Patients with diabetic foot ulcers usually suffer from inefficient epithelisation and angiogenesis accompanied by chronic wound healing. Diabetic foot ulcers remain a major challenge in clinical medicine; however, traditional treatments are incapable of transdermal drug delivery, resulting in a low drug delivery rate. We report the development of Ti₂C₃ MXenes-integrated poly-γ-glutamic acid (γ-PGA) hydrogel microneedles to release asiaticoside (MN-MXenes-AS). Asiaticoside was loaded into PGA-MXenes hydrogel to facilitate cell proliferation while regulating angiogenesis. The characterisation and mechanical strength of the microneedles were investigated in vitro, and the wound-healing efficacy of the microneedles was confirmed in diabetic mice. MXenes significantly improved the mechanical strength of microneedles, while γ-PGA hydrogels provided a moist microenvironment for wound healing. Mice treated with MN-MXenes-AS demonstrated obvious improvements in wound healing process. We successfully fabricated an MXenes-integrated microneedle that possesses sufficient rigidity to penetrate the cuticle for subcutaneous drug delivery, thereby accelerating diabetic wound healing. We demonstrated that MN-MXenes-AS is effective in promoting growth both in vivo and in vitro. Collectively, our data show that MN-MXenes-AS accelerated the healing of diabetic foot ulcers, supporting the use of these microneedles in the treatment of chronic wounds. Graphical Abstract

Traumatic brain injury (TBI) is the leading cause for high morbidity and mortality rates in young adults, survivors may suffer from long-term physical, cognitive, and/or psychological disorders. Establishing better models of TBI would further our understanding of the pathophysiology of TBI and develop new potential treatments. A multitude of animal TBI models have been used to replicate the various aspects of human TBI. Although numerous experimental neuroprotective strategies were identified to be effective in animal models, a majority of strategies have failed in phase II or phase III clinical trials. This failure in clinical translation highlights the necessity of revisiting the current status of animal models of TBI and therapeutic strategies. In this review, we elucidate approaches for the generation of animal models and cell models of TBI and summarize their strengths and limitations with the aim of exploring clinically meaningful neuroprotective strategies.

Glioblastoma (GBM) is the most common type of primary malignant brain tumor. Molecular hydrogen has been considered a preventive and therapeutic medical gas in many diseases including cancer. In our study, we sought to assess the potential role of molecular hydrogen on GBM. The in vivo studies were performed using a rat orthotopic glioma model and a mouse subcutaneous xenograft model. Animals inhaled hydrogen gas (67%) 1 h two times per day. MR imaging studies were performed to determine the tumor volume. Immunohistochemistry (IHC), immunofluorescence staining, and flow cytometry analysis were conducted to determine the expression of surface markers. Sphere formation assay was performed to assess the cancer stem cell self-renewal capacity. Assays for cell migration, invasion, and colony formation were conducted. The in vivo study showed that hydrogen inhalation could effectively suppress GBM tumor growth and prolong the survival of mice with GBM. IHC and immunofluorescence staining demonstrated that hydrogen treatment markedly downregulated the expression of markers involved in stemness (CD133, Nestin), proliferation (ki67), and angiogenesis (CD34) and also upregulated GFAP expression, a marker of differentiation. Similar results were obtained in the in vitro studies. The sphere-forming ability of glioma cells was also suppressed by hydrogen treatment. Moreover, hydrogen treatment also suppressed the migration, invasion, and colony-forming ability of glioma cells. Together, these results indicated that molecular hydrogen may serve as a potential anti-tumor agent in the treatment of GBM.

Oxidative stress and chronic inflammation have been implicated in the pathophysiology of metabolic diseases, including diabetes mellitus (DM), metabolic syndrome (MS), fatty liver (FL), atherosclerosis (AS), and obesity. Molecular hydrogen (H₂) has long been considered a physiologically inert gas. In the last two decades, accumulating evidence from pre-clinical and clinical studies has indicated that H₂ may act as an antioxidant to exert therapeutic and preventive effects on various disorders, including metabolic diseases. However, the mechanisms underlying the action of H₂ remain unclear. The purpose of this review was to (1) provide an overview of the current research on the potential effects of H₂ on metabolic diseases; (2) discuss the possible mechanisms underlying these effects, including the canonical anti-oxidative, anti-inflammatory, and anti-apoptotic effects, as well as suppression of ER stress, activation of autophagy, improvement of mitochondrial function, regulation of gut microbiota, and other possible mechanisms. The potential target molecules of H₂ will also be discussed. With more high-quality clinical trials and in-depth mechanism research, it is believed that H₂ will eventually be applied to clinical practice in the future, to benefit more patients with metabolic disease.

The heterogeneous and highly plastic cell populations of macrophages are important mediators of cellular responses during all stages of wound healing, especially in the inflammatory stage. Molecular hydrogen (H₂), which has potent antioxidant and anti-inflammatory effects, has been shown to promote M2 polarization in injury and disease. However, more in vivo time series studies of the role of M1-to-M2 polarization in wound healing are needed. In the current study, we performed time series experiments on a dorsal full-thickness skin defect mouse model in the inflammatory stage to examine the effects of H₂ inhalation. Our results revealed that H₂ could promote very early M1-to-M2 polarization (on days 2–3 post wounding, 2–3 days earlier than in conventional wound healing), without disturbing the functions of the M1 phenotype. Time series analysis of the transcriptome, blood cell counts, and multiple cytokines further indicated that peripheral blood monocytes were a source of H₂-induced M2 macrophages and that the functions of H₂ in macrophage polarization were not only dependent on its antioxidant effects. Therefore, we believe that H₂ could reduce inflammation in wound care by shifting early macrophage polarization in clinical settings.

Background: To investigate the potential effects of molecular hydrogen on ovarian cancer. Methods: The in vivo study was performed using Hs38.T xenografted BALB/c nude mice. The mean tumor volume was monitored during 6 weeks of hydrogen inhalation. Immunohistochemistry (IHC) staining was performed to determine the Ki67 and CD34 expression. The in vitro effects of molecular hydrogen on proliferation of Hs38.T and PA-1 were determined by Cell Counting Kit (CCK)-8 assay. Matrigel invasion assay was performed to determine the cell invasion ability. Wounding assay was employed to examine the motile nature of ovarian cancer cells. Colony formation assay was performed to investigate the effect of molecular hydrogen in tumorigenicity. To further investigate the effects of molecular hydrogen on cancer stem cells (CSCs) properties, we performed sphere-formation assays. Results: The in vivo study demonstrated that 6 weeks of hydrogen inhalation significantly inhibited tumor growth, as evidenced by decreased mean tumor volume (32.30%) and Ki67 expression (30.00%). Hydrogen treatment decreased the expression of CD34 (74.00%) demonstrating its anti-angiogenesis effects. The in vitro study showed that hydrogen treatment significantly inhibits cancer cell proliferation, invasion, migration and colony formation both in Hs38.T and PA-1 cells. An important finding in this study was that molecular hydrogen could also markedly inhibit sphere-forming ability of both PA-1 and Hs38.T cells. Conclusions: Molecular hydrogen may exert anti-tumor role in ovarian cancer through suppressing the proliferation of CSCs-like cells and angiogenesis.