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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., (© 2019 Wiley Periodicals, Inc.)

The osteochondral interface functions as a structural barrier between cartilage and bone, maintaining tissue integrity postinjury and during homeostasis. Regeneration of this calcified cartilage region is thus essential for integrative cartilage healing, and hydrogel-ceramic composite scaffolds have been explored for calcified cartilage formation. The objective of this study is to test the hypothesis that Ca/P ratio of the ceramic phase of the composite scaffold regulates chondrocyte biosynthesis and mineralization potential. Specifically, the response of deep zone chondrocytes to two bioactive ceramics with different calcium-phosphorus ratios (1.35 ± 0.01 and 1.41 ± 0.02) was evaluated in agarose hydrogel scaffolds over two weeks in vitro. It was observed that the ceramic with higher calcium-phosphorus ratio enhanced chondrocyte proliferation, glycosaminoglycan production, and induced an early onset of alkaline phosphorus activity, while the ceramic with lower calcium-phosphorus ratio performed similarly to the ceramic-free control. These results underscore the importance of ceramic bioactivity in directing chondrocyte response, and demonstrate that Ca/P ratio is a key parameter to be considered in osteochondral scaffold design. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2694-2702, 2017., (© 2017 Wiley Periodicals, Inc.)

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., (© 2017 Wiley Periodicals, Inc.)

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., (© 2016 Wiley Periodicals, Inc.)

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 β-tricalcium phosphate (β-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 β-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., (Copyright © 2015. Published by Elsevier Ltd.)

We have developed a calcium phosphate glass (CPG) doped with Zn(2+) or F(-) or combined Zn(2+) and F(-) ions, which are naturally found in the human body and play a dual role in bone formation and antibacterial activity. Previously, we have demonstrated that this family of CPGs has superior osteoconductive and resorbable properties in vivo. This study aimed to investigate the antibacterial property of CPGs incorporating Zn(2+) and/or F(-) . We used Streptococcus mutans as a model organism because it is one of the major human oral pathogens and an early colonizer, and it has been associated with several oral infections, such as dental caries, periodontitis, and peri-implantitis. CPGs of 0.01 and 0.05 g were incubated with S. mutans for 0, 2, 4, and 6 h. Serial dilutions were plated in triplicate and colony forming units were determined. The antimicrobial effect of CPG incorporating Zn(2+) or F(-) was greater than CPG incorporating both these ions. CPG without doping produced a moderate antimicrobial effect. This family of CPGs, previously shown to promote new bone formation in vivo, is demonstrated to have superior bactericidal properties., (Copyright © 2013 Wiley Periodicals, Inc.)

3D porous scaffolds are relevant biomaterials to bone engineering as they can be used as templates to tissue reconstruction. The aim of the present study was to produce and characterize in vitro 3D magnesium-carbonate apatite/collagen (MCA/col) scaffolds. They were prepared by using biomimetic approach, followed by cross-linking with 0.25% glutaraldehyde solution (GA) and liofilization. Results obtained with Fourier-transform infrared spectroscopy (FT-IR) confirmed the type-B carbonate substitution, while by X-ray diffraction (XRD), a crystallite size of ~10nm was obtained. Optical and electron microscopy showed that the cylindrical samples exhibited an open-porous morphology, with apatite nanocrystals precipitated on collagen fibrils. The cross-linked 3D scaffolds showed integrity when immersed in culture medium up to 14 days. Also, the immersion of such samples into an acid buffer solution, to mimic the osteoclastic resorption environment, promotes the release of important ions for bone repair, such as calcium, phosphorus and magnesium. Bone cells (SaOs2) adhered, and proliferated on the 3D composite scaffolds, showing that synthesis and the cross-linking processes did not induce cytotoxicity., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Objectives: This study aimed to determine the efficacy of experimental calcium phosphate-based solutions (sCaP) containing fluoride (F), with and without zinc (Zn) ions on reducing susceptibility to acid dissolution and Streptococcus mutans (S. mutans) colonization of dentin surfaces., Methods: Dentin sections were treated with double distilled water (control) and with sCaP solutions differing in pH and in F(-) and/or Zn(2+) ion concentrations. Solutions A (pH 7); B, C, and D (pH 5.5); solution C, twice Zn(2+) and F(-) ion concentration compared to B; solution D is similar to C but without Zn(2+). The dentin surfaces were characterized using scanning electron microscopy (SEM), x-ray diffraction, and Fourier Transform Infrared spectroscopy. Dissolution was determined in acidic buffer. Bacterial (S. mutans) attachment and growth were evaluated using SEM and Bioquant. Statistical analyses applied analysis of variance (ANOVA) and Duncan's multiple Range test., Results: Compared to control, dentin surfaces treated with sCaP solutions showed: (a) occluded dentin tubules; (b)reduced susceptibility to acid dissolution; and (c) Zn(2+) ions were more effective than F(-) ions in inhibiting bacterial colonization., Significance: Acidic sCaP containing both F and Zn ions have mineralizing, acid resistance, and antibacterial effects and may be potentially useful as a strategy against dentin caries formation and progression.

