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Biomedical subjects

R Z LeGeros

Publications and source records attributed to R Z LeGeros.

At least 19 recordsLinked to original sources

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.

Apatites↗

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.

Alkaline Phosphatase↗

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.

Animals↗

Biphasic calcium phosphate bioceramics: preparation, properties and applications.

Biphasic calcium phosphate (BCP) bioceramics belong to a group of bone substitute biomaterials that consist of an intimate mixture of hydroxyapatite (HA), Ca(10)(PO(4))(6)(OH)(2), and beta-tricalcium phosphate (beta-TCP), Ca(3)(PO(4))(2), of varying HA/beta-TCP ratios. BCP is obtained when a synthetic or biologic calcium-deficient apatite is sintered at temperatures at and above 700 degrees C. Calcium deficiency depends on the method of preparation (precipitation, hydrolysis or mechanical mixture) including reaction pH and temperature. The HA/beta-TCP ratio is determined by the calcium deficiency of the unsintered apatite (the higher the deficiency, the lower the ratio) and the sintering temperature. Properties of BCP bioceramics relating to their medical applications include: macroporosity, microporosity, compressive strength, bioreactivity (associated with formation of carbonate hydroxyapatite on ceramic surfaces in vitro and in vivo), dissolution, and osteoconductivity. Due to the preferential dissolution of the beta-TCP component, the bioreactivity is inversely proportional to the HA/beta-TCP ratio. Hence, the bioreactivity of BCP bioceramics can be controlled by manipulating the composition (HA/beta-TCP ratio) and/or the crystallinity of the BCP. Currently, BCP bioceramics is recommended for use as an alternative or additive to autogeneous bone for orthopedic and dental applications. It is available in the form of particulates, blocks, customized designs for specific applications and as an injectible biomaterial in a polymer carrier. BCP ceramic can be used also as grit-blasting abrasive for grit-blasting to modify implant substrate surfaces. Exploratory studies demonstrate the potential uses of BCP ceramic as scaffold for tissue engineering, drug delivery system and carrier of growth factors.

Journal Article↗

Formation and transformation of calcium phosphates: relevance to vascular calcification.

Only one phase, carbonate apatite, is present in the mineral phase of normal calcification while different calcium phosphate phases are found in the mineral phases of pathological calcifications. In vascular calcification, the mineral phase has been described as hydroxyapatite; carbonate apatite, amorphous calcium phosphate. It has also been speculated that octacalcium phosphate may be a precursor for the apatite. This paper provides a brief review of: (1) the different calcium phosphates in normal and pathological calcifications; (2) the conditions for the formation and transformation of biologically relevant calcium phosphates in synthetic systems and a concise discussion on the relevance of results from synthetic systems to the calcium phosphate phase(s) involved in vascular calcification.

Apatites↗

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.

Adolescent↗

Biphasic calcium phosphates: influence of three synthesis parameters on the HA/beta-TCP ratio.

Hydroxyapatite (HA) contents measurements were conducted on eight biphasic calcium phosphate (BCP) samples obtained by sintering calcium-deficient apatite formed previously by hydrolyzing a dicalcium phosphate dihydrate (DCPD) powder. We evaluated the influences and interactions of three synthesis factors: alkalinity, process duration, and concentration of the water suspension in DCPD. Those parameters were varied simultaneously between two limit levels. Experiments used a factorial design method (FDM) allowing optimization of the number of samples as well as statistical analysis of results. FDM showed that HA content, in a defined experimental area, can be described by a first-order polynomial equation in which the initial alcalinity and the DCPD/water ratio are the major influences. Experiment prove that pH measured at the end of the hydrolysis was predictive of the HA content in the final BCP. This study leads up to an isoresponse line diagram which will allow the synthesis of some BCP with fitted HA/beta-tricalcium phosphate ratios.

Biocompatible Materials↗

Ultrastructural properties of bone mineral of control and tiludronate-treated osteoporotic rat.

Bisphosphonates have been widely used in the treatment of human bone pathologies including osteoporosis. In this case, bisphosphonates have been shown to reduce bone resorption, thereby increasing the mass and mechanical resistance of bone. Determining the effects of these molecules on the properties of the bone apatite crystals could provide a better insight into the mechanism of bisphosphonate/bone interaction. The aim of this study was to determine the ultrastructural effects of a third generation bisphosphonate (tiludronate) on the morphology, size, distribution, chemical composition, and structure of apatite crystals in bone (trabecular) in a rat osteoporotic model. Four groups of rats were studied: (1) sham operated, (2) untreated ovariectomized (OVX), (3) OVX rats which received 35 mg/kg of tiludronate, (4) OVX rats which received 160 mg/kg of tiludronate. The rats of groups 3 and 4 received tiludronate orally in 2 consecutive days every week for 1 year. Scanning electron microscopy (SEM), high and low resolution transmission electron microscopy (TEM), and electron microprobe analysis (EDX) were used for the ultrastructural characterization of the bone mineral. This study demonstrated that tiludronate slightly increased the width of bone apatite crystals without changing any other crystal characteristics.

