Apatite crystallites: effects of carbonate on morphology.

Carbonate is a substituent in the apatite structure; when present, it limits the size of the growing apatite crystals and so influences their shape that they grow more equiaxed than needle-like. The tendency for carbonate apatites to be equiaxed is related to the nature of the chemical bonds formed in the crystal. The interference of carbonate with the good crystallization of apatite, and its weakening effect on the bonds in the structure, increase the dissolution rate and the solubility, thereby presumably contributing to the susceptibility to caries of dental apatites containing carbonate.

Adaptive crystal formation in normal and pathological calcifications in synthetic calcium phosphate and related biomaterials.

Mineralization and crystal deposition are natural phenomena widely distributed in biological systems from protozoa to mammals. In mammals, normal and pathological calcifications are observed in bones, teeth, and soft tissues or cartilage. We review studies on the adaptive apatite crystal formation in enamel compared with those in other calcified tissues (e.g., dentin, bone, and fish enameloids) and in pathological calcifications, demonstrating the adaptation of these crystals (in terms of crystallinity and orientation) to specific tissues that vary in functions or vary in normal or diseased conditions. The roles of minor elements, such as carbonate, magnesium, fluoride, hydrogen phosphate, pyrophosphate, and strontium ions, on the formation and transformation of biologically relevant calcium phosphates are summarized. Another adaptative process of crystals in biology concerns the recent development of calcium phosphate ceramics and other related biomaterials for bone graft. Bone graft materials are available as alternatives to autogeneous bone for repair, substitution, or augmentation. This paper discusses the adaptive crystal formation in mineralized tissues induced by calcium phosphate and related bone graft biomaterials during bone repair.

Adherent apatite coating on titanium substrate using chemical deposition.

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.

Strategies to affect bone remodeling: osteointegration.

Osteointegration was defined as a "direct structural and functional connection between ordered living bone and the surface of a load-carrying implant." Although osteointegration was meant originally to describe a biologic fixation of the titanium dental implants, it is now used to describe the attachment of other materials used for dental and orthopedic applications as well. Analyses of material-bone interface showed that osteointegrated implants can have an intervening fibrous layer or direct bone apposition characterized by bone-bonding depending on the composition and surface properties of the biomaterial. This article reviews biologic (host tissue properties and response), biomechanical, and biomaterial factors affecting osteointegration. Biologic factors include the quality of bone. Biomaterial factors include the effect of material composition on the bone-material interface. Suggested areas for future research include determining the correlation between oral bone status and osteoporosis, the effect of gender, age, and endocrine status (e.g., osteoporosis) on implant success or failure, the effect of calcium phosphate coating composition and crystallinity on in vivo performance of implants, the factors contributing to accelerated osteointegration, and development of osteoinductive implants.

Chemical and crystallographic events in the caries process.

The chemical and crystallographic events associated with the caries process can be described based on the results from the following studies: (a) effects of carbonate, magnesium, fluoride, and strontium on the physico-chemical properties--lattice parameters, crystallinity (crystal size and strain); dissolution properties of synthetic apatites; (b) factors influencing the in vitro formation and transformation of DCPD, OCP, AP (Ca-deficient apatites), FAP, beta-TCMP (Mg-substituted), and CaF2; and (c) studies on properties (crystallinity, composition, chemical, and thermal stabilities) of enamel, dentin, and bone. The dissolution of CO3-rich/Mg-rich/F-poor dental apatite crystals and re-precipitation of CO3-poor/Mg-poor/F-rich apatite in the presence of F- ions in solution contribute to a more acid-resistant surface layer of the caries lesion. Fluoride promotes the formation of less Ca-deficient and more stable apatite crystals. The presence of Ca, P, and F in solution inhibits dissolution of apatite more than does the presence of F alone. Low levels of F in solution promote the formation of (F, OH)-apatite, even under very acid conditions; an increase in F levels causes the formation of CaF2 at the expense of DCPD or apatite, especially in acid conditions. F in apatite and/or in solution suppresses extensive dissolution of dental apatite and enhances the formation of (F, OH)-apatite crystals which are more resistant against acid-dissolution than are F-free apatite crystals.

The crystalline components of dental calculi: human vs. dog.

