Benfotiamine promotes diabetic wound healing by modulating Trisk95 and reducing oxidative stress.

Benfotiamine promotes diabetic wound healing by modulating Trisk95 and reducing oxidative stress - PMC Skip to main content An official website of the United States government Here's how you know Here's how you know Official websites use .gov A .gov website belongs to an official government organization in the United States. Secure .gov websites use HTTPS A lock ( Lock Locked padlock icon ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites. Search Log in Dashboard Publications Account settings Log out Search… Search NCBI Primary site navigation Search Logged in as: Dashboard Publications Account settings Log in Search PMC Full-Text Archive Search in PMC Journal List User Guide PERMALINK Copy As a library, NLM provides access to scientific literature. Inclusion in an NLM database does not imply endorsement of, or agreement with, the contents by NLM or the National Institutes of Health. Learn more: PMC Disclaimer | PMC Copyright Notice Diabetol Metab Syndr . 2026 Mar 7;18:101. doi: 10.1186/s13098-026-02134-y Search in PMC Search in PubMed View in NLM Catalog Add to search Benfotiamine promotes diabetic wound healing by modulating Trisk95 and reducing oxidative stress Shumail Maqbool Shumail Maqbool 1 Department of Biomedicine, Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences & Technology (NUST), Sector H-12, Islamabad, Pakistan Find articles by Shumail Maqbool 1 , Hussain M Wahedi Hussain M Wahedi 1 Department of Biomedicine, Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences & Technology (NUST), Sector H-12, Islamabad, Pakistan Find articles by Hussain M Wahedi 1, ✉ Author information Article notes Copyright and License information 1 Department of Biomedicine, Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences & Technology (NUST), Sector H-12, Islamabad, Pakistan ✉ Corresponding author. Received 2025 Aug 13; Accepted 2026 Feb 17; Collection date 2026. © The Author(s) 2026 Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/ . PMC Copyright notice PMCID: PMC13081506  PMID: 41794782 Abstract Impaired wound healing in diabetic patients is a major clinical challenge, often associated with mitochondrial dysfunction and oxidative stress. Excessive ROS production increases inflammation and disrupts tissue regeneration. Current wound treatments primarily target infection control and wound closure, but do not address the underlying factors. Therefore, there is a critical need for therapies that target the underlying mechanisms sustaining impaired healing in diabetic wounds. This study investigates the therapeutic potential of benfotiamine in promoting diabetic wound healing by modulating Trisk95 expression and reducing oxidative stress. Molecular docking for benfotiamine-Trisk95 interaction and KEGG pathway analysis of benfotiamine-targeted genes were performed, which are related to diabetic complications. In vivo experimentation was conducted in diabetic mouse models, and benfotiamine was applied topically to the wounds to examine its effect on wound healing rate. The expression of Trisk95, IL-6, and IL-10 genes was analyzed, and antioxidant effects were evaluated by analyzing ROS levels. In in silico studies, molecular docking showed stable binding of benfotiamine to Trisk95, indicating a strong ligand–protein interaction. The KEGG analysis showed that benfotiamine-targeted genes are crucial mediators of oxidative stress and inflammation in diabetic complications and are linked to mitochondrial function. In vivo studies confirmed that Trisk95 expression is upregulated in diabetic mice as compared to non-diabetic mice. Further experiments showed that benfotiamine accelerates wound closure by significantly downregulating Trisk95 and IL-6, while upregulating IL-10 expression. Furthermore, benfotiamine increased the activity of antioxidant enzymes, including superoxide dismutase and Catalase, resulting in reduced ROS levels in wound tissues. In conclusion, these findings suggest that benfotiamine may be a potential candidate for promoting mitochondrial function and skin regeneration, which ameliorate wound healing in diabetic mice by modulating Trisk95 and exhibiting anti-inflammatory and antioxidant effects. Keywords: Trisk95, Diabetic wound healing, Antioxidant, Anti-inflammatory, ROS Background Diabetes Mellitus, a silent killer that has claimed millions of lives annually, serves as a stark reminder of the global health crisis and is a critical health emergency of the twenty-first century [ 1 ]. In 2021, over 537 million individuals worldwide were reported to have diabetes, and projections estimate a further increase to 643 million by 2030 and 784 million by 2045 [ 2 ]. Diabetes mellitus leads to impaired wound healing, which frequently results in amputation [ 3 , 4 ]. It is estimated that out of half a billion diabetic individuals worldwide one-third are likely to develop diabetic foot ulcer which leads to amputation in 17% individuals [ 5 ]. The continuous elevation of blood glucose levels impairs mitochondrial function, increases reactive oxygen species (ROS) levels, and induces damage to mitochondrial DNA, thereby contributing to a delayed wound-healing process [ 6 , 7 ]. Hyperglycemia also results in endoplasmic reticulum (ER) stress and mitochondrial dysfunction in diabetic wounds, contributing to cell dysfunction [ 8 ]. Hence, Diabetic wound healing is primarily influenced by chronic hyperglycemia-induced mitochondrial dysfunction, oxidative stress, and impaired calcium homeostasis [ 9 ]. A critical player in this process is Trisk95, a transmembrane protein that regulates excitation–contraction coupling [ 10 ] and is involved in the controlled calcium release from the sarcoplasmic reticulum (SR) [ 11 ]. Recent studies highlight that Trisk95 expression is upregulated in response to elevated glucose levels, independently of insulin signaling. Overexpression of Trisk95 affects the amount of calcium in the ER store. The upregulated Trisk95 form Trisk95/CSQ-2 complex binds to the ryanodine receptor (RyR-1) channels and closes them, thereby increasing the calcium storage in the ER. Increased ER calcium triggers mitochondrial changes by increasing the mitochondrial calcium uniporter (MCU) and MICU1 expression, activating the MCU complex in the skin cells, and facilitating calcium uptake [ 12 ]. Since elevation in intracellular Ca 2+ is the first damage signal [ 13 , 14 ] and is important in regulating mitochondrial function [ 15 , 16 ]. Increased calcium levels can affect mitochondrial function, altering mitochondrial metabolism and morphology [ 17 ]. These disruptions propagate oxidative stress and inflammatory cascades that compromise the tissue repair