High-Performance Liquid Chromatography Method for Rich Pharmacokinetic Sampling Schemes in Translational Rat Toxicity Models With Vancomycin.

A translational need exists to understand and predict vancomycin-induced kidney toxicity. We describe: (i) a vancomycin high-performance liquid chromatography (HPLC) method for rat plasma and kidney tissue homogenate; (ii) a rat pharmacokinetic (PK) study to demonstrate utility; and (iii) a catheter retention study to enable future preclinical studies. Rat plasma and pup kidney tissue homogenate were analyzed via HPLC for vancomycin concentrations ranging from 3-75 and 15.1-75.5 µg/mL, respectively, using a Kinetex Biphenyl column and gradient elution of water with 0.1% formic acid: acetonitrile (70:30 v/v). Sprague-Dawley rats (n = 10) receiving 150 mg/kg of vancomycin intraperitoneally had plasma sampled for PK. Finally, a catheter retention study was performed on polyurethane catheters to assess adsorption. Precision was <6.1% for all intra-assay and interassay HPLC measurements, with >96.3% analyte recovery. A two-compartment model fit the data well, facilitating PK exposure estimates. Finally, vancomycin was heterogeneously retained by polyurethane catheters.

Selected physicochemical properties of the experimental endodontic sealer.

This study explores water sorption, hygroscopic expansion, mechanical strength and ion release from the experimental amorphous calcium phosphate (ACP) composites formulated for application as endodontic sealers. Light-cure (LC) and dual-cure (DC; combined light and chemical cure) resins comprised urethane dimethacrylate (UDMA), 2-hydroxyethyl methacrylate (HEMA), methacryloyloxyethyl phthalate (MEP) and a high molecular mass oligomeric co-monomer, poly(ethyleneglycol)-extended UDMA (PEG-U) (designated UPHM resin). To fabricate composites, a mass fraction of 60% UPHM resin was blended with a mass fraction of 40% as-made (am-) or ground (g-) ACP. Glass-filled composites were used as controls. Both DC and LC ACP UPHM composites exhibited relatively high levels of water sorption accompanied by a significant hygroscopic expansion. The latter may potentially be useful to offset high polymerization stresses that develop in these materials. Ion release profiles of the experimental materials confirmed their potential for regeneration of mineral-deficient tooth structures. Their moderate to low mechanical strength after 3 months of aqueous immersion did not diminish the enthusiasm for the proposed use as endodontic sealers. For that application, DC g-ACP composites appear to be the most adequate, but micro-leakage and quantitative leachability studies are needed to fully establish their suitability.

AMORPHOUS CALCIUM PHOSPHATE COMPOSITES AND THEIR EFFECT ON COMPOSITE-ADHESIVE-DENTIN BONDING.

This study evaluates the bond strength and related properties of photo-polymerizable, remineralizing amorphous calcium phosphate (ACP) polymeric composite-adhesive systems to dentin after various periods of aqueous aging at 37 °C. An experimental ACP base and lining composite was made from a photo-activated resin comprising 2,2-bis[p-(2'-hydroxy-3'-methacryloxypropoxy)phenyl]propane (Bis-GMA), triethylene glycol dimethacrylate (TEGDMA), 2-hydroxyethyl methacrylate (HEMA) and zirconyl dimethacrylate (ZrDMA); designated BTHZ. An experimental orthodontic composite was formulated from a photo-activated resin comprising ethoxylated bisphenol A dimethacrylate (EBPADMA), TEGDMA, HEMA and methacryloxyethyl phthalate (MEP); designated ETHM. In both composite series three fillers were compared: 1) freshly precipitated zirconium-modified ACP freshly precipitated (as-prepared Zr-ACP), 2) milled Zr-ACP and 3) an ion-leachable fluoride glass. In addition to the shear bond strength (SBS), work to fracture and failure modes of the orthodontic composites were determined. The SBS of the base and lining ACP composites appeared unaffected by filler type or immersion time. In the orthodontic ACP composite series, milled ACP composites showed initial mechanical advantages over as-prepared ACP composites, and produced higher incidence of a failure mode consistent with stronger adhesion. After six months of aqueous exposure, 80 % of specimens failed at the dentin-primer interface, with a 42 % overall reduction in bond strength. BTHZ and ETHM based ACP composites are potentially effective anti-demineralizing-remineralizing agents with possible clinical utility as protective base-liners and orthodontic cements, respectively. The analysis of the bond strength and failure modalities suggests that milled ACP composites may offer greater potential in clinical applications.