Dental calculus from the dog was found to consist principally of the calcite form of calcium carbonate mixed with small amounts of apatite; other calcium phosphates, consistently present in human calculus, were not present in dog calculus. Precipitable calcium salts from human saliva were mainly apatite; for the dog the principal precipitated salt was calcium carbonate (calcite form).

Surface spectroscopy of apatitic materials: limitations and concerns.

The use of x-ray photoelectron spectroscopy for the study of biological materials has provided mixed results. The experimental difficulties associated with the application of this technique to the study of biological materials include the effects of ultra-high vacuum, x-ray exposure, surface charging, and, for insulating materials, the inherent lack of an intrinsic energy reference. Since most biological materials are insulators, the lack of an intrinsic energy reference complicates one's ability to draw direct comparisons between different studies. In this paper, the results from two different apatites are compared with the results obtained from the use of two conventional techniques for referencing binding energies. The results presented here are referenced only to the Pt Fermi level; no further energy modifications are performed. A comparison with the results of other investigators indicates a large difference in binding energies for all involved core levels. These results indicate that, in the study of insulating materials with x-ray photoelectron spectroscopy, it is essential that all data be acquired under exacting conditions on the same experimental system.

Possible physico-chemical processes in human dentin caries.

This study correlated ultrastructural observations on the presence of beta-tricalcium phosphate (beta-TCP) in arrested dentin caries with physico-chemical observations on the in vitro formation of Mg-substituted beta-TCP. The ultrastructural studies were made using high-resolution transmission electron microscopy (TEM) with the capability of microdiffraction and microanalysis on sites less than 10 nm in diameter. Mg-substituted beta-TCP was obtained, by a precipitation method, from solutions with Mg/Ca molar ratios of 5/95 and higher. Such correlations led to the postulation of a possible chronological sequence of physico-chemical events occurring at the crystal level during the progress and arrest of caries in human dentin. It is suggested that the initial mechanism for the observed occurrence of large crystals of Mg-substituted beta-TCP and of apatite in the tubule lumen is due to the dissolution of the dentin mineral (a CO3- and Mg-rich calcium OH-apatite) and reprecipitation of Mg-substituted beta-TCP, followed by that of CO3- and Mg-poor apatite.

Ultrastructural properties of laser-irradiated and heat-treated dentin.

Previous studies using scanning electron microscopy and infrared absorption spectroscopy reported that laser irradiation causes compositional changes in enamel. The purpose of this study was to evaluate the ultrastructural and compositional changes in dentin caused by irradiation with a short-pulse laser (Q-switched Nd:YAG). The irradiated and non-irradiated areas of the lased dentin samples were investigated by scanning (SEM) and transmission electron microscopy (TEM), micro-micro electron diffraction, and electron microprobe analysis of dispersive energy (EDX). Heat-treated dentin was similarly investigated. This study demonstrated that laser irradiation resulted in the recrystallization of dentin apatite and in the formation of additional calcium phosphate phases consisting of magnesium-substituted beta-tricalcium phosphate, beta-TCMP, beta-(Ca,Mg)3(PO4)2, and tetracalcium phosphate, TetCP, Ca4(PO4)O. TEM analyses of the modified and unmodified zones of the irradiated areas showed two types of crystal populations: much larger crystals from the modified zone and crystals with size and morphology similar to those of dentin apatite in the unmodified zone. The morphology of crystals in the modified zones in the irradiated dentin resembled those of dentin sintered at 800 or 950 degrees C. In the irradiated areas (modified and unmodified zones), the Ca/P ratio was lower compared with that in the non-irradiated dentin. The Mg/Ca ratio in the modified zones was higher than that in the unmodified zones and in the non-irradiated dentin. In sintered dentin, the Mg/Ca ratio increased as a function of sintering temperature. The ultrastructural and compositional changes observed in laser-irradiated dentin may be attributed to high temperature and high pressure induced by microplasma during laser irradiation. These changes may alter the solubility of the irradiated dentin, making it less susceptible to acid dissolution or to the caries process.

In vitro caries-like lesion formation in F-containing tooth enamel.

