Search

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.

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). 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.

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.

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.

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.

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.

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.

Synthetic apatites are widely used both in the dental and the orthopaedic fields due to their similarity in composition with the inorganic phase of hard tissues. Biologic apatites are not pure hydroxyapatite (HA), but are calcium-deficient apatites with magnesium and carbonate as minor but important substituents. The aim of the present study was to produce a more soluble biomaterial through the simultaneous substitution of magnesium and carbonate in the apatite structure to accelerate the degradation time in the body. The physico-chemical and dissolution properties of unsintered magnesium and carbonate-substituted apatite (MCAp) with similar Mg/Ca molar ratio (0.03) and varying C/P molar ratio were evaluated. The resultant powders were characterised using several techniques, such as FTIR, TGA, XRD, ICP and SEM, while the release of calcium ions in a pH 6 solution was monitored using a Ca-ion selective electrode. The results showed a decrease of crystallite size and an increase in the release of calcium to the medium as the carbonate content in the samples increased.

Differences in structural and crystallinity between the lateral and medial regions of the central femoral diaphysis in low-mineral-diet-fed ovariectomized rats were investigated using polarization microscopy and Raman spectral analysis. Eighteen 19-week-old female Wistar rats were divided into 3 groups (Group 1: sham ovariectomy + normal diet group, Group 2: ovariectomy + normal diet group, Group 3: ovariectomy + low-mineral-diet group). Measurements were performed in the lateral and medial regions of the femoral sagittal cross-section at 24 weeks of feeding. On polarization microscopy, tubular structures of haversian and Volkmanns canals and osteocytes were observed in the femur in Groups 1 and 2. In Group 3, the number of these tubular structures in the femur was decreased compared to those in Groups 1 and 2. Moreover, the bone width of the femur decreased in Group 3 compared to those in Groups 1 and 2. On Raman spectral analysis, the peaks of organic and inorganic components were observed in the lateral and medial regions of the femur in all groups. In magnified views of the peak of ν1PO43-spectrum, the peaks in the lateral and medial regions overlapped in Group 1, but the peak of the medial region was about 0.3 and 0.8 cm-1shifted toward the low wavenumber side compared to those of the lateral region in Groups 2 and 3, respectively. Based on these findings, the influences of reduced estrogen secretion and the low-mineral diet were clarified as differences in the femoral structural and crystallinity on polarization microscopy and Raman spectral analysis. Since highly sensitive detailed point analysis can be performed within a short time using Raman spectral analysis, it may be useful for high-dimensional structural analysis.

Ovariectomized rats were fed a diet containing minerals at high concentrations, such as Ca, P, and F (high-mineral diet), and changes in the femoral diaphysis were investigated after 24 weeks. The femur was mainly red and partially orange on the color scale of the 3D-map in Groups A and B, showing a high BMD. The region adjacent to the marrow cavity was yellow, showing a lower BMD than that in the outer region of the femur. In Group C, the red area was small in the outer region and the inner region was mainly yellow and green on the color scale. The inner region adjacent to the marrow cavity showed a view of unevenly resorbed bone, and the BMD was lower than those in Groups A and B. Incorporation of F into the body influences the apatite crystal structure and crystal growth, which subsequently influences adsorption of F to crystals and structural changes. Therefore, it is important to ingest F at the optimum concentration.

Bone is a composite of approximately 65% inorganic phase (carbonate apatite, CHA) and a 35% organic phase (mostly collagen). To date, several commercial composites consisting of natural or synthetic polymers and calcium phosphates ( hydroxyapatite, tricalcium phosphate, biphasic calcium phosphates) are recommended for use in bone repair. Objective: The aim of this study was to compare the physico-chemical properties of gelatin/carbonate apatite composites with that of bovine bone. Native (Gel) or cross-linked (Gel*) was used. Methods: The CHA was prepared by hydrolysis method. The gelatin (denatured collagen) was cross-linked using Genipin. The gelatin/CHA composite were prepared by mixing of 35% gelatin and 65% CHA and freeze-drying. The composites were characterized using x-ray diffraction (XRD), FT-IR spectroscopy, scanning electron microscopy (SEM) and thermogravimetry (TGA). Dissolution properties were determined in acidic buffer (0.1M KAc, pH 6, 37°C). Mechanical strength was determined using 3-point bend test. Bovine bone was similarly characterized for comparison. Results: The composition and crystallite size of the CHA were similar to that of the bone mineral. The Gel/CHA and Gel*/CHA composites showed several physico-chemical properties (crystallinity, composition, thermal stability, mechanical strength, dissolution rate) similar to that of bone. Gel*/CHA compared to Gel/CHA composites showed lower elastic modulus, flexural strength, dissolution rate, swelling and higher porosity. Conclusion: The Gel*/CHA composites presented several properties similar to those of bovine bone and may have potential as bone substitute materials.

