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In Vitro and In Vivo Biocompatibility Of ReOss® in Powder and Putty Configurations

Pintor, Andréa Vaz Braga; Resende, Rodrigo Figueiredo de Brito; Neves, Adriana Terezinha Novelino; Alves, Gutemberg Gomes; Coelho, Paulo G; Granjeiro, José Mauro; Calasans-Maia, Monica Diuana
This study evaluated comparatively two configurations (powder and putty) of a composite biomaterial based on PLGA (Poly(lactide-co-glycolide)/nanoescale hydroxyapatite (ReOss®, Intra-Lock International) through microscopic morphology, in vitro cytotoxicity, biocompatibility and in vivo response as a bone substitute. SEM and EDS characterized the biomaterials before/after grafting. Cytocompatibility was assessed with murine pre-osteoblasts. Osteoconductivity and biocompatibility were evaluated in White New Zealand rabbits. Both configurations were implanted in the calvaria of eighteen animals in non-critical size defects, with blood clot as the control group. After 30, 60 and 90 days, the animals were euthanized and the fragments containing the biomaterials and controls were harvested. Bone blocks were embedded in paraffin (n=15) aiming at histological and histomorphometric analysis, and in resin (n=3) aiming at SEM and EDS. Before implantation, the putty configuration showed both a porous and a fibrous morphological phase. Powder revealed porous particles with variable granulometry. EDS showed calcium, carbon, and oxygen in putty configuration, while powder also showed phosphorus. After implantation EDS revealed calcium, carbon, and oxygen in both configurations. The materials were considered cytotoxic by the XTT test. Histological analysis showed new bone formation and no inflammatory reaction at implant sites. However, the histomorphometric analysis indicated that the amount of newly formed bone was not statistically different between experimental groups. Although both materials presented in vitro cytotoxicity, they were biocompatible and osteoconductive. The configuration of ReOss® affected morphological characteristics and the in vitro cytocompatibility but did not impact on the in vivo biological response, as measured by the present model.
PMID: 29898056
ISSN: 1806-4760
CID: 3154792

Effect of Sonic Resin Composite Delivery on Void Formation Assessed by Micro-computed Tomography

Hirata, R; Pacheco, R R; Caceres, E; Janal, M N; Romero, M F; Giannini, M; Coelho, P G; Rueggeberg, F A
OBJECTIVES/OBJECTIVE:The aim of this study was to quantify the internal void volume formation in commercially available, resin composites inserted using conventional or sonic insertion methods, and analyzed using three-dimensional (3D) micro-computed tomography (μCT). METHODS AND MATERIALS/METHODS:Four resin composites were evaluated: one conventional (Herculite, Ultra, Kerr Corporation, Orange, CA, USA), one flowable bulk fill (SureFil SDR Flow, Dentsply International, York, PA, USA), and two packable bulk fill (SonicFill, Kerr Corporation, and Tetric EvoCeram Bulk Fill, Ivoclar Vivadent Inc, Schaan, Liechtenstein). Eight groups were evaluated according to each resin composite type and insertion method (conventional or sonic; n=5). Forty ABS 3D-printed cylindrical molds, 5.0 mm in diameter and 4.0 mm in depth, were fabricated. For the conventional resin composite, the mold was filled incrementally (two layers), while for bulk-fill resin composites, insertion was performed in a single increment. The sonic insertion method was performed using a specific handpiece (SonicFill Handpiece, Kerr Corporation). Resin composites were light cured using a multipeak light-emitting diode light-curing unit (VALO, Ultradent Products Inc, South Jordan, UT, USA) in its regular mode. Samples were evaluated by μCT, and data were imported into software (Amira, version 5.5.2, VSG, Burlington, MA, USA) for 3D reconstruction, from which the percentage of void volume was calculated. Data were analyzed using two-way analysis of variance and Tukey post hoc test at a preset alpha of 0.05. RESULTS:The conventional insertion method resulted in reduced porosity, compared with sonic insertion, for SureFil SDR Flow and Tetric EvoCeram bulk fill. The sonic insertion method did not demonstrate any influence on void formation for Herculite Ultra or SonicFill. CONCLUSION/CONCLUSIONS:Results suggest that the sonic insertion method might increase void formation during resin composite delivery, depending on restorative material brand.
PMID: 29394140
ISSN: 1559-2863
CID: 3039692

Histo-morphologic characteristics of intra-osseous implants of WE43 Mg alloys with and without heat treatment in an in vivo cranial bone sheep model