In vitro caries-like lesion formation in F-containing tooth enamel (shark) showed a lower depth of penetration compared to that in human enamel (42 vs. 100%). After longer periods of exposure, the depth of penetration in shark enamel is only 33% relative to that in human enamel. The extent of dissolution in acetate buffer for powdered human enamel and for synthetic F-free calcium-carbonated-apatites was greater than for powdered shark enamel and for synthetic F-containing apatites. These results suggest that the cariostatic action of fluoride may be explained in terms of a combination of two effects: (a) the stabilizing effect against acid dissolution (caries) when fluoride is incorporated in the tooth mineral (apatite), and (b) a greater degree of remineralization (repair) in the presence of F solution.

Some physico-chemical properties of deciduous enamel of children with and without pre-natal fluoride supplementation (PNF).

Physico-chemical investigations of enamel from deciduous teeth of a small number of children with and without pre-natal fluoride supplementation (PNF) exhibited the following differences between the PNF and non-PNF enamel, with the former showing: (a) more homogeneous and less extensive patterns of acid-etching; (b) denser crystal populations in intra-prismatic regions; (c) larger prism dimensions; (d) greater total mineral density (wt%ash); (e) higher degree of crystallinity; (f) smaller a-axis dimensions; and (g) more fluoride and less carbonate contents. On the basis of these preliminary findings, further studies with larger numbers of samples would seem appropriate.

Preparation of different forms of titanium oxide on titanium surface: effects on apatite deposition.

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.

Effect of enamel matrix proteins on the phenotype expression of periodontal ligament cells cultured on dental materials.

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.

Effect of enamel matrix proteins on the periodontal connective tissue-material interface after wound healing.

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.

A Raman and infrared spectroscopic investigation of biological hydroxyapatite.

Raman spectra were acquired on ox femur samples treated with hydrazine to remove the organic components of bone. A large increase in the signal-noise ratio of the mineral spectrum resulted from the exposure of the mineral surface and the removal of fluorescent components of the organic matrix. The effect of hydrazine treatment of the mineral matrix has been reinvestigated and shown to be slight on the basis of second derivative FTIR data. This is the first time that this high resolution technique has been applied to biological minerals.

Tissue response to biphasic calcium phosphate ceramic with different ratios of HA/beta TCP in periodontal osseous defects.

The purpose of this study was to determine the optimal ratio of calcium hydroxyapatite (HA) to beta tricalcium phosphate (beta TCP) in a biphasic porous calcium phosphate (BCP) ceramic for effective repair of periodontal osseous defects. Defects were surgically produced in beagle dogs and made chronic for 4 months to simulate periodontal disease. Mucoperiosteal periodontal flaps were reflected, followed by osseous defect debridement and root planing. Specially prepared ceramic with different HA/beta TCP ratios were implanted into the prepared defects. The sites were allowed to heal for 6 months, animals were euthanized, and site-blocks were removed for histological study. During the follow-up phase, scaling and polishing were done once a month, and standardized probing attachment levels were recorded pre- and 6-months postoperatively. The Duncan's multiple range test showed that all the treatments produced statistically significant higher gain in probing attachment levels than the control group (0HA/0 beta TCP) (P < 0.05). Among the 7 "active" treatment groups, 2 (65/35 and 85/15) had significantly higher gain in probing attachment levels than those in 3 groups (50/50, 100/0, and 0/100) (P < 0.05). Histologically, higher HA ratio (but not 100% HA) showed accelerated new bone formation and new attachment levels. Based on histological results, the 85HA/15 beta TCP ratio appears to demonstrate greater gain in attachment level and bone regeneration in the treatment of periodontal osseous defects.

Dental caries experience, tooth loss, and factors associated with unmet needs of Haitian immigrants in New York City.

OBJECTIVE: The aim of this study was to characterize the dental caries experience, tooth loss, and unmet need of a group of Haitian immigrant residents of New York City. METHODS: A purposive sample of 523 adults was obtained through community outreach activities during 1997-98. Clinical examinations were performed by calibrated examiners, according to NIDCR criteria. A comprehensive survey also was administered to all the participants. RESULTS: For the whole group, the mean number of missing teeth was 2.64 (SD = 4.12), the mean DMFT = 6.05 (SD = 5.26), the mean DMFS = 18.80 (SD = 21.04), and the mean DFS = 5.58 (SD = 6.17). Seventeen percent of the subjects had all their teeth sound, 59 percent had at least one tooth missing, 60 percent had at least one decayed tooth, and only 38 percent had restorations. Multivariate analyses showed that age, sex, education, dental insurance, frequency of dental visits, and dental floss use were predictors of unmet need. CONCLUSIONS: Although results showed a relatively low caries experience among this group of Haitian immigrants, the unmet need was very high. Furthermore, the tooth loss experience was relatively high for all age groups, further denoting a lack of access to preventive and restorative services.

