J Pediatr Endocrinol Metab 2018; aop

Ramank K. Marwahaa,*, M.K. Garga, Sushil Gupta, Mohd Ashraf Ganie, Nandita Gupta,
Archna Narang, Manoj Shukla, Preeti Arora, Annie Singh, Aditi Chadha and Ambrish Mithal

Association of insulin-like growth factor-1 and

IGF binding protein-3 with 25-hydroxy vitamin D
in pre-pubertal and adolescent Indian girls
https://doi.org/10.1515/jpem-2017-0275 Results: The mean age, BMI and serum 25OHD of girls
Received July 16, 2017; accepted December 1, 2017 were 11.5 ± 3.2  years, 18.7 ± 4.8 kg/m2 and 9.9 ± 5.6 ng/mL,
Abstract respectively. VDD was observed in 94.6% of girls. Unad-
justed serum IGF-1 levels and IGF-1/IGFBP-3 molar ratio were
Background: There is a high prevalence of vitamin D defi- significantly higher in girls with severe VDD as compared
ciency (VDD) in India. Molecular mechanisms suggest a to girls with mild-to-moderate VDD. However, these differ-
strong relationship between vitamin D and growth factors. ences disappeared when adjusted for age, height or sexual
However, there is a paucity of literature with regard to a maturation. The serum IGF-1 and IGFBP-3 levels increased
relationship between insulin-like growth factor-1 (IGF-1), significantly post supplementation with vitamin D.
insulin-like growth factor binding protein-3 (IGFBP-3) and Conclusions: There were no differences in serum IGF-1 lev-
vitamin D particularly in subjects with VDD. The objec- els and the IGF-1/IGFBP-3 molar ratio among VDD catego-
tive of the study was to assess the relationship between ries when adjusted for age, height and sexual maturation in
growth factors and serum vitamin D-parathormone (PTH) girls. Vitamin D supplementation resulted in a significant
status in school girls and study the impact of vitamin D increase in serum IGF-1 levels in VDD pre-pubertal girls.
supplementation on growth factors in pre-pubertal girls
Keywords: adolescents; insulin-like growth factor (IGF-1);
with VDD.
insulin-like growth factor binding protein-3 (IGFBP-3);
Methods: Our study subjects were apparently healthy
vitamin D deficiency.
school girls aged 6–18 years. The baseline height, weight,
body mass index (BMI), pubertal status, serum 25-hydroxy
vitamin D (25OHD), PTH, IGF-1 and IGFBP-3 were assessed
in 847 girls aged 6–18 years and in 190 pre-pubertal girls
with VDD following supplementation.
Introduction
Vitamin D deficiency (VDD) prevails in epidemic propor-
tions in the Indian sub-continent with a prevalence of
a
Ramank K. Marwaha and M. K. Garg are joint first authors 70%–90% in the general population including children
*Corresponding author: Maj Gen (Retd) Dr. Ramank K. Marwaha, and adolescents. This is prevalent in both urban and rural
Senior Consultant Endocrinology and Scientific Advisor (Projects),
settings and across all socio-economic and geographic
ILSI-India, Flat No. 17, Gautam Apartments, Gautam Nagar, New
Delhi 110049, India, Phone: +91 9810296820, Fax: +91 11 26523477, data [1–6]. Poor vitamin D synthesis in Indians is mainly
E-mail: [email protected] attributed to poor sun exposure due to cultural avoidance
M.K. Garg: Officer’s Training College, Lucknow, India of skin exposure, crowded houses with limited sun expo-
Sushil Gupta: Department of Endocrinology, Sanjay Gandhi Post sure and work culture of staying indoors. In addition, dark
Graduate Institute, Lucknow, India
skin complexion, atmospheric pollution, vegetarian foods
Mohd Ashraf Ganie and Nandita Gupta: Department of
Endocrinology and Metabolism, AIIMS, New Delhi, India
habits, sedentary life style, lack of vitamin D food fortifi-
Archna Narang: Department of Medicine, Sur Homeopathic College, cation program and poor intake of vitamin D supplements
New Delhi, India are additional causative factors [6–9].
Manoj Shukla: Sanjay Gandhi Post Graduate Institute, Lucknow, VDD, besides causing rickets in children, is also associ-
India ated with stunted growth primarily due to its effect on bone
Preeti Arora and Annie Singh: Central Council of Research in
mineralization. Insulin-like growth factor-1 (IGF-1) which
Homeopathy, Ministry of Ayush, New Delhi, India
Aditi Chadha: BHMS, Sur Homeopathic College, New Delhi, India is synthesized by the liver under the influence of growth
Ambrish Mithal: Department of Endocrinology and Metabolism, hormone (GH) circulates in association with insulin-like
Medanta Hospital, Gurugram, Haryana, India growth factor binding protein-3 (IGFBP-3), and acts through