We show that statistical modeling of analytical results is useful in providing insights into metabolism and disease in bioarcheology. Our results also imply that during the Renaissance in Europe widespread pollution of the biosphere with heavy metals such as mercury and lead affected the Italian nobility at that time. The activity of biologic clocks which control metabolism and autonomic nervous system (ANS) function can be gleaned from the analysis of hair. This provides a means of assessing the health of individuals who lived some six centuries before the present and allows the reconstruction of disease from archived tissues such as hair.

Monetite is a promising biomaterial for restoring the function of the compromised skeletal structures due to its superior osteoconductive and resorption properties. However, its potential as a coating for orthopaedic implants in load bearing applications has not yet been investigated. The aim of this study was to prepare monetite coating on Ti substrate in mild conditions, which may allow incorporation of protein-based drugs during the coating deposition. Coatings were prepared using chemical deposition of monetite on Ti substrate in acidic conditions at 75 °C for 24 h. Coatings were characterised by scanning electron microscope (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). Adhesion of the coatings to Ti substrate was measured using tensile tests. EDS confirmed the calcium phosphate nature of the coating and XRD and FTIR confirmed the monetite phase of the coatings. SEM revealed that the coatings were formed of densely packed plate-like monetite crystals, with the crystal size approximately 20 × 10 × 5 μm. Adhesion tests showed cohesive failure of the monetite coatings and the tensile strength of the coating was in the range of 15.43 ± 0.20 MPa. Mechanical tests showed that monetite coatings were stable and could be considered for use with Ti orthopaedic implants.

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 Zn2+ ion concentrations. Solutions A (pH 7); B, C, and D (pH 5.5); solution C, twice Zn2+ and F− ion concentration compared to B; solution D is similar to C but without Zn2+. 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) Zn2+ 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.

Due to the lack of macroporosity in current available Calcium Phosphate cement used in osteoarticular surgery, Micro and Macroporous Biphasic CaP Cement (MCPC™) was developed. The MCPC™ concept was the association of a settable and a fast resorbable matrix and a sieved fraction of microporous biphasic calcium phosphate (BCP) granules, recognized for the high osteoconductive and osteogenic properties. During the resorption of the matrix, a porous structure is created and the osteoconductive effect of the granules promotes the bone ingrowth. A goat preclinical study was realized to evaluate the efficacy of MCPC™ for C3 and C4 vertebral body filling defects during 6 months. Bone remodelling was evidenced demonstrating bone ingrowth at the expense of the cement and surrounding the residual BCP granules. Bone trabeculae were observed coming from the spongious bone to the implant site. Human vertebral body filling cases demonstrated the biocompatibility and the safety of MCPC™ for bone reconstruction. Results of this study demonstrated the importance of special combination of calcium phosphate granules in the MCPC™ to provide macroporosity and scaffolding for newly formed bone.

Both natural and synthetic materials have been utilized to provide three dimensional scaffold environments ideal for bone repair. The biomechanical and biocompatibility characteristics of these scaffolds play a vital role in successful tissue engineering constructs. Polymer/carbonate apatite (CHA) composites have shown to improve cell adhesion and proliferation on the scaffold as well as increase elastic modulus, toughness and strength. The aim of this study is to prepare CHA- polylactic-co-glycolide (PLGA) composites in the form of microsphere, scaffold and disc and evaluate their physico-chemical properties, mechanical properties and in vitro bioactivity. 3-D porous cylindrical composite scaffolds were prepared using PLGA/CHA composites with varying PLGA/CHA ratios (30:70 and 50:50). The CHA was prepared by hydrolysis method and characterized using x-ray diffraction (XRD) and Fourier Transform Infrared spectroscopy (FTIR). The physico-chemical and mechanical properties of the composite scaffolds were evaluated using scanning electron microscopy (SEM), micro-computed tomography (μCT), XRD, FTIR, and thermogravimetry (TGA). Flexural strength was determined using Instron. In vitro bioactivity was determined by the formation of apatite on composite disc surfaces after immersion in simulated body fluid (SBF). SEM and μCT analyses showed high porosity and interconnectivity between microspheres in the composite scaffolds. In vitro bioactivity was observed by the development of an apatite layer on the surfaces of the composite scaffolds after immersion in simulated body fluid. The mechanical strength of the scaffolds was to be dependent on the PLGA-CHA ratio. The elastic modulus, toughness and strength values obtained for the composites were similar to those of reported bone substituted materials. Results from this study provided information on the fabrication of PLGA-CHA scaffolds and their properties that may be useful for their potential application in bone repair and as scaffolds in tissue engineering for bone regeneration.

