Background: Chitosan-based hydrogels exhibit excellent temperature-sensitive properties and are widely used as skin dressings. However, several challenges remain, such as long gelation times and difficulties releasing insoluble drugs, which limit their application in skin wound healing. In this study, we developed a novel sulfobutyl-β-cyclodextrin/quercetin@chitosan/hyaluronic acid hydrogel (Qe/SBE@CS/HA Gel). In this gel, SBE not only encapsulates Qe to form inclusion complexes, thereby enhancing the solubility of Qe, but also shortens the gelation time of thermosensitive gels through electrostatic adsorption with chitosan. Methods: Qe/SBE was prepared using the saturated solution method, while Qe/SBE@CS/HA gel was fabricated via electrostatic adsorption. The performance of the gels was evaluated using antibacterial, antioxidant, compatibility, and skin infection damage models. Results: The Qe/SBE@CS/HA Gel exhibits both thermosensitivity and acid sensitivity, releasing 91.9% of Qe in a medium with a pH of 5.0. This gel displays notable antibacterial activity and antioxidant characteristics. Furthermore, it shows excellent biocompatibility, as evidenced by hemolytic and in vivo degradation tests. The gel has the capacity to modulate chronic inflammation and facilitate angiogenesis and collagen synthesis, thereby significantly accelerating wound healing in wound and infection models. Conclusions: This multi-responsive and multifunctional gel shows potential as a therapeutic strategy for bacterial infection wounds.

Background: Non-alcoholic fatty liver disease (NAFLD) is triggered by gut-liver axis disorders; enzyme-ultrasound assisted extraction (EAUE) better retains bioactive mulberry polysaccharides (MP) than hot water extraction. This work optimized EAUE by RSM and GA-BP to compare the gut-hepatic protective capacity of two MP preparations and reveal their microecological regulatory mechanism. Methods: We optimized EAUE conditions and characterized the structural features of EAUE-derived MP (MPEU). A 30-day concurrent modeling and MP-administration mouse trial was conducted with biochemical detection and 16S rRNA sequencing to assess MPEU and hot water extraction MP (MPHW) in vivo activity. Results: RSM yielded predictions closer to experimental values, and its optimal extraction conditions were determined as extraction time 107 min, liquid-to-solid ratio 33:1, enzyme addition amount 350 mg, and extraction temperature 52 °C, yielding MP at 10.06 ± 0.03%. MPEU retained complete glycosidic structures and outperformed MPHW in mitigating hepatic steatosis, intestinal damage and inflammation, by restoring SCFA-producing commensal bacteria and suppressing pro-inflammatory flora. Conclusion: In this study, RSM is preferable for EAUE optimization. Structurally intact MPEU exerts stronger NAFLD relief via remodeling gut microecology, promising as nutritional functional food ingredients.

Background: The disruption of bone immune function and bone metabolism homeostasis is intricately linked to the onset and progression of glucocorticoid-induced osteoporosis (GIOP). From the perspective of indirect pharmacology, bone immune cells act as critical intermediate mediators that bridge systemic drug exposure and local bone tissue responses. Wubi Shanyao Pills (WBSY), a Chinese patent medicine, have been shown to modulate T lymphocyte levels in naturally aging mice and improve bone strength. However, whether WBSY exerts anti-GIOP effects through an indirect immunomodulatory pattern and its underlying intermediate mechanisms remain unclear. Purpose: This study aims to elucidate the effects and mechanisms of WBSY on GIOP in mice. Methods: Initially, the chemical constituents of WBSY were characterized using high-performance liquid chromatography (HPLC). Following this, a GIOP mouse model was developed through intraperitoneal administration of dexamethasone at a dosage of 10 mg/kg every three days. The mice were subsequently treated with WBSY at varying dosages—high (1.5 g/kg), medium (0.75 g/kg), and low (0.375 g/kg)—via gavage over a 20-week duration. Throughout the experimental period, general physiological parameters of the mice were systematically monitored. Bone mass alterations were subsequently assessed through bone strength measurements, micro-computed tomography analysis of bone tissue, and histopathological examination. Potential target pathways were identified via transcriptomic analysis. Furthermore, bone immunology-related parameters were evaluated using flow cytometry, including the proportion of T lymphocytes in the spleen and bone marrow, as well as the expression levels of the CXC chemokine ligand 10-chemokine receptor 3 (CXCL10-CXCR3) axis. Concurrently, key indicators of the bone metabolism pathway, such as the Receptor Activator of Nuclear Factor-κB (RANK), Receptor Activator of Nuclear Factor-κB ligand (RANKL), and osteoprotegerin (OPG), were further analyzed. Results: The results of the HPLC analysis demonstrated that the concentrations of echinacoside, acteoside, and schisandrol in WBSY were 7.82 mg/g, 2.18 mg/g, and 0.32 mg/g, respectively. Subsequently, the findings revealed that WBSY significantly enhanced general physiological parameters, including body weight, grip strength, and anal/back temperature, in GIOP mice. WBSY not only enhanced bone strength and mitigated pathological damage to the femur, but also significantly improved bone microarchitecture. This improvement was evidenced by an increase in bone volume/tissue volume, trabecular thickness, and trabecular number, alongside a reduction in trabecular separation and the structural model index. Furthermore, transcriptomic analysis indicated that WBSY may exert anti-GIOP effects by modulating key pathways, including osteoclast differentiation, interleukin-17A (IL-17A), and nuclear factor kappa B (NF-κB). At the immunological level, WBSY was shown to increase the proportion of T cells, B cells, and CD4⁺ T cells in the spleen, downregulate the expression of CXCL10 and CXCR3, and inhibit the activation of CD4⁺ Th17 cells in the femur. At the molecular level, WBSY modulated the expression of key molecules involved in the bone metabolism pathway, including RANK, RANKL, and OPG. It consistently downregulated the levels of femoral inflammation signaling molecules such as tumor necrosis factor receptor-associated factor 6 (TRAF6), phosphorylated IκB kinase β/IκB kinase β (p-IKKβ/IKKβ), and phosphorylated nuclear factor kappa B/nuclear factor kappa B (p-NF-κB/NF-κB). Additionally, WBSY reduced the concentrations of bone resorption markers, including tartrate-resistant acid phosphatase (TRAP) and β-C-telopeptide of type I collagen (β-CTX). Concurrently, it upregulated the expression of osteogenic molecules such as Runt-related transcription factor 2 (Runx2) and bone morphogenetic protein 2 (BMP2), leading to increased concentrations of bone formation markers, including bone-specific alkaline phosphatase (BALP), procollagen type I N-terminal propeptide (PINP), and osteocalcin (BGP). Conclusions: In summary, this study suggests that WBSY exerts anti-GIOP effects via an indirect pharmacological mechanism. WBSY may modulate the CXCL10-CXCR3 axis to inhibit aberrant T cell homing to the bone marrow, thereby reducing the excessive activation of bone marrow-resident Th17 cells and the secretion of the key bridging cytokine IL-17A. This indirect immunomodulation may subsequently restore the balance of the RANKL/RANK/OPG pathway, ultimately synergistically promoting bone formation and inhibiting bone resorption.