Diabetes mellitus (DM) is a major challenge for global public health where its following complications have become one of the leading causes of morbidity and death associated with a higher risk of stroke, cardiovascular disease, metabolic disease, autoimmune disease, obesity, hypertension, amputation, retinopathy, nephropathy, neuropathy, and skin lesions. Diabetes mellitus characterized by elevated blood glucose and numerous systemic manifestation that can result in several issues that lower the impacted people's quality of life. [1]

 Five to ten percent of all cases of diabetes are type 1 diabetes. It is among the most common long-term illnesses that affect toddlers. Approximately 40% of people with type 1 diabetes are under 20. Every year, there are over 30,000 new cases of type 1 [2] Adult-onset diabetes, also known as non-insulin-dependent diabetes mellitus, was the prior name for type 2 diabetes. Between 90% and 95% of all instances of diabetes that are diagnosed fall into this disease stage. Obesity, a family history of diabetes, and a history of gestational diabetes (temporarily induced diabetes during pregnancy but for some full-blown diabetes results), glucose intake disturbance, race, and physical inactivity are major risk factors for Type 2 diabetes (T2DM). (Simon, 2012) Peripheral tissue resistance to insulin action, a condition where cells fail to use insulin appropriately as insulin requirements rise, is the defining feature for type 2 diabetes. Insulin resistance is as impairment in the insulin receptors function on the target cells as liver cells, adipocytes cells and muscle cells to respond to the insulin, and this will be followed by hyperinsulinemia, glucose intolerance, low HDL cholesterol, high triglycerides, obesity, and hypertension. These abnormalities are brought about by mutations in the insulin receptor gene, which can alter the number, structure, and signaling function of the receptors and lead to insulin resistance. [3]

The diabetic Mellitus has systematic effect and the blood parameters specially the platelets indices one of these effects. Hematological changes associated with diabetes are intricately tied to inflammation and endothelial dysfunction. Alterations in the morphology, dimensions, and physiological functions of red blood cells, white blood cells, and platelets are often correlated with hyperglycemia and its related metabolic syndrome [4]. Additionally, oxidative stress causes tissue damage, platelet destruction, and RBC dysfunction, all of which can impact blood cell function and hemostatic parameters and result in a number of complications.. [5]. Therefore, the current study designed to assessment the effect of T2DM on these parameters.

Subjects

At first, written consent was obtained from all participates enrolled in current study. This study included (60) patient’s samples with diabetes mellitus type II of only Iraqis males and females, and (20) samples of apparently healthy Iraqi volunteers, who has no history of diabetes and blood diseases with normal blood tests, all with age range (45-65) years. The samples was collected from the private laboratory of AL-Iraq laboratory in Diyala province from patients who intended the Lab for routine checkup. Patients with blood diseases, smokers, who have antiplatelet drugs, and who are with diabetic complications, were excluded. Age, gender, were inconsideration when the statistical analysis was done.

 After patients and volunteers were selected according to the research conditions, and underwent the desired test (complete blood count (CBC), and random blood sugar) the results were obtained from the Lab management.

Glucose by COBAS INTEGRA 400 plus: 

Fully automated biochemical analyzer with secure system which integrates 4 measurement technologies (photometry, turbidimetry, ISE, fluorescence polarimetry). The glucose is measured by Hexokinase-based enzymatic reference technique. The phosphorylation of glucose to glucose-6- phosphate by ATP is catalyzed by hexokinase. When NADP is present, glucose-6- phosphate is oxidized by glucose-6-phosphate dehydrogenase to gluconate-6-phosphate. There is no oxidation of any other carbohydrate. The rate at which NADPH is formed during the reaction is photometrically monitored and is directly proportional to the concentration of glucose.

