Compound Miao medicine Jiuxian Luohan bone-setting decoction improves rat femoral fracture healing: Associations with osteogenic and apoptosis-related markers

Abstract: Background: Fracture healing is a dynamic biological process involving inflammation, callus formation, mineralization and remodeling. Osteogenic and apoptosis-related markers may reflect changes during repair, but tissue-level marker expression alone does not establish a causal apoptotic mechanism. Objectives: To evaluate whether Compound Miao medicine Jiuxian Luohan bone-setting decoction is associated with improved femoral fracture healing in rats and with altered expression of runt-related transcription factor 2 (Runx2), Osterix, procollagen type I N-terminal propeptide (PINP), Bcl-2-associated X protein (Bax) and Caspase-3 at the fracture site. Methods: Twenty-five male specific-pathogen-free (SPF) Sprague-Dawley rats were used to establish a right femoral fracture model and were randomized into five groups (n=5/group): model, low-, medium- and high-dose decoction and orthopedic bone-setting tablet groups. Treatments were initiated 24 h after modeling and administered by gavage twice daily for 30 days. Anteroposterior and lateral radiographs were obtained on day 15 and scored using the Lane-Sandhu system. Fracture-site tissues were collected on day 30 for reverse transcription quantitative polymerase chain reaction (RT-qPCR), Western blotting, immunohistochemistry and Alizarin Red staining. Results: Compared with the model group, the high-dose decoction group showed a higher Lane-Sandhu score at week 2 and greater callus formation. Runx2, Osterix and PINP mRNA/protein levels increased, whereas Bax and Caspase-3 mRNA/protein levels decreased in treated groups, particularly in the high-dose group. Alizarin Red staining indicated greater deposition of mineralized matrix in treated tissues. Conclusion: Compound Miao medicine Jiuxian Luohan bone-setting decoction was associated with improved radiographic healing and favorable changes in osteogenic and apoptosis-related markers in this rat femoral fracture model.