Fig. 3.

Abx treatment rescued OB dysfunction co-cultured with BM neutrophils from SCD mice. Calvarial OBs were digested from 3-day-old Ctrl mice. BM neutrophils were isolated using MojoSort Mouse Neutrophil Isolation Kit from single-housed 4-month-old Ctrl and SCD male mice treated with H₂O or Abx. (A-D) Fixed neutrophils were plated on top of confluence OBs in osteogenic medium. (A) ALP activity at day 7 of co-culture. (B) Ocn mRNA expression at day 14 of co-culture. (C) Representative images and (D) quantitation of Alizarin Red staining at day 14 of co-culture. (E) OBs were plated on the bottom of Transwells. Fixed BM neutrophils were plated on 0.4 µm pore polycarbonate membrane inserts in osteogenic media. ALP activity was measured on day 7 of culture. n = 4 mice/group. * p < 0.05 compared with Ctrl-H₂O; ^#p < 0.05 compared with corresponding H₂O group; two-way ANOVA (A, B,D); T-test (E).