Takashi Hayami

Anisotropy of osteoporotic cancellous bone

To investigate the mechanism underlying femoral neck fracture, it is necessary to determine the various mechanical properties, including the bone strength, of the primary compressive group. We investigated the mechanical anisotrophy of the primary compressive group by comparing differences in its mechanical properties, depending on the loading direction. Twenty-three femoral heads of 20 female and 3 male patients with femoral neck fracture were studied. The mean age of these patients was 79.9 years (range, 63-98 years). A total of 82 cubic specimens (6.5 mm in length) were obtained (one to six specimens from each femoral head). The specimens obtained from each femoral head were randomly assigned into two groups: parallel and perpendicular. The parallel group included 43 specimens, and the perpendicular group included 39 specimens. A compressive load was applied either parallel or perpendicular to the primary compressive group of the specimens in each respective group. Three parameters were obtained: compressive stiffness, maximum stress, and maximum energy. We calculated the regression of three parameters against the square of the apparent dry density. These mechanical properties were compared between the two groups by testing the difference of the slopes in two regression lines by using analyses of covariance, in which two main effects of group (nominal value) and the square of the apparent dry density (continuous value) and an interaction between two factors were modeled. Three parameters were significantly correlated with the square of the apparent dry density in both groups. In all three measurements, the difference of the slopes between two regression lines was significantly different. This means that all three measurements decreased in the parallel group more than in the perpendicular one, as apparent dry density decreased. We consider that the bone strength of the proximal femur decreases more when stress is applied in the longitudinal direction (as in walking) and less when stress is applied in the transverse direction (as in a fall) when bone density decreases.

Control of proliferation and differentiation of osteoblasts on apatite-coated poly(vinyl alcohol) hydrogel as an artificial articular cartilage material.

One of the key challenges in employing biomaterials is determining how to fix them into the surrounding tissue. To enhance the interaction with surrounding bone, amorphous hydroxyapatite (HA) was coated onto the surface of the bio-inert poly(vinyl alcohol) hydrogel (PVA-H), as an artificial cartilage material, by a pulsed laser deposition technique. Next we examined the binding effects of the HA thin film (300 nm thick) to the underlying bone using osteoblast proliferation and differentiation. A mouse osteoblast cell line, MC3T3E1, was cultured on the HA-coated and noncoated PVA-H with a water content of 33% or 53% for 3 weeks. Cell proliferation, alkaline phosphatase (ALP) activity, and levels of osteocalcin were evaluated for biocompatibility and differentiation. HA coating enhanced the cell proliferation, the ALP activity, and the levels of osteocalcin on both low and high water-content PVA-Hs. The cell growth rates on the PVA-H were lower than on tissue culture dishes even after the HA coating was added; however, osteoblastic differentiation was highly promoted by the HA coating on low water content PVA-H. These results suggested that the HA coating on the PVA-H enhanced the affinity between the bone and the PVA-H as an artificial cartilage material in surface replacement arthroplasty.

Antidegenerative effects of partial disc replacement in an animal surgery model

Study Design. In vivo degenerative changes of rabbit intervertebral discs after partial disc replacements were evaluated radiologically and histologically in a controlled trial. Objective. To demonstrate the therapeutic effects of partial disc replacement in an animal surgical model. Summary of Background Data. Although some authors reported that partial disc replacements have beneficial clinical outcomes, there are few controlled animal studies in which the therapeutic effects of this procedure have been demonstrated. Methods. The implants for partial disc replacements were made of poly (vinyl alcohol) hydrogel and rod-shaped. The L2–L3 or L3–L4 intervertebral discs of Japanese white rabbits were pierced with a 2.0-mm Kirschner wire and implants were inserted into the holes. For comparative purposes, the adjacent discs underwent sham treatments or control treatments in which the disc was pierced but no implant was inserted. Sixty discs from 30 rabbits were analyzed radiologically and histologically for degenerative changes at 1, 3, or 6 months after surgery. Results. Radiologic analysis revealed that significantly less disc height was lost with the replacement treatment than with the control treatment. Change in disc height after the replacement treatment was not significantly different from that after the sham treatment. Histologic degeneration of the replaced discs was delayed in comparison with that of the control discs but progressed with time. Conclusions. The antidegenerative effects of partial disc replacement surgery were demonstrated by quantitative radiologic and histologic analyses. Degeneration of the anulus fibrosus after the replacement treatment was delayed by preserving disc height and occupying the space of the nucleus pulposus. Properly designed implants and minimally invasive techniques are necessary for long-term success.

Osteoconduction of a stoichiometric and bovine hydroxyapatite bilayer‐coated implant

OBJECTIVE To impart rapid and durable osteoconductivity to implants, a commercial titanium screw implant was coated with stoichiometric hydroxyapatite (HA; 50 nm thick), and then with bovine hydroxyapatite (B-HA; 300 nm thick) using the pulsed laser deposition technique. As control specimens, a commercial implant coated with HA (20 μm thick) using the flame spraying method (sprayed implant) and a simple titanium implant (basic implant) was used. METHODS The osteoconductivities of these three types of implant, after implantation for 4-24 weeks, were histologically evaluated. RESULTS Peeling of HA from the sprayed implant was observed by scanning electron microscopic observation. In the fourth postoperative week, the bilayered implant was already closely adhered to bone. On the other hand, the basic implant was surrounded by a gap containing connective tissue. With the sprayed implant, the bone adhered to the thick HA coating. CONCLUSION The bilayer deposition technique supplies quick and long-term fixation of implants to bone, because the B-HA film dissolves to aid osteoconduction right immediately after implantation and the HA thin film maintains osteoconduction without dissolution. Neither of the thin-film fractures easily compared with thick coatings.

