acid-phosphatase and octacalcium-phosphate

The present study was designed to investigate whether three sodium hyaluronic acid (HyA) medical products, Artz(®), Suvenyl(®) and a chemically modified derivative of sodium HyA Synvisc(®), can be used as suitable vehicles for an osteoconductive octacalcium phosphate (OCP). OCP granules (300-500 μm diameter) were mixed with these sodium HyAs with molecular weights of 90 × 10(4) (Artz(®)), 190 × 10(4) (Suvenyl(®)) and 600 × 10(4) (Synvisc(®)) (referred to as HyA90, HyA190 and HyA600, respectively). OCP-HyA composites were injected using a syringe into a polytetrafluoroethylene ring, placed on the subperiosteal region of mouse calvaria for 3 and 6 weeks, and then bone formation was assessed by histomorphometry. The capacity of the HyAs for osteoclast formation from RAW264 cells with RANKL was examined by TRAP staining in vitro. Bone formation was enhanced by the OCP composites with HyA90 and HyA600, compared to OCP alone, through enhanced osteoclastic resorption of OCP. HyA90 and HyA600 facilitated in vitro osteoclast formation. The results suggest that the osteoconductive property of OCP was accelerated by the HyAs-associated osteoclastic resorption of OCP, and therefore that HyA/OCP composites are attractive bone substitutes which are injectable and bioactive materials.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Bone Regeneration; Calcium Phosphates; Cell Line; Humans; Hyaluronic Acid; Implants, Experimental; Isoenzymes; Male; Mice; Mice, Inbred ICR; Molecular Weight; Osteoblasts; Osteocalcin; Osteoclasts; Osteogenesis; RANK Ligand; Skull; Spectroscopy, Fourier Transform Infrared; Tartrate-Resistant Acid Phosphatase; Viscosity; X-Ray Diffraction

The present study was designed to investigate whether the granule size of synthetic octacalcium phosphate (OCP) and the resultant intergranular spaces between the granules formed by the filling affect its osteoconductive and biodegradable characteristics in a mouse calvaria critical-sized defect up to 10 weeks after implantation. Mercury intrusion porosimetry showed that OCP granules having distinct diameter sizes ranging from 53 to 300 (S-OCP), 300 to 500 (I-OCP) and 500 to 1000 microm (L-OCP) produced distinct intergranular spaces between OCP granules ranging from 28.8 to 176.6 microm. The dissolution rate of OCP, estimated by the phosphate concentration in the culture medium, was the highest in S-OCP, followed by I-OCP and L-OCP, while the specific surface area of OCP decreased. Histological and histomorphometric analyses showed that bone formation around the implanted granules increased significantly with increasing granule size coupled with activating the appearance of TRAP- and cathepsin K-positive osteoclastic cells. The rate of new bone formation formed with L-OCP was two times higher than that formed with S-OCP at 10 weeks after implantation. The results indicated that the osteoconductive and biodegradable properties of OCP can be augmented by increasing the granule size, most probably by thus providing enough spaces between the granules, suggesting that the intergranular spaces formed by the granules may work similarly to pores, as reported in porous ceramic materials. It seems likely that the enhancement of bone formation by OCP is accompanied by simultaneous activation of osteoclastic resorption of OCP.

Topics: Acid Phosphatase; Animals; Bone Regeneration; Calcium Phosphates; Cathepsin K; Giant Cells; Humans; Isoenzymes; Mice; Mice, Inbred ICR; Microscopy, Electron, Scanning; Osteogenesis; Paraffin Embedding; Particle Size; Phosphates; Prosthesis Implantation; Spectroscopy, Fourier Transform Infrared; Tartrate-Resistant Acid Phosphatase; X-Ray Diffraction

