P3796

The rats in the Sham and MM + veh control groups (groups 1 and 2) received vehicle (sterile water used for injections) at 1 mL/kg subcutaneously (sc) 5 days/week, starting on the day of surgery. The rats in group 3 received zoledronic acid (Zol; Sargent Pharmaceuticals 25021-801, Schaumburg, IL, USA) at 100 μg/kg, sc, twice/week [18 (link)], and the rats in group 4 received human PTH (hPTH 1-34; Sigma-Aldrich P3796, St. Louis, MO, USA) at 40 μg/kg, sc, five times/week [25 (link)], starting on the day of surgery. To label actively mineralized bone surfaces, all of the rats in the study received calcein (Sigma-Aldrich Cat. No. C-0875, St. Louis, MO, USA) at 10 mg/kg (3.3 mL/kg) 13 days before necropsy and alizarin (Sigma-Aldrich Cat. No. A-5533, St. Louis, MO, USA) at 30 mg/kg (3.0 mL/kg) 3 days before necropsy.

HUVECs were purchased from ScienCell Research Laboratories (catalog number: 8000, USA) and were cultured as previously described. All experiments were performed using cells at passages 3 to 5. Cells (HUVECs) were then cultured in the presence of PTH (P3796, Sigma, 10^‐7 mol/L) for 48 hours in order to identify the effects of PTH with/without DAPT (10 ng/mL, Sigma Aldrich) according to the phenotypic transition and differentiation capability of ECs that undergoing EndMT.