Methods : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [2]
The ligations were maintained for 2 weeks, and after which the mice were euthanized by CO2 inhalation and the maxilla were harvested. The gingival tissues of half group of mice were isolated and collected for mRNA expression study. The skulls left were defleshed by beetles for 1 week. Briefly, in beetle’s chamber, freshly dissected skull was put in a paper cup with 0.5 cm diameter holes at the bottom, so beetles can move in with relatively controlled numbers. After that, the skulls were bleached by H2O2 (3%) for 4 h. Bone resorption was measured by microscope imaging analysis and μCT scan analysis. The skulls of the other half of the group were fixed in formalin overnight at 4 °C followed by EDTA decalcification for 3 weeks with agitation. After complete demineralization, implants were removed manually by rotating counterclockwise. All the decalcification samples were embedded into paraffin and cut in 5 μm sections along the mesial-distal plane and then subjected to H&E staining and TRAP staining.
The two dimension (2D) bone resorption measurements were assessed under a microscope (Nikon SMZ745T, Nikon Instruments Inc.) and analyzed by software ImageJ (NIH) on buccal and palatal surfaces for each segment, and a standard calibrator was used for calibration at the same magnification as previously described [36]. The bone resorption area was enclosed coronally by the CEJ of the molars, laterally by the exposed distal root of the first molar and the exposed mesial root of the third molar, and apically by the alveolar crest. The results are presented in square millimeters.
Mice maxillae were scanned with a high-resolution scanner (mCT-40, Scanco Medical). Samples were exposed to polychromatic X-rays on a rotating stage at a steep angle of 0.18° over 360°. Measurements were taken at an operating voltage of 70 kVp and 114 mA current and 6 mm isotropic voxel resolution, with an exposure time of 200 ms and five frames averaged per view. Quantitative three dimension (3D) measurements of the bone resorption were performed using Seg3D software as previously described [5].
Serial posts:
- Abstract : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [1]
- Abstract : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [2]
- Background : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [1]
- Background : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [2]
- Methods : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [1]
- Methods : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [2]
- Methods : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [3]
- Results : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [1]
- Results : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [2]
- Discussion : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [1]
- Discussion : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [2]
- Conclusions : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- Availability of data and materials : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- Abbreviations : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- References : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [1]
- References : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [2]
- References : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [3]
- References : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [4]
- Acknowledgements : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- Funding : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- Author information : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [1]
- Author information : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling [2]
- Ethics declarations : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- Additional information : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- Supplementary information : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- Rights and permissions : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- About this article : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through TLR2/4 signaling
- Table 1 Success rate (SR) of osseointegrated implants 4 weeks after implant placement : RANKL blockade alleviates peri-implant bone loss and is enhanced by anti-inflammatory microRNA-146a through
- Fig. 1. Mouse model of ligature-induced experimental peri-implantitis. (a) Tooth extraction: left maxillary first and second molars extracted at 4 weeks old and the tooth extraction socket healed well with smooth gingiva surface after 6 weeks post-extraction. Implant placement: implant was put in alveolar bone without flap elevation. Ligature placement: at 4 weeks post-implant, 7-0 ligatures were applied under the fixture head. Gingival injection: injections for animals were administered three times on days 3, 6, and 9 during 14 days ligation period. Sample collection: 14 days post-ligation, the gingival tissues and the skulls were collected. (b) Images depicting processing steps of the experimental design (scale bar, 500 μm) : RANKL blockade alleviates peri-implant
- Fig. 2. Anti-RANKL and anti-RANKL+miR-146a treatments decreased ligature-induced bone resorption with different patterns in experimental peri-implantitis of WT and TLR2/4 KO mice. Buccal side images of the defleshed skulls were taken of the control (non-ligation) group, ligation (non-treatment) group, ligation with anti-RANKL antibody (ligation+AR) treatment group, and ligation with anti-RANKL antibody + miR-146a (ligation+A+MiR) treatment group in WT mice and TLR2/4 KO mice (a) (scale bar, 500 μm). The bone resorption area based on these images was measured and analyzed for WT mice (b) and TLR2/4 KO mice (c) (mean ± SD, n = 6, *p < 0.05, **p < 0.01, SEM, standard error of difference between two means). Three dimension (3D) images from μCT were collected and analyzed for WT mice (d) and TLR4 KO mice (e) (mean ± SD, n = 6, *p < 0.05, **p < 0.01) : RANKL blockade alleviates peri-implant
- Fig. 3. Anti-RANKL and anti-RANKL+miR-146a treatments decreased TRAP-positive cell quantities with different patterns in experimental peri-implantitis of WT and TLR2/4 KO mice. TRAP-positive cells (red color) with 3 or more nuclei were considered osteoclasts and were shown in the control group, ligation group, ligation with anti-RANKL antibody treatment group, and ligation with anti-RANKL antibody + miR-146a treatment group in WT mice and TLR2/4 KO mice (a) (Im, implant; Av, alveolar bone; scale bar, 100 μm). The quantities of TRAP-positive cells were analyzed in each group of WT mice (b) and TLR2/4 KO mice (c) (mean ± SD, n = 6, **p < 0.01) : RANKL blockade alleviates peri-implant
- Fig. 4. Anti-RANKL and anti-RANKL+miR-146a treatments decreased the inflammatory cell infiltration of the implant gingival tissues with different patterns in experimental peri-implantitis of WT and TLR2/4 KO mice. HE staining of the gingival tissue around implants were performed in the control group, ligation group, ligation with anti-RANKL antibody treatment group, and ligation with anti-RANKL antibody + miR-146a treatment group in WT mice and TLR2/4 KO mice (a) (scale bar, 100 μm). Inflammatory cell numbers were measured and analyzed in each group of WT mice (b) and TLR2/4 KO mice (c) (mean ± SD, n = 6, **p < 0.01) : RANKL blockade alleviates peri-implant
- Fig. 5. Anti-RANKL and anti-RANKL+miR-146a treatments decreased gingival mRNA expression of TNF-α and RANKL with different patterns in experimental peri-implantitis of WT and TLR2/4 KO mice. Gingival tissues around ligatured implants and non-ligation implants were excised and processed for RT-qPCR analysis to determine mRNA level of TNF-α of WT mice (a) and TLR2/4 KO mice (b) (mean ± SD, n = 6, *p < 0.05, **p < 0.01) and mRNA level of RANKL of WT mice (c) and TLR2/4 KO mice (d) (mean ± SD, n = 6, *p < 0.05, **p < 0.01). : RANKL blockade alleviates peri-implant