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J Gandhara Med Dent Sci
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ORIGINAL ARTICLE
:
:
ASSOCIATION OF OSTEOPONTIN (OPN) LEVEL IN DIABETIC NEPHROPATHY PATIENTS AS
AN EARLY DIAGNOSTIC MARKER
Anwar Shahzad
1
, Khalil Ur 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 of
Osteopontin (Opn) Level in Diabetic
Nephropathy Patients as an Early
Diagnostic Marker. J Gandhara Med
Dent Sci. 2025;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, Pakistan Kidney Center,
Abbottabad
3
Consultant, Department of
Pathologist, Pakistan Kidney Center,
Abbottabad
4
Consultant, Department of
Pathologist, Pakistan Kidney Center,
Abbottabad
5
Consultant, Department of
Pathologist, Pakistan Kidney Center,
Abbottabad
Correspondence
1
Anwar Shahzad, Consultant, Research
Analyst, Pakistan Kideny Center,
Abbottabad
+92-314-2244567
redline.shahzad@gmail.com
ABSTRACT
OBJECTIVES
The study aimed to nd 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 were measured to conrm 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 square methods. The p-value of ≤0.05 was deemed
statistically signicant.
RESULTS
Osteopontin has a signicant relation with diabetes duration only. Serum
biomarkers of the DM show signicant osteopontin levels with 0.00 p values
for HBA1C and all biomarkers of Diabetes. The least-square distribution of
the dierent biomarkers with osteopontin level at 95 % condence interval
and standardized coecients show lower and upper bound and signicant
levels. Osteopontin levels show 0.29 and 0.38 lower and upper bound at a
95% condence 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 signicance 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 positively correlated in positive controls and low levels in
negative control and regular diabetic patients with no nephropathy.
KEYWORDS: Association, ANOVA, DM (Diabetes Mellitus), OPN
(Osteopontin), Nephropathy
INTRODUCTION
Diabetes Mellitus (DM) results from a prolonged
elevation of blood glucose levels, either an absolute or
relative insulin insuciency.
1
Diabetes Mellitus (DM)
is a widespread health condition worldwide, aecting
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 Organization, Diabetes Mellitus is the 4
th
most crucial non-communicable disease aecting the
population worldwide.
2
Diabetes Mellitus aects
almost all parts of the body, especially the urogenital
system, cardiovascular system, and eyesight due to
microangiopathies. 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 suer proteinuria, perhaps due to modications
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 kidney 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 diabetic 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 lters urine in
/10.37762/jgmds.668
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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 osteopontin 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 the 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 bearing
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 nephropathy.
Patients with post-menopausal women, Osteomalacia
and patients having inammatory conditions. Duration
of Diabetes mellitus in the cases was recorded on pre-
structured proforma along with other demographic data.
Fasting blood sugar, HbA1c, serum urea 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 signicant. Investigated data
was displayed on charts and a tabletop.
RESULTS
Table 1 Demonstrates ANOVA grades of the dierent
demographic variables based on serum osteopontin
levels. Diabetes duration is signicantly related to
osteopontin level with p values 0.00 and no signicant
relation with other demographic variables. Table 2
shows that the dierent serum biomarkers of Diabetes
mellitus changed due to uctuating blood sugar levels
in the body variable based on serum osteopontin levels.
It shows that osteopontin level shows 0.00 p values for
HBA1C, with osteopontin level, which means
Osteopontin has only a signicant relation with all
biomarkers of Diabetes. Table 3 shows the serum
osteopontin level in three dierent study groups,
including positive control groups, negative control
groups and diabetic patients in the study groups. Table
4 shows the serum osteopontin level in three dierent
study groups, including positive control groups,
negative control groups and diabetic patients in the
study groups in which signicant levels were shown in
diabetic nephropathic patients.
Table 1: The Dierent Variables Based on 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 Dierent 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
Blood
Urea
(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
ethical no: EC/03/24. dated: 21-06-2024. Sample Size
51
J Gandhara Med Dent Sci
April - June 2025
Table 4: Comparison of Adverse Eects 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
inammatory role of Osteopontin in diabetic
nephropathy, with elevated levels of Osteopontin being
consistently observed.
16
Numerous studies have
explored the link between osteopontin levels and the
impact of Diabetes mellitus in aected individuals.
17
Various investigations have indicated the inammatory
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 ve years
exhibited signicantly 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 with 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 nephropathy 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 signicant 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 signicantly
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 ndings 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 calcication and plays a
role in mineral metabolism. Osteopontin is a crucial
element in the development of calcication and
dysfunction in vascular epithelial structures, resulting in
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 diabetic
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 inuence of
medications on Osteopontin, necessitating conrmation
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
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CONTRIBUTORS
1. Anwar Shahzad - Concept & Design; Data
Analysis/Interpretation
2. Khalil Ur Rahman- Data Acquisition; Drafting Manuscript;
Supervision
3. Yasir Humayun – Data Acquisition; Drafting Manuscript;
Critical Revision; Final Approval
4. Henna Khalid - Data Acquisition; Supervision
5. Mariam Riaz - Drafting Manuscript; Critical Revision