Unlabelled: Osteoporosis affects the craniofacial and oral structures and has been associated with periodontal bone loss, tooth loss and reduced jaw bone mass., Objective: This study aimed to test the therapeutic efficacy of synthetic bone mineral (SBM) in minimizing alveolar bone loss induced by mineral deficiency in a rat model. SBM consists of a calcium carbonate apatite (similar to bone apatite) matrix incorporating magnesium, zinc, and fluoride ions., Design: Thirty female Sprague Dawley rats (2 months old) were randomly distributed into 3 groups (10 rats per group): GA (control), on basic diet; GB, on mineral deficient (MD) diet; and GC, on MD+SBM. The rats were sacrificed after 3 months, the jawbones were isolated and the soft tissues removed. Bone density was determined using X-ray radiography (Faxitron); mandibular cortical width, panoramic mandibular index, and alveolar resorption degree (M/M ratio) using BioquantOsteo; and bone micro-architecture micro-computed tomography and scanning electron microscopy., Results: Compared to control (GA), the rats on MD diet (GB) experienced significant mandibular bone loss while the rats on MD+SBM diet (GC) experienced significantly less bone loss compared to the GB group., Conclusion: SBM, administered orally, may have the potential as an osteoporosis therapeutic agent in minimizing or preventing alveolar bone loss induced by mineral deficiency., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Objective: The aim of this study was to evaluate the effects of ZnCl(2) on plaque-growth and vitality pattern of dental biofilm and to determine the optimum zinc concentration for the inhibition of plaque formation., Design: Data were collected from nine volunteers for whom a special-designed acrylic appliance was prescribed after a careful dental check up. The volunteers rinsed twice daily for 2min with ZnCl(2) of 2.5, 5, 10, 20mM as treatment and double distilled water (DDW) as control in respective assigned test weeks. The plaque index (PI) was assessed after 48h of appliance wearing. The glass discs with the adhered biofilm were removed from the splints and stained with two fluorescent dyes. The biofilm thickness (BT) and bacterial vitality of the whole biofilm as well as the mean bacterial vitality (BV) of the inner, middle and outer layers of biofilm were evaluated under confocal laser scanning microscope (CLSM)., Results: PI, BT and BV of biofilms treated by various concentrations of ZnCl(2) were reduced significantly when compared with the DDW group (p<0.05). PI, BT and BV of the 2.5mM ZnCl(2) group was significantly higher than groups of 5, 10, 20mM ZnCl(2) (p<0.05). The mean BV of the 3 layers (inner, middle and outer layers) showed that 2.5mM ZnCl(2) was the lowest concentration to inhibit BV in the outer layer, 5mM was the lowest concentration to extend this inhibition of BV to the middle layer, and none of the concentrations investigated in this study has shown any effect on bacteria inhibition in the inner layer., Conclusion: Zinc ions exhibited possible inhibitory effects on plaque formation, and have a promising potential to be used as an antibacterial agent in future dentifrices and mouthrinses., (Copyright © 2011. Published by Elsevier Ltd.)

The aim of this study was to evaluate differences in bones quality between newly formed bone and cortical bone formed around titanium alloy implants by using X-ray photoelectron spectroscopy. As a result of narrow scan measurement at 4 weeks, the newly formed bone of C1s, P2p, O1s, and Ca2p were observed at a different peak range and strength compared with a cortical bone. At 8 weeks, the peak range and strength of newly formed bone were similar to those of cortical bone at C1s, P2p, and Ca2p, but not O1s. The results from this analysis indicate that the peaks and quantities of each element of newly formed bone were similar to those of cortical bone at 8 weeks, suggestive of a strong physicochemical resemblance.

The success of implants in orthopaedic and dental load-bearing applications crucially depends on the initial biological fixation of implants in surrounding bone tissues. Using hydroxyapatite (HA) coating on Ti implant as carrier for bone morphogenetic proteins (BMPs) may promote the osteointegration of implants; therefore, reduce the risk of implant failure. The goal of this study was to develop an HA coating method in conditions allowing the incorporation of protein-based drugs into the coating materials, while achieving a mechanical stable coating on Ti implant. HA coatings were deposited on six different groups of Ti surfaces: control (no pretreatment), pretreated with alkali, acid, heat at 800°C, grit blasted with Al₂O₃, and grit blasted followed by heat treatment. HA coating was prepared using a two-step procedure. First step was the chemical deposition of a monetite coating on Ti substrate in acidic condition at 75°C and the second step was the hydrolysis of the monetite coating to HA. Coatings were characterized by scanning electron microscope (SEM), energy-dispersive X-ray spectroscopy (EDS), and X-ray diffraction (XRD). The roughness of substrates and coatings was measured using profilometry technique. The mechanical stability of the coatings deposited on the pretreated substrates was assessed using scratch test. The coatings deposited on the grit-blasted Ti surface demonstrated superior adhesive properties with critical shearing stress 131.6 ± 0.2 MPa., (Copyright © 2011 Wiley Periodicals, Inc.)