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2      Marwaha et al.: IGF-1, IGFBP-3 and 25OHD

IGF receptors [10]. Although it is involved in various physi- WS045, Narang Medical Limited, Delhi, India) with subjects standing
ological processes, it is primarily associated with growth. straight with their head held in the Frankfurt plane. The weight with-
out shoes and with light clothes on was measured to the nearest 0.1
It has been proposed that IGF-1 increases 1α-hydroxylase
kg, using an electronic scale (EQUINOX Digital weighing machine,
activity and increases the production of 1,25-dihydroxy- Model EB6171, Equinox Overseas Private Limited, New Delhi, India).
vitamin D from 25-hydroxy vitamin D (25OHD) [11, 12]. The BMI, defined as the ratio of body weight to height square, was
Besides, IGF-1 is also known to increase both – bone for- expressed in kg/m2. The pubertal staging was carried out by trained
mation and resorption [13]. Conversely, vitamin D has been professionals of the same sex based on the Tanner criteria [27].
The blood samples were collected in the fasting state between
implicated in IGF-1 production and secretion from the liver
8 and 9 AM. Serum was separated on site, put in three aliquots and
[14] and 1,25-dihydroxy-vitamin D for regulating the expres-
transported under dry ice to the respective laboratories in Delhi
sion of IGF-1 and IGFBP production and their receptors [15– and Lucknow. The serum samples were kept at −20 °C till analysis
18]. The reported significant increase in serum IGF-1 levels was undertaken. All samples were analyzed at one time. The serum
in infants and children with vitamin D deficient rickets fol- calcium, serum phosphate and serum alkaline phosphatase (SAP)
lowing treatment with vitamin D suggests an association were measured by a commercially available kit using an automated
biochemistry analyzer Hitachi 902 (Roche Diagnostics, Mannheim,
between serum 25OHD and IGF-1 levels [19, 20].
Germany) as reported in our earlier article [28]. The normal range
There is paucity of scientific literature with regard to for serum total calcium was 8.8–10.5  mg/dL (analytical sensitivity
the relationship between IGF-1, IGFBP-3 and vitamin D 0.2  mg/dL), inorganic phosphorus was 2.7–4.5  mg/dL (analytical
[19, 21–25]. Studies both in children and adults show con- sensitivity 0.3  mg/dL) and SAP was for females: <240 U/L and for
trasting results with regard to the relationship between males: <270 U/L (analytical sensitivity 5 IU/L). The serum 25OHD
was assayed using the chemiluminiscence method (Diasorin, Still-
vitamin D and IGF-1 and IGFBP-3. Studies in growth
water, MN, USA) and PTH (reference range: 10–65  pg/mL, ana-
hormone deficiency (GHD) children and infants and chil-
lytical sensitivity 0.7  pg/mL) using a electrochemiluminiscence
dren with nutritional rickets showed positive correlation assay (Roche diagnostics, GMDM-Manheim, Germany), respec-
in contrast to inverse correlation in normal adolescent tively. Intra- and inter-assay coefficients of variation were 3.5% and
children [14, 19, 23]. Due to absence of studies from India, 5% for serum 25OHD and 2.4% and 3.6% for serum PTH. A serum
the present study was undertaken to (a) assess the rela- 25OHD level of <20 ng/mL was defined as VDD [29]. VDD was fur-
ther classified as severe (25OHD <5 ng/mL), moderate (25OHD
tionship between these growth factors and serum vitamin
<10 ng/mL) and mild (25OHD <20 ng/mL) [30].
D-parathormone (PTH) status for correct interpretation IGF-1 and IGFBP-3  were estimated using a Immulite-1000 kit