mechanisms critical for efficient wound healing [ 18 ]. Furthermore, hyperglycemia also results in increased ROS production, leading to oxidative damage [ 19 , 20 ]. This occurs due to a higher influx of reducing equivalents into the electron transport chain of mitochondria [ 21 – 23 ]. In dysregulated conditions, ROS increases inflammation [ 24 ] and promotes inflammatory factor production by altering fibroblasts, endothelial cells, and keratinocytes [ 25 – 27 ]. This may result in inflammatory damage, cellular senescence, and cell death, impairing the healing process [ 28 ]. Despite the recognized importance of oxidative stress in diabetic wounds, the direct molecular mechanisms linking Trisk95 to wound healing impairment remain unknown. Benfotiamine (BFT) is a synthetic derivative of thiamine [ 29 ] that has anti-inflammatory, anti-oxidative, and other therapeutic effects. It has been widely studied and used for treating diabetic complications, neurodegenerative diseases, and inflammatory conditions [ 30 ]. Most importantly, BFT reduces ROS production and functions as a direct antioxidant [ 31 ]. In high glucose levels, BFT regulates the intracellular glucose and counteracts the detrimental effects of hyperglycemia [ 32 – 35 ]. Hence, BFT effectively alleviates inflammation and oxidative stress [ 36 ]. Thus, it makes the BFT a potential candidate for diabetic wound healing by affecting underlying mechanisms . Despite advances in diabetic wound healing research, the specific involvement of the calcium-handling protein Trisk95 remains inadequately characterized. Its dysregulation and subsequent effects on mitochondrial function and oxidative stress in diabetic skin have not been thoroughly investigated. Moreover, although BFT is known for its antioxidant and anti-inflammatory properties, its influence on Trisk95 and associated pathways in diabetic wound healing has not been reported. This study addresses this critical gap by investigating how BFT regulates Trisk95 and evaluating its impact on oxidative stress and skin regeneration in diabetic wounds. In this regard, the study aimed to explore the effect of Trisk95 downregulation in improving delayed healing in diabetic wounds. It was hypothesized that BFT may downregulate the Trisk95 level and reduce oxidative stress and pro-inflammatory response, thereby restoring mitochondrial function at the wound site, which may lead to improved wound healing in diabetic mice. Methods Molecular docking of BFT with Trisk95 Molecular docking studies were performed to evaluate the binding interaction between BFT and Trisk95. The 3D structure of Trisk95 was determined from AlphaFold3 and was refined by the Galaxyweb server, as the structure was not available on the PDB database. The BFT ligand structure was taken from PubChem and prepared for docking using the UCSF Chimera software. Then, Protein–Ligand docking simulations were carried out using the SwissDock software. Docking scores and binding energies were recorded, and ligand–protein interactions such as hydrogen bonds, hydrophobic interactions, and π-π stacking were analyzed. In silico functional enrichment analysis of BFT-targeted genes Additionally, KEGG pathway enrichment was performed to identify signaling pathways potentially affected by BFT and involved in diabetic wound repair. BFT-targeted genes were identified from existing literature, and genes closely related to diabetic complications were shortlisted, which may play a role in mitochondrial functioning and diabetic wound healing. To elucidate the function of these BFT-targeted genes, downstream-targeted genes were identified. The sources used to identify these genes include PubMed, Google Scholar, KEGG, and Wiki Pathways. To further explore the functional implications of these genes, KEGG pathway analysis and Functional Enrichment GO Biological Process were performed using ShinyGO0.77 software, which maps these genes to specific signaling pathways. Diabetic mouse model development for in vivo experiments The experiment was performed on Balb/c 6–7 weeks old male mice (bought from the National Institute of Health (NIH), Islamabad), weighing 25-35g. All animal experiments were conducted according to the ethical guidelines outlined in the NIH publication #85–23 (Revised 1985) on the Care and Use of Laboratory Animals and were approved by the NUST Institutional Review Board (IRB) (IRB No.: 11–2023-ASAB-01/01). Hypoglycaemic conditions that are close to Type 1 diabetes (T1D) were induced by a single intraperitoneal injection of 200mg/kg Alloxan (Sigma-Aldrich, USA) in normal saline. Blood glucose was measured using a handheld glucometer (Accu-Chek, Roche Diagnostics, USA) with blood obtained from the tail vein. Mice having blood glucose levels above 300mg/dL a few days post-injection were considered diabetic and were included in the experiment. Trisk95 expression analysis in diabetic and non-diabetic mice Following diabetes induction, the pilot study was conducted to assess the expression of Trisk95 in the skin tissue of diabetic and non-diabetic (normal control) mice. For this, mice were divided into two groups, each group containing 3 mice (n = 3). The hair was removed from the dorsal posterior region. Then, mice were sacrificed by overdose of Chloroform, followed by cervical dislocation as a secondary method to collect the skin tissue samples for further analysis. Wound induction and drug application For further experiments, diabetes was induced in the Balb/c mice, and the wounds were created by anesthetizing the mice with Ketamine (1mg/kg body weight) injections. Then the hair was removed from the dorsal posterior region, and the area was cleaned with 70% ethanol. Two cutaneous wounds were created employing a 6mm Biopsy Punch. The mice were randomly divided into four groups, each containing 7 mice (n = 7): Control (no treatment), Vehicle (0.5% CMC), 0.05% BFT, and 0.1% BFT groups. 