49 J Gandhara Med Dent Sci April - June 2025 ORIGINAL ARTICLE : : ASSOCIATION OF OSTEOPONTIN (OPN) LEVEL IN DIABETIC NEPHROPATHY PATIENTS AS AN EARLY DIAGNOSTIC MARKER Anwar Shahzad 1 , Khalil U r Rahman 2 , Yasir Hamayun 3 , Henna Khalid 4 , Mariam Riaz 5 How to cite this article Shahzad A, Rahman KU, Hamayun Y, Khalid H, Riaz M . Association o f Osteopontin (Opn) Level in Diabetic Nephropathy Patients as an Early Diagnostic Marker . J Gandhara Med Dent Sci. 202 5 ; 12( 2 ): 49-52.http://doi.org Date of Submission: 14 - 01 - 2025 Date Revised : 06 - 02 - 2025 Date Acceptance: 03 - 03 - 2025 2 Consultant, Department of Nephrologist, P akistan K id n e y C enter, Abbottabad 3 Consultant, Department of Pathologist , P akistan K id n e y C enter, Abbottabad 4 Consultant, Department of Pathologist, P akistan K id n e y C enter, Abbottabad 5 Consultant, Department of Pathologist, P akistan K idne y C enter, Abbottabad Correspondence 1 Anwar Shahzad , Consultant, Research Analyst, P akistan K ideny C enter, Abbottabad +92 - 314 - 2244567 redline.shahzad@gmail.com ABSTRACT OBJECTIVES The study aimed to find an association between OPN levels and diabetic nephropathy as a serum marker in DM patients for early detection. METHODOLOGY A cross - sectional study was conducted for 6 months on 277 diagnosed cases of DM with 10 positive and 10 negative controls collected through non - probability sampling. Fasting blood sugar, HbA1c, serum urea and creatinine levels of known diabetic patients we re measured to confirm their disease status and OPN using an ELISA Kit and compared in DM patients and controls. Statistical analysis was performed using SPSS v22.0. DM patients and negative controls were compared using unpaired one - way ANOVA and least squ are methods. The p - value of ≤0.05 was deemed statistically significant. RESULTS Osteopontin has a significant relation with diabetes duration only. Serum biomarkers of the DM show significant osteopontin levels with 0.00 p values for HBA1C and all biomar kers of Diabetes. The least - square distribution of the different biomarkers with osteopontin level at 95 % confidence interval and standardized coefficients show lower and upper bound and significant levels. Osteopontin levels show 0.29 and 0.38 lower and upper bound at a 95% confidence interval, with a 0.79 considerable level and 0.25 t distribution, with 0.01 beta values for diabetes duration. Blood creatinine level is 0.02 and 0.05 lower and upper bound, with 0.36 significance level for Osteopontin and 0 .02 standard error. Osteopontin levels 11ng/dl to 20 ng/dl have been found in 88 cases, while 09 in positive controls and 12 in negative controls. Meanwhile, only two positive controls were found>20 ng/dl osteopontin level. CONCLUSION Serum (OPN) was posit ively correlated in positive controls and low levels in negative control and regular diabetic patients with no nephropathy. KEYWORDS: Association, ANOVA, DM ( D iabetes M ellitus), OPN (Osteopontin), Nephropathy INTRODUCTION Diabetes Mellitus (DM) results from a prolonged elevation of blood glucose levels, either an absolute or relative insulin insufficiency. 1 Diabetes Mellitus (DM) is a widespread health condition worldwide, affecting about 8.5% of the population worldwide and 16% in Pakistan. This includes newly diagnosed cases and previously existing diabetic patients. 9 According to the World Health Organiza tion, Diabetes Mellitus is the 4 th most crucial non - communicable disease affecting the population worldwide. 2 Diabetes Mellitus affects almost all parts of the body, especially the urogenital system, cardiovascular system, and eyesight due to microangiop athies. Renal involvement of Diabetes mellitus may result in chronic renal failure requiring dialysis, which is a very costly procedure and treatment. 4 Current studies have shown the role of the RAS (Renin angiotensin) protein system in developing chronic kidney diseases, especially in diabetic patients. 5 Patients suffer proteinuria, perhaps due to modifications in the RAS (Renin Angiotensin) protein system responsible for kidney protein preservation. Once the the substance found in bone, the part Diabetes is established, it may result in kidne y failure if left untreated. 6 Some studies suggest that people with both Type - I and Type - II diabetes may have fewer podocytes in their glomeruli. This decrease in podocytes may happen simultaneously with the start of protein in the urine. 7 Control of diabe tic nephropathy mainly depends on early detection and prompt treatment. Early detection of diabetic nephropathy can be done using new biomarkers like Osteopontin. 8 Osteopontin (OPN) is a protein that sticks to calcium, that filters urine in /10.37762/jgmds.668