Torroni, Andrea; Xiang, Chongchen; Witek, Lukasz; Rodriguez, Eduardo D; Flores, Roberto L; Gupta, Nikhil; Coelho, Paulo G
WE43 Mg alloy, composed of Mg, Yttrium, Rare Earth elements, and Zirconium, has proved to be a suitable candidate for production of resorbable osteosynthesis implants in both clinical and experimental settings. In a previous study we tested biocompatibility and degradation properties of untreated (as-cast) and artificially aged (T-5) WE43 Mg-alloys as subperiosteal implants on a maxillofacial sheep model. Both the alloy compositions showed excellent biocompatibility, however, with respect to degradation rate, the as-cast form showed increased degradability compared with the T-5. In the present study, we tested the same alloy composition (i.e. as-cast and T-5) to assess their biological behavior and degradation pattern when implanted as endosteal implants on a calvarial bone sheep model. Six implants in form of cylindrical discs were tested in 6 sheep, one per composition of each disc was placed in two monocortical cranial defect created with high speed trephine bur in the parietal bone. After euthanasia at 6 weeks histomorphological analysis of the bone/implant specimens was performed. WE43-as cast showed higher degradation rate, increased bone remodeling, gas pockets formation and osteolysis compared with the T5 alloy. WE43-T5 showed greater bone/implant interface stability, and seemed to be more suitable for fabrication of endosteal bone screws.
PMID: 29398208
ISSN: 1878-4119
CID: 2947982

Metabolic imaging of fatty kidney in diabesity: validation and dietary intervention

Jonker, Jacqueline T; de Heer, Paul; Engelse, Marten A; van Rossenberg, Evelien H; Klessens, Celine Q F; Baelde, Hans J; Bajema, Ingeborg M; Koopmans, Sietse Jan; Coelho, Paulo G; Streefland, Trea C M; Webb, Andrew G; Dekkers, Ilona A; Rabelink, Ton J; Rensen, Patrick C N; Lamb, Hildo J; de Vries, Aiko P J
Background: Obesity and type 2 diabetes have not only been linked to fatty liver, but also to fatty kidney and chronic kidney disease. Since non-invasive tools are lacking to study fatty kidney in clinical studies, we explored agreement between proton magnetic resonance spectroscopy ( 1 H-MRS) and enzymatic assessment of renal triglyceride content (without and with dietary intervention). We further studied the correlation between fatty kidney and fatty liver. Methods: Triglyceride content in the renal cortex was measured by 1 H-MRS on a 7-Tesla scanner in 27 pigs, among which 15 minipigs had been randomized to a 7-month control diet, cafeteria diet (CAF) or CAF with low-dose streptozocin (CAF-S) to induce insulin-independent diabetes. Renal biopsies were taken from corresponding MRS-voxel locations. Additionally, liver biopsies were taken and triglyceride content in all biopsies was measured by enzymatic assay. Results: Renal triglyceride content measured by 1 H-MRS and enzymatic assay correlated positively ( r = 0.86, P < 0.0001). Compared with control diet-fed minipigs, renal triglyceride content was higher in CAF-S-fed minipigs (137 +/- 51 nmol/mg protein, mean +/- standard error of the mean, P < 0.05), but not in CAF-fed minipigs (60 +/- 10 nmol/mg protein) compared with controls (40 +/- 6 nmol/mg protein). Triglyceride contents in liver and kidney biopsies were strongly correlated ( r = 0.97, P < 0.001). Conclusions: Non-invasive measurement of renal triglyceride content by 1 H-MRS closely predicts triglyceride content as measured enzymatically in biopsies, and fatty kidney appears to develop parallel to fatty liver. 1 H-MRS may be a valuable tool to explore the role of fatty kidney in obesity and type 2 diabetic nephropathy in humans in vivo .
PMID: 28992141
ISSN: 1460-2385
CID: 2732032

Residual stress of porcelain-fused to zirconia 3-unit fixed dental prostheses measured by nanoindentation