Automatic hematology analyzer (Diagon Ltd): 

Complete blood count is done by 3-part hematology analyzer. It is specific for measuring the parameters of RBC, MCV, HCT, RDW-SD, RDW-CV, HGB, MCH, MCHC, PLT, MPV, and PCT. It can perform Up to 60 samples/h and give the results with minutes. It measures the electrical resistance across one or more microchannels. A brief increase in resistance (when cells that have greater resistance than the electrolyte solution that the cells are suspended in) when the cells passed between the channels. Coulter counter can detect this changes 

Statistical analysis 

The Statistical Package for Social Sciences (SPSS) version 26.0 and Graph pad prism v.6. were used to analyze the data. To define the variables, statistical information like mean and standard deviation was measured. Two groups were compared using student t test. Others parameters describes as frequency and percentage and Pearson-Chi-square tests were used to see whether there were significantly difference in percentage. The results were considered statistically significant when the p value was less than 0.05, high significant when the p value was less than 0.001.

Demographic characteristics of the study groups 

In this study, a total of 80 individuals participated, consisting of 60 patients with type 2 diabetes mellitus (T2DM) and 20 controls. The mean ages for the T2DM patients and healthy controls were 56.86 ± 1.65 and 51.00 ± 4.35 years, respectively. There was not a significant difference between the two groups in terms of age (P = 0.614) (Table 1)

Table (1): The mean age of study groups.

Blood Glucose levels among the study groups

The results revealed significant differences between the patients diagnosed with Type 2 Diabetes Mellitus (T2DM) and the healthy controls. Among patients with T2DM, the mean blood glucose level was 208.40 ± 14.56 mg/dL, while in the healthy control group, it was notably lower at 106.75 ± 4.81 mg/dL. These results imply that patients with T2DM have significantly higher blood glucose levels compared to healthy individuals, as illustrated in Figure 1. with a p-value of 0.001.

Figure 1 : Blood Glucose Levels Among Patients with T2DM and Healthy Control.

4.2. Complete Blood Count (CBC) parameters across the study groups

The results showed that in T2DM patients, the White Blood Cell count (WBC) was found to be higher at 8.6742 ± 0.47564, while in Healthy Controls it was 7.6580 ± 0.26437 (P = 0.06), Conversely, Mean Corpuscular Hemoglobin Concentration (MCHC) was significantly lower in T2DM patients (31.9183 ± 0.18246) compared to Healthy Controls (32.7067 ± 0.27403) (P = 0.024), as shown in table 2.

Table 2: demonstrates the comparison of Complete Blood Count (CBC) parameters between Patients with Type 2 Diabetes Mellitus (T2DM) and Healthy Controls. A p-value of < 0.05 is considered statistically significant. Data are presented as mean ± standard error (SE).

Moreover, Red Blood Cell count (RBC), Hemoglobin (HGB), Mean Corpuscular Volume (MCV), and Hematocrit (HCT) levels did not reveal significant differences between the two groups (P = 0.929, P = 0.513, P = 0.996, and P = 0.911 respectively). Similarly, Mean Corpuscular Hemoglobin (MCH), Platelet count (PLT), Platelet Crit (PCT), did not exhibit significant differences between T2DM patients and Healthy Controls (P > 0.05 for all). However, Mean Platelet Volume (MPV), increased significantly (P < 0.001) between the two groups.Red Cell Distribution Width - Coefficient of Variation (RDWCV) shown a significant difference between the study groups (P < 0.001). figure 3 a, and b shown the platelets in blood smear stained with Giemsa stain. 

 Figure 2a, and b. shown the platelets in blood smear stained with Giemsa stain.