of serum IGF-1 and IGFBP-3 in apparently healthy Indian (Siemens Healthcare GmbH, Henkestr. Erlangen, Germany) by a
school girls and (b) study the impact of vitamin D supple- solid-phase, enzyme-labelled chemiluminescent immunomet-
mentation on growth factors in pre-pubertal girls. ric assay. The analytical sensitivity for IGF-1 and IGFBP-3  were
20 ng/mL and 0.1 µg/mL, respectively. Intra- and inter-assay coeffi-
cients of variation were <5% and <10% for both. The calculation of the
IGF-I to IGFBP-3 molar ratio was performed after conversion to nmol/L
Materials and methods (IGF-I: ng/mL * 0.1307 = nmol/L; IGFBP-3: ng/mL * 0.03478 = nmol/L).
To better illustrate the relative abundance of IGF-I and IGFBP-3  mole
We had previously evaluated school girls in the age group of cules in blood, the resulting molar ratio was expressed as a percent-
6–18  years studying in fee paying schools located in four different age (ratio IGF-I [nmol/L] to IGFBP-3 [nmol/L] * 100) [31].
geographical zones of Delhi for IGF-1, IGFBP-3 and their relation with In view of a high prevalence of VDD reported in Indian school
thyroid functions [26]. A total of 847 girls, whose consent had been children and vitamin D being known to be involved in the production
obtained, were recruited for the study. One school from each zone and secretion of IGF-1 from the liver, we decided to undertake a pilot
was selected on the basis of permission granted by the school author- study to assess the impact of vitamin D supplementation on serum
ities to provide a representative sample for the study. The study pro- IGF-1, IGFBP-3 and their molar ratio in pre-pubertal girls as these
tocol was approved by the Ethics Committee of the All India Institute growth factors are greatly influenced by the pubertal changes [26].
of Medical Sciences, New Delhi. At the time of initiating the study, These pre-pubertal girls with VDD, aged ≤10  years of age, received
the parents of children were sent a detailed study protocol and their 60,000 IU of micellized vitamin D under supervision once a month
interaction with principal investigator was arranged by the school for a period of 6 months. Of the 348 pre-pubertal girls, 190 consented
authorities to clear their doubts before obtaining their consent. This to be a part of the study. Three girls who had entered puberty at the
study was conducted according to the guidelines laid down in the end of supplementation were excluded from the study and three oth-
Declaration of Helsinki. ers dropped out. One hundred and eighty-four girls, who completed
Girls with clinically overt hepatic, renal, neoplastic, gastrointes- the study, were re-evaluated for all the parameters within 3 days of
tinal, dermatological and endocrine and systemic infective disorders the last supplementation dose.
and steroid intake were excluded. The entire cohort of school girls Statistical analysis was carried out using the SPSS version 20.0
underwent an assessment for height, weight, body mass index (BMI), (Chicago, IL, USA). Data were presented as mean ± SD (95% confi-
pubertal status and blood test for 25OHD, PTH, IGF-1 and IGFBP-3. dence interval) or number (%) unless specified. IGF-1 levels and the
The height was measured to the nearest 0.1 cm using a portable wall- IGF-1/IGFBP-3  molar ratio were first normalized by conversion to
mounted stadiometer (Holten’s Stadiometer, 200 cm/78 inches, Model log (natural), and the values were expressed after converting back

