Characterization of the chemically deposited hydroxyapatite coating on a titanium substrate.

Bioactive hydroxyapatite (HA) coating on titanium (Ti) implant can be used as a drug delivery device. A controlled release of drug around the implant requires the incorporation of drug into the coating material during the coating process. HA coating was prepared using a two-step procedure in conditions suitable for simultaneous incorporation of the protein-based drug into the coating material. Monetite coating was deposited on Ti substrate in acidic condition followed by the transformation of the monetite coating to HA. X-ray diffraction (XRD) confirmed the formation of the monetite phase at the first step of the coating preparation, which was transformed into HA at the second step. Fourier transform infrared spectroscopy demonstrated typical bands of a crystallized carbonated HA with A- and B-type substitution, which was confirmed by the XRD refinement of the structural parameters. Scanning electron microscope was used to observe the morphology of monetite and HA coatings. Adhesion of the coatings was measured using a scratch tester. The critical shearing stress was found to be 84.20 ± 1.27 MPa for the monetite coating, and 44.40 ± 2.39 MPa for the HA coating.

Calcium phosphate-based solutions promote dentin tubule occlusions less susceptible to acid dissolution.

PURPOSE: To determine the efficacy supersaturated calcium phosphate (CaP) solutions containing fluoride (F) and zinc (Zn) ions in occluding dentin tubules with precipitates less susceptible to acid dissolution and to compare the performance of these solutions with the oxalate solutions containing calcium (Ca) or phosphate (P) ions. METHODS: Dentin sections from human molars divided into groups: Group A - control (treated with double distilled H2O), Groups A1, A2 and A3 were treated with experimental solutions supersaturated with respect to F and Zn-substituted calcium phosphates. Solutions A1 and A2 were similar in composition but differed in pH values (A1, pH 7; A2, pH 5.5). Solutions A2 and A3 were similar in pH (pH 5.5) but the A3 solution had twice the concentrations of F and Zn2+ ions compared to A2. Another group of dentin sections were treated with A3 solution, oxalate solution containing Ca (OX/Ca) and OX solution containing P (OX/P). The control and treated dentin sections were characterized using scanning electron microscopy. RESULTS: All treated dentin sections showed occluded dentin tubules; with the group A3 showing the highest percent of occluded dentin tubules. The precipitates in the dentin tubules treated with A3 remained while those treated with OX/Ca or OX/P dissolved after exposure to an acidic buffer.

In vivo efficacy of calcium phosphate-based synthetic-bone-mineral on bone loss resulting from estrogen and mineral deficiencies.

Osteopenia and osteoporosis affect over 40 million US adults 50 years and older. Both diseases are strongly influenced by estrogen and nutritional-mineral deficiencies. This study investigates the efficacy of orally delivered synthetic-bone-mineral (SBM), a newly developed calcium phosphate based biomaterial, on reversing bone loss induced by these two critical deficiencies. Thirty 3-month-old female rats were randomly allocated to either control-sham surgery on normal diet; or one of the four experimental groups: Sham surgery on a low mineral diet (LMD), ovariectomized (OVX) on LMD, OVX on LMD with SBM with/without fluoride (F). The rats were sacrificed after 6 months, at 9-month-old. After 6 months, although all groups lost bone mineral density relative to controls, the supplemented OVX rats showed higher bone mineral density than their unsupplemented counterparts. The 2 SBM supplemented groups improved bone loading capacity by 28.1 and 35.4% compared to the OVX LMD group. Bones from supplemented rats exhibited higher inorganic/organic ratios. The addition of F did not have a significant influence on bone loss. Our findings suggest that SBM supplement is effective in maintaining bone health and offsetting the deleterious effects of estrogen and/or mineral deficiencies on bone density, microarchitecture, and strength.

Inhibitory effect of Zn2+ in zinc-containing beta-tricalcium phosphate on resorbing activity of mature osteoclasts.

