Efficacy comparison and mechanism study of a novel single-stage dosing regimen versus traditional three-stage regimen for fracture healing
WANG Lin-jue,GU En-peng,GUO Rui
Abstract:
Objective To compare the promotive effects of the novel single-stage administration regimen and the traditional three-stage regimen on fracture healing, and to preliminarily investigate their roles in activating the BMP/Smad signaling pathway. Methods A rat tibial fracture model was established and randomly divided into four groups: blank control, model control, single-stage treatment (administered a comprehensive formula throughout the experiment), and three-stage treatment (administered Fuyuan Huoxue decoction on days 1-7, Heying Zhitong decoction on days 8-28, and Jianbu Huqian pill after day 28). Post-surgery, X-ray imaging was used to assess the degree of fracture healing. Levels of calcium (Ca), phosphorus (P), alkaline phosphatase (ALP), and acid phosphatase (ACP) were measured. Histological examination via HE staining was performed to observe callus tissue healing. Immunohistochemistry was used to detect the protein levels of BMP2, Runx2, and Smad1. RT-qPCR was employed to quantify the mRNA expression levels of BMP2, Runx2, and Smad1. Results Compared with the model control group, both the single-stage and three-stage treatment groups showed significantly accelerated fracture healing, with the single-stage group exhibiting the most pronounced effect. Alkaline phosphatase (ALP) levels in the model control group were (165±4.7)U/L. However, the single-stage administration and traditional three-stage regimen groups showed ALP levels of (233±12.2)U/L and (174±20.3)U/L, respectively. The significant elevation of ALP in the single-stage administration suggests that its effect on promoting fracture repair was markedly superior to both the model control group and the traditional three-stage regimen. The acid phosphatase (ACP) level in the single-stage administration was (0.192±0.027), which was significantly lower than that in the model control group (0.340±0.017) and the traditional three-stage regimen (0.265±0.014). Hematoxylin-eosin (HE) staining revealed a marked reduction in local inflammatory cell infiltration and abundant formation of cartilage and new bone in the single-stage administration; the traditional three-stage regimen also exhibited reduced inflammation and increased collagen/cartilage tissue. Immunohistochemical analysis showed that the expression level of Runx2 in the single-stage administration was (57.1±5.9)%, significantly higher than that in the model control group (41.6±3.1)% and the traditional three-stage regimen (48.3±4.0)%. Levels of Smad and BMP2 were also elevated to varying degrees, and RT-PCR results further corroborated this finding. Conclusion The single-stage comprehensive treatment regimen effectively promotes fracture osteogenesis and remodeling by activating the BMP/Smad signaling pathway, and its effect is superior to the three-stage treatment approach.