50 J Gandhara Med Dent Sci April - June 2025 the kidneys and the cells that cover blood vessels. 9 The severity of Diabetic nephropathy largely depends on the serum osteopontin level. A study conducted by Yamaguchi et al. revealed a relationship between nephropathy and osteopon tin levels in 301 diabetic nephropathy patients. They observed that plasma osteopontin concentration was raised compared to other biomarkers inside the urinary sample of diabetic patients compared to the control group. 10 The current study is done to mark t he presence of osteopontin levels in diabetic nephropathic patients as an early detection marker as compared to other biomarkers. 11 METHODOLOGY A cross - sectional study with positive and negative control was conducted at various tertiary care Hospitals, including Pakistan Kidney Center Abbottabad, King Abdullah Teaching Hospital Mansehra and Ayub Medical College Abbottabad for 6 months. Sample Size was collected for 277 diagnosed cases of DM with 10 positive controls (established cases of diabetic nephropathy patients) and 10 negative controls (normal individuals) having no history of Diabetes collected through non - probability consecutive sampling. Patients with diabetic nephropathy were taken as the cases, while the positive control were the patients with no diabetic nephropathy despite a positive history of Diabetes. Negative controls were the normal individuals with no diabetes and diabetic nephro pathy. Patients with post - menopausal women, Osteomalacia and patients having inflammatory conditions. Duration of Diabetes mellitus in the cases was recorded on pre - structured proforma along with other demographic data. Fasting blood sugar, HbA1c, serum ur ea and creatinine levels of known diabetic patients were measured to confirm their disease status. According to the manufacturer’s instructions, Osteopontin was measured using an ELISA Kit (BioAssay USA). Osteopontin levels of the DM patients and controls were compared. Statistical analysis was performed using (SPSS) v22.0. DM patients and negative controls were compared by using unpaired one - way ANOVA. The p - value of ≤0.05 was deemed statistically significant. Investigated data was displayed on charts and a tabletop. RESULTS Table 1 Demonstrates ANOVA grades of the different demographic variables based on serum osteopontin levels. Diabetes duration is significantly related to osteopontin level with p values 0.00 and no significant relation with other demographic variables. Table 2 shows that the different serum biomarkers of Diabetes mellitus changed due to fluctuating blood sugar levels in the body variable based on serum osteopontin levels. It shows that osteopontin level shows 0.00 p values for HBA1 C, with osteopontin level, which means Osteopontin has only a significant relation with all biomarkers of Diabetes. Table 3 shows the serum osteopontin level in three different study groups, including positive control groups, negative control groups and di abetic patients in the study groups. Table 4 shows the serum osteopontin level in three different study groups, including positive control groups, negative control groups and diabetic patients in the study groups in which significant levels were shown in d iabetic nephropathic patients. Table 1: The Different Variables Based o n Serum Osteopontin Levels Total of Squares df Total Mean F Sig. Gender Cases 15.81 72 0.22 0.88 0.73 Controls 50.90 204 0.25 Total 66.71 276 Diabetes Duration Cases 53.62 72 0.74 1.70 0.00 Controls 89.12 204 0.43 Total 142.75 276 Age in Years Cases 8345.41 72 115.90 0.90 0.68 Controls 26093.00 204 127.90 Total 34438.419 276 Squares Table 2: ANOVA Results of Different Variables Like Serum Total Squares Df Mean of Squares F Sig. HBA1c Cases 20626.14 72 286.47 69.18 0.00 Controls 844.67 204 4.14 Total 21470.82 276 Micro - albumin level mg/dl Cases 1210522.13 72 16812.80 11.10 0.00 Controls 308797.81 204 1513.71 Total 1519319.95 276 Blood Creatini ne level mg/dl Cases 7787.92 72 108.16 7.83 0.00 Controls 2815.22 204 13.800 Total 10603.14 276 BloodUrea(mg/dl) Cases 498599.94 72 6924.99 5.97 0.00 Controls 236505.72 204 1159.34 Total 735105.66 276 Fasting Blood Sugar(m g/dl) Cases 492982.47 72 6846.97 2.05 0.00 Controls 680352.43 204 3335.06 Total 1173334.90 276 Table 3: ANOVA Test Comparisons among All Studied Groups Positive Control Group Negative Control Group Diabetic Group Test Valu e P - valu e 10 10 277 S.OP N(ng/ /ml) Mean ± SD 71.80 ± 20.37 169.12 ±20.56 278.42 ± 48.83 229.3 01 0.00 1 Ra nge 41.8 – 101.4 135.7 – 214.7 214.8 – 379.8 Association of Osteopontin (OPN) Level in Diabetic Nephropathy