Long term effect of the growing instability of the bone-implant interface due to bone resorption at the interface is a problem for the implants, including bioactive ceramics. Zn2+ -containing tricalcium phosphate (ZnTCP) is a material which may overcome this problem. The present study aims to clarify whether Zn2+ -containing tricalcium phosphate (ZnTCP) ceramics with a Zn2+ content of 0.316 (ZnTCP316) and 0.633 (ZnTCP633) wt % suppress resorption by mature osteoclasts in vitro. Suppression would be due to an increase in the number of apoptotic osteoclasts and the inhibition of the resorbing activity of osteoclasts, the latter being the major mechanism of the suppression. The number of apoptotic osteoclasts was significantly 6.3 times higher with ZnTCP633 than with tricalcium phosphate ceramic (TCP) after 24-h culture. The net contribution to resorption of this change in apoptotic cell numbers is much smaller than that of the change in resorbing activity. The osteoclasts cultured on ZnTCP formed fewer actin rings than those cultured on the TCP. The mRNA expression of CAII and cathepsin K/OC2 in the osteoclasts on ZnTCP633 was downregulated 0.5-fold and 0.6-fold, respectively, compared with that on the TCP. The volume of resorption pits was downregulated 0.4-fold in the ZnTCP633 than that in TCP. These results suggest that ZnTCPs suppressed the resorbing activity of mature osteoclasts probably through a local increase in the level of Zn2+. Bone substitutes or coating layers containing ZnTCP would be promising biomaterials from the viewpoint of counteracting osteoclastic bone resorption at the bone-implant interface.

Efficacy of the injectable calcium phosphate ceramics suspensions containing magnesium, zinc and fluoride on the bone mineral deficiency in ovariectomized rats.

The purpose of this study was to evaluate the therapeutic efficacy of a new calcium phosphate (CaP)-based formulation in improving the bone mineral deficiency in ovariectomized (OVX) rats. The ions release experiments for CaP preparations (G2: 0.46% Mg, 5.78% Zn, and 2.5% F; G3:3.1% Mg, 0.03% Zn, and 3.01% F; G4: 1.25% Mg, 1.77% Zn, 1.35% F) and of a Zn-TCP (G1: 6.17% Zn) powders, the initial Mg and Zn ion release rates of MZF-CaPs were performed in acetate buffer at pH 4.5 (37 degrees C). Wistar rats were divided into six groups including a normal (not OVX) group (GN) and a control, OVX group (GC). Rats in groups GC, G1, G2, G3, G4 were OVX. Suspensions consisting of CaP preparations (G2, G3, G4) and of a Zn-TCP (G1) powders were injected in the right thighs of OVX rats in all groups except for GN and GC, once a week for 4 weeks. GN and GC rats were injected with saline solutions. Plasma was analyzed for Zn land alkaline phosphatase levels. The bone mineral density (BMD) was measured using DEXA and the bone (femur) strength determined using three-point-bending analysis. G1 and G2 groups showed high plasma Zn levels. The area under the curve of plasma Zn was significantly greater in the G1, G2, and GN groups than in the G3, G4, and GC groups (p < 0.05). The BMD and bone mechanical strength of the right femur were significantly higher in the G1, G2, G3, and G4 groups than GC group on day 28. The right femur had significantly greater BMD and bone mechanical strength than the left femur in G1, G2, G3, and G4 groups. However, there was no significant difference in the BMD of the right femur between the G1, G2, G3, and G4 groups. Results indicate that the new injectable CaP formulations are effective in improving bone properties of OVX rats and may be useful in osteoporosis therapy.

Bioactive materials in endodontics.

Endodontic treatment in dentistry is a delicate procedure and many treatment attempts fail. Despite constant development of new root canal filling techniques, the clinician is confronted with both a complex root canal system and the use of filling materials that are harmful for periapical tissues. This paper evaluates reported studies on biomaterials used in endodontics, including calcium hydroxide, mineral trioxide aggregate, calcium phosphate ceramics and calcium phosphate cements. Special emphasis is made on promising new biomaterials, such as injectable bone substitute and injectable calcium phosphate cements. These materials, which combine biocompatibility, bioactivity and rheological properties, could be good alternatives in endodontics as root canal fillers. They could also be used as drug-delivery vehicles (e.g., for antibiotics and growth factors) or as scaffolds in pulp tissue engineering.

