Fig 2: Cotreatment of NF-?B signaling pathway inhibitor BAY117082 and RR-L containing serum showed no better effect than BAY117082 along on MSCs in vitro. No differences were observed between RR-L containing serum and BAY117082 in (a) osteogenesis staining; (b) adipogenesis staining, and (c-e) NF-?B signaling pathway activation of MSCs. (f–h) Statistical results of Alizarin red, Oil red staining, and IF analysis. (i) Representative western blots image of PPAR?, RUNX2, OCN, p65, and ß-actin. (j–m) Statistical results of western blot via Image Lab software. (n–q) mRNA expressions of PPAR?, RUNX2, OCN, and p65. #p < 0.05 versus ctrl, NS, p > 0.05 versus BAY117082 group.

Fig 3: The effect of LPCs in vascularization and osteocyte generation. (A) Representative three-dimensional microcomputed tomography angiography (µCTA) images in the femoral head in control group and LPCs-treated group. (B) HE staining and immunohistochemical staining for Osteocalcin for both vehicle and LPCs cells treated groups. The black arrow shows osteocyte and lacunae. (C–D) Quantitative analysis of empty lacunae number and positive area of OCN. **p = 0.0023, *p = 0.0389, n = 3.

Fig 4: RR-L reversed TNF-a-induced NF-?B signaling pathway activation related to MSC differentiation alterations. (a) RR-L containing serum inhibited TNF-a-induced osteogenesis inhibition of MSCs. (b) RR-L containing serum inhibited TNF-a-induced adipogenesis activation of MSCs. (c–e) RR-L containing serum inhibited TNF-a-induced NF-?B signaling pathway activation of MSCs. (f–h) Statistical results of Alizarin red, Oil red staining, and IF analysis. (i) Representative western blots image of PPAR?, RUNX2, OCN, p65, and ß-actin. (j–m) Statistical results of western blot via Image Lab software. (n–q) mRNA expressions of PPAR?, RUNX2, OCN, and p65. #p < 0.05 versus ctrl, *p < 0.05 versus TNF-a.