Original Article
Loss of Bcl-3 delays bone fracture healing through activating NF-κB signaling in mesenchymal stem cells

https://doi.org/10.1016/j.jot.2022.07.009Get rights and content
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Abstract

Background

Bone fracture healing is a postnatal regenerative process in which fibrocartilaginous callus formation and bony callus formation are important. Bony callus formation requires osteoblastic differentiation of MSCs.

Materials and methods

The formation of callus was assessed by μCT, Safranin-O, H&E and Masson trichrome staining. Osteogenesis of MSCs was analyzed by ALP staining, ARS staining, qRT-PCR and WB. And we also used IF and TOP/FOP Flash luciferase reporter to assess the nuclear translocation of PP65.

Results

In this study, we found Bcl-3 showed a significant correlation with bone fracture healing. Results of μCT showed that loss of Bcl-3 delays bone fracture healing. Safranin-O, H&E and Masson trichrome staining confirmed that loss of Bcl-3 impacted the formation of cartilage and woven bone in callus. Further experiments in vitro manifested that Bcl-3-knockdown could inhibit MSCs osteoblastic differentiation through releasing the inhibition on NF-κB signaling by Co-IP, IF staining and luciferase reporter assay.

Conclusions

We unveiled that loss of Bcl-3 could lead to inhibited osteogenic differentiation of MSCs via promoting PP65 nuclear translocation.

The translational potential of this article

Our data demonstrated that overexpression of Bcl-3 accelerates bone fracture healing, which serves as a promising therapeutic target for bone fracture treatment.

Keywords

Bcl-3
Fracture healing
Mesenchymal stem cells
NF-κB

Abbreviations

marrow mesenchymal stem cell
MSCs
B-cell lymphoma 3
Bcl-3
Runt-related transcription factor 2
Runx2
peroxisome proliferator-activated receptor gamma
Ppar-γ
inhibitor of κB
IκB
phosphorylation of P65
PP65
wild type
WT
quantitative reverse transcriptase polymerase chain reaction
qRT-PCR
Micro-computed tomography
μCT
bone volume per tissue volume
BV/TV
Co-immunoprecipitation
Co-IP
Alkaline Phosphatase
ALP
Alizarin Red S
ARS

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1

These authors contributed equally to this work