Bone quality in zebrafish vertebrae improves after alendronate administration in a glucocorticoid-induced osteoporosis model.

Glucocorticoid-induced osteoporosis (GIOP) changes the microarchitecture of bones and often leads to the reduction of bone-mineral density (BMD) and increased fracture rates. Zebrafish has been used as an alternative model for GIOP, however, the interaction of GIOP, and its treatment, with zebrafish bone morphometrics and mechanical properties, remains a challenge. Thus, this study aimed to evaluate the effects of prednisolone and alendronate on the properties of zebrafish vertebrae. Adult 7-month-old zebrafish were distributed into four groups: control (CTRL), prednisolone-only (PN), alendronate-only (ALN), and the sequential use of both medicines (PN + ALN). Fish skeletons were scanned via micro-tomography (n = 3) to obtain vertebra morphometrics (e.g., BMD). Bone morphology was assessed using scanning electron microscopy (n = 4) and the biomechanical behaviour with nanoindentation technique (n = 3). The BMD decreased in PN (426.08 ± 18.58 mg/cm3) and ALN (398.23 ± 10.20 mg/cm3) groups compared to the CTRL (490.43 ± 41.96 mg/cm3) (p < 0.001); however, administering the medicines in sequence recovered the values to healthy levels (495.43 ± 22.06 mg/cm3) (p > 0.05). The bone layered structures remain preserved in all groups. The vertebrae of the groups that received ALN and PN + ALN, displayed higher modulus of elasticity (27.27 ± 1.59 GPa and 25.68 ± 2.07 GPa, respectively) than the CTRL (22.74 ± 1.60 GP) (p < 0.001). ALN alone increased the hardness of zebrafish vertebrae to the highest value among the treatments (1.32 ± 0.13 GPa) (p < 0.001). Conversely, PN + ALN (1.25 ± 0.11 GPa) showed unaltered hardness from the CTRL (1.18 ± 0.13 GPa), but significantly higher than the PN group (1.08 ± 0.12 GPa) (p < 0.001). ALN administered after GIOP development, rescued osteoporotic condition by recovering the BMD and bone hardness in zebrafish vertebrae.

Influence of Prednisolone and Alendronate on the de novo Mineralization of Zebrafish Caudal Fin.

Dysregulated balance between bone resorption and formation mediates the onset and progression of osteoporosis. The administration of prednisolone is known to induce osteoporosis, whereas alendronate is commonly used to reverse the process. However, the assessment of the effects of such medicines on the nanostructure of bone remodeling and mechanical properties remains a major technical challenge. The aim of this study was to apply various analytical approaches to evaluate the compositional, morphological, and mechanical properties of regenerative zebrafish caudal fin bony rays affected by prednisolone and alendronate. Adult wild-type AB strain zebrafish were first exposed to 125µM of prednisolone for 14 days to develop glucocorticoid-induced osteoporosis. Fish fins were then amputated and let to regenerate for 21 days in tank water containing 30µM of alendronate or no alendronate. The lepidotrichia in the proximal and distal regions were evaluated separately using confocal microscope, scanning electron microscope, electron-dispersive spectroscopy, Raman spectroscopy, atomic force microscopy, and a triboindenter. As expected, prednisolone led to significant osteoporotic phenotypes. A decrease of Ca/P ratio was observed in the osteoporotic subjects (1.46 ± 0.04) as compared to the controls (1.74 ± 0.10). Subsequent treatment of alendronate overmineralized the bony rays during regeneration. Enhanced phosphate deposition was detected in the proximal part with alendronate treatment. Moreover, prednisolone attenuated the reduced elastic modulus and hardness levels (5.60 ± 5.04 GPa and 0.12 ± 0.17 GPa, respectively), whereas alendronate recovered them to the pre-amputation condition (8.68 ± 8.74 GPa and 0.34 ± 0.47 GPa, respectively). As an emerging model of osteoporosis, regrowth of zebrafish caudal fin was shown to be a reliable assay system to investigate the various effects of medicines in the de novo mineralization process. © 2020 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Incorporation of amoxicillin-loaded microspheres in mineral trioxide aggregate cement: an in vitro study.

OBJECTIVES: In this study, we investigated the potential of amoxicillin-loaded polymeric microspheres to be delivered to tooth root infection sites via a bioactive reparative cement. MATERIALS AND METHODS: Amoxicillin-loaded microspheres were synthesized by a spray-dray method and incorporated at 2.5% and 5% into a mineral trioxide aggregate cement clinically used to induce a mineralized barrier at the root tip of young permanent teeth with incomplete root development and necrotic pulp. The formulations were modified in liquid:powder ratios and in composition by the microspheres. The optimized formulations were evaluated in vitro for physical and mechanical eligibility. The morphology of microspheres was observed under scanning electron microscopy. RESULTS: The optimized cement formulation containing microspheres at 5% exhibited a delayed-release response and maintained its fundamental functional properties. When mixed with amoxicillin-loaded microspheres, the setting times of both test materials significantly increased. The diametral tensile strength of cement containing microspheres at 5% was similar to control. However, phytic acid had no effect on this outcome (p > 0.05). When mixed with modified liquid:powder ratio, the setting time was significantly longer than that original liquid:powder ratio (p < 0.05). CONCLUSIONS: Lack of optimal concentrations of antibiotics at anatomical sites of the dental tissues is a hallmark of recurrent endodontic infections. Therefore, targeting the controlled release of broad-spectrum antibiotics may improve the therapeutic outcomes of current treatments. Overall, these results indicate that the carry of amoxicillin by microspheres could provide an alternative strategy for the local delivery of antibiotics for the management of tooth infections.