Natural bone constitutes of an inorganic phase (a biological nanoapatite) and an organic phase (mostly type I collagen). The challenge is to develop a material that can regenerate lost bone tissue with degradation and resorption kinetics compatible with the new bone formation. The aim of this study was to prepare self-organized magnesium and carbonate substituted apatite/collagen scaffolds, cross-linked with glutaraldehyde (GA). Bovine tendon was submitted to alkaline treatment resulting in a negatively charged collagen surface. The scaffolds were prepared by precipitation: simultaneous dropwise addition of solution containing calcium (Ca) and magnesium (Mg) ions and collagen into a buffered solution containing carbonate and phosphate ions in reaction vessel maintained at 37 °C, pH=8. The reaction products were cross-linked with 0.125 and 0.25% (v/v) glutaraldehyde (GA) solution and freeze-dried. The samples were characterized by Fourier-transformed infrared spectroscopy (FTIR). In vitro cytotoxicity (based on three parameters assays) and scaffolds degradation in culture medium and osteoblastic cells culture were performed in the cross-linked materials. No cytotoxic effects were observed. The cross-linked samples with the lower GA concentration showed a lower stability when placed in contact with culture medium. Human osteoblasts attached on the scaffolds surface cross-linked with 0.25% GA, forming a continuous layer after 14 days of incubation. These results showed potential application of the designed scaffolds for bone tissue engineering.

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. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2011.

The aim of this study was to monitor changes in bone density and bone quality (microarchitecture) in the femoral epiphyseal region in rats with bone loss induced by oestrogen deficiency and accelerated by mineral deficiency. Forty-eight Wistar rats were divided into 3 groups (n = 16 per group): Group 1, sham-operated rats on a normal diet; Group 2, ovariectomised rats on a normal diet; and Group 3, ovariectomised rats on a mineral-deficient diet. Observations were conducted 4, 8, 16, and 24 weeks after the operation (sham or ovariectomy), while the rats were on their respective diets. Microscopic computed tomography of the femoral epiphyseal region was used to generate 3-dimensional bone density distribution maps, and to compute bone density values. Densities of cortical and cancellous bone in Groups 2 and 3 were significantly less than densities in Group 1, and densities in Group 3 were significantly less than densities in Group 2 at nearly every post-operative time interval.This study demonstrated that: a) ovariectomy in rats induces bone loss; b) a mineral-deficient diet accelerates bone loss that has been induced by oestrogen deficiency; and c) 3-dimensional bone density distribution maps may be useful in diagnosing osteoporosis and other diseases causing bone loss. Microscopic computed tomography is a powerful analytical tool for measuring spatial variations in bone density, and temporal changes in the patterns of 3-dimensional bone density distribution maps.

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.

CO 3 - and/or F-substituted apatites have been considered as potential bone substitution material for dental and orthopedic applications. The objective of this study was to compare physico-chemical properties and in vitro bioactivity in fetal bovine serum (FBS) of apatites containing CO 3 and/or F. The results showed that CO 3 and F in apatites have opposite effects on crystallinity and solubility. Calcium deficient hydroxyapatite (HA) and F-substituted apatite (FA) partially transformed to beta-tricalcium phosphate (β-TCP) at temperature 950 °C. After immersion in FBS for 10 days, calcium deficient HA, FA, and CO 3 -substituted apatite (CHA) and CO 3 - and F-substituted apatite (CFA) pellet surfaces all showed formation of apatite.