Fardin, Vinicius P; Bonfante, Estevam A; Coelho, Paulo G; Janal, Malvin N; Tovar, Nick; Witek, Lukasz; Bordin, Dimorvan; Bonfante, Gerson
OBJECTIVE:To evaluate the residual stress (nanoindentation based on hardness) of fatigued porcelain-fused to zirconia 3-unit fixed dental prostheses (FDP) with different framework designs. METHODS:Twenty maxillary 3-unit FDP replacing second-premolar (pontic) were fabricated with conventional framework-design (even-thickness of 0.5mm and 9mm2 connector area) and modified framework-design (thickness of 0.5mm presenting lingual collar connected to proximal struts and 12mm2 connector area). Connector marginal ridges were loaded and the fractured and suspended FDPs were divided (n=3/each) into: (1) Fractured zirconia even-thickness (ZrEvenF); (2) Suspended zirconia even-thickness (ZrEvenS); (3) Fractured zirconia with modified framework (ZrModF); (4) Suspended zirconia with modified framework (ZrModS); (5) Non-fatigued FDP with conventional framework design (Control). The FDPs were nanoindented at 0.03mm (Region of Interest (ROI) 1), 0.35mm (ROI 2) and 1.05mm (ROI 3) distances from porcelain veneer outer surface with peak load 4000μN. The Linear Mixed Analysis of Variance (ANOVA) Model on ranks and Least Significant Difference Test on ranks (95%) were used. RESULTS:Highest rank hardness values were found for Control group and ZrModS, whereas the lowest values were found in ZrModF. Statistical differences (p=0.000) were found among all groups except for comparison between ZrModS and Control group (p=0.371). Hardness between ROIs were statistically significant different (p<0.001) where ROI 1 presented the lowest values. SIGNIFICANCE/CONCLUSIONS:Framework-design modification did not influence the residual stress of porcelain-fused to zirconia fatigued 3-unit FDP. Whereas fractured FDPs showed the highest residual stress compared to suspended and control FDPs. Residual stress increased as nanoindented away from framework.
PMID: 29258696
ISSN: 1879-0097
CID: 2893802

Dipyridamole enhances osteogenesis of three-dimensionally printed bioactive ceramic scaffolds in calvarial defects

Bekisz, Jonathan M; Flores, Roberto L; Witek, Lukasz; Lopez, Christopher D; Runyan, Christopher M; Torroni, Andrea; Cronstein, Bruce N; Coelho, Paulo G
PURPOSE/OBJECTIVE:The objective of this study was to test the osteogenic capacity of dipyridamole-loaded, three-dimensionally printed, bioactive ceramic (3DPBC) scaffolds using a translational, skeletally mature, large-animal calvarial defect model. MATERIALS AND METHODS/METHODS:Custom 3DPBC scaffolds designed to present lattice-based porosity only towards the dural surface were either coated with collagen (control) or coated with collagen and immersed in a 100 μM concentration dipyridamole (DIPY) solution. Sheep (n = 5) were subjected to 2 ipsilateral trephine-induced (11-mm diameter) calvarial defects. Either a control or a DIPY scaffold was placed in each defect, and the surgery was repeated on the contralateral side 3 weeks later. Following sacrifice, defects were evaluated through microcomputed tomography and histologic analysis for bone, scaffold, and soft tissue quantification throughout the defect. Parametric and non-parametric methods were used to determine statistical significance based on data distribution. RESULTS:No exuberant or ectopic bone formation was observed, and no histologic evidence of inflammation was noted within the defects. Osteogenesis was higher in DIPY-coated scaffolds compared to controls at 3 weeks (p = 0.013) and 6 weeks (p = 0.046) in vivo. When bone formation was evaluated as a function of defect radius, average bone formation was higher for DIPY relative to control scaffolds at both time points (significant at defect central regions at 3 weeks and at margins at 6 weeks, p = 0.046 and p = 0.031, respectively). CONCLUSION/CONCLUSIONS:Dipyridamole significantly improves the calvarial bone regeneration capacity of 3DPBC scaffolds. The most significant difference in bone regeneration was observed centrally within the interface between the 3DPBC scaffold and the dura mater.
PMCID:5803375
PMID: 29292126
ISSN: 1878-4119
CID: 2899422

Micro-computed tomography evaluation of volumetric polymerization shrinkage and degree of conversion of composites cured by various light power outputs

Atria, Pablo J; Sampaio, Camila S; Caceres, Eduardo; Fernandez, Jessica; Reis, Andre F; Giannini, Marcelo; Coelho, Paulo G; Hirata, Ronaldo
This study evaluated the influence of different light-curing modes on the volumetric polymerization shrinkage and degree of conversion of a composite resin at different locations using micro-computed tomography and Fourier transform infrared spectroscopy (FTIR). Specimens were divided into 4 groups based on the light-curing mode used (Bluephase 20i): 1 -High (1,200 mW/cm2); 2 -Low (650 mW/cm2); 3 -Soft-start (650-1,200 mW/cm2); and 4 -Turbo (2,000 mW/cm2). Degree of conversion was calculated by the measurement of the peak absorbance height of the uncured and cured materials at the specific wavenumbers, and was performed by FTIR 48 h after curing resin samples. Degree of conversion was analyzed using two-way ANOVA. No significant differences were observed independent of the region of the restoration investigated (p>0.05). Different curing modes did not influence volumetric shrinkage neither degree of conversion of class I composite resin restorations.
PMID: 29081446
ISSN: 1881-1361
CID: 2766152