Mineral deposition promoted by resin-based sealants with different calcium phosphate additions.

The aim of this study was to evaluate the influence of different calcium phosphates (CaPs) on the physical, biological, and remineralizing properties of experimental resin-based sealants (RBSs). Triethylene-glycol dimethacrylate (90wt%) and bisphenol A-glycidyl methacrylate (10wt%) were used to produce resin-based sealants. Hydroxyapatite (SHAp), α-tricalcium phosphate (Sα-TCP) and octacalcium phosphate (SOCP) were added to the sealants in a 10wt% concentration. One group without CaPs was used as the control group (SCG). The degree of conversion (DC) was assessed with Fourier-transformed infrared spectroscopy, whereas cytotoxicity was tested with the HaCaT keratinocyte cell line. The ultimate tensile strength (UTS) was used to assess the mechanical strength of the experimental RBSs. Sealed enamel was used for colorimetric assay. Mineral deposition was assessed with Raman spectroscopy after 7, 14, and 28 days of sample immersion in artificial saliva. Scanning electron microscopy was used to analyze the surface morphology after 28 days of immersion. The addition of 10wt% of fillers significantly reduced the DC of sealants. SOCP groups showed reduced cell viability. Higher UTS was found for Sα-TCP and SHAp. The color analysis showed that SGC and demineralized teeth presented higher mismatches with the sound tissue. Mineral deposition was observed for SHAp and Sα-TCP after 7 days, with increased phosphate content and mineral deposits for SHAp after 28 days. RBS with the addition of 10% HAp promoted increased mineralization in vitro after 28 days, and did not affect cell viability, DC, mechanical properties, or RBS color in the enamel.

Boron Nitride Nanotubes as Filler for Resin-Based Dental Sealants.

The aim of this study was to evaluate the influence of boron-nitride nanotubes (BNNTs) on the properties of resin-based light-curing dental sealants (RBSs) when incorporated at different concentration. RBSs were formulated using methacrylate monomers (90 wt.% TEGDMA, 10 wt.% Bis-GMA). BNNTs were added to the resin blend at 0.1 wt.% and 0.2 wt.%. A Control group without filler was also designed. Degree of conversion, ultimate tensile strength, contact angle, surface free energy, surface roughness and color of the RBSs were evaluated for the tested materials. Their cytotoxicity and mineral deposition ability (Bioactivity) were also assessed. A suitable degree of conversion, no effect in mechanical properties and no cytotoxic effect was observed for the experimental materials. Moreover, the surface free energy and the surface roughness decreased with the addition of BNNTs. While the color analysis showed no difference between specimens containing BNNTs and the control group. Mineral deposition occurred in all specimens containing BNNTs after 7d. In conclusion, the incorporation of BNNTs may provide bioactivity to resin-based dental sealants and reduce their surface free energy.

Calcium phosphates as fillers for methacrylate-based sealer.

OBJECTIVES: The aim of this study was to evaluate the mineral deposition, push-out bond strength, radiopacity, the degree of conversion, film thickness, flow, calcium ion release, and pH of experimental endodontic sealers containing hydroxyapatite (HAp), aflfa-ticalcium phosphate (α-TCP), or octacalcium phosphate (OCP) particles. MATERIALS AND METHODS: Fifty single straight root human premolars were instrumented and divided into five groups (n = 10). Experimental endodontic sealers were formulated by 70 wt% urethane dimethacrylate (UDMA), 15 wt% of glycerol-1,3-dimethacrylate (GDMA), 15 wt% of ethoxylated bisphenol A glycol dimethacrylate (BISEMA), camphorquinone (CQ), N,N-dihydroxyethyl-para-toluidine (DHEPT), and benzoyl-peroxide. 10 wt% of each HAp, α-TCP, or OCP were added to the resin and its properties were assessed. RESULTS: After 7 days, the degree of conversion ranged from 44.69% (GOCP) to 50.74% (Gcontrol) and no statistical difference were observed (p < 0.05). GAHplus showed the highest push-out bond strength 4.91 (± 2.38) MPa at 28 days of analysis (p < 0.05). Film thickness and pH were not statistically different (p > 0.05). Statically lower values of flow were found for GHAp, GOCP, and Gα-TCP (p < 0.05). Calcium deposition values were higher for GHAp at 28 days. CONCLUSIONS: Bond strength, degree of conversion, and film thickness of endodontic sealers with phosphates showed similar results compared with AHplus, but displayed higher amounts of Ca2+ release. CLINICAL RELEVANCE: Phosphate fillers improve the performance of endodontic sealers after 28 days of simulated body fluid.