Physico-chemical-mechanical and in vitro biological properties of calcium phosphate cements with doped amorphous calcium phosphates.

Calcium phosphate cements (CPCs) are successfully used as bone substitutes in dentistry and orthopaedic applications. This study investigated the physico-chemical-mechanical properties of and in vitro biological properties (cell response) of CPCs prepared with amorphous calcium carbonate phosphate (ACCP) doped with magnesium (ACCP-Mg), zinc (ACCp-Zn) or fluoride (ACCP-F) ions. The experimental CPC consisted of alpha-TCP, doped ACCP, and MPCM powders as matrix and biphasic calcium phosphate (BCP) granules. X-ray diffraction analysis showed that the matrix converted to apatite with poor crystallinity (reflecting small crystal size) after setting for 24 h, while BCP remained apparently unchanged. Cements with ACCP-F (F-CPC) had shorter setting times and greater compressive strength compared to cements with ACCP-Mg (Mg-CPC) or ACCP-Zn (Zn-CPC). Scanning electron microscopy (SEM) showed that crystals set on Mg-CPC and Zn-CPC were smaller compared to those on F-CPC. The total porosity of Mg-CPC was greater compared to Zn-CPC or F-CPC. Osteoblast-like cells, MC3T3-E1, remained viable and maintained their ability to express alkaline phosphatase in contact with the CPCs with doped ACCPs.

Antibacterial and bioactive coatings on titanium implant surfaces.

Various surface modifications have been tried for enhancing osseointegration of the dental implants like mechanical and/or chemical treatments and deposition of calcium phosphate coatings. The objective of this research was to develop calcium-phosphate based thin coatings with antibacterial and bioactive properties for potential application in dental implants. Titanium (Ti) discs were immersed in different calcifying solutions: CaP (positive control), F-CaP, Zn-CaP, and FZn-CaP and incubated for 24 h. Negative control was uncoated Ti discs. Coated surfaces were characterized using X-ray diffraction, scanning electron microscopy and energy dispersive spectroscopy. Antibacterial properties were tested using Porphyromonas gingivalis because of its strong association with periodontal and peri-implant infections. Bacterial adhesion and colonization were studied at different timepoints. The coated surfaces had compositional characteristics similar to that of bone mineral and they inhibited the growth, colonization and adherence of P. gingivalis, resulted in reduced thickness of biofilms and bacterial inhibition in the culture medium as compared to the positive and negative controls (p < 0.05). There was no significant difference between the experimental groups (p > 0.05). It has been previously demonstrated that these coatings have excellent in vitro bioactivity (formed carbonate hydroxyapatite when immersed in a simulated body fluid). Such coatings can enhance osseointegration and prevent infection in implants, thereby improving the success rates of implants. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2218-2227, 2017.

Biphasic calcium phosphate: a scaffold for growth plate chondrocyte maturation.

While skeletal development can occur by either intramembranous or endochondral bone formation, all current tissue engineering approaches for bone repair and regeneration try to mimic intramembranous ossification. In this study, we propose to create an in vitro cartilage template as the transient model for in vivo endochondral bone formation. The goals of this study are to (1) establish a method of growing chondrocytes in a well-characterized macroporous biphasic calcium phosphate (MBCP) scaffold and (2) induce maturation of chondrocytes grown in the MBCP scaffold. Chondrocytes isolated from chick embryonic tibia were grown on MBCP particles and treated with retinoic acid to induce chondrocyte maturation and extracellular matrix deposition. Chondrocytes were observed to attach and proliferate on the MBCP scaffold. The thickness of the chondrocyte and extracellular matrix layer increased in the presence of the retinoid. Alkaline phosphatase activity and expression, proteoglycans synthesis, cbfa1 and type I collagen mRNA levels also increased in the presence of retinoic acid. These results demonstrated for the first time the proliferation, maturation of chondrocytes, and matrix deposition on MBCP, suggesting the potential for such scaffold in tissue engineering via the endochondral bone formation mechanism.