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 Marwaha et al.: IGF-1, IGFBP-3 and 25OHD      3

with exponential to log natural. A p-value for trend was used to test

for trend
p-Value

<0.0001
<0.0001
<0.0001
<0.0001
0.169
<0.0001
0.001
<0.0001
<0.0001
differences between vitamin D categories. A post-hoc analysis was
used to compare the significance level between two groups within
each parameter. Pearson’s correlation coefficient was calculated to
assess the strength of relationship between IGF-1 and IGFBP-3  with
various parameters. Age, height and pubertal status were entered as
covariate separately to adjust the effect of these parameters on IGF-1,

Vitamin D sufficient 

44 (5.2%) 
11.3 ± 3.5 (10.3–12.4) 
1.43 ± 0.16 (1.38–1.48) 
37.7 ± 14.1 (33.4–42.0) 
17.7 ± 3.7 (16.5–18.8) 
9.4 ± 0.6 (9.2–9.6) 
4.8 ± 0.7 (4.5–5.0) 
245 ± 87 (216–274) 
27.9 ± 5.8 (26.2–29.8) 
33 ± 13 (32, 13–60) 
(>20 ng/mL)
IGFBP-3 and the IGF-1/IGFBP-3 molar ratio. A paired t-test was used to
assess the various parameters pre- and post-vitamin D supplementation.

Results
The mean age of the study subjects was (11.5 ± 3.2  years,
range 5.8–18  years), height (1.44 ± 0.15  m, range 1.02–
1.81  m), weight (40.5 ± 16.1 kg, range 14.5–98.8 kg) and

Mild VDD (25OHD 

241 (28.5%) 
10.7 ± 3.1 (10.3–11.0) 
1.40 ± 0.15 (1.39–1.42) 
36.0 ± 14.4 (34.1–37.8) 
17.6 ± 4.3 (17.0–18.1) 
9.4 ± 0.7 (9.3–9.5) 
5.0 ± 0.7 (4.9–5.1) 
249 ± 75 (239–259) 
13.4 ± 2.6 (13.1–13.7) 
43 ± 18 (41, 15–134) 
>10 to ≤ 20 ng/mL)
BMI (18.7 ± 4.8 kg/m2, range 10.46–39.08 kg/m2), respec-
tively. Out of 847 girls, 348 (41.1%) were pre-pubertal and
the rest had entered puberty. VDD was observed in 94.8%
of girls and the mean baseline serum 25OHD levels were
9.9 ± 5.6 ng/mL.

Relation of growth factors to vitamin D

The basic characteristics of the study population accord-

Moderate VDD (25OHD 

445 (52.5%) 
11.5 ± 3.1 (11.2–11.7) 
1.44 ± 0.15 (1.43–1.46) 
41.0 ± 16.4 (39.5–42.6) 
18.9 ± 5.0 (18.5–19.4) 
9.4 ± 0.6 (9.4–9.5) 
5.0 ± 0.7 (4.9–5.0) 
250 ± 84 (242–258) 
7.7 ± 1.4 (7.6–7.8) 
56 ± 38 (48, 14–409) 
>5 to ≤ 10 ng/mL)

ing to vitamin D status are given in Table  1. Unadjusted

serum IGF-1 levels and the IGF-1/IGFBP-3 molar ratio were
significantly higher in girls with severe VDD as compared
to girls with mild-to-moderate VDD. However, these differ-
ences disappeared when adjusted for age, height or sexual
maturation (Table  2). No differences were observed in
Table 1: Basic characteristics of study population according to vitamin D status.

serum IGFBP-3 among the different vitamin D categories.