Effect of microstructure on tensile and compressive behavior of WE43 alloy in as cast and heat treated conditions

Xiang, Chongchen; Gupta, Nikhil; Coelho, Paulo; Cho, Kyu
The influence of heat treatment on the microstructure, tensile and compressive properties and failure mechanisms of WE43 alloy is studied. The eutectic phase is dissolved into the α-Mg phase and the grain size is refined in the heat treated alloy. Heat treatment improves the tensile yield and ultimate strengths by 40% and 53%, respectively. The compressive yield and ultimate strengths of heat treated alloy are also 55% and 23%, respectively, higher compared to the as-cast alloy. Compressive characterization is also conducted at high strain rates. The energy absorption capability of WE43-T5 specimens is higher than the as-cast WE43 specimens at all strain rates investigated in this work. Failure initiates with cracks in the eutectic phase in the as-cast alloy. These cracks grow through the grain to result in transgranular fracture. The absence of eutectic mixture in heat treated alloy results in grain boundary sliding and crack initiation at triple junctions. The crack propagation is delayed in the absence of eutectic precipitates, which improves the mechanical properties of the heat treated alloy.
SCOPUS:85032345142
ISSN: 0921-5093
CID: 2795642

Effect of implant placement depth on the peri-implant bone defect configurations in ligature-induced peri-implantitis: An experimental study in dogs

Huang, B; Zhang, L; Xu, L; Zhu, W; Witek, L; Tovar, N; Coelho, P-G; Meng, H
BACKGROUND:The subcrestal placement of implant platform has been considered a key factor in the preservation of crestal bone, but the influence of implant placement depth on bone remodeling combined with peri-implantitis is not fully understood. The aim of this study was to assess the effect of the crestal or subcrestal placement of implants on peri-implant bone defects of ligature-induced peri-implantitis in dogs. MATERIAL AND METHODS/METHODS:Eight weeks after tooth extraction in six beagle dogs, two different types of implants (A: OsseoSpeed(TM), Astra, Molndal, Sweden; B: Integra-CP(TM), Bicon, Boston, USA) were placed at either crestal or subcrestal (-1.5 mm) positions on one side of the mandible. Ligature-induced peri-implantitis was initiated four weeks after the installation of the healing abutment connections. After 12 weeks, tissue biopsies were processed for histological analyses. RESULTS:Supra-alveolar bone loss combined with a shallow infrabony defect was observed in crestal level implants while deep and wide infrabony defects were present in subcrestal level groups. Subcrestal groups showed significantly greater ridge loss, depths and widths of infrabony defects when compared to crestal groups (P <0.001). CONCLUSIONS:Within the limitations of the animal study, it can be stated that the implants at subcrestal position displayed greater infra-osseous defect than implants at crestal position under an experimental ligature-induced peri-implantitis.
PMCID:5822536
PMID: 29274159
ISSN: 1698-6946
CID: 2961202

Kinetics of conversion of brushite coatings to hydroxyapatite in alkaline solution

Da Rocha, Daniel Navarro; Da Silva, Marcelo H.Prado; De Campos, José Brant; Marçal, Rubens L.Santana Blazutti; Mijares, Dindo Q.; Coelho, Paulo G.; Cruz, Leila Rosa
This work describes the kinetics of conversion of brushite coatings, produced by chemical deposition, to hydroxyapatite. The conversion was performed in alkaline solution at three temperatures: 50 °C, 55 °C, and 60 °C. The evolution of the transformation was assessed by X-ray diffraction, and the phase fractions were determined by Rietveld refinement. The Johnson-Mehl-Avrami equation was applied to the data in order to describe the conversion mechanisms. The value of 42 kJ/mol obtained for the conversion activation energy is of the same order as found in similar surface-controlled dissolution processes. The Avrami exponent was n = 3.5, which is consistent with an interface-controlled growth of three-dimensional HA crystals with decreasing nucleation rate.
SCOPUS:85044425670
ISSN: 2238-7854
CID: 3859342