The horizontal alveolar bone resorption (ABR) area and the presence of periodontal intrabony defects were measured by histomorphometry as described previously [37] (link), . Briefly, the maxilla and mandible (n = 10−15) were immersed in 3% (vol/vol) hydrogen peroxide overnight after autoclaving and defleshing the maxillae and mandibles and stained in an aqueous solution of 0.1% methylene blue to delineate the cemento-enamel junction (CEJ) [37] (link), [38] (link), [39] (link). Digital images of both buccal and lingual root surfaces of all molar teeth were captured under a 10×stereo dissecting microscope (SteReo Discovery V8; Carl Zeiss Microimaging, Inc, Thornwood, NY), after superimposition of buccal and lingual cusps to ensure reproducibility and consistency. The line tool was used to measure the horizontal alveolar bone resorption from the CEJ to the alveolar bone crest (ABC). The surface perimeters of CEJ and ABC were traced using the calibrated line tool (AxioVision LE 29A software version 4.6.3.). Two blinded examiners performed all measurements twice at separate times. The means of the measurements were obtained for each of the two quadrants. Periodontal intrabony defects were detected under a 10×stereo dissecting microscope (SteReo Discovery V8) by an experienced periodontist (JL). The maxillae and mandibles were tilted and stabilized with dental wax to verify the presence of the intrabony defects in buccal and lingual surfaces. Only the presence or absence of intrabony defects was detected because the crevasses in the mouse jaw are too small to measure depth and width.
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