The purpose of this study was to investigate changes in bone quality associated with the mineralization of new bone forming around implants. Eight 18-week-old male New Zealand white rabbits and implants fabricated from Ti-15%Zr-4%Nb-4%Ta alloy were used in the study. Characteristics of new bone formed around implants at 4 and 8 weeks and cortical bone were compared using X-ray photoelectron spectroscopy (XPS) for elemental qualitative and quantitative analysis, polarized microscopy (PM) for histological examination, and Fourier transform infrared Imaging (FTIR Imaging) to characterize molecular distributions in new bone and cortical bone. Results from XPS analysis showed changes in the chemical bonding in new bone over time (4 weeks vs. 8 weeks). PM showed changes in collagen birefringence, orientation and intensity in new bone over time, becoming more similar to those of cortical bone after 8 weeks. FTIR Imaging showed that in new bone close to cortical bone, amounts of PO 4 3- and CO 3 2- ions (representing mineral or inorganic components), and amounts of amide I (representing the organic component) increased over time as mineralization progressed. In new bone more distant from cortical bone, PO 4 3- , CO 3 2- , and amide I distributions also increased over time, but to a lesser extent to that observed near cortical bone. Our findings clearly showed changes in the quality of new bone forming around implants, and these changes were related to mineralization.

The purpose of this study was to determine the maximum substitution of magnesium, Mg 2+ , for calcium, Ca 2+ sites in a beta-tricalcium phosphate (Mg-β-TCP) powder using the Rietveld method. Ca-deficient apatite with a theoretical 0.2 Mg/Ca ratio was synthesized and the resultant powder sintered at 1000 °C for 4 h. The maximum Mg 2+ substitution of nearly 15 mol% in β-TCP structure was deduced from the occupancy factors. The analysis indicated that the maximum substitution of Mg 2+ on the M(4) and M(5) sites was, approximately, 4.21 and 10.72 mol%, corresponding to the Ca 2.61 (Mg1 0.28, Mg2 0.11 )(PO 4 ) 2 stoichiometric formula. A small displacement of the Ca atoms position was observed around the Mg1(5)PO 4 octahedral, and their increasing shift to the PO 4 triangles corners.

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 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).

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.

Silk has been used in biomedical applications for centuries. The potential of silk for application in tissue engineering is currently being explored. The purpose of this study was to develop new method of mineralizing silk with carbonate apatite and determine cell response. The cocoons were placed in sodium bicarbonate solution and heat treated. The treated silk fibers were mineralized with carbonate apatite (CHA) using precipitation and new microwave methods. The mineralized and non-mineralized silks were characterized using SEM, EDS, XRD, FT-IR, and TGA. Cell response to mineralized silk and non-mineralized silk was determined using human osteoblast like cells (MG-63). The microwave method was more efficient than the precipitation method in terms of the amount of minerals incorporated with the silk and time required for mineralization. EDS, FT-IR and XRD identified CHA in the mineralized silk. In terms of cell response, greater production of type 1 collagen was observed with CHA mineralized silk compared with non-mineralized silk.

–Objectives: The aim of this preliminary study was to compare the perception of oral health among subgroups of Asian-American residents of New York City, USA. Methods: A close-ended questionnaire was administered to 255 Chinese, 134 Indian and 84 Pakistani adults, aged 18–65 years, during 1994–95. A comprehensive dental and oral examination was also performed. The associations of demographic and oral health variables with perceived oral health were evaluated using multivariate ordinal regression models. Results: When data were analyzed in a multivariate context, only ethnicity and income were significant predictors of perceived oral health, after adjusting for DMFT. The within-group multivariate analysis of the three ethnic subgroups’ results were as follows: Among the Chinese there were no significant predictors, only income was strongly suggestive; among the Indians, number of missing teeth and number of years in the USA were significant predictors; and within the Pakistani group, DMFT was the only significant predictor. Conclusions: Results suggest that there are ethnic differences in the perception of oral health status even after adjusting for clinical variables as well as for demographic variables in this particular group of Asian-American residents of New York City. Predictors associated with the perception of oral health are different for each ethnic group. When designing oral health promotion activities to diverse ethnic groups, the cultural characteristics of each subgroup should be considered.

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.