200µL solution of BFT (Acmec Biochemical Co., Ltd., China) was applied topically to the wounds daily, dissolved in 0.5% carboxymethylcellulose (CMC) (Sigma-Aldrich, USA). The wounds of all groups were monitored and photographed daily. Mice were sacrificed after a 10-day experiment on days 3, 7, and 10. Histological analysis For histological analysis, tissue samples from the wound on the 3rd, 7th, and 10th day were fixed in a 10% formalin solution and then embedded in paraffin wax. Then, tissues were sliced into 4 μm-thick sections and stained with hematoxylin and eosin (H&E) and Mason’s trichrome staining. The stained sections were visualized and photographed by a light microscope, and the images were analysed using ImageJ software. H&E slides were also analyzed for white cell infiltrate, and the assessment was based on qualitative scoring of subepithelial mixed white cell infiltration (1 = absent, 2 = mild, 3 = moderate, 4 = marked, 5 = exuberant). Gene expression analysis Total RNA was isolated from wounded skin tissue samples by utilizing the Trizole (Invitrogen, USA) method, and cDNA was synthesized using cDNA synthesis kit (Thermo Scientific, USA). RT-PCR was performed on the same biological sample three times (technical triplicates) for gene expression analysis of Trisk95 and inflammatory cytokines, IL-6 and IL-10. Gene expression was normalized to GAPDH and analyzed using the 2^–ΔΔCt method. Biochemical profiling RIPA buffer was added to the skin tissue to prepare a protein lysate based on weight. The tissue was minced and centrifuged at 14,000 rpm at 4°C for 10 min. After centrifugation, the supernatant was collected and stored at -20 °C. ROS measurement The ROS assay was conducted on the same biological sample three times by following the established protocol [ 37 ]. Reagent 1 (R1) was formulated at 100 µg/ml concentration using N, N-diethyl-para-phenylendiamine (DEPPD) (Fluka Analytical) in a 0.1 M sodium acetate (VWR International Ltd, England) buffer. Reagent 2 (R2) was made by dissolving 0.5% FeSO4 (Sigma Aldrich, USA) in the same buffer solution. R1 and R2 were subsequently combined in a 1:25 ratio. For the assay, 5 µl of the sample and 140 µl of buffer were added to each well of a 96-well plate, then incubated at room temperature for 5 min. After the incubation period, 100 µL of the R1: R2 mixture was added to each well, and the absorbance was measured at 505 nm using an Agilent UV–Visible spectrophotometer (UK). A standard calibration curve for the buffer was utilized to determine the levels of ROS. Finally, the ROS levels were adjusted to the total protein concentration. SOD activity The measurement of SOD activity was conducted on the same biological sample three times, according to an established protocol [ 38 ]. A solution was made using potassium phosphate (Sigma Aldrich, USA) buffer, L-methionine (Scharlau, Spain), Nitroblue tetrazolium salt (NBT) (Thermo Scientific, USA), and Triton X-100 (Solarbio, China). Following this, 5 µl of the protein lysate was added to each well of a 96-well plate, together with 243 µl of the reaction mixture. The plate was then exposed to a fluorescence lamp for 7 min. After this exposure, the plate was allowed to incubate at room temperature for 5 min. Subsequently, 3 µl of chilled riboflavin (BDH Laboratory Supplies, England) was introduced, and the plate was incubated at room temperature for 8 min. After the second incubation period, three absorbance measurements were taken at 540 nm at 1-min intervals using an Agilent UV–Visible spectrophotometer (UK). Catalase measurement Catalase was measured on the same biological sample three times by using H 2 O 2 as a substrate [ 38 ]. For catalase measurement, 8.09 µl sample, 161µl 50mM potassium phosphate buffer pH 7, and 80.9µl 5.9mM H 2 O 2 (E. Merck, Germany) were added to the 96-well plate. The 3 absorbance readings were recorded at 240nm at 30-s intervals using an Agilent UV–Visible spectrophotometer (UK). Statistical analysis Tissue samples from each animal were processed individually, and each assay was performed on the same biological sample three times (technical triplicates). Data is presented as the mean ± standard deviation. The statistical analysis was performed using GraphPad Prism (GraphPad Software, USA). For comparisons between two groups, an unpaired t-test is used. For comparisons among three or more groups, a two-way analysis of variance (ANOVA) followed by Tukey’s multiple comparisons test is used to compare more than two groups. The probability values of P < 0.05 were considered statistically significant. Results Molecular docking of BFT with Trisk95 and KEGG pathway enrichment analysis elucidate mechanistic role of BFT in diabetic wound healing BFT showed a favorable binding affinity to the Trisk95 active site with a docking score of -6.028 kcal/mol (Table 1 ), indicating a strong ligand–protein interaction. The molecular docking analysis revealed key hydrogen bonds formed between BFT and amino acid residues ASP72, ASP69, and THR99 of Trisk95, along with hydrophobic interactions that stabilize the complex. Visualization confirmed that BFT occupies a pocket critical for Trisk95 function, suggesting potential modulation of its activity (Fig. 1 A). Table 1. Molecular docking models and binding affinity scores for BFT-Trisk95 interaction Model Calculated affinity (kcal/mol) 1 -6.028 2 -5.583 3 -5.428 4 -5.426 5 -5.256 6 -5.212 7 -5.183 8 -5.154 9 -5.123 10 -5.061 11 -5.048 12 -5.027 13 -5.003 14 -4.930 15 -4.903 16 -4.849 17 -4.797 18 -4.734 19 -4.647 20 -4.544 Open in a new tab Fig. 1. Open in a new tab Molecular docking of BFT with Trisk95 and KEGG pathway enrichment analysis of BFT-targeted genes linked to diabetic complications. ( A ) 3D representation of the predicted different binding poses of BFT at the Trisk95 binding pocket after molecular docking. ( B ) , ( C ) The enriched pathways of BFT targeted genes. ( D ), ( E ) The enriched pathways of downstream targeted genes. The chart shows the top 10 enriched pathways and biological functions in which these genes are involved. The color gradient corresponds to the -log10 FDR values, where red represents the highest significance and blue represents the lowest significance The KEGG Analysis map shows that BFT-targeted genes are predominantly involved in EGFR tyrosine kinase inhibitor resistance and the AGE-RAGE signaling pathway in diabetic complications (Fig. 1 B). The Functional Enrichment GO Biological Process shows that these genes are mainly involved in response to ROS, chemical stress, apoptosis, and programmed cell death (Fig. 1 C). To further understand how these initial gene targets translate into cellular outcomes, a second KEGG analysis was conducted, focusing on the downstream targets of these genes. Results identify that the downstream genes are significantly involved in platinum drug resistance, measles, and apoptosis signaling pathways (Fig. 1 D). The Functional Enrichment GO Biological Process of downstream genes shows that they play a critical role in mitochondrial membrane permeabilization (Fig. 1 E), which is a vital aspect of cellular health, predominantly in the context of diabetic wounds. List of BFT-Trisk95 docking conformations and their estimated binding energies (kcal/mol) from molecular docking simulations. Trisk95 expression is elevated in diabetic mice skin and topical application of BFT improves wound healing in diabetic mice The mRNA expression of Trisk95 of diabetic and normal control mice showed a significant upregulation in diabetic skin tissues (Fig. 2 A). This observation aligns with previous reports indicating