Immobilized rhBMP-2/succinylated type I atelocollagen gene expression of intracellular signaling molecules on ST2 cells.

Recombinant human bone morphogenetic protein-2 (rhBMP-2) chemically-bonded to succinylated type I atelocollagen, a biomaterial carrier with a porous structure, was reported to augment cellular activity of ST2 cells. The Smad protein family has been suggested to play an important role in the intracellular signaling pathway of BMP by its binding to receptors on target cells. However, there has been no study analyzing the downstream genes of the rhBMP-2 induced intracellular signal transduction pathway. The purpose of this study was to examine the effect of immobilized rhBMP-2 on gene expression of intracellular signaling molecules on ST2 cells. Our study showed two expression patterns of downstream genes of rhBMP-2 intracellular signal transduction pathway. In the first pattern, BMPR-IA, Smad 1, and Smad 5 genes showed high basic expression before the addition of rhBMP, and the high level of gene expression continued for long period and decreased in the late stage when rhBMP-2 was immobilized. In the second pattern, Smad 6, Smad 7, and Smad 8 genes showed low basic expression before the addition of rhBMP-2 and a continuous increase from the beginning was followed by a decrease in the late stage when rhBMP-2 was immobilized. Our results also showed that intracellular signaling continued for prolonged period when rhBMP-2 was immobilized to succinylated type I atelocollagen. This study indicated that immobilizing rhBMP-2 is an efficient method to increase bone induction.

Preservation and promotion of bone formation in the mandible as a response to a novel calcium-phosphate based biomaterial in mineral deficiency induced low bone mass male versus female rats.

Calcium and other trace mineral supplements have previously demonstrated to safely improve bone quality. We hypothesize that our novel calcium-phosphate based biomaterial (SBM) preserves and promotes mandibular bone formation in male and female rats on mineral deficient diet (MD). Sixty Sprague-Dawley rats were randomly assigned to receive one of three diets (n = 10): basic diet (BD), MD or mineral deficient diet with 2% SBM. Rats were sacrificed after 6 months. Micro-computed tomography (µCT) was used to evaluate bone volume and 3D-microarchitecture while microradiography (Faxitron) was used to measure bone mineral density from different sections of the mandible. Results showed that bone quality varied with region, gender and diet. MD reduced bone mineral density (BMD) and volume and increased porosity. SBM preserved BMD and bone mineral content (BMC) in the alveolar bone and condyle in both genders. In the alveolar crest and mandibular body, while preserving more bone in males, SBM also significantly supplemented female bone. Results indicate that mineral deficiency leads to low bone mass in skeletally immature rats, comparatively more in males. Furthermore, SBM administered as a dietary supplement was effective in preventing mandibular bone loss in all subjects. This study suggests that the SBM preparation has potential use in minimizing low peak bone mass induced by mineral deficiency and related bone loss irrespective of gender. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1622-1632, 2016.

Effects of immobilized recombinant human bone morphogenetic protein-2/succinylated type I atelocollagen on cellular activity of ST2 cells.

The use of recombinant human bone morphogenetic protein-2 (rhBMP-2) to induce ectopic bone formation requires a carrier. Type I atelocollagen, a biomaterial with a porous structure, excellent operational features, and biocompatibility, is an effective carrier for rhBMP-2. However, the conventionally used lyophilized rhBMP-2/collagen mixture does not necessarily give adequate bone-induction effect. In the present study, we examined the effect of immobilizing rhBMP-2 to type I atelocollagen on the cellular activity of ST2 cells. The following results were obtained: (1) rhBMP-2 was effectively immobilized to succinylated type I atelocollagen, indicating the usefulness of succinylated type I atelocollagen in immobilization; (2) studies of alkaline phosphatase activity confirmed the effectiveness of rhBMP-2 immobilized on succinylated atelocollagen in augmenting cellular activity.

