Background: Infected bone defect poses a great challenge for orthopedists because it is difficult to cure. Tissue-engineered bone based on the human mesenchymal stem cells (hMSCs), has currently taken a promising treatment protocol in clinical practice. In a previous study, a porous hydroxyapatite/fibronectin/alginate (PHA/FN/ALG) composite scaffold displayed favorable biological properties as a novel scaffold, which was considered better than single-material scaffolds. In addition, Wnt11 has been demonstrated to play an important role in the development of osteoblasts, but until recently, its role in the osteogenic differentiation of hMSCs in infectious environment remained unclear. Methods: In this study, we constructed a PHA/FN/ALG composite scaffold with layer-by-layer technology. Furthermore, we also constructed Wnt11-silenced (RNAi) and -overexpressing hMSCs by lentiviral transduction. The gene transduction efficacy was confirmed by quantitative PCR assay and Western blot analysis. Tissue-engineered bone was constructed with hMSCs and PHA/FN/ALG composite scaffolds, and then was implanted into an infected bone defect model for evaluating the osteogenic capacity by quantitative PCR, gross observation, micro-CT and histology analysis. Results: All those cells showed similar adhesion abilities and proliferation capacities in scaffolds. After tissue-engineered bone implantation, there were high levels of systemic inflammatory factors in vivo, which significantly declined three days after antibiotic therapy. One or two months after implantation, the results of osteogenic-related gene analyses, gross observation, micro-CT and histology consistently showed that the Wnt11 over-expression hMSC group displayed the strongest osteogenesis capacity, whereas the Wnt11-RNAi hMSC group displayed inferior osteogenesis capacity, when compared with the other cell-containing groups. However, the blank control group and the only composite scaffold without cell implantation group both showed extremely weak osteogenesis capacity. Conclusion: Our results revealed that the Wnt11 gene plays an important role in hMSCs for enhancing the osteogenesis in an infectious environment.

Objectives: The present work studied the mechanism of action of Wen-Dan decoction in treating steroid-induced necrosis of the femoral head (SINFH) via network pharmacology and in vivo experiments. Methods: The active compounds of Wen-Dan decoction and SINFH-related target genes were identified through public databases. Based on the results, further network pharmacological analysis was conducted to explore the potential key active compounds, core targets and biological processes of Wen-Dan decoction in SINFH. Subsequently, the potential mechanisms of action of Wen-Dan decoction in the treatment of SINFH obtained by network pharmacology analyses were validated through in vivo experiments. Results: A total of 608 DEGs (230 upregulated and 378 downregulated) in SINFH were identified. A GO functional enrichment analysis revealed that the SINFH-related genes were mainly involved in neutrophil activation and the immune response, and a KEGG pathway analysis showed that the SINFH-related genes were mainly associated with cytokine receptor interactions, lipids and atherosclerosis, and tuberculosis. In addition, a total of 147 active ingredients of Wen-Dan decoction were identified; of these, the core ingredient was quercetin, and licorice was an active ingredient. Moreover, 277 target genes in the treatment of SINFH with Wen-Dan decoction were identified, and NCF1, PTGS2, and RUNX2 were selected as core target genes. A quantitative real-time polymerase chain reaction (qRT–PCR) assay of peripheral blood from SINFH patients and control individuals showed that the patients exhibited highest levels of PGTS2 and NCF1 in blood. The qRT–PCR analysis of peripheral blood and femoral bone tissue from a mouse model of SINFH and control mice treated with quercetin showed that the PGTS2 and NCF1 levels revealed lower levels in the experimental animals compared with the controls, which was consistent with the bioinformatics analysis results. HE staining, immunohistochemistry and TUNEL staining confirmed significant improvement in hormone-induced femoral head necrosis in the mice treated with quercetin. Conclusions: This study provides new insights into the genes and related pathways involved in SINFH and shows that PTGS2 and NCF1 could be used as potential drug targets in SINFH. Quercetin could improve SINFH by promoting osteogenesis and inhibiting apoptosis.

Purpose: Segmental tibial diaphyseal bone defects after open fracture are difficult to treat and require timely, coordinated reconstruction. We aimed to develop evidence-based surgical guidelines based on meta-analysis and consensus recommendations from an expert committee of orthopedic surgeons. Methods: Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline, we searched 5 databases (1989 to 2024) for studies evaluating surgical treatment of post-traumatic tibial diaphyseal segmental defects. Random-effects models pooled union rate, union time, and complication rate across 4 techniques (Ilizarov bone transport [IBT], free vascularized bone grafting [FVBG], nonvascularized autografting [AG], and the Masquelet technique [MT]). Consensus recommendations were developed using a Delphi process. Results: Twenty-eight studies, including 824 patients (824 tibias) were analyzed: IBT (11 studies; n=361), FVBG (4; n=68), AG (10; n=164), and MT (6; n=67). Pooled union rates were 96% (IBT), 95% (FVBG), 95% (AG), and 81% (MT). Mean pooled union times were 33.04 weeks (IBT), 37.81 (FVBG), 29.73 (AG), and 29.74 (MT). Pooled complication rates were 82% (IBT), 94% (FVBG), 44% (AG), and 53% (MT). Conclusion: On the basis of the evidence and expert consensus, early operative management with prompt stabilization and antimicrobial therapy, supported by multidisciplinary care, is recommended. For large or complex defects, IBT and FVBG are generally preferred, while AG and MT remain options in selected scenarios. Outcomes depend on early intervention, appropriate technique selection, and structured rehabilitation; further high-quality studies are needed to validate these recommendations.