Animals↗

Three-dimensional finite element analyses of four designs of a high-strength silicon nitride implant.

The effects of implant shape and size on the stress distribution around high-strength silicon nitride implants under vertical and oblique forces were determined using a three-dimensional finite element analysis. Finite element models were designed using as a basis the serial sections of the mandible. Using Auto-CAD software, the model simulated the placement of implants in the molar region of the left mandible. Results of the analyses demonstrated that mainly the implant root shape and the directions of bite forces influence the stress distributions in the supporting bone around each implant. Implant size is a lesser factor. The serrated implants presented a larger surface area to the bone than either the cylindrical or tapered implants, which resulted in lower compressive stress around the serrated implants. With increasing implant diameter and length, compressive stress decreased. The mean compressive stress distribution on the serrated implants was more flat (platykurtic) than on either the cylindrical or tapered implants. Results of studies on two load directions (vertical and oblique) showed that, in either case, the compressive stress in the cortical bone around the neck of the implant was higher than in the cancellous bone along the length of the implant. The most extreme principal compressive stress was found with oblique force. This study provides the first information on the relationship between shape of the silicon nitride implant and stress on the supporting bone.

Alveolar Process↗

Early events associated with periodontal connective tissue attachment formation on titanium and hydroxyapatite surfaces.

Endosseous dental implants can support at least three types of biomaterial/connective tissue interfaces: osseointegration, fibro-osseous integration, and periodontal connective tissue attachment. Although a periodontal connective tissue attachment offers distinct advantages, only osseointegration and fibro-osseous integration are at present clinically achievable. Recent studies indicate a periodontal regeneration-competent cell population and an appropriate biomaterial substrate both are required for periodontal connective attachment formation on biomaterial surfaces. We therefore have developed an in vitro model to characterize the effects of various biomaterial substrates on the early events of periodontal connective tissue attachment formation. Primary cultures of periodontal ligament and gingival connective tissue cells were cultured on uncoated (control) and coated (titanium- and hydroxyapatite-coated) tissue culture plastic, and the level of cell proliferation, collagen, and noncollagen protein synthesis, alkaline phosphatase activity, and expression of a 42 kD cementum extracellular matrix protein were measured over 5, 7, and 9 days in culture. While all three substrates supported cell attachment, proliferation, and protein synthesis, only uncoated and titanium-coated tissue culture plastic supported expression of the cementum extracellular matrix protein after 9 days of culture. In addition, the levels of cell proliferation and collagen and noncollagen protein synthesis for cells grown on hydroxyapatite-coated surfaces lagged behind cells cultured on the control or titanium-coated surfaces at each of the three time points. These data suggest that biomaterial substrates markedly can influence the temporal sequence of extracellular matrix proteins associated with periodontal connective tissue attachment formation. In addition to surface composition (titanium versus hydroxyapatite), surface properties (e.g., topography) also may have an effect on periodontal connective tissue attachment formation. This model may be of use in designing biomaterials to support the formation of periodontal connective tissue attachment in vivo.

Animals↗

XRD, SEM-EDS, and FTIR studies of in vitro growth of an apatite-like layer on sol-gel glasses.

A glass with a composition (in mole %) of: SiO2 (70), CaO (26), and P2O5 (4) was obtained using a sol-gel method. The in vitro bioactivity of the glass was assessed by determining the changes in surface morphology and composition after soaking in simulated body fluid (SBF) for periods of up to 14 days at 37 degrees C. X-ray diffraction, scanning electron microscopy, X-ray energy dispersive spectroscopy, and FTIR analyses of the glass surface after different soaking periods in SBF demonstrated the growth of an apatite-like layer on the glass surface. In the first stage, an amorphous calcium phosphate layer was formed; after 7 days this surface consisted of spheres, with diameters ranging between 2 and 15 microm, composed of needle-like apatite crystallites (250 x 100 nm) with a crystallinity similar to that of a biological apatite.

Apatites↗

Cellular responses to whitlockite.

Whitlockite crystals have been observed in both degenerating and normal articular cartilages. To determine their potential for inducing cartilage degeneration, we studied their ability to induce mitogenesis and synthesis and secretion of metalloproteases in vitro. Whitlockite crystals were found to stimulate cell proliferation and to stimulate synthesis and secretion of stromelysin and collagenase. However, they were less stimulatory than crystals that contained calcium (Ca) and phosphate without magnesium substitution for Ca. Whitlockite crystals elicit biologic cellular responses that suggest potential pathogenicity in arthritis, but are less potent than Ca phosphate crystals without magnesium.