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.

Biological apatites are characterized by the presence of minor constituents such as magnesium (Mg), chloride (Cl), or fluoride (F) ions. These ions affect cell proliferation and osteoblastic differentiation during bone tissue formation. F-substituted apatites are being explored as potential bonegraft materials. The aim of the present study is to investigate the mechanism of bone formation induced by fluoride-substituted apatite (FAp) by analyzing the effect of FAp on the process of in vivo bone formation. FAps containing different F concentrations (l-FAp: 0.48 wt%, m-FAp: 0.91 wt%, h-FAp: 2.23 wt%) and calcium-deficient apatite (CDA), as positive control, were implanted in rat tibia and bone formation was evaluated by histological examination, immuhistochemistry, in situ hybridization and tartrate-resistant acid phosphatase examinations. The results showed that l-FAp, m-FAp, h-FAp, and CDA biomaterials allowed migration of macrophages, attachment, proliferation, and phenotypic expression of bone cells leading to new bone formation in direct apposition to the particles. However, the l-FAp preparation allowed faster bone conduction compared to the other experimental materials. These results suggest that FAp with low F concentration may be an efficient bonegraft material for dental and medical application.

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.

BCP are considered the most promising biomaterials for bone reconstruction. This study aims at analyzing the outcomes of patients who received BCP as bone substitutes in orthopaedic surgeries. Sixty-six patients were categorized according to the etiology and morphology of the bone defects and received scores after clinical and radiographic evaluations. The final results corresponded to the combination of both parameters and varied from 5 (excellent result) to 2 or lower (poor result). Most of the patients who presented cavitary defects or bone losses due to prosthesis placement or revision, osteotomies, or arthrodesis showed good results, and some of them excellent results. However, patients with segmental defects equal or larger than 3 cm in length were classified as moderate results. This study established clinical parameters where the BCP alone can successfully support the osteogenic process and where the association with other tissue engineering strategies may be considered.

Our previous study demonstrated that calcium-bonded titanium surface (Ca-Ti) can be obtained by hydrothermal reaction between titanium (Ti) and CaCl(2) and that bone-apatite like formation was observed after immersion in simulated body fluid. The purpose of the study was to determine the in vivo response to Ca-Ti surface using a rodent tibia model. Cylinders of commercially pure Ti were divided into three groups: (1) untreated group; (2) NaOH+hTi group: soaked in 5 mol/L NaOH solution at 60 degrees C then heated at 400 degrees C for 1 h; and (3) Ca-Ti group: hydrothermally treated in the presence of 10 mmol/L CaCl(2) at 200 degrees C for 24 h. The cylinders implanted in surgically created defects in tibias of 8-week old male Wistar rats were retrieved after 1, 2, and 4 weeks. Histomorphometric evaluations were made on stained decalcified thin sections. Results showed that at 1, 2, and 4 week after implantation, respectively, bone contact was 55.2 +/- 16.4%, 88.1 +/- 9.9%, and 96.1 +/- 4.8% for Ca-Ti implants, 5.7 +/- 5.3%, 19.9 +/- 1.2%, 57.4 +/- 4.8% for untreated; and 27.2 +/- 0.7%, 70.9 +/- 7.7%, and 96.0 +/- 5.1% for NaOH+hTi implants. These results suggest that hydrothermal treatment with CaCl(2) provides a bioactive Ca-Ti bonded surface that allows bone formation greater than that obtained with NaOH+heat treated Ti surfaces., (Copyright 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2010.)

Bone repair and regeneration is one of the most extensively studied areas in the field of tissue engineering. All of the current tissue engineering approaches to create bone focus on intramembranous ossification, ignoring the other mechanism of bone formation, endochondral ossification. We propose to create a transient cartilage template in vitro, which could serve as an intermediate for bone formation by the endochondral mechanism once implanted in vivo. The goals of the study are (1) to prepare and characterize type I collagen sponges as a scaffold for the cartilage template, and (2) to establish a method of culturing chondrocytes in type I collagen sponges and induce cell maturation. Collagen sponges were generated from a 1% solution of type I collagen using a freeze/dry technique followed by UV light crosslinking. Chondrocytes isolated from two locations in chick embryo sterna were cultured in these sponges and treated with retinoic acid to induce chondrocyte maturation and extracellular matrix deposition. Material strength testing as well as microscopic and biochemical analyzes were conducted to evaluate the properties of sponges and cell behavior during the culture period. We found that our collagen sponges presented improved stiffness and supported chondrocyte attachment and proliferation. Cells underwent maturation, depositing an abundant extracellular matrix throughout the scaffold, expressing high levels of type X collagen, type I collagen and alkaline phosphatase. These results demonstrate that we have created a transient cartilage template with potential to direct endochondral bone formation after implantation., ((c) 2010 Wiley Periodicals, Inc.)

