Pietra Colacrai Arikita
Research Mentor(s): Hsiao Hsin Sung
Mentor Department: Orthopaedic Research Lab (ORL)
Authors: Pietra Colacrai Arikita, Vanini Pimpalwar, Hsiao Sung, Kenneth Kozloff
Session: Session 6 (3:00pm – 3:50pm)
Presentation Type: Poster 46
Abstract
Osteogenesis imperfecta (OI) is a congenital disorder marked by low bone mass and craniofacial deformities. Sclerostin antibody (SclAb), known for its anabolic effects on bone, holds potential for improving bone health, yet its influence on alveolar bone inflammation and resorption in OI requires further exploration. This study examines the impact of SclAb on inflammation and resorption in the alveolar bone in an OI mouse model. Wild-type (WT) and Brtl/+ OI mice, aged three months, received subcutaneous injections of SclAb (25 mg/kg) or a vehicle control, twice weekly for five weeks. After treatment, alveolar bone tissues were collected, snap-frozen, and stored at -80 °C for protein extraction. Proteins from alveolar bone were extracted using RIPA lysis buffer, and concentrations were measured with a BCA kit. Samples were heated at 95 °C for 5 minutes in 4× Nupage buffer before analysis. Proteins (20 µg per sample) were separated on a 4%–12% polyacrylamide gel and transferred onto a PVDF membrane. Ponceau Red staining confirmed equal sample loading. Blocking was performed with 5% non-fat dry milk and 0.05% Tween 20, followed by overnight incubation with primary antibodies against IL-1ß, IL-6, IL-17, and TNFa. Membranes were then incubated with secondary antibodies for 30 minutes at room temperature. Detection was via SuperSignal West Pico PLUS Chemiluminescent Substrate and captured using the ImageQuant LAS 4000 system. Band intensities were quantified using an XRS+ Chemidoc system and Image Lab software, with ß-actin as a loading control. We hypothesize that PBS-treated Brtl/+ OI mice will show higher inflammatory markers in alveolar bone compared to WT and SclAb-treated Brtl/+ mice. We also expect that SclAb- treated Brtl/+ mice will exhibit inflammatory marker levels approaching those of WT controls.