In vitro remineralization of enamel by polymeric amorphous calcium phosphate composite: quantitative microradiographic study.

OBJECTIVE: This study explores the efficacy of an experimental orthodontic amorphous calcium phosphate (ACP) composite to remineralize in vitro subsurface enamel lesions microradiographically similar to those seen in early caries. METHODS: Lesions were artificially created in extracted human molars. Single tooth sections a minimum of 120microm thick were cut and individually placed in holders exposing only the carious enamel surface. The exposed surfaces were either left untreated (control) or coated with a 1mm thick layer of the experimental ACP composite (mass fraction 40% zirconia-hybridized ACP and 60% photo-activated resin), or a commercial fluoride-releasing orthodontic cement. The composite-coated sections were then photo-cured and microradiographic images were taken of all three groups of specimens before the treatment. Specimens were then cyclically immersed in demineralizing and remineralizing solutions for 1 month at 37 degrees C to simulate the pH changes occurring in the oral environment. Microradiographs of all specimens were taken before and after treatment. RESULTS: Quantitative digital image analysis of matched areas from the contact microradiographs taken before and after treatment indicated higher mineral recovery with ACP composites compared to the commercial orthodontic F-releasing cement (14.4% vs. 4.3%, respectively), while the control specimens showed an average of 55.4% further demineralization. SIGNIFICANCE: Experimental ACP composite efficiently established mineral ion transfer throughout the body of the lesions and restored the mineral lost due to acid attack. It can be considered a useful adjuvant for the control of caries in orthodontic applications.

Degree of vinyl conversion, polymerization shrinkage and stress development in experimental endodontic composite.

This study explores degree of vinyl conversion (DVC), polymerization shrinkage (PS) and shrinkage stress (PSS) of the experimental amorphous calcium phosphate (ACP) composites intended for use as an endodontic sealer. Light-cure (LC), chemical cure (CC) or dual-cure (DC; combined light and chemical cure) resins comprised urethane dimethacrylate (UDMA), 2-hydroxyethyl methacrylate (HEMA), methacryloyloxyethyl phthalate (MEP) and a high molecular mass oligomeric co-monomer, poly(ethyleneglycol)-extended UDMA (PEG-U) (designated UPHM resin). To fabricate composites, a mass fraction of 60 % UPHM resin was blended with a mass fraction of 40 % as-made (am-ACP) or ground ACP (g-ACP). DVC values of copolymer (unfilled UPHM resin) and composite specimens were determined by infrared spectroscopy. Glass-filled composites were used as controls. PS and PSS of composites were determined by dilatometry and tensometry, respectively. LC copolymers attained extraordinary high DVC values at 24 h post-cure (95.7 %), compared to CC (52 %) and DC (79.3 %) copolymer specimens. While the DVC values of LC and DC am-ACP composites were reduced between 5 and 10 %, DVC values of DC g-ACP composites increased almost 8 % compared to the corresponding copolymers. High DVC attained in LC composites was, expectedly, accompanied with high PS values (on average 7 vol%). However, PSS developed in LC and especially DC composites did not exceed PSS values seen in other UDMA-based composites. Based on this initial evaluation, it is concluded that, DC, g-ACP filled UPHM composite shows promise as an endodontic sealer. However, further physicochemical evaluations, including water sorption, mechanical stability and ion release as well as a leachability studies need to be performed before this experimental material is tested for cellular responses and, eventually recommended for clinical utility.

Light-cured dimethacrylate-based resins and their composites: comparative study of mechanical strength, water sorption and ion release.