Recently, dental implants have gained much attention for a great number of researchers in dental application. Until now, there are few reports about the properties and metabolism related to the bite force response of the newly bone formed surrounding the implants, which is poorly understood. For that reason, the purpose of this study was to elucidate the new bone formation surrounding the implants and its process of maturation by Micro-CT, polarizing microscope and FTIR analysis. Thus, the implants were inserted in the tibia of rabbits. After 4 and 8 weeks of surgical procedure, three-dimensional image of the implant and bone of non-decalcified samples was observed by Micro-CT analysis; bone structure and its maturity were seen by polarizing microscope; and finally, the PO 4 3- , CCO 3 2- and Amide I elements included in the interface bone- implant and in the new bone during the course time were examined by FTIR analysis. The results of this study, analyzed by this 3 methods, demonstrated that: 1) It was observed a qualitative difference in the new bone formation and the existing bone at 4 and 8 weeks of implantation; 2) There was also a qualitative difference in the newly bone formed at 4 and 8 weeks; 3) The maturation of the new bone started from the exhibiting bone; and 4) None of the methods used in current study destroyed the samples, and reproducibility results were obtained. In conclusion, we suggest a possible qualitative evaluation of new bone formation surroundings the Ti implant.

A newly developed titanium alloy, Ti-15%Zr-4%Nb-4%Ta (Ti15Zr4Nb4Ta), has been shown to have excellent mechanical characteristics, corrosion fatigue strength, anticorrosion, cytocompatibility, and biocompatibility. The purpose of this study was to compare the bone mineral density (BMD) and area of newly formed bone around Ti15Zr4Nb4Ta implants to that around Ti-6%Al-4%V alloy (Ti6Al4V) implants. We inserted 2 types of implants (surface treat of the machine and the blast implant) in New Zealand white rabbit femurs under general anesthesia. The rabbit was done perfusion fixation of 4, 8, 16, 24, and 48 weeks later, and the femurs with the implants were collected and we observed the surrounding bone by an implant drawing test. We measured the BMD and the area of the newly formed bone surrounding the implant inserted part using image analysis software Image-Pro PLUS Ver. 4.0 ® which it developed newly in micro focus computed tomography (micro CT). The results showed the machine and the blast implant together, which the BMD and area around the Ti15Zr4Nb4Ta implants were comparable to or greater than those around the Ti6Al4V implants. These results suggested that the new alloy, Ti15Zr4Nb4Ta, might be useful for orthopedic or dental implants with the added advantage of superior biologic safety and mechanical properties than the current alloy, Ti6Al4V.

Since 1952 when Branemark first reported osseointegration of titanium (Ti) with bone, many academic and industrial research activities have endeavored to improve the efficacy of Ti or Ti alloy (Ti6Al4V) by modifying the chemistry, topography and design of the implant surface. Strong bonding between implant and host tissue minimize the micromovements that promote fibrous tissue formation at the implant interface that may lead to implant failure. Surface design include lateral holes perpendicular to the implant axis, grooves, variations of spacings between ridges, etc. Physico-mechanical means of surface modification is by grit-blasting with various abrasives (alumina, silica, apatitic abrasive), laser ablation, spark discharge, etc. Chemical modifications include: acid etching, treatment with alkali, treatment with fluoride, coating with titanium or with calcium phosphate (by plasma spray, electromagnetic sputtering, electrochemical deposition). A review of studies on Ti or Ti alloy implants with different surfaces showed the following methods to enhance osseointegration and greater bone formation: (1) grit-blasting with apatitic abrasive; (2) acid-etching with mixed acids; (3) adjusting plasma-spray parameter to get a higher HA/ACP ratio in the coating; (4) employing electrochemical deposition (with pulse modulation) or precipitation to obtain thin coating with homogeneous composition; and/or (5) Ftreatment.

The purpose of this study was to determine the effect of sintering conditions on microporosity of and cell proliferation and bone ingrowth on biphasic calcium phosphate (BCP) bioceramics. Discs were prepared from a calcium-deficient apatite preparation that upon sintering at 1050oC and above, results in a BCP with 60% hydroxyapatite (HA)/ 40% beta-tricalcium phosphate (β-TCP) ratio. The discs were divided into groups which were sintered under different conditions of heating rate (programmed vs. non-programmed) and temperature (1050°C vs. 1200°C). The discs were characterized in terms of composition (HA/β-TCP ratio), surface morphology, surface area, surface microporosity, per cent microporosity, and dissolution properties. The in vitro effect of sintering conditions on cell proliferation was determined using an established mouse fibroblast cell line (L929). Results demonstrated the following: (a) the HA/β-TCP ratio remained 60/40 regardless of sintering conditions; (b) the % microporosity, surface microporosity, surface area of the BCP and cell proliferation on the BCP significantly decreased with increasing sintering temperature, and (c) the extent of dissolution decreased with decreasing per cent microporosity. The in vivo study indicated no tissue adverse reaction and direct bone contact with the implant surface, confirming the biocompatibility of the BCP bioceramics. Resorption of the BCP and bone ingrowth was directly related to the sintering temperature: the higher the temperature, the lower the resorption and the bone ingrowth. Results of this study indicate that per cent microporosity of the BCP bioceramics affects its dissolution properties and cell response. The study demonstrates that optimum per cent microporosity elicits optimum cell response and should be considered to provide osteogenic/osteoinductive property to bioceramics.