elevated Trisk95 expression in response to hyperglycemia, independent of insulin signaling pathways [ 12 ]. Fig. 2. Open in a new tab Expression of Trisk95 in diabetic and non-diabetic mouse skin samples and Topical application of BFT on wound healing in diabetic mice. ( A ) Graphical representation of mRNA expression of Trisk95, demonstrating significant upregulation in diabetic skin samples compared to controls (95% CI = 6.202 to 9.890, p = 0.0003). The data were statistically analyzed by an unpaired t-test and are presented as mean ± SD. ***p < 0.001 vs. control. ( B ) Photographic representation of wound healing processed on Days 0, 3, 7, and 10 after the puncture. Scale bar set at 6mm. ( C ) Graphical representation of average wound area on days 0, 1, 3, 5, 7, 9, 10 (n = 7) with Significant differences compared to control, 95% CI = 3.039 to 18.68, P value = 0.0024. ( D ) Graphical representation of wound closure rate (%) on days 3, 5, 7, 9, 10 (n = 7). All the data were statistically analysed by an unpaired t-test (A), two-way ANOVA, and are presented as mean ± SD. **p < 0.01, *p < 0.05 vs control The pictorial representation of mouse wounds showed faster wound closure and dermal regeneration in BFT-treated mice than in the control and vehicle mice. Specifically, the wound size decreased significantly in 0.1% of BFT-treated mice than in other groups (Fig. 2 B). On days 7 and 10, the 0.1% BFT-treated group displayed a notable reduction in wound size, with areas of 8.9 mm 2 and 3.9 mm 2 , respectively, resulting in closure rates of 71.2% and 88%. In contrast, the wound sizes in the other groups did not exhibit significant contraction during this period (Fig. 2 C, D). BFT promotes skin tissue development and collagen deposition H&E staining demonstrates that wounds of diabetic mice, treated by BFT, formed complete and uniform dermal and epidermal structures on day 10 (Fig. 3 A, B). Figure 3 C shows prolonged inflammation in the control and vehicle groups and significantly accelerated resolution in the 0.1%BFT group. Mason’s Trichrome staining showed more collagen deposition in the BFT-treated groups (Fig. 4 ). However, the 0.1%BFT-treated group showed well-defined epidermal and dermal layers and better cellular arrangement, and the mice in the remaining groups showed irregular and incomplete structures of the dermis and epidermis. Fig. 3. Open in a new tab BFT enhances epidermal regeneration in diabetic mice at the wound site. ( A ) Epidermal regeneration was assessed through H&E staining on days 3, 7, and 10 of wounded skin samples of all groups. Images were captured at 10X, scale bar set at 100 μm. ( B ) Graphical representation of recovered epidermis area on Days 3, 7, and 10 (n = 7) with Significant differences compared to control on day 10, 95% CI = -35.28 to -24.32, P < 0.0001. ( C ) Qualitative scoring of white cell infiltration on days 3, 7, and 10. Black arrows = Recovered Epidermis, Blue arrows = White Cell Infiltration. All the data was statistically analyzed by two-way ANOVA and is presented as mean ± SD. ****p < 0.0001 vs. control Fig. 4. Open in a new tab BFT promotes collagen deposition in diabetic mice at the wound site. ( A ) Collagen deposition was assessed through Masson’s trichrome staining on days 3, 7, and 10 of wounded skin samples of all groups of mice. Scale bar set at 100 µm ( B ) Graphical representation of collagen deposition on Days 3, 7, and 10 (n = 7) with Significant differences compared to control on day 10, 95% CI = -25.97 to -20.32, P < 0.0001. Red arrows = Collagen Deposition. All the data was statistically analyzed by two-way ANOVA and is presented as mean ± SD. ****p < 0.0001 vs. control BFT downregulates Trisk95 and reduces hyperglycemia-induced oxidative stress The impact of BFT was evaluated on the gene expression of Trisk95, which is involved in Ca 2+ signaling (Fig. 5 A). BFT significantly downregulated the Trisk95 expression, thereby improving mitochondrial function. Fig. 5. Open in a new tab BFT downregulates Trisk95 and reduces hyperglycemia-induced oxidative stress. Graphical representation of ( A ) Trisk95, ( B ) ROS, ( C ) SOD, and ( D ) Cat on Days 3, 7, and 10 of all groups (n = 3) with Significant differences compared to vehicle on day 10 of Trisk95, 95% CI = 16.27 to 21.51, P value =  < 0.0001. All the data was statistically analyzed by two-way ANOVA and is presented as mean ± SD. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05 vs control Then, it was assessed whether BFT could reduce the high level of ROS at the wound site and exert an antioxidant effect that prevents cellular damage by catalyzing the conversion of detrimental peroxides to neutral compounds and improve mitochondrial function in diabetic wounds (Fig. 5 B, C, D). Results show that BFT could reduce the ROS production. Further analysis shows that BFT employs an antioxidant effect by enhancing the activity of antioxidant enzymes, Cat and SOD. BFT reduces inflammatory response in diabetic wounds As ROS is closely linked to inflammation in the wound microenvironment, mRNA expression of IL -6 and IL-10 cytokines was examined in wounded skin tissues (Fig. 6 A, B). BFT significantly downregulated the IL-6 expression and upregulated the IL-10 expression. Fig. 6. Open in a new tab BFT reduces inflammatory response in diabetic wounds. Graphical representation of mRNA expression of ( A ) IL-6, and ( B ) IL-10 on days 3, 7, and 10 of all groups (n = 3) with Significant differences compared to vehicle on day 10, 95% CI of IL6 = 7.520 to 10.15, 95% CI of IL10 = -21.45 to -10.22, P value =  < 0.0001. All the data was statistically analyzed by two-way ANOVA and is presented as mean ± SD. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05 vs control Discussion The increasing evidence shows significant oxidative stress and mitochondrial dysfunction in diabetic wounds [ 39 ]. The persistent rise in blood glucose levels impairs mitochondrial function and increases ROS production, which in turn affects the wound healing process adversely [ 6 , 7 ]. In any case, proper mitochondrial functioning and ROS levels are considered crucial in promoting wound healing and preventing other diseases such as Alzheimer’s and aging. This study employed BFT to investigate its wound-healing properties in diabetic mice by regulating Trisk95, oxidative stress, and inflammation at the wound site. BFT shows antioxidant and anti-inflammatory capabilities that help improve diabetic complications. [ 30 , 40 ]. Molecular docking of BFT and Trisk95 indicated a stable ligand–protein interaction. These findings suggest that BFT may directly interact with Trisk95, influencing its role in calcium handling and mitochondrial regulation, which is further correlated with the observed in vivo improvements