Background: Few studies have examined utilization of oral health care services among immigrants. The authors examined the determinants of utilization of oral health care among a diverse group of immigrants in New York City., Methods: The authors examined and interviewed 1,417 foreign-born people, aged 18 to 65 years, who were residents of New York City. They conducted examinations by using criteria established by the National Institute of Dental and Craniofacial Research, Bethesda, Md. The authors used unconditional logistic regression to estimate odds ratios (ORs) and 95 percent confidence intervals for having visited a dentist within the previous year for each of the independent variables., Results: More than 70 percent of the participants lacked dental insurance and only about 31 percent reported that they had visited a dentist within the previous year. Flossing (OR = 1.18), dental insurance (OR = 1.58), having a regular source of dental care (OR = 4.76) and more filled teeth (1.33) were independent predictors of utilization of services., Conclusions: Having a regular source of dental care and having dental insurance are important predictors of immigrants' utilization of oral health care services in New York City., Clinical Implications: The study results suggest the importance of establishing affordable, culturally appropriate, community-based oral health care services to improve the oral health of vulnerable populations.

The purpose of this study was to prepare and characterize a three-dimensional scaffold consisting of porous nonwoven silk fibroin net/nano-apatite composite. The silk fibroin net was mineralized with apatite by alternative soaking in calcium and phosphate solutions. The scaffold exhibited a porous microstructure with open porosity (70-78%), with an average pore size of about 163 +/- 40.0 microm. The swelling ratio and water uptake were 4.55% and 81.93%, respectively, indicating excellent hydrophilicity. The nano-sized hydroxyapatite crystals were needle like, with a length of approximately 100-300 nm and a diameter of approximately 20-60 nm. The results of in vitro cell culture study using rat osteoblast cells demonstrated that the nonwoven silk fibroin net/nano-apatite composite showed excellent cytocompatibility for the growth of osteoblasts and had the capability to improve the viability of osteoblasts.

The mineral in bone is an impure hydroxyapatite, with carbonate as the chief minor substituent. Fluoride has been shown to stimulate osteoblastic activity and inhibit osteoclastic resorption in vitro. CO(3)- and F-substituted apatite (CFA) has been considered as potential bone graft material for orthopedic and dental applications. The objective of this study was to determine the effects of simultaneously incorporated CO(3) and F on the crystallographic physico-chemical properties of apatite. The results showed that increasing CO(3) and Na content in apatites with relatively constant F concentration caused a decrease in crystallite size and an increase in the extent of calcium release; increasing F content in apatites with relatively constant CO(3) concentration caused an increase in crystallite size and a decrease in the extent of Ca release. These findings suggest that CFAs as bone graft materials of desired solubility can be prepared by manipulating the relative concentrations of CO(3) and F incorporated in the apatite.

The specific aim of this study was to evaluate the efficacy of well-characterized Mg/Zn/F-CaP preparations (administered by injection) in preventing bone mineral deficiency in ovariectomized (OVX) rats. Donryu rats (4 weeks old, average weight 70 g) were divided into six experimental groups: GN (normal), GC (control, OVX), and OVX rats injected with suspensions of MZF-CaPs (G2, G3, and G4) or with Zn-containing tricalcium phosphate (ZnTCP, G1). The composition of the preparations was G1: 34.1 wt % Ca; 19.5 wt % P; and 6.17 wt % Zn; G2 (#51): 23.7 wt % Ca; 13.6 wt % P; 1.1 wt % Mg; 2.9 wt % Zn, and 1.1 wt % F; G3 (#68): 22.4 wt % Ca; 14.1 wt % P; 2.6 wt % Mg, 2.6 wt % Zn, and 2.3 wt % F; G4 (#76): 28.5 wt % Ca; 15.5 wt % P; 1.9 wt % Mg, 1.8 wt % Zn, and 3.0 wt % F. The suspensions (10 mg/0.2 mL) were injected into the right thigh once a week for 12 weeks. Bone mineral density (BMD) was measured by X-ray computed tomography and bone mechanical strength (BMS) was measured as femoral three-point bending strength. BMD and BMS were significantly higher in the femurs from groups G1, G2, G3, and G4 than from GC. No significant difference was observed in BMD or BMS between the left and right femurs for any group. The results indicate that the injected Mg/Zn/F-CaP compounds were effective in preventing bone loss induced by ovariectomy in rats and suggest that these compounds have potential use for treating osteoporosis., ((c) 2009 Wiley Periodicals, Inc.)

The final aim of our study is to develop a novel calcium phosphate cement based on zinc-containing α-tricalcium phosphate (αZnTCP) and evaluate its potential as bonegraft material in vivo. In the present study, in vivo efficacy of zinc in hardened bodies of αZnTCP was explored. The hardened bodies prepared from αZnTCP with zinc content of 0.00, 0.04, 0.08, 0.11 and 0.19 wt % were prepared by mixing pure αTCP or αZnTCP powder with 12 wt% sodium succinate solution at a solid-to-liquid ratio of 2.0. Due to the release of zinc ions into the physiological salt solution during curing, the zinc content in the hardened bodies was calculated to be 0.00, 0.03, 0.06, 0.10 and 0.18 wt%, respectively. The hardened bodies were implanted in the femora and tibia of white rabbits for 4 weeks. Histological and histomorphometric evaluation showed that the hardened body containing 0.03 wt% zinc, significantly promoted more new bone formation without evoking adverse tissue reactions than that without zinc. The hardened bodies containing 0.06 and 0.10 wt% zinc also resulted in the increase in numbers of active osteoblasts surrounding the new bone but caused inflammation at the implant sites. Results of this study indicate that the hardened body prepared with αZnTCP is superior to that prepared with αTCP in promoting new bone formation due to the release of zinc ions. This study also indicates that the optimum amount of zinc in the hardened body is about 0.03 wt % to avoid inflammatory reaction.

