Authors: E. Gemelli, Christiane Xavier Resende, Carlos M. Lepienski, Gloria Dulce de Almeida Soares
Abstract: In this study we report on the microstructure and its mechanical behavior of a Ca-P
coating produced on bioactive titanium by immersion in a simplified simulated body fluid (S-SBF).
The coating was probed by nanoindentation in several point times up to the formation of
octacalcium phosphate (OCP). Amorphous calcium phosphate, formed after 1h of immersion in SSBF,
presented the highest values of hardness (H) and elastic modulus (E). Nucleation of OCP was
observed after 2-2,5h of immersion in S-SBF. From this stage on, lower values of H and E were
obtained, probably due to the low dense structure of the coating.
365
Authors: Takashi Kumagai, Takahisa Anada, Yoshitomo Honda, Masamichi Takami, Ryutaro Kamijyo, Hidetoshi Shimauchi, Osamu Suzuki
Abstract: The present study was designed to investigate whether bone morphogenetic protein-2
(BMP-2) adsorbed onto octacalcium phosphate (OCP) and hydroxyapatite (HA) surfaces influences
osteoblastic cell differentiation. Osteoblastic cell line, UAMS32, were cultured on plastic plate that
was coated of synthetic OCP and HA. BMP-2 was pre-adsorbed on these calcium phosphates at 4°C
or room temperature for 24h. The cells more proliferated on the BMP-2 pre-adsorbed OCP and HA
than that of the controls (OCP and HA coating without BMP-2). ALP activities were higher on HA
than on OCP when BMP-2 were pre-adsorbed at 4°C than at room temperature. The results suggest
that pre-adsorption of BMP-2 in different temperature affects osteoblastic cell differentiation, most
probably through different adsorption state of BMP-2 on this calcium phosphate.
1025
Authors: Osamu Suzuki, Shinji Kamakura, Takahisa Anada
Abstract: The present study was designed to investigate the mechanism of in vivo conversion from
synthetic octacalcium phosphate (OCP) into hydroxyapatite (HA) at ultrastructural level, where the
implanted OCP is enhancing bone regeneration in mouse calvarial bone defect. OCP granules were
implanted into the subperiosteal area of the calvaria of 7-week-old BALB/c mice for 3 weeks.
Transmission electron microscopy of undecalcified frontal sections, obtained from the acrylic
resin-embedded skull specimens showed that the bone crystals in newly formed bone directly bonded
to the OCP particles implanted. The morphological characteristic of original plate-like OCP particles
was remained unchanged even after the implantation, whereas a number of de novo nano-particles
were also directly formed onto the plate-like OCP particles. Some of OCP particles were linked with
other OCP particles through these nano-particles. The results suggest that the OCP-apatite conversion,
involving the enhanced bone regeneration, advances via topotaxial conversion without changing the
original OCP morphology, accompanied by solution-mediated de novo nano-apatite formation, in the
vicinity of the implanted OCP particles.
993
Authors: Yoshitomo Honda, Shinji Kamakura, Takashi Kumagai, Osamu Suzuki
Abstract: Bone regeneration by calcium phosphates has been known to be intricately dependent on
material properties or implanted milieu of host animals, such as site and species. Critical sized
calvarial defects of mouse were recently used as the model for investigating bone regeneration ability
and the mechanisms. The purpose of the present study is to investigate whether the critical sized
mouse calvarial defects can be utilized to examine bone regeneration with synthetic octacalcium
phosphate (OCP). OCP , prepared by wet synthesis methods, was sieved 0.3 ~ 0.5 mm in diameter and
used for the animal experiment. At 14 days after surgery, histological examination showed that
implantation of OCP grafted defects significantly enhanced bone formation compared with the
control defect. OCP tended to convert to hydroxyapatite with time. The tartrate-resistant acid
phosphatase (TRAP) positive osteoclastic cells were observed around the OCP particles. The results
suggest that the mouse critical sized calvarial bone defects are useful model to investigate the bone
formation by the OCP implantation.
1253
Authors: Christiane Xavier Resende, Gustavo Mendes Platt, Jean Dille, Ivan Napoleão Bastos, Gloria Dulce de Almeida Soares
Abstract: In this work, a solution able to precipitate calcium phosphate in titanium samples was
studied. At first, a thermodynamic analysis of the proposed solution was conducted using a
computational simulator that considers most of chemical reactions and evaluates parameters such as
activity of species. After this procedure, experimental tests were performed in order to confirm this
precipitation. With the use of TRIS at concentration of 50mM, the deposits were basically
composed of octacalcium phosphate, as confirmed in some characterization techniques. The deposit
presents a thickness of approximately 15μm after a 7-day exposure in the designed solution.
665
Authors: Shinji Kamakura, Kazuo Sasaki, Yoshitomo Honda, Taisuke Masuda, Takahisa Anada, Tadashi Kawai, Aritsune Matsui, Keiko Matsui, Seishi Echigo, Osamu Suzuki
Abstract: Our previous studies showed that synthetic octacalcium phosphate (OCP) enhances bone
regeneration more than hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP). Also, a synthetic
bone substitute constructed of synthetic octacalcium phosphate (OCP) and porcine atelocollagen
sponge (OCP/Col) showed stable bone regeneration. The present study was designed to investigate
the difference of bone regeneration by OCP/Col and other calcium phosphate-collagen composites.