in wound healing. Subsequently, KEGG pathway enrichment analysis was conducted to explore broader signaling cascades associated with BFT-targeted genes involved in diabetic complications. KEGG analysis showed that BFT-targeted genes were predominantly involved in the AGE-RAGE signaling pathway in diabetic complications. This includes Ca 2+ signaling, ROS, IL-6, TNF-α, and RAGE, which are crucial mediators of inflammation and oxidative stress in diabetic complications. The KEGG analysis of the downstream targets of these genes showed their intricate link to mitochondrial function, a critical aspect of cellular health, particularly in the context of diabetic wounds. In diabetic wounds, mitochondrial dysfunction is observed [ 39 ]. Given the connection among these pathways and mitochondrial function and ROS, the observed gene alterations may ultimately impact mitochondrial health and ROS levels, affecting diabetic wound healing. Pathways that are linked to mitochondrial dysfunction likely contribute to impaired wound healing under hyperglycemic conditions [ 8 ]. In in-vivo experimentation, elevated Trisk95 level in diabetic mice ascompred to the non-diabetic mice establish a foundational understanding of Trisk95 dynamics in normal versus diabetic conditions, providing crucial context for subsequent evaluation of BFT’s modulatory effects in the diabetic wound models. BFT showed faster wound closure, increased epidermal regeneration, and collagen deposition at the wound site. Increased cases show that delayed chronic diabetic wound healing is mainly linked to ER stress, mitochondrial dysfunction, oxidative stress, and chronic inflammation [ 41 ]. Increased skin glucose upregulates the Trisk95 level, which causes mitochondrial damage [ 12 ]. It was observed that BFT downregulated the level of Trisk95 at the wound site in diabetic mice, as it is evident that BFT in hyperglycaemic conditions markedly enhances glucose oxidation and promotes the absorption of glucose [ 42 ]. BFT reduces SR Ca 2+ load and reverses the hyperglycemia-induced disruptions in Ca 2+ homeostasis [ 43 ]. This suggests that, by downregulating Trisk95, BFT may restore calcium-related mitochondrial dysfunction and alleviate oxidative stress in diabetic wounds . Our findings raise the possibility that BFT-mediated modulation of Trisk95 may intersect with calcium signaling and mitochondrial integrity, processes intimately linked with wound repair. However, further mechanistic studies are needed to confirm these associations. Furthermore, BFT triggers a crucial biochemical pathway in hyperglycemic conditions that prevents AGE formation and oxidative stress [ 40 , 44 ]. Results indicate that BFT, especially at higher doses, effectively modulated oxidative stress by regulating ROS levels and enhancing the key antioxidant enzymes, including SOD and Cat. Under normal physiological conditions, ROS is essential to overcome microbial invasion and to regulate intracellular signaling pathways, which alter wound healing [ 45 ]. However, excessive oxidative stress can damage the structure and DNA of cells, which may ultimately cause cell death and subsequent tissue damage [ 46 ]. Also, ROS has harmful effects on cellular homeostasis, including increased redox imbalance [ 47 ]. It also triggers an antioxidant effect, with the help of SOD, Cat, and glutathione reductase enzymes [ 48 ]. SOD catalyzes the conversion of superoxide radicals into hydrogen peroxide [ 49 ]. Likewise, Cat catalyzes the breakdown of H 2 O 2 into water and oxygen, further contributing to the reduction of oxidative stress [ 50 ]. Jha et al. (2025) highlighted that balanced upregulation of antioxidant enzymes, SOD, together with downstream scavengers like Cat and GSH, is more effective in reducing oxidative stress and supporting wound healing than changes in a single enzyme [ 51 ]. This aligns with our findings, as BFT promoted coordinated increases in SOD and Cat, suggesting a coordinated antioxidant response that not only lowers oxidative burden but may also restore redox-sensitive signaling important for tissue repair. In hyperglycemic conditions, BFT also shows anti-inflammatory properties [ 48 ]. This study found that BFT significantly reduced the pro-inflammatory cytokine IL-6 after day 7, as proinflammatory cytokines are essential for initiating wound healing [ 52 ]. IL-6 is also crucial for timely wound healing [ 53 , 54 ], because it stimulates the release of pro-inflammatory cytokines and also produces anti-inflammatory cytokines like IL-10 [ 55 ]. BFT upregulated the expression of anti-inflammatory cytokine IL-10. In postnatal cutaneous dermal wounds, overexpression of IL-10 promotes scarless tissue repair by regulating inflammation and promoting extracellular wound matrix formation [ 56 ]. This showed that BFT provides the proper resolution of inflammation and tissue repair. It is crucial since dysregulated inflammatory cytokine signaling can lead to a prolonged proliferative phase and hypertrophic scar formation [ 57 ]. Such dysregulation is commonly observed in various diabetic complications, where altered proinflammatory cytokine levels contribute to the chronic inflammation associated with non-healing diabetic ulcers [ 58 ]. Our study demonstrates that BFT accelerates diabetic wound healing, evidenced by improved wound closure rates and favorable modulation of oxidative stress markers and inflammation. These findings align well with recent reports on antioxidant therapies for improving diabetic wound repair. Majie et al. (2024a, 2024b) investigated phenylethanoid glycosides (PhGs) from Clerodendrum glandulosum and demonstrated, through in silico and in vivo studies, that these compounds exhibit potent antioxidant, anti-inflammatory, and antimicrobial effects, which led to enhanced wound closure and extracellular matrix formation in diabetic models [ 59 , 60 ]. Notably, antioxidants like N-acetylcysteine (NAC) primarily restore glutathione levels and reduce oxidative injury, thereby promoting angiogenesis and tissue repair [ 61 , 62 ], and resveratrol improves wound healing through sirtuin-1 activation and modulation of inflammatory signaling [ 63 , 64 ]. These findings underscore the therapeutic potential of targeting oxidative stress and inflammation through diverse molecular mechanisms to optimize diabetic wound healing. Notably, our study also identified the effect of BFT on Trisk95 expression. This suggests that BFT may influence additional pathways beyond classical antioxidant defenses, potentially involving calcium signaling and mitochondrial stability. Despite these valuable insights, some limitations should be acknowledged. First, our work was conducted in a murine model, which, although widely used in preclinical wound healing