This study explored how resin type affects selected physicochemical properties of complex methacrylate copolymers and their amorphous calcium phosphate (ACP)-filled and glass-filled composites. Two series of photo-polymerizable resin matrices were formulated employing 2,2-bis[p-(2'-hydroxy-3'-methacryloxypropoxy)phenyl]propane (Bis-GMA) or an ethoxylated bisphenol A dimethacrylate (EBPADMA) as the base monomer, Unfilled copolymers and composites filled with a mass fraction with 40 %, 35 % and 30 %, respectively, of ACP or the un-silanized glass were assessed for biaxial flexure strength (BFS), water sorption (WS) and mineral ion release upon immersion in HEPES-buffered saline solution for up to six months. Substituting EBPADMA for Bis-GMA significantly reduced the WS while only marginally affected the BFS of both dry and wet copolymers. Independent of the filler level, both dry and wet ACP composites formulated with either BTHM or ETHM resins were mechanically weaker than the corresponding copolymers. The BFS of ACP composite specimens after 1 month in saline did not further decrease with further aqueous exposure. The BFS of glass-filled composites decreased with the increased level of the glass filler and the time of aqueous exposure. After 6 months of immersion, the BFS of glass-filled BTHM and ETHM composites, respectively, remained 58 % and 41 % higher than the BFS of the corresponding ACP composites. Ion release data indicated that a minimum mass fraction of 35 % ACP was required to attain the desired solution supersaturation with respect to hydroxyapatite for both the BTHM and ETHM derived composites.

ILLUMINATING THE ROLE OF AGGLOMERATES ON CRITICAL PHYSICOCHEMICAL PROPERTIES OF AMORPHOUS CALCIUM PHOSPHATE COMPOSITES.

Water sorption (WS), mechanical strength, and ion release of polymeric composites formulated with 40 % as-made or milled amorphous calcium phosphate (ACP) are compared after 1, 2 and 3 months of aqueous exposure. Ethoxylated bisphenol A dimethacrylate, triethylene glycol dimethacrylate, 2-hydroxyethyl methacrylate and methacryloxyethyl phthalate comprised the resin. The WS (mass %) peaked at 3 months. WS of as-made ACP composites was significantly higher than WS of milled ACP composites and copolymers. Both composite groups experienced decreases in biaxial flexural strength (BFS) with water aging, with milled ACP composites retaining a significantly higher BFS throughout immersion. Ion release was moderately reduced in milled ACP composites, though they remained superior to as-made ACP composites due to significantly lower WS and higher BFS after prolonged aqueous exposure.

Amorphous calcium phosphate composites with improved mechanical properties.

Hybridized zirconium amorphous calcium phosphate (ACP)-filled methacrylate composites make good calcium and phosphate releasing materials for anti-demineralizing/remineralizing applications with low mechanical demands. The objective of this study was to assess the effect of the particle size of the filler on the mechanical properties of these composites. Photo-curable resins were formulated from ethoxylated bisphenol A dimethacrylate, triethylene glycol dimethacrylate, 2-hydroxyethyl methacrylate and methacryloxyethyl phthalate. Camphorquinone and ethyl-4-N,N-dimethylaminobenzoate were utilized as components of the photoinitiator system. After 2 h of mechanical milling in isopropanol, an approximate 64 % reduction in the median particle diameter was observed [27.48 µm vs. 9.98 µm] for unmilled and milled wet ACP, respectively. Dry ACP showed a 43 % reduction in particle size from pre- to post-milling. As well as dry composites, those that had been immersed in aqueous media were evaluated for their Young's Modulus, water sorption, biaxial tensile, three-point flexural and diametral tensile strength. Mechanically milling the filler increased the volume of fine particles in the composite specimens, resulting in a more homogeneous intra-composite distribution of ACP and a reduction in voids. In turn, less water diffused into the milled composites upon aqueous exposure, and they showed a marked improvement in biaxial flexure strength and a moderate improvement in flexural strength over composites with unmilled ACP. The demonstrated improvement in the mechanical stability of milled Zr-ACP composites may help in extending their dental applicability.