Zinc effect on the in vitro formation of calcium phosphates: relevance to clinical inhibition of calculus formation.

PURPOSE: To determine the effect of zinc on the in vitro formation of calcium phosphates and its relevance to calculus inhibition. MATERIALS AND METHODS: Different types of calcium phosphate phases (amorphous calcium phosphate, ACP; dicalcium phosphate dihydrate, DCPD; octacalcium phosphate, OCP; and carbonate hydoxyapatite, CHA) were precipitated from solutions containing increasing concentrations of zinc (Zn) ions. The precipitates were characterized using x-ray diffraction, infrared absorption spectroscopy, and scanning electron microscopy. RESULTS: The presence of Zn ions affected the type and amount of calcium phosphate phases formed. Zn, even at concentrations as low as 0.1 mM/L, inhibited the crystal growth of DCPD, OCP and AP; and, at higher concentrations (0.5 mM to 2 mM/L), promoted the formation of amorphous calcium phosphate, ACP, or Zn-substituted tricalcium phosphate (beta-TCP) depending on the reaction pH and temperature.

Crystallographic and chemical properties of Mg-containing apatites before and after suspension in solutions.

Mg-containing synthetic apatites were prepared and then incubated in unbuffered 0.9% NaCl solution and in buffered 0.1 M CH3COOK solution (pH 7.4) at 37 degrees C to examine the changes in their crystallographic and chemical properties after suspension in these solutions. After one month of incubation, the magnesium (Mg) content in the samples decreased greatly, especially in samples with higher Mg content. The a and c axis dimensions of the samples changed with the decrease of Mg content. On the other hand, Mg concentrations in both solutions increased more than the Ca concentrations. Mg-substituted beta-tricalcium phosphate (beta-TCP), or whitlockite, formed from solutions with high Mg concentration. These results show that Mg-containing apatites dissolve to a greater extent than Mg-free apatites with the subsequent reprecipitation of Mg-poor apatites. In addition, the high Mg/Ca in solution favoured the reprecipitation of Mg-containing beta-TCP.

Intra-oral comparison and evaluation of the ability of fluoride dentifrices to promote the remineralization of caries-like lesions in dentin and enamel.

The effect of three types of fluoride dentifrices and one non-fluoride dentifrice were evaluated and compared intra-orally for their ability to promote the remineralization of caries-like lesions in thin sections of human dentin and enamel. The three fluoride dentifrices were silica based. One dentifrice contained 0.76% sodium monofluorophosphate (MFP), another contained 0.247% sodium fluoride (NaF), and the third contained 0.247% NaF plus 1.3% pyrophosphate and 1.5% polyvinylmethyl ether/maleic acid (PVM/MA) copolymer (denoted NaF/PPi). The fourth dentifrice (placebo) was also silica based, but contained no fluoride, and served as a negative control. Two week treatment resulted in the following percent mineral changes for the enamel specimens: +21 +/- 16 for MFP, +10 +/- 30 for NaF, +16 +/- 15 for NaF/PPi, and -30 +/- 30 for the placebo. The respective percent mineral change values for the dentin specimens were +28 +/- 20 for MFP, +22 +/- 19 for NaF, +28 +/- 18 for NaF/PPi, and -34 +O/- 61 for the placebo. A statistical analysis revealed that the three fluoride dentifrices were significantly better (p<0.05) than the placebo at promoting remineralization for both types of specimens. No significant differences were found among the three fluoride dentifrices. The following can be concluded from this study: 1) silica based dentifrices containing MFP and NaF are effective at remineralizing caries-like lesions in both enamel and dentin; 2) there was no statistical difference between NaF and MFP in their ability to promote remineralization; and 3) pyrophosphate does not interfere with the remineralizing effects of NaF.