We have developed a new Ti alloy, Ti-15%Zr-4%Nb-4%Ta alloy (Ti-15-4-4) that showed higher biological safety and mechanical properties than the currently used Ti-6%Al-4%V alloy. The purpose of this study is to determine the biological performance of the new alloy. Ti-15-4-4 implants (machined or blasted) were placed in surgically created defects in rabbit femurs. The rabbits were sacrificed after 4, 8, 16, 24 and 48 weeks. Bone mineral density (BMD) and area of newly formed bone around the implants were measured using micro-CT. Results showed that the Ti-15-4-4 alloy is biocompatible and forms new bone around the Ti-15-4-4 implant, regardless of the surface treatment. The BMD and area of newly formed bone around the blasted implant surfaces were significantly greater than those around the machined surfaces. These results indicate that the new Ti-15-4-4 alloy has a potential for use as implants and has the advantage of improved mechanical properties described in earlier studies.

To clarify new bone tissues, radiography with a newly developed tunable wavelength and highly parallel Parametric X-ray, PXR, was applied. Methods: PXR was generated by a LINAC at LEBRA, Nihon University (Hayakawa et al., 2005). X-ray wavelength was tuned from 7 KeV, 0.177 nm, to 16 KeV, 0.0775 nm. Coated or grit-blasted Ti-alloy implants modified with coating or blasting were implanted in surgically created defects in rabbit tibia. Undecalcified polished thin sections were prepared from the implant/bone areas 1 and 4 weeks after implantation. Results: PXR radiography showed a distinct difference between the newly formed bone and the compact bone. Color-mapping of the images showed an increase in the total amount of bone formation with time. Conclusion: Application of LEBRA-PXR, a high-powered, highly monochromatized and highly parallel oriented X-ray allowed easy and accurate radiographic analysis of new bone formation around the implant.

The aim of this study was to deposit an adherent apatite coating on titanium substrate using a two-step chemical deposition method. First, titanium substrates were immersed in an acidic solution containing calcium and phosphate ions, resulting in the deposition of a monetite (CaHPO4) coating. Second, the monetite crystals were converted 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. 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 was 13.1N. In conclusion, this study showed the potential of a two-step chemical deposition method for depositing an adherent coating of apatite at low temperatures.

Osteoporosis is a ‘silent’ disease characterized by thinning cortical bone and disorganized trabecular architecture causing bone fragility leading to fracture. Osteoporosis results when the rate of bone resorption far exceeds the rate of bone formation. Current pharmaceutical interventions (estrogen therapy, bisphosphonate-based drugs) focus on inhibiting bone resorption. However, some of these therapies have serious side effects (e.g., cancer risk from estrogen therapy; osteonecrosis of the jaw and delayed fracture healing from bisphosphonate-based drugs). The long term objective of the study was to develop a novel material for potential osteoporosis therapy, prevention and fracture repair. This novel material MZF-CaP or synthetic bone mineral, SBM) incorporates Mg, Zn and F ions in a calcium phosphate matrix. Separately, magnesium (Mg), zinc (Zn) and fluoride (F) ions have been associated with biomineralization and osteoporosis therapy in human and in animals. MZF-CaP or SBM was prepared by a modified hydrolysis method previously described and characterized using x-ray diffraction, FT-IR spectroscopy, inductive coupled plasma and dissolution in acidic buffer. Separately, male and female Sprague-Dawley rats were randomly assigned to the following groups depending on the diet: GA: normal on basic diets; GB: on mineral deficient diets (md); GC: on md + Mg-CaP; GD: on md + Zn-CaP; GE: md+F-CaP; and GF: md+MZF-CaP. The rats were sacrificed after 3 months and the femur bones separated, cleaned of extraneous soft tissues and stored until needed for analyses. Femur bones were analyzed using microradiography (Faxitron), scanning electron microscopy (SEM) and microCT. Results: SEM, Faxitron and microCT analyses showed thinning of cortical bone and disorganized trabecular bone architecture for osteoporotic rats on mineral deficient diet (GB) and prevention of bone loss in rats receiving the supplemented diets (GC,GD,GE,GF). Conclusion: These results indicate that the novel material, MZF-CaP or SBM had a potential for osteoporosis therapy and prevention. Studies to demonstrate the use of SBM in reversing (recovering) bone loss are in progress.