Tricalcium phosphate (TCP) is recognized as a promising bone replacement material due to its high bioactivity and resorbable properties. To mimic biological apatites, incorporation of magnesium (Mg) in TCP was proposed. Mg-substituted TCP (beta-TCMP) and beta-TCP dense tablets were obtained by pressing and sintering at 1,000 degrees C Mg-substituted calcium deficient apatite (Mg-CDA) and commercial TCP, respectively. The materials were characterized using X-ray diffraction, infrared spectroscopy and electron microscopy. Human osteoblast cells (SaOs2) were seeded onto the sintered tablets for 4 h, 24 h and 7 days. Results showed that Mg-CDA was completely transformed into beta-TCMP. Moreover, beta-TCMP stimulated adhesion and proliferation of human osteoblast cells. Consequently, the magnesium incorporation on calcium deficient apatites followed by sintering at 1,000 degrees C seems to be a useful path to obtain biocompatible and non cytotoxic dense tablets with TCP structure with potential application on bone engineering.

The equilibrium solubility of Mg-containing beta-tricalcium phosphate (betaMgTCP) with various magnesium contents was determined by immersing betaMgTCP powder for 27 months in a CH(3)COOH-CH(3)COONa buffer solution at 25 degrees C under a nitrogen gas atmosphere. The negative logarithm of the solubility product (pK(sp)) of betaMgTCP was expressed as pK(sp)=28.87432+1.40348C-0.3163C(2)+0.04218C(3)-0.00275C(4)+0.0000681659C(5), where C is the magnesium content in betaMgTCP (mol.%). The solubility of betaMgTCP decreased with increasing magnesium content owing to the increased structural stability and possible formation of a whitlockite-type phase on the surface. As a result, betaMgTCP with 10.1 mol.% magnesium had a lower solubility than that of hydroxyapatite below pH 6.0. betaMgTCP was found to be more soluble than zinc-containing beta-tricalcium phosphate given the same molar content of zinc or magnesium. The solubility of betaMgTCP and release rate of magnesium from betaMgTCP can be controlled by adjusting the Mg content by selecting the appropriate pK(sp).

Bone consists of a mineral phase (carbonate apatite) and an organic phase (principally collagen). Cancellous bone is characterized by interconnecting porosity necessary for tissue ingrowth and nourishment of bone cells. The purpose of the present study was to fabricate macroporous carbonate apatite (CAP) blocks with interconnecting porosity as potential bone substitute biomaterials by hydrothermal conversion of alpha-TCP foam in carbonate solution. The fabrication of the macroporous CAP was accomplished in two steps: (1) preparation of alpha-TCP foams using polyurethane foams as templates, and (2) hydrothermal conversion at 200 degrees C of alpha-TCP foam in the presence of ammonium carbonate solutions of different concentrations. The maximum carbonate content of the resultant CAP foam was approximately 7.4 wt %. The mean porosity of the CAP foam was as high as 93 vol %. The macroporous CAP blocks or granules prepared in this manner has properties similar to that of bone in mineral composition and in having interconnecting macroporosity necessary for osteoconductivity and tissue ingrowth. On the basis of composition and interconnecting macroporosity, the CAP foam materials could be ideal biomaterials for bone repair and as scaffolds for tissue engineering., (2008 Wiley Periodicals, Inc.)

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.

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., ((c) 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2008.)

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., ((c) 2007 Wiley-Liss, Inc.)

Purpose: It is now well known that implant surface properties affect osseointegration. Grit-blasting with abrasives and coating by plasma are methods to modify implant surfaces. This study aimed to compare the direction of new bone formation associated with three types of surfaces., Materials and Methods: Titanium (Ti) alloy rods grit-blasted with alumina abrasive (Group 1, G1), with apatitic abrasive (Group 2, G2), and with apatitic abrasive and plasma-sprayed with hydroxyapatite (Group 3, G3) were implanted in surgically created defects in tibias of New Zealand white rabbits for 2 and 4 weeks. After sacrifice, the implants and surrounding bones were obtained and analyzed using back scattered imaging., Results: Differences in patterns of bone formation among the groups were observed: originating from the cortical bone towards the implant surface (Type A), surrounding the implant (Type B) and originating from the medullary cavity (Type C). G1 and G3 showed Types A and B while G2 exhibited Types A, B and C. After 4 weeks, greater amount of new bone was observed in G2 group compared with those in G1 and G3 groups., Conclusions: This study demonstrated that patterns of bone formation are influenced by methods of surface modification.