There was no correlation noted between 25OHD levels and
IGF-1 (r = −0.038, p = 0.270), IGFBP-3 (r = −0.029, p = 0.405)
Severe VDD 

117 (13.8%) 
13.3 ± 2.7 (12.8–13.8) 
1.51 ± 0.13 (1.49–1.53) 
48.6 ± 15.8 (45.7–51.5) 
20.8 ± 4.9 (20.0–21.7) 
9.3 ± 0.7 (9.2–9.4) 
4.7 ± 0.7 (4.5–4.8) 
283 ± 118 (261–306) 
4.5 ± 0.5 (4.4–4.6) 
108 ± 85 (71, 18–460)a 
(25OHD ≤ 5 ng/mL)

and their molar ratio (r = 0.018, p = 0.595).

Effect of vitamin D supplementation

on growth factors

The pre- and post-supplementation parameters of 184

girls evaluated for the study are given in Table 3. A signifi-
cant improvement in heights, weights and BMI observed
Serum PTH values (median, range).
 

 
 
 
 
 
 
 
Serum alkaline phosphatase, U/L  
 
 

was probably a result of normal growth pattern. A sig-

nificant increase in serum calcium and 25OHD along with
Serum phosphates, mg/dL

a marked decline in serum PTH was noted following

Serum calcium, mg/dL

vitamin D supplementation. All girls achieved vitamin D

Serum PTH, pg/mL

sufficiency with a mean serum 25OHD levels of 35.0 ± 16.5

25OHD, ng/mL

ng/mL. Serum IGF-1 and IGFBP-3 levels increased signifi-

Parameters

BMI, kg/m2
Weight, kg

cantly post supplementation with vitamin D. This increase,

Age, years
Height, m
Number

though, was more in girls with moderate-to-severe VDD

compared to girls with mild VDD (28.4 ± 91.9  ng/mL vs.
a

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4      Marwaha et al.: IGF-1, IGFBP-3 and 25OHD

Table 2: IGF-1 and IGFBP-3 among vitamin D deficient girls.

Parameters   Severe VDD  Moderate VDD  Mild VDD  p-Value

(25OHD ≤5 ng/mL) (25OHD >5 to ≤10 ng/mL) (25OHD >10 to ≤20 ng/mL) for trend

Number   117  445  241 

IGF-1, ng/mLa
 Unadjusted   352 ± 1.1 (264–393)  298 ± 1.0 (279–318)  265 ± 1.0 (244–288)  <0.0001
 Adjusted for age   282 ± 1.1 (254–312)  299 ± 1.0 (284–315)  294 ± 1.0 (274–316)  0.588
 Adjusted for height   295 ± 1.1 (266–327)  296 ± 1.0 (281–311)  293 ± 1.1 (273–315)  0.977
 Adjusted for puberty   287 ± 1.1 (259–318)  297 ± 1.0 (282–313)  294 ± 1.1 (274–316)  0.828
IGFBP-3, µg/mL
 Unadjusted   4.2 ± 0.19 (3.8–4.6)  4.2 ± 0.09 (4.0–4.4)  4.1 ± 0.13 (3.8–4.3)  0.340
 Adjusted for age   4.1 ± 0.19 (3.7–4.5)  4.2 ± 0.09 (4.0–4.4)  4.1 ± 0.13 (3.9–4.4)  0.699
 Adjusted for height   4.2 ± 0.19 (3.8–4.6)  4.2 ± 0.09 (4.0–4.4)  4.1 ± 0.13 (3.8–4.3)  0.806
 Adjusted for puberty   4.0 ± 0.18 (3.7–4.4)  4.2 ± 0.09 (4.0–4.4)  4.2 ± 0.13 (3.9–4.4)  0.644
IGF-1/IGFBP-3 molar ratio
 Unadjusted   35.2 ± 1.1 (31.3–39.7)  29.7 ± 1.0 (28.2–31.6)  27.3 ± 1.0 (25.1–29.6)  0.033
 Adjusted for age   28.9 ± 1.1 (25.9–32.1)  29.8 ± 1.0 (28.2–31.4)  29.9 ± 1.0 (27.7–32.2)  0.912
 Adjusted for height   30.1 ± 1.1 (27.1–33.5)  29.5 ± 1.0 (27.9–31.1)  29.8 ± 1.0 (27.7–32.0)  0.995
 Adjusted for puberty   29.7 ± 1.1 (26.6–33.1)  29.6 ± 1.0 (28.1–31.3)  29.7 ± 1.0 (27.6–32.0)  0.979