The present study was designed to investigate whether the stoichiometry of octacalcium phosphate OCP affects its osteoconductive and immune response characteristics in rat bone marrow. Those characteristics of synthetic, well-grown OCP but with a non-stoichiometric composition were compared with those of a slightly hydrolyzed OCP (low crystalline OCP: LC-OCP), the fully hydrolyzed apatitic product of OCP or biodegradable beta-tricalcium phosphate (beta-TCP) ceramic, by their implantation in rat tibia for 56 days. The physicochemical aspect of implants and biological responses were analyzed by X-ray diffraction, histomorphometry, immunohistochemistry and expression of mRNA around the implants. The remarkable findings were that: (1) the highest bone formation rate was obtained for beta-TCP whereas the lowest for LC-OCP at Day 14; (2) the rates were reversed and reached the highest for LC-OCP until Day 56; (3) the early expression of ostoeoclast markers TRAP and cathepsin-K was suppressed with LC-OCP; (4) the expression of inflammatory markers IL-beta1 and TNF-alpha was suppressed with LC-OCP. The results confirmed that the partially hydrolyzed OCP with Ca/P molar ratio 1.37 (LC-OCP) enhances bone formation most, suppressing early osteoclast activity and reducing inflammation.

Topics: Acid Phosphatase; Animals; Bone Regeneration; Calcium Phosphates; Crystallization; Cytokines; Gene Expression Profiling; Gene Expression Regulation; Hydrolysis; Immunohistochemistry; Implants, Experimental; Inflammation Mediators; Isoenzymes; Male; Osteocalcin; Osteoclasts; Osteogenesis; Osteopontin; Rats; Rats, Sprague-Dawley; RNA, Messenger; Surface Properties; Tartrate-Resistant Acid Phosphatase; Tibia; Time Factors

The present study was designed to investigate whether the microstructure of synthetic octacalcium phosphate (OCP) affects its intrinsic bone regenerative properties as a scaffold and its conversion process into hydroxyapatite (HA). Our previous studies indicated that an agregate of OCP crystals, consisting of randomly oriented plate-like crystals, are capable of enhancing both osteoblastic cell differentiation in vitro and bone regeneration. While the transformation of OCP into HA has been considered in relation to the stimulatory capacity of OCP in bone regeneration, little is known about the effect of the microstructure of OCP granules on these capabilities. Two types of OCP granules, with identical diameters (300-500 microm) but composed of crystals with distinct crystal dimensions (4.0 and 26.6 microm length), were prepared (hereafter referred to as fine OCP granules [F-OCP] and coarse OCP granules [C-OCP], respectively). The intergranule distances and the porosity, including the intergranule spaces, were 108.5 microm and 93.7% for F-OCP, and 67.5 microm and 95.7% for C-OCP, as estimated by mercury intrusion. The OCP granules were implanted in mouse critical-sized calvarial defects for up to 14 days. Histological examination demonstrated that osteoblastic cells aligned on the surface of F-OCP at day 7 and formed new bone around the granules up to day 14. On the other hand, cells around C-OCP were sparse at day 7, and resulted in only slight bone formation around the granules at day 14. X-ray diffraction showed that both OCP granules tended to be converted to an apatite structure with similar conversion velocity by the implantation. Adhesion of mouse bone marrow stromal ST-2 cells was markedly inhibited on C-OCP compared to F-OCP in vitro. These results suggested that the microstructure consisting of plate-like crystals of OCP controls cell adhesion on the crystal surfaces and their resultant bone regenerative properties as well as the physicochemical effect associated with the transitory nature of OCP previously reported.

Topics: Acid Phosphatase; Animals; Bone and Bones; Bone Regeneration; Calcium Phosphates; Cell Adhesion; Crystallization; Hydrolysis; Isoenzymes; Mice; Tartrate-Resistant Acid Phosphatase; Time Factors; X-Ray Diffraction

Previous studies showed that synthetic octacalcium phosphate (OCP) enhances bone formation coupled with its own osteoclastic biodegradation more than non-biodegradable hydroxyapatite (HA), including sintered HA ceramic, when implanted in animal bone defects. The present study was designed to investigate whether synthetic OCP in granule form has biodegradable characteristics when implanted in the subperiosteal area of mouse calvaria in comparison with non-sintered stoichiometric HA, especially in relatively short periods after implantation. OCP crystals exhibited plate-like morphology, whereas HA crystals had a sphere-like structure. Both crystals had large pore volumes >75% in total, with micropores within the granules. Direct bonding of newly formed bone was discernible in HA until 35 days after implantation by element analysis for calcium and phosphorus. However, histomorphometric analysis demonstrated that bone formation was facilitated on OCP surfaces with greater alkaline phosphatase activity than on HA up to 21 days. The surfaces attacked by tartrate-resistant acid phosphatase positive osteoclast-like cells were significantly greater than those of HA. OCP became encapsulated and replaced with new bone with prolonged implantation periods up to 180 days. The results suggest that the enhanced bone formation in mouse calvaria could be associated with the biodegradable nature of OCP, and that OCP could be used in augmenting intramembranous bone volume.