Fluoride (F-)-substituted B-type carbonate hydroxyapatite (CHAP) powders were prepared for application as bone substitute materials having the ability to enhance bone formation and to suppress bone resorption due to the therapeutic effect of F-. F- was adsorbed on CHAP in a sodium fluoride solution followed by heating at 700 degrees C in carbon dioxide flow to substitute F- for the hydroxyl ion in the CHAP structure. The F- contents in the F-substituted CHAP powders were 16-22 times greater than that in normal adult human bones. The carbonate ion contents in the F-substituted CHAP powders corresponded to or were higher than that in normal adult human bones. F-substituted CHAP powder with CO3(2-) and F- contents of 11.03 and 0.66 wt%, respectively, slowly released F- in a physiological salt solution to a sufficiently high F- level. The F- concentration slowly increased and reached 67.20 +/- 4.81 microg l(-1), which was 1.5-9.3 times higher than that in the body fluid of normal adult humans, near the therapeutic window of F-, and far lower than the estimated toxic level. Therefore, the F-substituted CHAP can promote bone formation. The present F-substituted CHAP has the advantage of slow F- release over sodium fluoride and sodium monofluorophosphate which are highly soluble salts and cannot be sintered into a ceramic body.

Purpose: Current commercial guided bone regeneration membranes are susceptible to bacterial colonization, leading to premature membrane degradation. The purpose of this research is to modify current resorbable guided bone regeneration membranes with antibacterial property by mineralizing with zinc phosphate., Materials: Resolut Adapt LT (Gore-Tex; W.L. Gore & Associates, Inc., Flagstaff, AZ), composed of copolymer PGA/TMC, and BioMend Extend (Zimmer Dental, Carlsbad, CA), composed of bovine type 1 collagen, were used. The membranes were mineralized with zinc phosphate. The mineralized membranes were characterized using scanning electron microscopy, energy dispersive spectroscopy, x-ray diffraction, Fourier transform infrared spectroscopy, inductive coupled plasma, and thermogravimetry. Antibacterial property of zinc phosphate mineralized and nonmineralized membranes were determined using Actinobacillus actinomycetemcomitans standard strain ATCC 29522., Results: Scanning electron microscopy, energy dispersive system, and Fourier transform infrared identified zinc phosphate in the zinc phosphate mineralized membranes. Zinc phosphate mineralized membranes showed significant reduction in bacterial colony, forming units compared to nonmineralized membranes., Conclusion: Results of this study suggest that the use of zinc phosphate mineralized membranes can inhibit oral bacterial colonization and prevent inflammation due to membrane exposure. This antibacterial property may help achieve the optimal goal of guided bone regeneration.

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.

Statement of Problem: Ceramic restorations have been known to cause wear of opposing enamel., Purpose: The purpose of this study was to evaluate enamel wear caused by 3 ceramic substrates in the glazed and polished conditions., Material and Methods: Sixty ceramic discs (10 x 2 mm)-20 each of Finesse, All-Ceram, and IPS-Empress-were prepared and glazed. Each group of 20 was divided into 2 groups of 10. The surfaces of one group were ground and polished using a porcelain polishing kit (Dialite). The remaining 10 were left as glazed. Ten specimens of a type III gold alloy were cast into rectangular shapes of 10 x 12 x 2 mm and polished. Seventy human cusps were prepared from sound, caries-free, extracted teeth and abraded against the substrates in a wear machine for a total of 10,000 cycles. The cusp height loss was traced before and after the wear test using a profile projector. Mean surface roughness (R(a)) values for the substrates were also recorded with a profilometer before testing. Differences in R(a) were evaluated using 1- and 2-way ANOVA and the Scheffe post hoc test (alpha = .05)., Results: One-way ANOVA indicated that enamel height loss was significantly different by material (P < .001) and surface condition (glazed and polished or glazed; P < .05). Gold, polished Finesse, and polished All-Ceram were the least abrasive, whereas glazed IPS-Empress was the most abrasive. There was no significant interaction effect between substrate type and surface condition. Significant differences were found when R(a) of the substrate condition was compared with enamel wear (P < .01)., Conclusions: Gold, polished Finesse, and polished All-Ceram caused the least enamel wear, whereas IPS-Empress caused the most wear. Cast gold was significantly different than glazed IPS-Empress (P < .05), whereas other groups overlapped. There was significant correlation between R(a) and enamel wear (P < .01).