research, does not fully recapitulate the complexity of human diabetic wounds. Second, the direct mechanistic link between BFT treatment and Trisk95 modulation is correlative rather than causative. Though, we assessed Trisk95 levels, but we did not directly evaluate downstream functional pathways such as calcium signaling, mitochondrial activity, or apoptosis, leaving important mechanistic questions unresolved. Additionally, potential pharmacological interactions between BFT and medications commonly prescribed to diabetic patients were not addressed and warrant future investigation. Future research should therefore focus on establishing a direct relation through approaches such as Trisk95 knockdown or overexpression and delineating the precise molecular pathways linking BFT with calcium homeostasis, mitochondrial function, and apoptosis, as well as testing whether combining BFT with other antioxidant compounds produces additive or synergistic effects. Ultimately, controlled clinical trials will be required to establish whether the benefits observed in animal models translate into meaningful improvements in human diabetic wound management. Conclusion Thus, this study demonstrates that BFT significantly regulates Trisk95 expression, contributing to reduced oxidative stress and improved skin regeneration in diabetic wounds (Fig. 7 ). These findings highlight Trisk95 as a promising novel therapeutic target for enhancing diabetic wound healing due to its critical role in calcium homeostasis and modulation of mitochondrial function in skin cells. Targeting Trisk95 could lead to the development of more effective treatment strategies aimed at restoring calcium balance and mitochondrial health, thereby reducing oxidative damage and promoting efficient skin regeneration. Our findings with BFT’s regulation of Trisk95 further underscore its potential as a molecular intervention to enhance antioxidant defenses and improve healing outcomes in diabetic wounds. Future studies should focus on fully elucidating the signaling pathways involved and validating Trisk95-targeted therapies in clinical settings. Fig. 7. Open in a new tab Schematic summary of the beneficial effects of BFT on diabetic wound healing Acknowledgements The authors acknowledge the infrastructure and financial support provided by the National University of Sciences and Technology, Islamabad. The authors acknowledge the technical support provided by the cell biology lab at Quaid-i-Azam University, Islamabad. Abbreviations AGE Advanced glycation end products BFT Benfotiamine Ca 2+ Calcium Cat Catalase CMC Carboxymethylcellulose DEPPD N, N-diethyl-para-phenylendiamine GSH Glutathione H 2 O 2 Hydrogen peroxide IL-6 Interleukin-6 MCU Mitochondrial calcium uniporter NAC N-acetylcystein NBT: Nitroblue tetrazolium salt PhGs Phenylethanoid glycosides ROS Reactive oxygen species RyR Ryanodine receptor SOD Superoxide dismutase SR Sarcoplasmic reticulum T1D Type 1 diabetes TNF Tumor necrosis factor Author contributions Hussain M. Wahedi: conceptualized the study and provided technical supervision for experiments and manuscript preparation. Shumail Maqbool: performed the experiments and prepared the manuscript. Funding The National University of Sciences and Technology funded this study. Data availability The data that support the findings of this study are available from the corresponding author upon reasonable request. Declarations Ethics approval and consent to participate All animal practices conformed to the National Institutes of Health Guidelines on the Use of Laboratory Animals and were approved by the NUST Ethical Review Committee. Consent for publication Not applicable. Competing interests The authors declare no competing interests. Footnotes Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. References 1. Geng K, Ma X, Jiang Z, Huang W, Gu J, Wang P, et al. High glucose-induced STING activation inhibits diabetic wound healing through promoting M1 polarization of macrophages. Cell Death Discov. 2023;9(1):136. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 2. Jan A, Zakiullah AS, Muhammad B, Arshad A, Shah Y, et al. Decoding type 2 diabetes mellitus genetic risk variants in Pakistani Pashtun ethnic population using the nascent whole exome sequencing and MassARRAY genotyping: A case-control association study. PLoS ONE. 2023;18(1):e0281070. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 3. Falanga V. Wound healing and its impairment in the diabetic foot. Lancet. 2005;366(9498):1736–43. [ DOI ] [ PubMed ] [ Google Scholar ] 4. Peppa M, Stavroulakis P, Raptis SA. Advanced glycoxidation products and impaired diabetic wound healing. Wound Repair Regen. 2009;17(4):461–72. [ DOI ] [ PubMed ] [ Google Scholar ] 5. Kilic M, Olgun N, Dündar M, Advan SC, Kücük FZ, Okcuoglu S, et al. Prevalence, risk level and risk factors of diabetic foot ulcer among adult individuals with diabetes in the Southeastern Anatolia Region of Turkiye. J Tissue Viability. 2025;34(1):100839. [ DOI ] [ PubMed ] [ Google Scholar ] 6. Cano Sanchez M, Lancel S, Boulanger E, Neviere R. Targeting oxidative stress and mitochondrial dysfunction in the treatment of impaired wound healing: a systematic review. Antioxidants. 2018;7(8):98. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 7. Zhang Z, Huang Q, Zhao D, Lian F, Li X, Qi W. The impact of oxidative stress-induced mitochondrial dysfunction on diabetic microvascular complications. Front Endocrinol. 2023;14:1112363. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 8. Wang H, Gong J, Chen W, Sun Q, Zhang T, Lin Y, et al. Tetrahedral framework nucleic acids’ role in facilitating chronic diabetic wound repair via the endoplasmic reticulum-mitochondrial pathway. Nano Today. 2024;56:102252. [ Google Scholar ] 9. Pan Y, Chen L, Chen Y, Thomas ER, Zhou S, Yang Y, et al. Mitochondrial dysfunction in diabetic ulcers: pathophysiological mechanisms and targeted therapeutic strategies. Front Cell Dev Biol. 2025;13:1625474. 10.3389/fcell.2025.1625474. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 10. Oddoux S, Brocard J, Schweitzer A, Szentesi P, Giannesini B, Brocard J, et al. Triadin deletion induces impaired skeletal muscle function. J Biol Chem. 2009;284(50):34918–29. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 11. Wium E, Dulhunty AF, Beard NA. A skeletal muscle ryanodine receptor interaction domain in triadin. Plosone. 2012. 10.1371/journal.pone.0043817. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 12. Ali N, Rezvani HR, Motei D, Suleman S, Mahfouf W, Marty I, et al. Trisk 95 as a novel skin mirror for normal and diabetic systemic glucose level. Sci Rep. 2020;10(1):12246. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 13. Lansdown AB. Calcium: a potential central regulator in wound healing in the skin. Wound Repair Regen. 