The osteoconductive property of calcium phosphate (CaP) biomaterials allow attachment, proliferation, migration, and phenotypic expression of bone cells leading to formation of new bone. The purpose of research is to develop new method of mineralizing commercial GBR membranes with calcium phosphate (CaP) and determine cell response. Resolut Adapt LT (Gore-tex) composed of co-polymer PGA/TMC and Biomend Extend (Zimmer) composed of bovine collagen were used. Membranes were mineralized with CaP using precipitation and new microwave methods. The mineralized and non-mineralized membranes were characterized using SEM, EDS, XRD, FT-IR, and TGA. Cell response to mineralized and non-mineralized membranes was determined using human osteoblastlike cells (MG-63). Microwave method was more efficient in terms of amount of minerals incorporated with membranes and time required. SEM, EDS, and FT-IR identified carbonate apatite in the mineralized membranes. No significant difference in cell proliferation was observed between mineralized and non-mineralized membranes. Greater production of type 1 collagen was observed with CaP mineralized membranes.

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.

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.

The linguloid brachiopod shell family has been the focus of several studies because of the similarity in the composition of the mineral phase of these shells to that of human bone. However, ultrastructural features of Lingula shells have not yet been fully demonstrated at high magnification using Transmission Electron Microscopy (TEM) and Electron Diffraction. Ultrastructural characterization of the mineral phase in Lingula shells will improve our understanding of the biomineralization processes and mineral/organic interaction in more complex systems such as in bone or in other human mineralized tissues. In this study, the mineral phase of Lingula adamsi was characterized using a combination of ultrastructural and crystallographic techniques. The results showed that L. adamsi shells consist of apatite crystals of varying size, shape, and orientation in different areas of the shell. The c-axis of apatite was parallel to the shell surface and crystals were organized in different laminae. Compared to trabecular bovine bone, L. adamsi shells demonstrated a higher crystallinity and a lower amount of carbonate and organic compounds. This study therefore demonstrated how dissimilar organic matrix between L. adamsi shell and trabecular bone can modify the ultrastructural characteristics of apatite crystals in these two biomineralized tissues.

This study examined the possibility of synthesis of biphasic calcium phosphate by sintering a mixture of hydroxyapatite and calcium phosphate glass. The effect of the concentration of calcium phosphate glass in a mixture on the proliferation and differentiation of MG-63 preosteoblast-like cells in a hydtoxyapatite scaffold was investigated. The addition of 5 wt% of calcium phosphate glass significantly improved the level of attachment, proliferation and differentiation of MG-63 cells onto the hydroxyapatite scaffolds, particularly when the surface was modified with 2% bovine serum albumin (p

Osteoclasts isolated from rabbits were cultured on zinc-containing tricalcium phosphate (ZnTCP) disks with zinc contents of 0.316 and 0.633 wt%, and on β-tricalcium phosphate (TCP) disks with nearly identical porosities, grain sizes and surface roughnesses. ZnTCPs directly suppressed the resorption activity of mature osteoclasts by enhancing apoptosis. We hypothesized that resorbing osteoclasts attached to ZnTCP locally accumulate zinc ions within the space defined by the clear zone during resorption, which in turn leads to apoptosis induction, even though the change in chemical composition of the culture medium is very small.

The purpose of this study was to compare the bone regenerative effect of calcium phosphate glass according to the particle size in vivo. We prepared two different sizes, that is 400 μm and 40 μm, of calcium phosphate glass powder using the system CaO-CaF2-P2O5-MgO-ZnO. Critical-sized calvarial 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 histological as well as histomorphometrical studies. The particle size of the calcium phosphate was crucial; 400 μm particles promoted new bone formation, while 40 μm particles inhibited it because of severe inflammation.