The dissolution rates of ceramic tricalcium phosphate (TCP) and zinc-containing tricalcium phosphate (ZnTCP) ceramics at pH 5.5 decrease with increasing zinc content. The relative dissolution rates of ceramic ZnTCP with zinc contents of 0.5 and 1.0 mol% are more than three times higher than the relative resorbed volumes of these ZnTCP ceramics which were reported previously. The dissolution rates are likely controlled by only one mechanism, that is polynucleation, in the zinc content range from 0 to 1.0 mol%. Since the surface area of ceramic TCP or ZnTCP is much lower than that of powdered TCP or ZnTCP, the initial dissolution flux of ceramic TCP or ZnTCP remains unchanged for a longer period than that of powdered TCP. As a result, no change in dissolution kinetics is expected during one cycle of osteoclastic resorption. Since neither the dissolution rate nor the change in dissolution kinetics accounts for the previously reported drastic reduction in osteoclastic resorption associated with ZnTCP, it is concluded that the drastic reduction in resorption arises from the suppressive effects of ZnTCP on osteoclast formation or osteoclastic activity.

The purpose of this study was to investigate the bone regenerative effect of calcium phosphate glass in vivo. We prepared two different sizes of calcium phosphate glass powder using the system CaO-CaF2-P2O5-MgO-ZnO; the particle size of the powders were 400 microm and 40 microm. 8 mm calvarial critical-sized defects were created in 60 male Sprague-Dawley rats. The animals were divided into 3 groups of 20 animals each. Each defect was filled with a constant weight of 0.5 g calcium phosphate glass powder mixed with saline. As controls, the defect was left empty. The rats were sacrificed 2 or 8 weeks after postsurgery, and the results were evaluated using radiodensitometric and histological studies; they were also examined histomorphometrically. When the bigger powders with 400 microm particle were grafted, the defects were nearly completely filled with new-formed bone in a clean healing condition after 8 week. When smaller powders with 40 microm particle were transplanted, new bone formation was even lower than the control group due to a lot of inflammatory cell infiltration. It was concluded that the prepared calcium phosphate glass enhanced the new bone formation in the calvarial defect of Sprague-Dawley rats and it is expected to be a good potential materials for hard tissue regeneration. The particle size of the calcium phosphate was crucial; 400 microm particles promoted new bone formation, while 40 microm particles inhibited it because of severe inflammation.

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.

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.

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., ((c) Wiley Periodicals, Inc.)

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.

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., (Copyright (c) 2005 Wiley Periodicals, Inc.)

The influence of temperature and aging time on the morphology and mechanical properties of nano-sized hydroxyapatite (HA) synthesized by a hydrothermal method is reported here. The pre-mixed reactants were poured into a stirred autoclave and reacted at temperatures between 25-250 degrees C for 2-10 h. HA powders thus obtained were examined using X-ray diffraction (XRD), high-resolution field emission scanning electron microscopy (FESEM) and a particle size analyzer. It was found that the aspect ratio of the particles increased with the reaction temperature. The length of the HA particles increased with the reaction temperature below 170 degrees C, but it decreased when the temperature was raised above 170 degrees C. The agglomerates of HA particles were formed during synthesis, and their sizes were strongly dependent on reaction temperatures. As the reaction temperature increased, the agglomerate size decreased (p = 0.008). The density of the discs pressed from these samples reached 85-90% of the theoretical density after sintering at 1200 degrees C for 1 h. No decomposition to other calcium phosphates was detected at this sintering temperature. A correlation existed (p = 0.05) between the agglomerate sizes of HA particles synthesized at various conditions and their sintered densities. With the increase of the agglomerate size, the sintered density of the HA compact decreased. It was found that both the sintered density and flexural strength increased with increasing aging time and reaction temperature. A maximum flexural strength of 78 MPa was observed for the samples synthesized at 170 degrees C for 5 h with the predicted average at these conditions being 65 MPa. These samples attained an average sintered density of 88%.

Plasma-sprayed "HA" coatings on commercial orthopedic and dental implants consist of mixtures of calcium phosphate phases, predominantly a crystalline calcium phosphate phase, hydroxyapatite (HA) and an amorphous calcium phosphate (ACP) with varying HA/ACP ratios. Alternatives to the plasma-spray method are being explored because of some of its disadvantages. The purpose of this study was to deposit an adherent apatite coating on titanium substrate using a two-step method. First, titanium substrates were immersed in acidic solution of calcium phosphate resulting in the deposition of a monetite (CaHPO4) coating. Second, the monetite crystals were transformed to apatite by hydrolysis in NaOH solution. Composition and morphology of the initial and final coatings were identified using X-ray diffraction (XRD), Scanning Electron Microscopy, and Energy Dispersive Spectroscopy (EDS). The final coating was porous and the apatite crystals were agglomerated and followed the outline of the large monetite crystals. EDS revealed the presence of calcium and phosphorous elements on the titanium substrate after removing the coating using tensile or scratching tests. The average tensile bond of the coating was 5.2 MPa and cohesion failures were observed more frequently than adhesion failures. The coating adhesion measured using scratch test with a 200-microm-radius stylus was 13.1N. Images from the scratch tracks demonstrated that the coating materials were squashed without fracturing inside and/or at the border of the tracks until the failure point of the coating. In conclusion, this study showed the potential of a chemical deposition method for depositing a coating consisting of either monetite or apatite. This method has the advantage of producing a coating with homogenous composition on even implants of complex geometry or porosity. This method involves low temperatures and, therefore, can allow the incorporation of growth factors or biogenic molecules., (Copyright (c) 2005 Wiley Periodicals, Inc.)