Bone formation in calvarial defects of Sprague-Dawley rats by transplantation of calcium phosphate glass.

The purpose of this study was to investigate the bone-regenerative effect of calcium phosphate glass in vivo. We prepared amorphous calcium phosphate glass powder having a mean particle size of 400 microm in the system CaO-CaF2-P2O5-MgO-ZnO. Calvarial critical-sized defects (8 mm) were created in 60 male Sprague-Dawley rats. The animals were divided into an experimental group and control group of 30 animals each. Each defect was filled with a constant weight of 0.5 g calcium phosphate glass powder mixed with saline. As a control, the defect was left empty. The rats were sacrificed 2, 4, or 8 weeks postsurgery, and the results evaluated using radiodensitometric and histological studies; they were also examined histomorphometrically. When the calcium phosphate glass powders with 400-microm particles were grafted, the defects were nearly completely filled with new-formed bone in a clean healing condition after 8 weeks. It was observed that the prepared calcium phosphate glass enhanced new bone formation in the calvarial defect of Sprague-Dawley rats and could be expected to have potential for use as a hard tissue regeneration material.

Ultrastructural study of calculus-enamel and calculus-root interfaces.

UNLABELLED: The attachment of dental calculus to the tooth (enamel or cementum) surface affects the ease or difficulty of its removal. Understanding the ultrastructural features of the calculus-tooth interface will help in the development of efficient strategies for efficient removal of dental calculus. OBJECTIVE: The aim of this study was to determine the ultrastructural characteristics of the calculus-tooth interface in relation to the occurrence of calculus fracture. DESIGN: Investigation of the ultrastructural characteristics of the calculus-tooth interface was made on eight human molars with mature supragingival and subgingival calculus using scanning electron microscopy (SEM), transmission electron microscopy (TEM) and fourier transform infra-red (FT-IR) spectroscopy. RESULTS: Fractures were shown by SEM to consistently occur within the calculus itself, but not at the calculus-tooth interface. Higher magnification revealed that the enamel apatite crystals (in the case of supragingival calculus) or the cementum apatite crystals (in the case of subgingival calculus) appeared intimately connected with the calculus crystals at the calculus-enamel or calculus-cementum interface. TEM micrographs confirmed this intimate direct connection or fusion (epitaxial growth) of calculus crystals with enamel and cementum apatite crystals. FT-IR showed lower concentrations of organic phase attributed to microorganisms and higher concentrations of collagen at the calculus-cementum interface compared to that in the calculus away from the interface. CONCLUSION: Difficulty in complete calculus removal from tooth surfaces (especially from cementum or dentin) may be due in part to the intimate contact between the calculus and the tooth, due to the chemical bonding between the calculus crystals and the tooth apatite crystals and occasional fusion (i.e., epitaxial growth) of the calculus calcium phosphate crystals with the enamel, dentin or apatite crystals. This cohesive bonding results in fracture planes occurring within the calculus instead of at the calculus-tooth interface.

In vivo effect of fluoride-substituted apatite on rat bone.

Different types of calcium phosphate compounds are commercially available for medical and dental applications as bone substitute materials. Biological apatites contain several kinds of minor elements such as carbonate (CO3), magnesium (Mg), and fluoride (F) in enamel, dentin, and bone. It has been shown that F ion and F-substituted apatite promoted osteoblast proliferation and inhibited osteoclast cell activity. The purpose of this study was to investigate the in vivo rat tibia activity on F-substituted apatite (FAp). Apatites of unsintered calcium deficient apatite (CDA), and FAps, with low, medium, and high F concentrations, were implanted in rat tibia for 1 and 2 weeks. Implanted tissues were embedded in paraffin blocks, stained with hematoxylin-eosin and histomorphometrically observed. Results showed that low F concentration induced better and faster new bone formation in vivo compared to CDA. Therefore the results suggested that F as a minor element in bone rendered a suitable effect on bone formation in vivo.