2002;10(5):271–85. [ DOI ] [ PubMed ] [ Google Scholar ] 14. Xu S, Chisholm AD. C. elegans epidermal wounding induces a mitochondrial ROS burst that promotes wound repair. Dev Cell. 2014;31(1):48–60. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 15. Szymański J, Janikiewicz J, Michalska B, Patalas-Krawczyk P, Perrone M, Ziółkowski W, et al. Interaction of mitochondria with the endoplasmic reticulum and plasma membrane in calcium homeostasis, lipid trafficking and mitochondrial structure. Int J Mol Sci. 2017;18(7):1576. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 16. Romero-Garcia S, Prado-Garcia H. Mitochondrial calcium: transport and modulation of cellular processes in homeostasis and cancer. Int J Oncol. 2019;54(4):1155–67. [ DOI ] [ PubMed ] [ Google Scholar ] 17. Matuz-Mares D, González-Andrade M, Araiza-Villanueva MG, Vilchis-Landeros MM, Vázquez-Meza H. Mitochondrial calcium: effects of its imbalance in disease. Antioxidants. 2022;11(5):801. 10.3390/antiox11050801. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 18. Ukaegbu K, Allen E, Svoboda KK. Reactive oxygen species and antioxidants in wound healing: mechanisms and therapeutic potential. Int Wound J. 2025;22(5):e70330. 10.1111/iwj.70330. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 19. Li Q, Liang S, Lai Q, Shen L, Zhang Y, Guo R. Heme oxygenase‑1 alleviates advanced glycation end product‑induced oxidative stress, inflammatory response and biological behavioral disorders in rat dermal fibroblasts. Exp Ther Med. 2021;22(5):1–11. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 20. Teng M, Li Z, Wu X, Zhang Z, Lu Z, Wu K, et al. Development of tannin-bridged cerium oxide microcubes-chitosan cryogel as a multifunctional wound dressing. Colloids Surf B. 2022;214:112479. [ DOI ] [ PubMed ] [ Google Scholar ] 21. Brownlee M. Biochemistry and molecular cell biology of diabetic complications. Nature. 2001;414(6865):813–20. [ DOI ] [ PubMed ] [ Google Scholar ] 22. Nishikawa T, Edelstein D, Brownlee M. The missing link: a single unifying mechanism for diabetic complications. Kidney Int. 2000;58:S26–30. [ DOI ] [ PubMed ] [ Google Scholar ] 23. Suarez J, Hu Y, Makino A, Fricovsky E, Wang H, Dillmann WH. Alterations in mitochondrial function and cytosolic calcium induced by hyperglycemia are restored by mitochondrial transcription factor A in cardiomyocytes. Am J Physiol Cell Physiol. 2008;295(6):C1561–8. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 24. Qiao Y. Reactive oxygen species in cardiovascular calcification: role of medicinal plants. Front Pharmacol. 2022;13:858160. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 25. Hu R, Wang MQ, Ni SH, Wang M, Liu LY, You HY, et al. Salidroside ameliorates endothelial inflammation and oxidative stress by regulating the AMPK/NF-κB/NLRP3 signaling pathway in AGEs-induced HUVECs. Eur J Pharmacol. 2020;867:172797. [ DOI ] [ PubMed ] [ Google Scholar ] 26. Yang C, Ling H, Zhang M, Yang Z, Wang X, Zeng F, et al. Oxidative stress mediates chemical hypoxia-induced injury and inflammation by activating NF-κb-COX-2 pathway in HaCaT cells. Mol Cells. 2011;31:531–8. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 27. Maldaner DR, Azzolin VF, Barbisan F, Mastela MH, Teixeira CF, Dihel A, et al. In vitro effect of low-level laser therapy on the proliferative, apoptosis modulation, and oxi-inflammatory markers of premature-senescent hydrogen peroxide-induced dermal fibroblasts. Lasers Med Sci. 2019;34:1333–43. [ DOI ] [ PubMed ] [ Google Scholar ] 28. Xu Z, Liu Y, Ma R, Chen J, Qiu J, Du S, et al. Thermosensitive hydrogel incorporating Prussian blue nanoparticles promotes diabetic wound healing via ROS scavenging and mitochondrial function restoration. ACS Appl Mater Interfaces. 2022;14(12):14059–71. [ DOI ] [ PubMed ] [ Google Scholar ] 29. Volvert M-L, Seyen S, Piette M, Evrard B, Gangolf M, Plumier JC, et al. Benfotiamine, a synthetic S-acyl thiamine derivative, has different mechanisms of action and a different pharmacological profile than lipid-soluble thiamine disulfide derivatives. BMC Pharmacol. 2008;8:1–11. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 30. Bozic I, Lavrnja I. Thiamine and benfotiamine: focus on their therapeutic potential. Heliyon. 2023. 10.1016/j.heliyon.2023. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 31. Schmid U, Stopper H, Heidland A, Schupp N. Benfotiamine exhibits direct antioxidative capacity and prevents induction of DNA damage in vitro. Diabetes Metab Res Rev. 2008;24(5):371–7. [ DOI ] [ PubMed ] [ Google Scholar ] 32. Kohda Y, Kanematsu M, Kono T, Terasaki F, Tanaka T. Protein O-glycosylation induces collagen expression and contributes to diabetic cardiomyopathy in rat cardiac fibroblasts. J Pharmacol Sci. 2009;111(4):446–50. [ DOI ] [ PubMed ] [ Google Scholar ] 33. Hammes H-P, Du X, Edelstein D, Taguchi T, Matsumura T, Ju Q, et al. Benfotiamine blocks three major pathways of hyperglycemic damage and prevents experimental diabetic retinopathy. Nat Med. 2003;9(3):294–9. 10.1038/nm834. [ DOI ] [ PubMed ] [ Google Scholar ] 34. Oh S-H, Witek RP, Bae S-H, Darwiche H, Jung Y, Pi L, et al. Detection of transketolase in bone marrow–derived insulin-producing cells: benfotiamine enhances insulin synthesis and glucose metabolism. Stem Cells Dev. 2009;18(1):37–46. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 35. Beltramo E, Berrone E, Buttiglieri S, Porta M. Thiamine and benfotiamine prevent increased apoptosis in endothelial cells and pericytes cultured in high glucose. Diabetes Metab Res Rev. 2004;20(4):330–6. [ DOI ] [ PubMed ] [ Google Scholar ] 36. Bozic I, Savic D, Stevanovic I, Pekovic S, Nedeljkovic N, Lavrnja I. Benfotiamine upregulates antioxidative system in activated BV-2 microglia cells. Front Cell Neurosci. 2015;9:351. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 37. Hayashi I, Morishita Y, Imai K, Nakamura M, Nakachi K, Hayashi T. High-throughput spectrophotometric assay of reactive oxygen species in serum. Mutat Res. 2007;631(1):55–61. [ DOI ] [ PubMed ] [ Google Scholar ] 38. Ishtiaq A, Bakhtiar A, Silas E, Saeed J, Ajmal S, Mushtaq I, et al. Pistacia integerrima alleviated Bisphenol A induced toxicity through Ubc13/p53 signalling. Mol Biol Rep. 2020;47:6545–59. [ DOI ] [ PubMed ] [ Google Scholar ] 39. Jiang G, Jiang T, Chen J, Yao H, Mao R, Yang X, et al. Mitochondrial dysfunction and oxidative stress in diabetic wound. J Biochem Mol Toxicol. 2023. 10.1002/jbt.23407. [ DOI ] [ PubMed ] [ Google Scholar ] 40. Balakumar P, Rohilla A, Krishan P, Solairaj P, Thangathirupathi A. The multifaceted therapeutic potential of benfotiamine. Pharmacol Res. 2010;61(6):482–8. [ DOI ] [ PubMed ] [ Google Scholar ] 41. Jiang G, Jiang T, Chen J, Yao H, Mao R, Yang X, et al. Mitochondrial dysfunction and oxidative stress in diabetic wound. J Biochem Mol Toxicol. 