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.

Methods of preparing different types of titanium oxide (TiO(2)) and their effects on apatite deposition and adhesion on titanium surfaces were investigated. Forty-eight commercially pure titanium (Ti) discs were divided into four groups (12 per group) and each group was subjected to the following treatments: Group 1, heat treatment at 750 degrees C; Group 2, oxidation in H(2)O(2) solution followed by heat treatment; Group 3, dipping in rutile/gelatin slurry; and Group 4, dipping in anatase/gelatin slurry. Surface-treated Ti discs were immersed in a supersaturated calcium phosphate solution to allow apatite deposition. Results showed that the percentage of area covered by deposited apatite was highest in Group 4 compared to the other groups. Apatite deposited on Ti discs pretreated in H(2)O(2) solution (Group 2) demonstrated the highest adhesion to the titanium substrate. Results from this study indicated that surface treatment method affects the type of TiO(2) layer formed (anatase or rutile) and affects apatite deposition and adhesion on the Ti surface., ((c) 2004 Wiley Periodicals, Inc.)

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).

Cells within the periodontal ligament have the potential to regenerate a periodontal connective tissue attachment on pathologically exposed root surfaces as well as on several material surfaces including titanium. However, rather than a periodontal connective tissue attachment, a fibrous encapsulation or chronic inflammatory response has been reported at the material connective tissue interface for most dental materials. Cementum is the first tissue of the periodontal connective tissue attachment to develop and the secretion of enamel matrix related proteins on the newly mineralized dentin surface precedes and is thought to induce cementum formation. Enamel matrix-related proteins may also function in the adult because the application of an acid extract of porcine enamel protein matrix (Emdogain(R), EMD) on pathologically exposed root surfaces has been shown to result in cementum regeneration. Therefore, the objective of the present study was to determine whether the application of EMD to materials that do not normally support cementogenesis in vivo would alter the in vitro phenotype of periodontal ligament (PDL) cells including the synthesis of cementum-associated extracellular matrix proteins. Primary PDL cells were established from 21-day-old Sprague-Dawley rats, and were cultured on four materials commonly encountered in dental practice (gutta percha, calcium hydroxide, amalgam, and super EBA cement) with and without the application of EMD. After 7 or 14 days of culture, total-DNA content, collagen synthesis, alkaline phosphatase activity, and the synthesis of a 42-kDa cementum-associated extracellular matrix protein were determined. PDL cells cultured on all materials had decreased total DNA content. The application of EMD further decreased total DNA content. PDL cells cultured on gutta percha and calcium hydroxide with the application of EMD had similar levels of collagen synthesis and alkaline phosphatase activity but also expressed a 42-kDa cementum extracellular matrix-associated protein when compared to the other groups. These results suggest that EMD can alter the phenotype of PDL cells when cultured on these dental materials., (Copyright 2004 Wiley Periodicals, Inc. J Biomed Mater Res 69A: 172-179, 2004)

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., (Copyright 2004 Wiley Periodicals, Inc. J Biomed Mater Res 69A: 188-195, 2004)

The periodontal ligament has the potential to regenerate a complete periodontal connective tissue attachment, starting with the deposition of cementum, on pathologically exposed root surfaces as well as several materials including titanium oxide. However, most commonly used dental materials result in a fibrous encapsulation or a chronic inflammatory response after periodontal wound healing rather than the formation of a periodontal connective tissue attachment. Recently, an extract of porcine enamel matrix (Emdogain(R), EMD) has been reported inductive of cementum formation in both in vivo and in vitro studies. The aim of this study was to determine the effect of EMD, when applied to materials previously reported not supportive of periodontal connective tissue formation, on the periodontal connective tissue-material interface obtained with these materials in vivo. Bilateral osteotomies were performed on the mandible of a Yucatan minipig exposing the buccal root surface of four premolars. A series of four preparations were placed in each root surface that were subsequently filled with calcium hydroxide, gutta percha, mineral trioxide aggregate (MTA), or left unfilled. One side, in addition, received an application of EMD prior to surgical closure. A bioabsorbable surgical barrier membrane was placed over the osteotomy sites to exclude gingival connective tissue from the wound-healing environment. The mucoperiosteal flaps were then readapted and sutured in position. The animal was euthanized 10 weeks after the procedure, block sections obtained and prepared for light microscopy. Results demonstrated complete regeneration of alveolar bone and periodontal ligament in all four teeth from the EMD-treated side. Fibers from the periodontal ligament were observed to insert into a mineralized matrix consistent with cementum on all four root preparations. In contrast, massive root resorption without regeneration of alveolar bone was found on all teeth from the side not treated with EMD. The results of this pilot study suggest that the application of EMD to material surfaces that normally do not support periodontal connective tissue attachment formation can alter the type of periodontal connective tissue interface obtained with these materials., (Copyright 2004 Wiley Periodicals, Inc. J Biomed Mater Res 69A: 180-187, 2004)