51 J Gandhara Med Dent Sci April - June 2025 Table 4: Comparison of Adverse Effects in Both Groups Osteopontin level ng/dl Total < 1 to 10 11 to 20 >20 Study Populati on Cases 183 88 00 271 Positive Controls 02 09 02 13 Negative Controls 01 12 00 13 Total 186 109 02 297 DISCUSSION Osteopontin (OPN) is a multifunctional protein associated with Type II Diabetes and disorders affecting patients' vascular architecture. However, the specific role of Osteopontin in individuals with Type - 1 diabetes remains unclear. 14,15 Our study aims to investigate the role of Osteopontin in the progression of Diabetes mellitus, utilizing it as a biomarker for the study. Several studies have highlighted the inflammatory role of Osteopontin in diabetic nephropathy, with elevated levels of Osteopontin being consistently observed. 16 Numerous studies have explored the link between osteopontin levels an d the impact of Diabetes mellitus in affected individuals. 17 Various investigations have indicated the inflammatory role of Osteopontin in diabetic nephropathy, with higher concentrations of Osteopontin being observed. 17 Our study found that patients with diabetic nephropathies with a duration exceeding five years exhibited significantly higher osteopontin levels. 18 The investigation's results illustrate the serum levels of various diabetic indicators, with fasting blood sugar averaging 267 mg/dl. In line w ith our research, Yamaguchi and colleagues explored osteopontin levels, observing diabetic complications in microvascular scenarios among 229 patients with Type II Diabetes. This manifested as advanced retinopathy, noticeable neuropathy, and more evident n ephropathy in both plasma and urine. Notably, osteopontin levels showed a substantial and marked increase as nephropathy advanced. However, discernible changes in osteopontin development were not noted in retinopathy or neuropathy. 18 It has been determined that osteopontin levels are a significant predictor exclusively in end - stage disorders of renal origin, with no notable impact. In a study within a multiethnic cohort by Zhang, it was observed that Osteopontin levels were significantly higher in subjects with advanced established diabetic nephropathy (64.7 ng/mL) compared to diabetic patients without diabetic nephropathy (51.7 ng/mL; p<0.001). These elevated Osteopontin levels have been linked to the onset and severity of diabetic nephropathy, establishing Osteopontin as a potential biomarker for diabetic nephropathy. These findings align with the research conducted by El Dayem, which examined the correlation between high Osteopontin levels and diabetic nephropathy in eighty patients with Type I Diabetes. 19 Elevated OPN is intricately involved in severe and extensive vascular calcification and plays a role in mineral metabolism. Osteopontin is a crucial element in the development of calcification and dysfunction in vascular epithelial structures, resulting i n nephropathy. 20 Individuals with diabetic nephropathy exhibited a prolonged DM duration compared to those with Type II Diabetes without nephropathy. Additionally, a noteworthy increase in BMI was noted in both cases and controls. This aligns with existing research, indicating a predisposition to insulin resistance and diabetic nephropathy in individuals with higher BMI. 21 Consistent with earlier research, all diabetic patients in our study exhibited elevated osteopontin levels, particularly those with diab etic nephropathy, marked by positive microalbuminuria. 22 LIMITATIONS The limitation of this research stems from its single - centre focus. Moreover, assessing vascular dysfunction as a cause of microvascular issues relies solely on vascular markers, neglecting alternative factors. It’s crucial to acknowledge the potential influence of medications on Osteopontin, necessitating confirmation across diverse populations. CONCLUSIONS Serum (OPN) levels marked in diabetic nephropathy patients and positive controls were high as compared to low levels in negative control. Association of Osteopontin (OPN) Level in Diabetic Nephropathy CONFLICT OF INTEREST: None FUNDING SOURCES: None REFERENCES 1. 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