Vanini Pimpalwar
Research Mentor(s): Hsiao Hsin Sung
Mentor Department: Orthopaedic Research Lab (ORL)
Authors: Vanini Pimpalwar, Pietra Colacrai Arikita, Hsiao Sung, Kenneth Kozloff
Session: Session 6 (3:00pm – 3:50pm)
Presentation Type: Poster 47
Abstract
Osteogenesis imperfecta (OI) is a congenital skeletal dysplasia marked by low bone mass and craniofacial issues. Sclerostin antibody (SclAb), an anabolic bone medication, has shown potential in improving bone parameters, but its effects on alveolar bone inflammation and resorption in OI are not well understood. This study aims to evaluate the impact of SclAb on inflammatory response and resorption in the alveolar bone of an OI mouse model. Three-month-old wild-type (WT) and Brtl/+ OI mice were administered subcutaneous SclAb injections at 25 mg/kg or a vehicle control twice weekly for five weeks. Alveolar and femoral bone samples were collected and stored in RNAlater at -80°C. RNA extraction was performed using the TRIzol method (Invitrogen), followed by spectrophotometric assessment of RNA concentrations and purity using the NanoDrop 2000 (Thermo Fisher Scientific). A total of 600 ng of RNA was reverse transcribed into cDNA using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). The resulting cDNA, diluted to 5 ng/µL, underwent amplification with Power SYBR® Green PCR Master Mix and gene-specific primers in a QuantStudio 6 Pro system, adhering to manufacturer protocols. Relative expression levels of IL-1ß, IL-6, IL-17, and TNFa were normalized to Gapdh and calculated as fold changes over control using the 2-Ct method, and reported as mean ± standard deviation. Experiments were conducted in triplicate with controls ensuring that signals were not artifacts of DNA contamination or primer dimer formation. Melt curve analysis confirmed amplicon specificity. We hypothesize that PBS-treated Brtl/+ OI mice will show higher inflammatory marker expression in alveolar bone compared to femoral bone. We also anticipate that SclAb-treated Brtl/+ mice will exhibit gene expression levels nearing those of WT controls.