Effect of calcium phosphate coating and rhBMP-2 on bone regeneration in rabbit calvaria using poly(propylene fumarate) scaffolds.

Various calcium phosphate based coatings have been evaluated for better bony integration of metallic implants and are currently being investigated to improve the surface bioactivity of polymeric scaffolds. The aim of this study was to evaluate the role of calcium phosphate coating and simultaneous delivery of recombinant human bone morphogenetic protein-2 (rhBMP-2) on the in vivo bone regeneration capacity of biodegradable, porous poly(propylene fumarate) (PPF) scaffolds. PPF scaffolds were coated with three different calcium phosphate formulations: magnesium-substituted ß-tricalcium phosphate (ß-TCMP), carbonated hydroxyapatite (synthetic bone mineral, SBM) and biphasic calcium phosphate (BCP). In vivo bone regeneration was evaluated by implantation of scaffolds in a critical-sized rabbit calvarial defect loaded with different doses of rhBMP-2. Our data demonstrated that scaffolds with each of the calcium phosphate coatings were capable of sustaining rhBMP-2 release and retained an open porous structure. After 6weeks of implantation, micro-computed tomography revealed that the rhBMP-2 dose had a significant effect on bone formation within the scaffolds and that the SBM-coated scaffolds regenerated significantly greater bone than BCP-coated scaffolds. Mechanical testing of the defects also indicated restoration of strength in the SBM and ß-TCMP with rhBMP-2 delivery. Histology results demonstrated bone growth immediately adjacent to the scaffold surface, indicating good osteointegration and osteoconductivity for coated scaffolds. The results obtained in this study suggest that the coated scaffold platform demonstrated a synergistic effect between calcium phosphate coatings and rhBMP-2 delivery and may provide a promising platform for the functional restoration of large bone defects.

Oral cancer risk perception among participants in an oral cancer screening program.

Oral cancer screening provides an opportunity for early detection and for education to high-risk tobacco and alcohol users. To plan interventions that would motivate oral cancer screening and risk reduction, we surveyed oral cancer risk perception and risk behaviors among participants in a free oral-cancer screening. Participants (N = 803) were racially diverse; 43% had a history of smoking and 9%, a history of alcohol abuse. Current smokers and those with higher lifetime tobacco exposure perceived themselves to be at higher risk for cancer than nonsmokers or those with less lifetime tobacco exposure (all ps, <0.01). Alcohol use was unrelated to oral cancer risk perception (p > 0.05). Compared with women, men (p = 0.01) felt more at risk; compared with other racial groups, Asians (p < 0.05) felt less at risk. Demographic differences were explained by differences in risk behaviors. Current smoking status (beta, 0.196; p < 0.001) and level of tobacco exposure (beta, 0.100; p < 0.05) were the only significant independent predictors of heightened risk perception (R(2) = 0.09). Those with alcohol abuse histories, and older smokers, may need increased education. These findings support the need for health education materials that incorporate the oral cancer risk perception of high-risk individuals.

Adherent octacalciumphosphate coating on titanium alloy using modulated electrochemical deposition method.