Calcium Phosphates↗

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.

Calcium↗

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.

Calcium Phosphates↗

Fragmentation of the distal portion of Tomes' processes of secretory ameloblasts in the forming enamel of rat incisors.

In order to investigate enamel and dentin phospholipid metabolic pathways, two separate experiments were carried out. Firstly, rats were given chloroquine, a drug which induces a lipidosis-like disease. Extensive accumulation inside lysosomes was seen in all the groups of cells in the forming part of the rat incisor, except secretory ameloblasts which were unaffected by the drug. Secondly, the uptake and fate of 3H-choline were studied by radioautography on rats fed either normally or on an essential fatty acid deficient diet (EFAD). Four hours after the injection of the precursor, incorporation reached a maximum then decreased gradually. At 4 days the forming enamel displayed higher silver grain density than any other compartment. In EFAD rats 3H-choline incorporation was decreased drastically in each compartment except in the forming enamel which was not affected by the deficiency. The longer retention of the labeling in the forming enamel and the lack of lysosomal accumulation in chloroquine-treated secretory ameloblasts support the hypothesis that fragments of the distal Tomes' process are released during enamel formation. Disconnected from the cells, membrane remnants are neither reinternalized nor subjected to further degradation inside lysosomes.

Ameloblasts↗

pH and carbonate levels in developing enamel.

Our earlier studies showed that the surface of developing and calcifying enamel changes its pH alternatively along the tooth axis when stained with pH indicating dyes. Based on the pH conditions, the enamel at this stage was distinguished as neutral zone (N1 and N2) and acid zone (A1 and A2). The aim of the present study was to correlate changes of pH with proteolytic activity and crystal size of the calcifying bovine enamel. Specimens of developing bovine enamel were separated into four maturing stages using pH staining methods. Crystal chemistry of the developing enamel was investigated using thermogravimetry (TGA), ICP emission spectrometry, X-ray diffractometry (XRD) and infrared spectroscopy (IR). Previous biochemical analysis of proteolytic enzyme activity from enamel indicated that the optimal pH of the major protease was approximately pH 6.0, coinciding with the pH of the A1 zone. IR, TGA and XRD analyses showed that most of the organic components of the enamel decomposed at 580 degrees C. Higher levels of carbonate were observed in the secretory stages than in mature enamel. The Ca/P molar ratio of the enamel apatite was lower than the stoichiometric value of 1.67. These results suggest that growth and maturation of enamel apatite crystals is related to a decrease in the carbonate level and appear to be related to the alternative calcification and decomposition of enamel proteins.

Ameloblasts↗

Physicochemical characterization of the nucleational core of matrix vesicles.

While previous studies revealed that matrix vesicles (MV) contain a nucleational core (NC) that converts to apatite when incubated with synthetic cartilage lymph, the initial mineral phase present in MV is not well characterized. This study explored the physicochemical nature of this Ca2+ and Pi-rich NC. MV, isolated from growth plate cartilage, were analyzed directly by solid-state 31P NMR, or incubated with hydrazine or NaOCl to remove organic constituents. Other samples of MV were subjected to sequential treatments with enzymes, salt solutions, and detergents to expose the NC. We examined the NC using transmission electron microscopy, energy-dispersive analysis with x-rays, and electron and x-ray diffraction, Fourier transform-infrared spectroscopy, high performance thin-layer chromatographic analysis, and SDS-polyacrylamide gel electrophoresis. We found that most of the MV proteins and lipids could be removed without destroying the NC; however, NaOCl treatment annihilated its activity. SDS-polyacrylamide gel electrophoresis showed that annexin V, a phosphatidylserine (PS)-dependent Ca2+-binding protein, was the major protein in the NC; high performance thin-layer chromatographic analysis revealed that the detergents removed the majority of the polar lipids, but left significant free cholesterol and fatty acids, and small but critical amounts of PS. Transmission electron microscopy showed that the NC was composed of clusters of approximately 1.0 nm subunits, which energy-dispersive analysis with x-rays revealed contained Ca and Pi with a Ca/P ratio of 1.06 +/- 0. 01. Electron diffraction, x-ray diffraction, and Fourier transform-infrared analysis all indicated that the NC was noncrystalline. 1H-Cross-polarization 31P NMR indicated that the solid phase of MV was an HPO42--rich mixture of amorphous calcium phosphate and a complex of PS, Ca2+, and Pi. Taken together, our findings indicate that the NC of MV is composed of an acid-phosphate-rich amorphous calcium phosphate intermixed with PS-Ca2+-Pi, annexin V, and other proteins and lipids.

Animals↗

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.

Animals↗