Abstract
BackgroundGrowing evidence has implicated core-binding factor beta (Cbfb) as a contributor to osteoblast differentiation, which plays a key role in fracture healing. Herein, we aimed to assess whether Cbfb affects osteoblast differentiation after fibula fracture.MethodsInitially, we established a Cbfb conditional knockout mouse model for subsequent studies. Immunohistochemical staining was conducted to detect the expression of proliferating cell nuclear antigen (PCNA) and collagen II in the fracture end. Next, we isolated and cultured osteoblasts from specific Cbfb conditional knockout mice for BrdU analysis, alkaline phosphatase (ALP) staining, and von Kossa staining to detect osteoblast viability, differentiation, and mineralization, respectively. Western blot analysis and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) were used to detect the expression of osteoblast differentiation-related genes.ResultsThe Cbfb conditional knockout mice exhibited downregulated expression of PCNA and collagen II, reduced ALP activity, and mineralization, as well as diminished expression of osteoblast differentiation-related genes. Further, Cbfb knockout exerted no obvious effects on osteoblast proliferation.ConclusionsOverall, these results substantiated that Cbfb could promote fibula fracture healing and osteoblast differentiation and thus provided a promising therapeutic target for clinical treatment of fibula fracture.
Highlights
Growing evidence has implicated core-binding factor beta (Cbfb) as a contributor to osteoblast differentiation, which plays a key role in fracture healing
Construction of Cbfb conditional knockout mouse model Loxp was inserted at both ends of the Cbfb allele to construct Cbfb fluorine and oxygen (Flox) mice (Jackson Lab, CA, USA), which were hybridized with Dermo promoter Cre (Cyclization Recombination Enzyme) mice (Jackson Lab, CA, USA), and the first filial generation Cbfbf/f; Dermo-Cre mice were obtained
Histological observation of morphological characteristics of Cbfbf/f; Dermo-Cre mice At the 1st day after fracture, the staining results revealed that most fracture ends of Cbfbf/f; Dermo-Cre mice and wild type (WT) mice were filled with fibrous tissues
Summary
Growing evidence has implicated core-binding factor beta (Cbfb) as a contributor to osteoblast differentiation, which plays a key role in fracture healing. We aimed to assess whether Cbfb affects osteoblast differentiation after fibula fracture. Cbfb is essential for embryonic bone morphogenesis and postnatal cartilage and bone formation, through its control of the balance of chondrocyte differentiation and proliferation via regulation of Indian hedgehog expression and parathyroid hormone-related protein receptor [10]. Cbfb has been reported to modulate bone development through stabilizing runt-related transcription factor (Runx) family proteins, and Runx serves as a critical target in fracture healing [15,16,17]. We conducted this study to ascertain specific mechanistic actions of Cbfb in fibula fracture healing via regulation of osteoblast differentiation in an animal model
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