DISCUSSION

The term diabetes mellitus (DM) refers to a collection of metabolic diseases marked by chronic hyperglycemia brought on by deficiencies in either insulin action or secretion, or both. In diabetes, persistent hyperglycemia damages, malfunctions, and fails many organs, especially the kidneys, heart, blood vessels, nerves, and eyes. Type 2 diabetes mellitus (T2DM) is primarily a defect in carbohydrate metabolism, arising from significant insulin resistance combined with relative insulin deficiency, or a predominant defect in secretion of insulin, which may occur with or without insulin resistance. [6]

The current study found the WBCs count in patients group was higher than control, and this results are compatible with [7,8] .Epidemiological studies have shown that WBC as an indicator of inflammation could predict diabetes risk, in diabetes mellitus, a chronic inflammatory state brought on by insulin's effects on muscles, adipose tissue, and the liver promotes WBC differentiation and maturation through proinflammatory cytokines. [9]

Conversely, Mean Corpuscular Hemoglobin Concentration (MCHC) was significantly lower in T2DM patients when compared to healthy controls and this finding is consistent with a study achieved [10] Throughout their lives, erythrocytes were exposed to hyperglycemic circumstances, which led to a number of structural and functional alterations, including hypochromia, which is characterized by a decrease in MCHC and is frequently observed in iron deficiency anemia, thalassemia, and anemia of inflammation. [11]. It is widely acknowledged that diabetes mellitus is linked to chronic inflammation and elevated levels of inflammatory cytokines such as interleukin-1 and interleukin-6. Thus, by retaining iron in reticuloendothelial cells, inflammation-inducible cytokines and hepcidin contribute significantly to the development of microcytic hypochromic red blood cells.[12] However, Mean Platelet Volume (MPV), increased significantly in patients’ group when compared with control, this suggests that platelet size is larger in T2DM patients, which could indicate increased platelet activation and a higher risk of thrombotic complications associated with diabetes. These results are consistent with a research conducted [13,14]

Platelets with high MPV are more reactive and aggregable. They have denser granules and thus are more involved in prothrombotic events. Hyperglycemia can induce non-enzymatic glycation of platelet surface proteins. This leads to a decrease in membrane fluidity, which further leads to the activation of platelets. Platelets adhere to the endothelium and release mediators involved in thrombus formation and inflammation. [15,16]

Red Cell Distribution Width - Coefficient of Variation (RDW-CV) displayed significant increase in patients group by compare with control. A high level of anisocytosis, or variation in the size of the circulating erythrocytes, is indicated by a high RDW which is associated with distortion and degradation of erythropoiesis [17]. Representing a higher degree of oxidative damage and chronic inflammation. Anemia can also result in elevated RDW. Otherwise, factors such megaloblastic anemia and iron insufficiency, are linked to micro or macro cytosis, but such confounders are negligible because all the selected patients for the study were diabetes without complication. Patients with diabetes may have shorter red blood cell lifespans. Therefore, different disruptions in the hematopoietic milieu have an impact on RBCs. [18]

Another study [19] reported that higher RDW-CV levels in patients with type 2 diabetes mellitus compared to non-diabetic controls. The study also found that RDW-CV levels were positively correlated with HbA1c levels and fasting plasma glucose levels in patients with diabetes.

However, a study [20]reported no significant difference in RDW-CV levels between patients with type 2 diabetes mellitus and non-diabetic controls. This study did report a significant positive correlation between RDW-CV levels and HbA1c levels in patients with diabetes mellitus.

In conclusion, the association between RDW-CV levels and diabetes mellitus appears to be inconsistent across different studies. While some studies have reported higher RDW-CV levels in patients with diabetes mellitus, other studies have not found any significant difference. Further research is needed to clarify the relationship between RDW-CV levels and diabetes mellitus. 

The current study demonstrates that Type 2 Diabetes Mellitus (T2DM) significantly associated with specific blood indices, including increased WBC, MPV, and RDW-CV, and decreased MCHC when compared with control healthy group. These changes refer to the chronic inflammation and oxidative stress associated with hyperglycemia, contributing to complications like cardiovascular diseases. Monitoring these indices could provide valuable insights into the inflammatory state and potential risks in T2DM patients.

The authors declare that they have no conflict of interest

No funding sources

The study was approved by the University of Bilad Alrafidain, Diyala, Iraq.

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