After approval of the ethical committee of the Faculty of Medicine for Girls, Al-Azhar University, and before enrollment in our study, all participants were informed in details about our study aim and signed consent. Adult patients of both sexes with T2DM were enrolled in this study.

Study design

This is a case-control study.

Study participants

This study was conducted on 45 participants aged from 25 to 60 years of both sexes. Patients were recruited from the Ophthalmology Department at Al-Zahraa University Hospital during the period between March 2021 and October 2021.

Inclusion criteria

Diabetic patients were diagnosed according to the criteria established by the American Diabetes Association (ADA) which are fasting blood glucose (FBG) ≥ 126 mg/dL (7.0 mmol/L) or 2-h plasma glucose (2-HPG) ≥ 200 mg/dL (11.1 mmol/L) or, random plasma glucose ≥ 200 mg/dL (11.1 mmol/L) or, hemoglobin A1c (ΗbA1c) ≥ 6.5%.

Exclusion criteria

All patients known to have history of micro- and macro-vascular complications of DM other than DR. Also, patients suffer from any other ocular disease that may affect ocular circulation (e.g., glaucoma, age-related macular degeneration, and retinal vascular occlusion) or inherited macular disease. Patients with cardiovascular disease, stroke or transient ischemic attacks, liver disease, evidence of sepsis, and any autoimmune diseases as systemic lupus erythematosus were excluded.

The matching criteria

The controls were age- and sex-matched apparently healthy control.

All participants were subjected to the following:

  • Clinical examination, complete medical, and ophthalmological history including the history of ocular disease, previous ocular trauma, or intraocular surgery.

  • Best-corrected visual acuity (BCVA).

  • Slit-lamp biomicroscopic examination.

  • Fundus examination by slit-lamp biomicroscopy using + 90 D noncontact lens and indirect ophthalmoscopy. The presence or absence of DR was confirmed by fundus examination. All participants were subjected to following laboratory investigations.

Laboratory investigations included the following:

  1. a.

    Complete blood count (CBC) was performed using fully automated hematology analyzer (Sysmex KX21N, Kobe, Japan).

  2. b.

    Glycated hemoglobin (HbA1c), fasting blood sugar, and 2 h post-prandial (2HPP) were performed by fully automated chemistry analyzer Cobas c 311 (Germany).

  3. c.

    Serum Fetuin-A level was performed by enzyme-linked immunosorbent assay (ELISA) according to the manufacturer’s instructions using Human Fetuin-A ELISA Kit (Cat. NO.E1386Hu), supplied by the Bioassay Technology Laboratory (China). This method used to quantify the level of human Fatuin-A with a detection range of 0.75–15 μmol/l.

Statistical analysis

Data were coded and entered using the statistical package for the Social Sciences (SPSS) version 26 (IBM Corp., Armonk, NY, USA). Data were summarized using the mean and standard deviation (SD) for normally distributed quantitative variables or median and interquartile range for non-normally distributed quantitative variables. Comparisons between groups were done using unpaired t test or analysis of variance (ANOVA) while non-parametric Kruskal-Wallis test and Mann-Whitney test were used for non-normally distributed quantitative variables. Correlations between quantitative variables were done using Spearman correlation coefficient. P values ≤ 0.05 were considered as statistically significant, and ≤ 0.001 were considered highly significant.

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