Our aims in this study were (1) to develop an electrochemical method of depositing adherent octacalciumphosphate (OCP) and other calcium phosphate coatings on titanium alloy (Ti6Al4V) substrates of different shapes and surface preparations, (2) to determine the properties of the coating (composition, morphology, thickness, dissolution), and (3) to observe transformation of OCP to carbonatehydroxyapatite (CHA) in simulated body fluid (SBF). Titanium (Ti)-alloy plates, tensile bars with four types of surfaces (grit-blasted with apatitic abrasive, chemically textured, arc-deposited, and Co-Cr-beaded) and dissolution cylinders were electrochemically coated with the use of modulated pulse time electric fields programmed with a custom-made dual microprocessor. Modulated electrochemical deposition (MECD) was carried out with pH and temperature conditions favorable for OCP formation. Coatings were characterized using X-ray diffraction, FT-IR, scanning electron microscopy, tensile strength tests, and solubility tests. XRD and FT-IR analyses showed that pure, uniform OCP coatings were produced on Ti6Al4V surfaces with coating-to-substrate tensile strengths greater than 7,000 psi. Coatings on Ti arc-deposited surfaces, chemically textured surfaces, and Co-Cr-beaded surfaces all gave tensile strengths ranging from 5,000 to 7,000 psi, with no coating shadows in the crevices. Dissolution of OCP coating in 100 mL of 0.1 M Tris buffer solution was determined from the amount of calcium (Ca) released onto the buffer, which was 7.7 +/- 1.0 ppm Ca at pH 7.3 after 4 h, and 22 -/+ 1.4 ppm Ca at pH 3 after 2 h. We found that OCP crystal size can be controlled by the current density and relative pulse time modulation. Our study demonstrated the following: (1) Highly adherent calcium phosphate (e.g., OCP) coating of uniform compositions (e.g., OCP) on Ti-alloy substrates can be obtained at low temperatures with the use of MECD by optimizing pulse time modulation of the electric field, reaction pH, temperature, and electrolyte composition; and (2) OCP readily transforms to CHA when exposed to SBF.

In vitro response of osteoblast-like and odontoblast-like cells to unsubstituted and substituted apatites.

Different types of calcium phosphate compounds [calcium-deficient apatite (CDA); beta-tricalcium phosphate (beta-TCP); biphasic calcium phosphate (BCP)] are commercially available for medical and dental applications as bone substitute materials. Most of the reported in vitro studies on cell-material interactions have used osteoblast-like cells. The purpose of this study was to investigate the in vitro response of osteoblast-like (MC3T3-E1) and odontoblast-like (MDPC23) cells on unsubstituted (HA) and substituted (F-substituted) apatites. MC3T3-E1 and MDPC23 were cultured in alpha-modified medium containing 10% fetal bovine serum, ascorbic acid (50 microg/mL) and beta-glycerophosphate (2 mM). The cells were seeded on pellets made from HA, and FAp (with low, medium, and high F concentrations). Cell morphology was observed after 7 and 14 days using scanning electron microscopy (SEM). Cell attachment and differentiation were determined from the DNA content, alkaline phosphatase (ALP) activity, and total collagen content. Pellet surface composition was characterized by using Fourier Transform infrared spectroscopy. MC3T3-E1 and MDPC23 cells on HA were normal in shape and in fusion but not on FAp. Results of this study showed that the pattern of cell proliferation of osteoblast-like cells was different from that of the odontoblast-like cells. This study suggests that cell morphology, fusion, and proliferation on biomaterial surfaces depend on cell type (osteoblast-like vs odontoblast-like cell) and biomaterial composition (unsubstituted vs substituted F-apatites).

Proliferation, differentiation, and calcification of preosteoblast-like MC3T3-E1 cells cultured onto noncrystalline calcium phosphate glass.

The purpose of this study was to investigate calcium phosphate glass as a potential biomaterial for hard tissue repair. We prepared calcium phosphate glass using the system CaO-CaF(2)-P(2)O(5)-MgO-ZnO and cultured MC3T3-E1 cells onto the glass in alpha-MEM with beta-glycerophosphatase and ascorbic acid. Proliferation of the cells was determined to evaluate the biocompatibility of the prepared calcium phosphate glass. The alkaline phosphatase activity was measured to examine the osteoblast differentiation. Mineralization was evaluated by staining the calcium precipitates with Alizarin red. Culture onto the calcium phosphate glass exhibited no significant influence on cell proliferation compared to the polystyrene chosen as a control in this experiment (p > 0.05). The alkaline phosphatase activity in the experimental group, however, was enhanced by the calcium phosphate glass significantly at 10-18 days after incubation than that of the control group (p < 0.05). The promotion of bone-like tissue formation by the calcium phosphate glass was observed after 7 days and thereafter. The results of the present study indicate that the prepared calcium phosphate glass affects osteogenesis by increasing calcification of the extracellular matrix.