OCP/Col, β-TCP$-collagen% composite (β-TCP/Col), or HA$-collagen% composite (HA/Col) sponge
was prepared from pepsin-digested atelocollagen isolated from the porcine dermis and OCP, β-TCP,
or HA granules, respectively. A standardized critical-sized defect was made in the rat calvarium, and
various calcium phosphate-collagen composites were implanted into the defect. The rats were fixed
at four weeks after implantation and radiographic and histological examinations were performed by
undecalcified cross sections of implants. Radiographic examination showed that uniform radiopaque
masses were observed in the created defects treated with OCP/Col, whereas granulous and foggy
radiopacity was observed in β-TCP/Col and HA/Col. Histological examination showed that newly
formed bone was observed in the reticulum of OCP/Col and around the implanted OCP. The
regenerated bone by β-TCP/Col or HA/Col seemed to be less than that by OCP/Col and would not to
be nucleated by the granules of β-TCP or HA. The present study indicated that bone regeneration by
OCP/Col was different from those of β-TCP/Col and HA/Col. Application of OCP/Col would be
expected for clinical use in the future.
1229
Authors: Ana Maria Minarelli Gaspar, Sybele Saska, J. Jimenez, L.R. da Cunha, P.D.A. Bolini, Claudinete V. Leal, L. Morejón-Alonso, R. García Carrodeguas, A. López-Bravo, Cecília A.C. Zavaglia
Abstract: The biological behavior of a new bioactive material composed of calcium-deficient
hydroxyapatite, octacalcium phosphate, and β-tricalcium phosphate was investigated by in vitro
indirect and direct cytotoxicity, cell adhesion and proliferation tests, and by in vivo subcutaneous
and bone implantation in rats. The results of the in vitro studies showed that the material is
biocompatible and no cytotoxic. Slightly poorer initial cell adhesion and lower cell proliferation
than in control was observed, which were attributed to the reactivity and roughness of the material
surface. In vivo results showed that the material is biodegradable and bioactive in bone tissue, but
only biocompatible and partially biodegradable in soft tissue.
1127
Authors: Yusuke Orii, Hiroshi Masumoto, Takashi Goto, Yoshitomo Honda, Takahisa Anada, Keiichi Sasaki, Osamu Suzuki
Abstract: The surface oxide films were prepared by Electron Cyclotron Resonance (ECR)
plasma oxidation on Ti substrates. Octacalcium phosphate (OCP) and dicalcium phosphate
dihydrate (DCPD) peaks were formed after calcification by supersaturated calcium and
phosphate solutions. Calcification ability was enhanced with increasing the oxidation time and
the total pressure of ECR plasma treatment during oxidation. The results demonstrated that the
calcium phosphate nucleation and the deposition can be controlled by various ECR plasma
conditions.
725
Authors: Hiroshi Masumoto, Takashi Goto, Yusuke Orii, Yoshitomo Honda, Osamu Suzuki, Keiichi Sasaki
Abstract: Surface titanium oxide (TiO2) films were fabricated on implant titanium (Ti) at low
temperatures by electron-cyclotron-resonance (ECR) plasma oxidation. The relationship among
the oxidization conditions, crystal structure, morphology and osteoconductive property were
investigated. Although crystallized TiO2 film was not prepared by thermal oxidation at 300°C,
crystallized rutile-type TiO2 film was formed by ECR plasma oxidation at 300°C. Rough
morphology was observed in the substrate surface oxidized by ECR plasma. Mixtures of
octacalcium phosphate (OCP) and dicalcium phosphate dihydrate (DCPD) were observed after
calcification. The XRD peak intensities of the OCP and DCPD formed on the ECR plasma
oxidized Ti were larger than those of calcified on the thermal oxidized Ti. ECR plasma
oxidation at low temperature would induce osteoconductive calcium phosphate on implant Ti.
717
Authors: Takahisa Anada, Akihiro Araseki, Shou Matsukawa, Tomokazu Yamasaki, Shinji Kamakura, Osamu Suzuki
Abstract: Our previous studies suggested that synthetic octacalcium phosphate (OCP) enhances bone
regeneration more than hydroxyapatite (HA). However, the molecular mechanisms to induce
osteogenic phenotype in osteoblast by OCP have not been identified. OCP tended to convert into an
apatite structure in vivo and in vitro, and its process was accompanied by calcium consumption from
the surrounding solution and the release of phosphate ions into the solution at a physiological
condition. The present study was designed to investigate whether the dissolution of ionic products of
OCP affects on proliferation and differentiation of mouse bone marrow stromal ST-2 cells in vitro.
The number of cells treated with OCP-conditioned medium was slightly decreased in comparison to
that of control at day 7. On the other hand, the level of alkaline phosphatase activity increased in
OCP-conditioned medium. These results demonstrated that OCP is capable of inducing osteoblastic
cell differentiation in ST-2 cells.
31