Corn oil

Fractures were performed in 8–9-week-old animals. Mice were anesthetized with isoflurane and a closed transverse diaphyseal fracture on right femur was created using a drop‐weight blunt guillotine device, after inserting a 24G needle into the intramedullary canal to stabilize the fracture.²¹ To trigger overexpression in αSMACreNICD1 mice, tamoxifen (Sigma Aldrich, St. Louis, MO, USA) dissolved in corn oil (Acros Organics, Belgium) was injected intraperitoneally (ip) at a dose of 75 μg/g of body weight on 0, 2 and 4 days post fracture (DPF) to experimental and control groups. Buprenorphine (0.1 mg/kg) was given subcutaneously during the first 2 days of healing. Fractures were confirmed immediately after the procedure and the healing process was monitored by X-ray (Faxitron LX 60) on 7, 14, 21- and 42 DPF.

4-CB-2′,5′-HQ, and 4-CB-2′,5′-BQ were generously provided by Dr. Hans-Joachim Lehmler and Dr. Gregor Luthe. Lipopolysaccharide (LPS) from Escherichia coli 0111:B4 (Sigma Aldridge) was suspended in Hank’s balanced salt solution (HBSS). Corn oil was purchased from Acros Organics, Morris Plains, NJ. Dimethyl sulfoxide (DMSO) was purchased from Fisher Scientific, Pittsburgh, PA. PAMAM dendrimers (G5-NH₂), 20% w/w in water, were purchased from Dendritech, Midland, MI. MicroSprayer^® model 1A–1C (stainless steel, 1.25″ after 120° bend) equipped with FMJ-250 high-pressure (3000 psi) syringe for ITI was purchased from PennCentury, Wyndmoor, PA. Sep-Pak^® plus C18 cartridges (Cat.No.WAT020515) were purchased from Waters, Milford, MA. Prostaglandin E metabolite enzyme immunoassay (EIA) kit (Cat.No.514531) was purchased from Cayman Chemical, Ann Arbor, MI.

Male mice (older than eight weeks) were injected with carbon tetrachloride (CCl4; Sigma) for acute liver injury model. CCl4 was dissolved in corn oil (Fisher) at a final concentration of 20% (v/v) for intraperitoneal administration (1 ml/kg). Mice were sacrificed at various time points, and liver tissues were collected for further analyses.

Due to the poor solubility of BPA, we first dissolved 1 g of BPA (Milipore Sigma, Cat. No.239658–50G, USA) into 3 mL of pure ethyl alcohol (Sigma-Aldrich, Cat. No. E7023, USA). This mixture was then added to 97 mL of corn oil (Fisher Scientific, Cat. No S25271, USA) to make a stock solution at 10 μg/μL concentration of BPA and 3% ethanol. The stock solution was further diluted as follows: 5 mL of the stock was added to 95 mL of corn oil to make Dilution One at a BPA concentration of 0.5 μg/μL and 0.15% of ethanol. Then, 2.5 mL of Dilution One was added to 247.5 mL of corn oil to make Dilution Two (working solution), which was at a BPA concentration of 0.005 μg/μL and 0.0015% ethanol. The appropriate amount of Dilution Two was added into a 1/2 Honey Nut Cheerio (treat) (Honey Nut Cheerio, General Mills) to have a final exposure dose of 5 μg/kg/day of BPA. The control group also received a treat with corn oil and the same amount of ethanol (0.0015%) as the BPA-treated group. Both control and treatment groups were administered with the appropriate amount of BPA and corn oil in the treat, according to body weight, for 19–21 consecutive days.