Protein Adsorption on Patterned Hydroxyapatite Thin Films Fabricated by Pulsed Laser Deposition

Protein adsorption on hydroxyapatite (HAP) thin film was investigated before and after patterning. Hydroxyapatite thin film 100 nm thick was deposited by pulsed laser deposition. The film was patterned by photolithography and wet etching with HCl solution. Proteins (phospholyrase b, bovine serum albumin, and others) labeled with fluorescein isothiocyanate (FITC) were used as the reagent. After the HAP film was soaked in the reagent and washed with pure water, a conspicuous contrast in FITC was observed between the HAP pattern and the glass substrate (or photoresist). This behavior showed that the biocompatibility of the HAP thin film was not influenced by the patterning process. Our technique for HAP thin film is adaptable for applications involving biosensors as electronic devices and scaffolds for tissue culture.

Fabrication of a Large Hydroxyapatite Sheet

A large hydroxyapatite (HA) sheet of 50 mm diameter was constructed using a new fabrication process. The HA sheet was prepared by pulsed laser deposition (PLD) and then separated from the substrate. An intermediate layer of spin-coated photoresist between the PLD film and the substrate was dissolved in acetone to enable the separation. Sufficient crystallinity was obtained after postannealing. The area of the HA sheet prepared by the proposed process was approximately 20 times larger than that of the HA sheet prepared by the previous process. The proposed process can also be used for larger sheets. The HA sheet has promise in medical and dental applications.

Regeneration of Tooth Enamel by Flexible Hydroxyapatite Sheet

Before a tooth erupts, the ameloblasts are lost, which means that the tooth enamel does not regenerate itself after tooth eruption. In the present study, we attempt to regenerate the tooth enamel artificially using a flexible hydroxyapatite (HAp) sheet. First, a HAp film was deposited on a soluble substrate by pulsed laser deposition (PLD) using an ArF excimer laser. Next, the HAp film was collected as a freestanding sheet by dissolving the substrate using a solvent. The HAp sheet was adhered to the extracted human teeth using a calcium phosphate solution. The variation of the crystal structure of the HAp sheet with time was investigated by X-ray diffraction analysis. Furthermore, the variation in the mechanical characteristics with time between the HAp sheet and dental enamel were evaluated using tensile and scratch tests. The results suggest that the HAp sheet became fused to the tooth enamel within approximately one week.

Adhesive Strength between Flexible Hydroxyapatite Sheet and Tooth Enamel

Tooth enamel cannot be reconstructed once it is destroyed immoderately. Hydroxyapatite (HAp) thin sheet can potentially be used for a novel dental biomaterial to repair the enamel. Using a pulsed laser deposition (PLD) method, we have successfully created a flexible HAp sheet of less than a few micrometers in thickness. Due to its flexibility, the HAp sheet is tightly adhered on curved surfaces at the target site. In the present study, we newly developed double-layered sheets composed of HAp film coated with tricalcium phosphate (TCP) thin layer. The HAp/TCP sheet was adhered to the extracted human teeth using a calcium phosphate solution for 3 days. The adhesive strength between the HAp/TCP sheet and tooth enamel was evaluated by quasi-static tensile tests. Moreover, the interface structure between them was observed by a scanning electron microscopy. As a result of the mechanical evaluation, the adhesive strength was greater than approximately 2.5 MPa. The electron microscopic observation revealed that the sheet was partially fused with the enamel. These findings suggest the possibility that enamel defects are repaired using the HAp/TCP sheet for a short duration.

The Energy-Absorbing Function of Cancellous Bone and Its Influence on the Loosening of Artificial Joints

Deformation behavior of trabecular architecture (cancellous structure) was visualized using an ultrahigh-speed video recording camera during both static and impact compression tests of subchondral cancellous bone at nominal strain rates from 0.008 to 25 s−1. The mechanical energy-absorbing mechanisms of cancellous bone could be roughly classified into two types: viscoelastic hysteresis and release of elastic energy from microfractures of the trabeculae. The energy-absorbing ability of degenerated cancellous bone was approximately 50% that of normal bone. The osteoarthritic process results in a change in the cancellous structures such that a pronounced fabric direction is not evident. In the case of normal cancellous bone, the structural stiffness of the bone was maintained by each trabecula functioning as a long column in the cancellous structure. It was confirmed visually that, for the in vitro case, aseptic loosening of the prosthesis was caused by a decrease in the cancellous bone stock or reduction in the energy-absorbing abihty of the joints. Thus, the energy-absorbing properties of subchondral cancellous bone were dependent on the morphology and deformability of the cancellous structure.

C-Axis-Oriented Hydroxyapatite Film Grown Using ZnO Buffer Layer

A method of fabricating c-axis-oriented hydroxyapatite film on a quartz crystal microbalance (QCM) sensor was investigated. ZnO was used as a template to obtain a hexagonal hydroxyapatite crystal of uniaxial orientation. The ZnO was grown as a c-axis film on a Au/quartz with the surface structure of a QCM sensor. Under optimized conditions, hydroxyapatite was deposited by pulsed laser deposition. X-ray diffraction showed the hydroxyapatite film to be oriented along the c-axis. Because Au and ZnO are applied to many devices, the anisotropic properties of hydroxyapatite may be incorporated into these devices as well as QCM sensors.