Values are expressed as mean ± SE. aValues for IGF-1 and IGF-1/IGFBP-3 molar ratio are derived from converting it to log natural and then
converting back to a number using exponential after analysis.

Table 3: Changes in parameters pre- and post-vitamin D supplementation.

Parameters Pre-supplementation Post-supplementation Paired t-test

Height, m 1.30 ± 0.11 1.33 ± 0.11 <0.0001

Weight, kg 26.6 ± 8.0 28.9 ± 8.8 <0.0001
BMI, kg/m2 15.4 ± 3.2 16.1 ± 3.5 <0.0001
Serum calcium, mg/dL 9.4 ± 0.7 9.9 ± 0.6 <0.0001
Serum phosphates, mg/dL 5.2 ± 0.6 5.1 ± 0.5 0.028
Serum ALP, U/L 259 ± 68 264 ± 68 0.469
25OHD, ng/mL 10.3 ± 3.7 35.0 ± 16.5 <0.0001
Serum PTH, pg/mL 42 (15–157) 29 (7–87) <0.0001
IGF-1, ng/mL 145 ± 2 159 ± 2 0.044
IGFBP-3, µg/mL 3.7 ± 0.8 3.8 ± 1.1 0.037
IGF-1/IGFBP-3 molar ratio 15.35 ± 1.55 16.32 ± 1.62 0.11
a
Serum PTH values expressed as median (range). bValues for IGF-1 and IGF-1/IGFBP-3 molar ratio are derived from converting it to log natural
and then converting back to a number using exponential after analysis. ALP, alkaline phosphatase.

14.0 ± 83.1 ng/mL), it did not reach statistical significance A study carried out by Witkowska-Sędek et  al. [14]
(p = 0.275). The increase in IGF-1 levels also did not cor- to evaluate the relationship between serum 25OHD and
relate with the change in height (r = 0.052, p = 0.484), IGF-1 in children and adolescents with GHD observed a
weight (r = 0.019, p = 0.800), BMI (r = 0.023, p = 0.755), significant positive correlation between the two para-
25OHD (r = 0.125, p = 0.090) and PTH (r = −0.124, p = 0.095), meters. They concluded that VDD should be normal-
but significantly correlated with the change in IGFBP-3 ized before IGF measurements to obtain reliable IGF-1
(r = 0.365, p = <0.0001). values [14]. It will therefore be beneficial to assume that
with increasing levels of vitamin D, IGF-1 levels will also
increase because of increased IGF-1 production and secre-
Discussion tion from the liver. However, contrary to this assumption,
the serum IGF-1 levels and the IGF-1/IGFBP-3  molar ratio
In this study, we have evaluated the relationship between were higher in girls with severe VDD when compared with
growth factors and 25OHD in Indian school girls and those with mild-to-moderate VDD in the present study.
assessed the effect of vitamin D supplementation on the This difference, though, disappeared when serum values
growth factors among these girls with VDD. of growth factors were adjusted for age, height and sexual