2023;37(7):e23407. [ DOI ] [ PubMed ] [ Google Scholar ] 42. Fraser D, Hessvik NP, Nikolić N, Aas V, Hanssen KF, Bøhn SK, et al. Benfotiamine increases glucose oxidation and downregulates NADPH oxidase 4 expression in cultured human myotubes exposed to both normal and high glucose concentrations. Genes Nutr. 2012;7(3):459–69. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 43. Ceylan-Isik AF, Wu S, Li Q, Li S-Y, Ren J. High-dose benfotiamine rescues cardiomyocyte contractile dysfunction in streptozotocin-induced diabetes mellitus. J Appl Physiol. 2006;100(1):150–6. [ DOI ] [ PubMed ] [ Google Scholar ] 44. Thornalley PJ. The potential role of thiamine (vitamin B1) in diabetic complications. Curr Diabetes Rev. 2005;1(3):287–98. [ DOI ] [ PubMed ] [ Google Scholar ] 45. Jindam A, Yerra VG, Kumar A. Nrf2: a promising trove for diabetic wound healing. Ann Transl Med. 2017;5(23):469. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 46. Tabak O, Gelisgen R, Erman H, Erdenen F, Muderrisoglu C, Aral H, et al. Oxidative lipid, protein, and DNA damage as oxidative stress markers in vascular complications of diabetes mellitus. Clin Invest Med. 2011;34(3):E163–71. [ DOI ] [ PubMed ] [ Google Scholar ] 47. Long M, de la Rojo Vega M, Wen Q, Bharara M, Jiang T, Zhang R, et al. An essential role of NRF2 in diabetic wound healing. Diabetes. 2016;65(3):780–93. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 48. Erdogan M, Yalcin A. Protective effects of benfotiamine on irisin activity in methotrexate-induced liver injury in rats. Arch Med Sci. 2020;16(1):205–11. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 49. Zhang W, Chen L, Xiong Y, Panayi AC, Abududilibaier A, Hu Y, et al. Antioxidant therapy and antioxidant-related bionanomaterials in diabetic wound healing. Front Bioeng Biotechnol. 2021;9:707479. 10.3389/fbioe.2021.707479. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 50. Kurahashi T, Fujii J. Roles of antioxidative enzymes in wound healing. J Dev Biol. 2015;3(2):57–70. [ Google Scholar ] 51. Jha B, Majie A, Roy K, Lim WM, Gorain B. Glycyrrhizic acid-loaded poloxamer and HPMC-based in situ forming gel of acacia honey for improved wound dressing: formulation optimization and characterization for wound treatment. ACS Appl Bio Mater. 2024;8(1):310–28. 10.1021/acsabm.4c01212. [ DOI ] [ PubMed ] [ Google Scholar ] 52. Nirenjen S, Narayanan J, Tamilanban T, Subramaniyan V, Chitra V, Fuloria NK, et al. Exploring the contribution of pro-inflammatory cytokines to impaired wound healing in diabetes. Front Immunol. 2023;14:1216321. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 53. Lin Z-Q, Kondo T, Ishida Y, Takayasu T, Mukaida N. Essential involvement of IL-6 in the skin wound-healing process as evidenced by delayed wound healing in IL-6-deficient mice. J Leukoc Biol. 2003;73(6):713–21. [ DOI ] [ PubMed ] [ Google Scholar ] 54. Kishimoto T. The biology of interleukin-6. Blood. 1989;74(1):1. [ PubMed ] [ Google Scholar ] 55. Johnson BZ, Stevenson AW, Prêle CM, Fear MW, Wood FM. The role of IL-6 in skin fibrosis and cutaneous wound healing. Biomedicines. 2020;8(5):101. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 56. Short WD, Steen E, Kaul A, Wang X, Olutoye OO, Vangapandu HV, et al. IL-10 promotes endothelial progenitor cell infiltration and wound healing via STAT3. The FASEB J. 2022;36(7):e22298. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 57. Kondo T, Ohshima T. The dynamics of inflammatory cytokines in the healing process of mouse skin wound: a preliminary study for possible wound age determination. Int J Legal Med. 1996;108:231–6. [ DOI ] [ PubMed ] [ Google Scholar ] 58. Lee EG, Luckett-Chastain LR, Calhoun KN, Frempah B, Bastian A, Gallucci RM. Interleukin 6 function in the skin and isolated keratinocytes is modulated by hyperglycemia. J Immunol Res. 2019;2019(1):5087847. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 59. Majie A, Saha R, Sarkar B. Diabetic wound healing potentials of phenylethanoid glycosides from Clerodendrum glandulosum Lindl.: connecting the dots by integrating network pharmacology, molecular docking, and molecular dynamics simulation. S Afr J Bot. 2024;173:232–44. [ Google Scholar ] 60. Majie A, Saha R, Sarkar A, Bhowmik R, Karmakar S, Sharma V, et al. A novel chitosan-PEG hydrogel embedded with in situ silver nanoparticles of Clerodendrum glandulosum Lindl. extract: evaluation of its in vivo diabetic wound healing properties using an image-guided machine learning model. Biomater Sci. 2024;12(16):4242–61. [ DOI ] [ PubMed ] [ Google Scholar ] 61. Zhu Q, Liu X, Zhu Q, Liu Z, Yang C, Wu H, et al. N-acetylcysteine enhances the recovery of ischemic limb in type-2 diabetic mice. Antioxidants. 2022;11(6):1097. 10.3390/antiox11061097. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 62. Sahasrabudhe SA, Terluk MR, Kartha RV. N-acetylcysteine pharmacology and applications in rare diseases—repurposing an old antioxidant. Antioxidants. 2023;12(7):1316. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] 63. Beegum F, PV A, George KT, KP D, Begum F, Krishnadas N, et al. Sirtuins as therapeutic targets for improving delayed wound healing in diabetes. J Drug Target. 2022;30(9):911–26. [ DOI ] [ PubMed ] [ Google Scholar ] 64. Liang C, Xing H, Wang C, Xu X, Hao Y, Qiu B. Resveratrol protection against IL-1β-induced chondrocyte damage via the SIRT1/FOXO1 signaling pathway. J Orthop Surg Res. 2022;17(1):406. [ DOI ] [ PMC free article ] [ PubMed ] [ Google Scholar ] Associated Data This section collects any data citations, data availability statements, or supplementary materials included in this article. Data Availability Statement The data that support the findings of this study are available from the corresponding author upon reasonable request. Articles from Diabetology & Metabolic Syndrome are provided here courtesy of BMC ACTIONS View on publisher site PDF (2.6 MB) Cite Collections Permalink PERMALINK Copy RESOURCES Similar articles Cited by other articles Links to NCBI Databases Cite Copy Download .nbib .nbib Format: AMA APA MLA NLM Add to Collections Create a new collection Add to an existing collection Name your collection * Choose a collection Unable to load your collection due to an error Please try again Add Cancel Follow NCBI NCBI on X (formerly known as Twitter) NCBI on Facebook NCBI on LinkedIn NCBI on GitHub NCBI RSS feed Connect with NLM NLM on X (formerly known as Twitter) NLM on Facebook NLM on YouTube National Library of Medicine 8600 Rockville Pike Bethesda, MD 20894 Web Policies FOIA HHS Vulnerability Disclosure Help Accessibility Careers NLM NIH HHS USA.gov Back to Top