (ECM) in the serum deposits on the substrate; rounded cells then attach and spread upon it. Cells form focal adhesions and polarize, then start to migrate or proliferate to form colonies. Comparison of the attachment and behavior of osteoblastic cells on titanium (Ti) and hydroxyapatite (HA) revealed that more cells attached on HA and that these spread more rapidly than on Ti. In contrast, cells did not form good stress fibers or vinculin-positive focal adhesions on HA, whereas the cells on Ti possessed well-defined and polarized stress fibers. The initial attachment of OE cells to Ti was inferior to that on polystyrene culture dish or glass, and the OE cell migration area, indicated by the deposition of LN5, was smaller on Ti than on the other materials. This review summarizes data on the attachment and behavior of osteoblastic cells and OE cells on biomaterials, which may suggest future improvements in surface properties.

Animals↗

Direct attachment of fibronectin to tresyl chloride-activated titanium.

The aim of this study was to bind fibronectin directly to a titanium surface treated with tresyl chloride (2,2,2-trifluoroethanesulfonyl chloride) for the development of a strong connection of a dental implant to subepithelial connective tissues and/or peri-implant epithelia. Basic terminal OH groups of mirror polished titanium were allowed to react with tresyl chloride at 37 degrees C for 2 days. The tresylated titanium disk was then immersed into a fibronectin/phosphate-buffered saline solution for 24 h at 37 degrees C. The activation reaction of the basic OH of titanium with tresyl chloride was confirmed by S2p, F1s, and O1s spectra using X-ray photoelectron spectroscopy and -O-S-O2- bonds using Fourier transform infrared reflection-absorption spectroscopy. After the reaction of fibronectin with titanium, the X-ray photoelectron spectroscopy revealed the remarkable effect of the activation of terminal OH groups with the tresyl chloride treatment. The N1s peak derived from the attached fibronectin still remained after 60 s of argon-ion sputtering after tresyl chloride treatment. In contrast, the N1s peak of the specimen not treated with tresyl chloride almost disappeared after only 10 s of argon-ion etching. Fibronectin, a well-known cell-adhesive protein, could easily be attached to the titanium surface by use of the tresyl chloride activation technique.

Coated Materials, Biocompatible↗

The attachment and growth behavior of osteoblast-like cells on microtextured surfaces.

In previous studies, we showed that the application of microgrooves on a surface can direct cellular morphology and the deposition of mineralized matrix of osteoblast-like cells (Biomaterials 20 (1999) 1293; Clin. Oral Impl Res. 11 (2000) 325). In this study, we evaluated the attachment and growth behavior of these cells, using scanning- and transmission electron microscopy (SEM/TEM). Smooth and microgrooved polystyrene substrates were made (groove depth 0.5-1.5 microm, groove- and ridge width 1-10 microm). On these substrates, osteoblast-like cells were cultured for periods up to 16 days. SEM showed that the cells, and their extensions, closely followed the surface on smooth and wider grooved (>5 microm) substrates. In contrast, narrow grooves (<2 microm) were bridged. After 16 days of incubation, the matrix showed extensive deposition of collagen fibrils, and the formation of calcified nodules. With TEM it was shown that on the smooth and wider grooved substrates, focal adhesions were spread throughout the surface. However, on narrow grooves focal adhesions were always positioned on the edges of surface ridges only. Apparently, most extracellular matrix (ECM) was produced by the cells that directly adhered to the substrate. Deposition of ECM was seen in the surface grooves, as well as in between the cell layers. On basis of the current study and previous experiments, we conclude that microgrooves are able to influence bone cell behavior by (1) determining the alignment of cells and cellular extensions, (2) altering the formation and placement of cell focal adhesions, and (3) altering ECM production. Therefore, microgrooved surfaces seem interesting to be applied on bone-anchored implants.

Animals↗