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maturation. A similar study addressing the relationship of vitamin D (300,000 units) in infants and children with
between serum IGF-1, vitamin D and ferritin levels in Leb- nutritional rickets was carried out by Soliman et al. [19].
anese school children also showed an inverse relation- Besides reporting an increase in serum calcium, phos-
ship between the two parameters in both sexes. However, phates and a decrease in SAP and PTH, it also showed
after adjustment for age, height and BMI, the correlation an increase in the mean serum IGF-1 levels of 26 ng/mL
between IGF-1 and serum 25OHD disappeared [23]. post vitamin D supplementation, consistent with a rise of
The hypothesis put across for the above observation 22 ng/mL in the present study. A study among 22 infants
was that VDD creates an environment of acquired growth with rickets also revealed similar findings [20].
hormone (GH)/IGF-1 resistance, which leads to an increase Studies among adults with different clinical presen-
in serum IGF-1 levels with severe VDD. In one of the in-vitro tations showed contrasting results following vitamin D
studies, evaluating the effect of vitamin D on the gene supplementation. Ameri et al. [21] conducted a study in
expression of growth factors on human fetal epiphyseal adult subjects with GHD (39 subjects) who, when given
chondrocytes, vitamin D was shown to regulate human vitamin D supplementation daily for 3  months resulted
fetal epiphyseal growth by making it more sensitive to GH in an increase in serum IGF-1 levels by 31 ng/mL. In con-
and IGF-1 [32]. In an another experimental study, vitamin trast, vitamin D supplementation among VDD or vitamin
D and IGF-1  were observed to mutually regulate their D insufficient hypertensive adults (175 participants) for
respective receptors in growth plate chondrocytes, where 8 weeks did not result in any significant effect on IGF-1
low levels of serum 25OHD were shown to be stimulatory levels [22]. Another study among middle-aged overweight
and high levels inhibitory [33]. Similarly, the ligand acti- and obese subjects showed that vitamin D supplementa-
vated vitamin D receptor is known to negatively regulate tion for 1 year decreased the IGF-1/IGFBP-3  molar ratio
the growth factors in the presence of IGFBP-3 [34]. Hence, [25]. This difference can be explained by the inclusion
low serum 25OHD levels may up-regulate the production of many patients with vitamin D insufficiency and a
of growth factors in the presence of IGFBP-3. In the present higher level of mean 25OHD levels, which will alter the
study, there was no correlation seen between 25OHD and gene expression of growth factors [33]. This is further
serum IGFBP-3, though active vitamin D has been shown substantiated by the finding in our study that serum
to regulate the transcription of IGFBP-3 [16, 17]. IGF-1 response was higher with severe VDD as compared
Studies both in children and adults show contrast- to mild-to-moderate VDD. Another explanation given
ing results with regard to relationship between vitamin for the increase in IGF-1  was the associated increase in
D and IGF-1 and IGFBP-3. A study in GHD children and IGFBP-3 with vitamin D supplementation [16, 17], which
infants and children with nutritional rickets showed a would increase the bound form of IGF-1. In the present
positive correlation in contrast to an inverse correlation in study, there was a significant increase in serum IGFBP-3,
normal adolescent children [14, 19, 23]. The present study and increase in IGF-1 was only correlated with IGFBP-3,
in normal healthy girls, however, observed no correlation supporting the explanation.
between serum 25OHD and IGF-1, IGFBP-3 and their molar This study had the following limitations: (a) absence
ratio. Similarly, adult studies also showed either a positive of boys in the study because the permission given to
correlation [15, 35] or no correlation [22, 24, 36, 37]. The conduct the study was by schools enrolling only girls. (b)
studies which showed a positive correlation had vitamin D Recruitment of control subjects with VDD was not under-
sufficient population [15, 31, 35] as against subjects in the taken as it would have been unethical to hold treatment
present study who were mostly VDD. This further reiter- with vitamin D supplementation for that long and it would
ates our observation that the population with severe VDD have also been difficult to explain to the parents of these
alters the relationship between serum 25OHD and growth children as to why we did not treat their VDD girls once
factors by either acquired resistance or some unknown they had their serum vitamin D report.
mechanisms.