Topics: Absorbable Implants; Acid Phosphatase; Alkaline Phosphatase; Animals; Bone and Bones; Calcification, Physiologic; Calcium; Calcium Phosphates; Durapatite; Isoenzymes; Membranes; Mice; Mice, Inbred BALB C; Microscopy, Electron, Scanning; Osteogenesis; Phosphorus; Tartrate-Resistant Acid Phosphatase; X-Ray Diffraction

Synthetic octacalcium phosphate (OCP) has been shown to enhance bone formation and to biodegrade if implanted into bone defects. Here, we hypothesized that an OCP-atelocollagen complex (OCP/Col) is biodegradable and can induce bone formation in a thickness-dependent manner when implanted into the calvaria. OCP/Col disks (diameter, 9 mm; thickness, 1 or 3 mm) were implanted into a subperiosteal pocket in the calvaria of 12-week-old Wistar rats for 4, 8, and 12 weeks and subsequent bone formation was monitored. X-ray diffraction analysis and Fourier transform infrared spectroscopy showed that OCP in the OCP/Col implants was converted into a carbonate-rich apatite after 4 weeks. Although thinner disks tended to be replaced by new bone, thicker disks were progressively resorbed by osteoclast-like cells until 12 weeks, possibly via the increased mechanical load in the subperiosteal pocket. Therefore, OCP/Col can increase appositional intra-membranous bone formation if the appropriate size of the implant is applied.

Topics: Absorbable Implants; Acid Phosphatase; Animals; Apatites; Biomarkers; Bone Substitutes; Calcium Phosphates; Cathepsin K; Collagen; Drug Carriers; Isoenzymes; Male; Osteoclasts; Osteogenesis; Periosteum; Prosthesis Design; Rats; Rats, Wistar; Skull; Spectroscopy, Fourier Transform Infrared; Stress, Mechanical; Tartrate-Resistant Acid Phosphatase; Time Factors; X-Ray Diffraction

Octacalcium phosphate (OCP) is thought to be a precursor of the mineral crystals in biological apatite. Synthetic OCP has been shown to be converted into an apatite structure when implanted in murine calvarial bone, to enhance bone regeneration more than synthetic hydroxyapatite (HA), and to degrade faster than biodegradable beta-tricalcium phosphate. This study was designed to investigate whether OCP implantation enhances the formation and resorption of new bone (remodeling) concomitant with OCP degradation when implanted intramedullary in a rabbit femur for 12 weeks, compared to sintered HA ceramic. Histological and histomorphometric analyses using undecalcified specimens showed that the area of bone apposition was significantly higher on OCP than on HA between 2 and 3 weeks, whereas it subsequently became smaller on OCP than on HA. The area attacked by multinucleated giant cells, including tartrate-resistant acid phosphatase (TRAP)-positive cells, was significantly higher for OCP than for HA at 8 weeks. Radiography revealed resorption of OCP but not of HA. The results disclose some osteoconductive characteristics of synthetic OCP in the bone marrow space: (1) enhancement of bone regeneration at the initial bone apposition stage and (2) stimulation of resorption of the newly formed bone coupled with OCP biodegradation mediated by TRAP-positive osteoclast-like cells. These results suggest that synthetic OCP would be a more useful bone substitute than HA in implant applications where rapid bone formation and concomitant implant resorption are important considerations.

Topics: Acid Phosphatase; Animals; Bone Marrow Cells; Bone Regeneration; Calcium Phosphates; Drug Implants; Femur; Hydroxyapatites; Isoenzymes; Male; Microscopy, Electron, Scanning; Rabbits; Tartrate-Resistant Acid Phosphatase; X-Ray Diffraction