Search

Diabetic foot ulcers (DFU) is a complication associated with diabetes characterised by high morbidity, disability, and mortality, involving chronic inflammation and infiltration of multiple immune cells. We aimed to identify the critical genes in nonhealing DFU using single-cell RNA sequencing, transcriptomic analysis and machine learning. The GSE165816, GSE134431, and GSE143735 datasets were downloaded from the GEO database. We processed and screened the datasets, and identified the cell subsets. Each cell subtype was annotated, and the predominant cell types contributing to the disease were analysed. Key genes were identified using the LASSO regression algorithm, followed by verification of model accuracy and stability. We investigated the molecular mechanisms and changes in signalling pathways associated with this disease using immunoinfiltration analysis, GSEA, and GSVA. Through scRNA-seq analysis, we identified 12 distinct cell clusters and determined that the basalKera cell type was important in disease development. A high accuracy and stability prediction model was constructed incorporating five key genes (TXN, PHLDA2, RPLP1, MT1G, and SDC4). Among these five genes, SDC4 has the strongest correlation and plays an important role in the development of DFU. Our study identified SDC4 significantly associated with nonhealing DFU development, potentially serving as new prevention and treatment strategies for DFU., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Yuming Shen reports financial support was provided by National Key Research and Development Program of China. Yungang Hu reports financial support was provided by Jiangxi Provincial Natural Science Foundation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Exosomes isolated from platelet-rich plasma (PRP-exos) have been recently deemed as an optimized therapeutic strategy in diabetic foot ulcer (DFU) treatment. Herein, we aimed to explore whether MALAT1 participates in DFU wound healing by PRP-exos treatment and the related preliminary mechanism. Fibroblasts were isolated from healthy donors and DFU patients, and the expression of MALAT1, miR-374a-3p and DNMT3A were detected by RT-PCR. The effect of MALAT1 and miR-374a-3p on DFU fibroblast function was verified by gain/loss of function experiment. The targeted binding of MALAT and miRNA was verified by double luciferase reporter gene assay. PRP-exos were isolated from normal human blood and characterized, and then co-cultured with DFU fibroblasts. The MALAT1 expression was donwregulated while the miR-374a-5p expression was upregulated in DFU fibroblasts. Double luciferase reporter gene assay demonstrated the targeted binding of MALAT and miR-374a-5p. Overexpression of MALAT1 or knockdown of miR-374a-5p could increase viability and inhibit apoptosis and pyroptosis of DFU fibroblast. And overexpression of miR-374a-5p reversed the effect of PRR-exos or MALAT1 overexpression on cell viability, apoptosis and pyroptosis. Collectively, MALAT1 mediated signal axis participates in the role of PRP-exos in promoting DFU wound healing, which may help identify optimal targets and effective therapies for DFU treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Hypoxia inducible factor 1 alpha (HIF-1α) is a key regulator of the genes involved in the cellular response to hypoxia. This study aims to determine the HIF-1α gene polymorphism and its association with protein expression in diabetic subjects with and without diabetic foot ulcers (DFU). A total of 529 patients with T2DM (N = 185), DFU (N = 199) and Control (N = 145) were accounted for the study. PCR-RFLP experiment was carried out in order to find the allelic and genotypic comparison of HIF-1α gene in various groups of patients. There was a highly increased frequency of GA, RR value of 3.533(2.099-5.950) with p-value of 0.0001 on DFU patients when compared to that of control subjects with risk allele of GA, RR value of 1.756 (1.294-2.384) with p-value of 0.00001. Thus, we found that there was a significant association of HIF-1α polymorphism in exon 12 among DFU patients when compared to control groups. The circulatory HIF-1α protein expression study indicated a decreased expression in DFU levels when compared to T2DM and control. Overall, the study showed that there is an association of HIF-1α polymorphism (G1970A) in diabetes and DFU patients when compared to the healthy group., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Compared to other complications the genetics of diabetic foot ulcer is poorly studied. The Interleukin (IL)-6 (-174G > C/rs1800795), Tumor Necrosis Factor (TNF)-α (-308G > A/rs1800629) and (-238G > A/rs361525) and Stromal cell Derived Factor (SDF)-1 (+801G > A/rs1801157) are well characterized single nucleotide polymorphisms (SNPs) which were previously shown to be associated with Diabetic Foot Ulcer (DFU). In the present study, we looked at the association of these SNPs with foot microbial infection, Wagner's ulcer grade and treatment procedure, along with serum levels of these cytokines (intermediate phenotype) and other serum biomarkers (adiponectin, leptin, CRP and HOMA-IR) in subjects with DFU. Subjects with DFU (n = 270) were genotyped by PCR-RFLP and the serum levels of IL-6, TNF-α and SDF-1 were determined by ELISA. Microbial infections were determined by standard microbiological methods. Ulcer grade and treatment procedures were recorded. IL-6 (-174G > C), TNF-α (-308G > A) and SDF-1 (+801G > A) SNPs were associated with severe microbial infections. TNF-α (-308G > A) and (-238G > A) SNPs were associated with severe ulcer grades. SDF-1 (+801G > A) SNP was associated with major amputation even after adjusting for confounding variables. Identification of these SNPs in DFU subjects would help in identifying high risk individuals who need better treatment care., (Copyright © 2018. Published by Elsevier B.V.)

Lysyl oxidase (LOX) is an extra-cellular matrix-modifying enzyme that has been linked to cell proliferation, metastasis, angiogenesis and wound healing. This study was designed to examine the association of LOX gene polymorphism G473A, G>A, (rs1800449) located in exon 1 of the LOX gene in diabetic subjects with and without diabetic foot ulcers (DFU) and its impact of expression on DFU. Genotypic analysis of 906 samples showed a significant increase in the presence of 'A' allele in type 2 diabetes mellitus (T2DM) and DFU when compared to that of control subjects. Allele wise analysis showed a higher frequency of 'A' allele in the T2DM (36.23%, OR 1.069, P value 0.29) and DFU (41.69%, OR 1.195, P value 0.003) when compared to that of control subjects (33.17%). Interestingly, real time RT-PCR results showed significant increased transcript level of the LOX gene on the AA genotype of DFU when compared to that of the AA genotype of T2DM and control subjects. Our finding predicts that there is an association of LOX gene polymorphism (G473A) on diabetes and DFU patients when compared to that of healthy controls. Thus, this study merits further evaluation on a mechanistic approach of this gene., (Copyright © 2017 Elsevier B.V. All rights reserved.)