Conclusions
Effect of vitamin D supplementation
on growth factors Though the serum IGF-1 levels and the IGF-1/IGFBP-3 molar
ratio were apparently higher in girls with severe VDD when
There are limited studies evaluating the effect of vitamin D compared to those with mild-to-moderate VDD, but this
supplementation on growth factors [19–22, 25]. A study in difference disappeared when adjusted for age, height and
children assessing the impact of intramuscular mega dose sexual maturation. Vitamin D supplementation resulted

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6      Marwaha et al.: IGF-1, IGFBP-3 and 25OHD

in a significant increase in serum IGF-1 levels in VDD pre- 8. Marwaha RK, Sreenivas V, Talwar D, Yenamandra VK, Challa
pubertal girls. A, et al. Impact of solar UVB radiation (290–320) on vitamin
D synthesis in children with type IV and V skin. Br J Dermatol
2015;173:604–6.
Acknowledgments: The authors gratefully acknowledge the 9. Marwaha RK, Puri S, Tandon N, Dhir S, Agarwal N, et al. Effects
assistance and participation of the school principals, chil- of sports training & nutrition on bone mineral density in young
dren and their parents in carrying out this study. We deeply Indian healthy females. Indian J Med Res 2011;134:307–13.
appreciate the assistance of the following team mem- 10. Jones JI, Clemmons DR. Insulin-like growth factors and their
binding proteins: biological actions. Endocr Rev 1995;16:3–34.
bers from the Society of Endocrine Health Care of Elderly,
11. Nesbitt T, Drezner MK. Insulin-like growth factor-I regulation of
Adolescents and Children (SEHEAC) and students of SUR
renal 25-hydroxyvitamin D-1-hydroxylase activity. Endocrinology
Homeopathic Medical College: Mrs. Pamela Marwaha, Dr. 1993;132:133–8.
SK Mathur, Mrs. Neeru Gandhi, Dristhi Sharma, Archita 12. Bianda T, Hussain M, Glatz Y, Bouillon R, Froesch E, et al. Effects
Gupta and Shrijan Jaipuriar. The study was supported by a of short-term insulin-like growth factor-I or growth hormone
research grant from CCRH, Ministry of Ayush. treatment on bone turnover, renal phosphate reabsorption and
1,25 dihydroxy-vitamin D3 production in healthy man. J Intern
Author contributions: All the authors have accepted
Med 1997;24:1143–50.
responsibility for the entire content of this submitted 13. Locatelli V, Bianchi VE. Effect of GH/IGF-1 on bone metabolism
manuscript and approved submission. and osteoporosis. Int J Endocrinol 2014;2014:235060.
Research funding: Financial support was provided by 14. Witkowska-Sędek E, Kucharska A, Rumińska M, Pyrżak B.
Central Council of Research in Homeopathy, Ministry of Relationship between 25(OH)D and IGF-I in children and ado-
lescents with growth hormone deficiency. Adv Exp Med Biol
Ayush and Endocrine and Diabetes Foundation of India,
2016;912:43–9.
New Delhi.
15. Gómez JM. The role of insulin-like growth factor I compo-
Employment or leadership: None declared. nents in the regulation of vitamin D. Curr Pharm Biotechnol
Honorarium: None declared. 2006;7:125–32.
Competing interests: The funding organization(s) played 16. Malinen M, Ryynänen J, Heinäniemi M, Väisänen S, Carlberg
no role in the study design; in the collection, analysis, and C. Cyclical regulation of the insulin-like growth factor binding
protein 3 gene in response to 1alpha,25-dihydroxyvitamin D3.
interpretation of data; in the writing of the report; or in the
Nucleic Acids Res 2011;39:502–12.
decision to submit the report for publication. 17. Matilainen M, Malinen M, Saavalainen K, Carlberg C. Regula-
tion of multiple insulin-like growth factor binding protein
genes by 1alpha,25-dihydroxyvitamin D3. Nucleic Acids Res
2005;33:5521–32.
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