Induction of an autoimmune thyroid disease with nickel chloride in Brown Norway rats/ Kahverengi Norvec sicanlarinda nikel klorid ile otoimmun tiroid hastaliginin induklenmesi.
Abstract: Objective: Hazards caused by nickel exposure, due to its direct and autoimmune reaction, are a growing problem. The aim of this study is to explore whether nickel will induce autoimmune thyroid disease in immunosensitive rats.

Materials and Methods: In this study, twelve Brown Norway rats were selected from our previous study (Almogairen et al, Lupus 2009 April). The rats were sacrificed after the exposure to nickel and then thyroidectomized. Histological studies of the thyroid were performed in the group of six rats positively responding to nickel and were compared with the equal number of rats in the autoantibody-negative control group.

Results: Histologically, epithelial follicular proliferation in the thyroid gland was significant in the subcutaneous nickel group (p=0.05).

Conclusions: When correlating the above results with serum ANA response of the same rats, it might be concluded that nickel chloride might induce autoimmune thyroid disease in immnosensitive rats.

Key words: Autoimmune, thyroid, ANA, nickel

Amac: Nikele maruz kalmanin getirdigi tehlikeler, direkt ve otoimmun reaksiyon nedeniyle, buyumekte olan bir problemdir. Bu calismanin amaci, nikelin otoimmun tiroid hastaligini baslatip baslatmadigini arastirmaktir.

Gerec ve Yontemler: Bu calismada, daha onceki calismamizdan (Almogairen at al., Lupus, 2009 Nisan) 12 kahverengi Norvec sicani secildi. Sicanlar nikele maruz birakildiktan sonra olduruldu ve tiroidektomize edildi. Nikele pozitif cevap veren gruptaki 6 sican uzerinde histolojik incelemeler yapildi ve ayni sayidaki antikor negatif kontrol grubu sicanlari ile karsilastirildi.

Bulgular: Histolojik olarak, subkutan nikel uygulanan grubun tiroid bezinde epitelyal follikuler proliferasyon anlamliydi.

Sonuc: Yukaridaki sonuclar, ayni sicanlarin serum antinukleer antikor (ANA) cevabi ile iliskilendirildiginde, nikel klorurun immunosensitif sicanlarda otoimmun tiroid hastaliginin induklenmesinde onemli rol oynayabilecegi sonucuna varilabilir.

Anahtar kelimeler: Otoimmun, tiroid, ANA, nikel
Article Type: Report
Subject: Thyroid diseases (Risk factors)
Thyroid diseases (Diagnosis)
Thyroid diseases (Research)
Nickel compounds (Health aspects)
Nickel compounds (Research)
Antinuclear antibodies (Physiological aspects)
Antinuclear antibodies (Research)
Author: Mogairen, Sultan Al-
Pub Date: 12/01/2009
Publication: Name: Turkish Journal of Endocrinology and Metabolism Publisher: Galenos Yayincilik Audience: Academic Format: Magazine/Journal Subject: Health Copyright: COPYRIGHT 2009 Galenos Yayincilik ISSN: 1301-2193
Issue: Date: Dec, 2009
Topic: Event Code: 310 Science & research
Product: Product Code: 2819959 Nickel Compounds NAICS Code: 325188 All Other Basic Inorganic Chemical Manufacturing SIC Code: 2819 Industrial inorganic chemicals, not elsewhere classified
Geographic: Geographic Scope: Saudi Arabia Geographic Code: 7SAUD Saudi Arabia
Accession Number: 219520153
Full Text: Introduction

Nickel allergy is the most common cause of allergic contact dermatitis. It's prevalence in the general population is about 7-10% with predominance in females (female: male ratio of 7-14:1) (1). Nickel is used for making stainless steel which is used for production of equipments, fashion jewelry, coins, metal eyeglasses, prosthetic implants, as well as dental crown, bridges and implants (1-6).

One of the target organs to toxins and drugs is the thyroid gland, which dysfunction is manifested as hypothyroidism, hyperthyroidism, c-cell hyperplasia (7,8). Thyroid diseases are associated with drugs such as interferon-alpha, interleukin-2, lithium, cyclosporine and amiodarone, or toxins such as organochlorine (9-16).

Chronic interferon-alpha exposure leads to changes in thyroid gland, including enlarged follicles lined by thickened granular and oxyphilic thyrocytes with diffuse mononuclear infiltrates of the stroma (8). Organochlorine induces thyroid gland cysts and c-cell hyperplasia (7).

The objective of this experimental study is to demonstrate whether nickel would induce autoimmune thyroid disease in Brown Norway rats. This study is an extension of our previous study (Almogairen et al, Lupus 2009 April) 17,but examining the correlation between the autoimmune response and the histopathological changes in the thyroid gland.

Materials and Methods

Brown Norway rats (BN) were purchased from Charles Rivers Laboratories, Wilmington, U.S.A. They were kept in polycarbonate metrolon plastic cages covered with stainless steel cover in the animal house in the College of Medicine, King Saud University, Riyadh, Saudi Arabia. They were maintained under 12-hr dark: 12-hr light cycles and were kept under observation for three weeks. No evidence of sickness was observed. All rats were 8-11 weeks old at the onset of the experiment. There were a total number of twelve rats with an average weight of 157 g. There were four groups: the first and the second groups (six rats) were called subcutaneous nickel group (three rats) and oral nickel group (three rats). The third and the fourth groups (six rats) were called subcutaneous normal saline control group (three rats) and oral normal saline control group (three rats).

All the above groups were selected for thyroid histological studies from our previous study, where the first and the second nickel groups were autoantibody-positive for serum ANA, anti-dsDNA, anti-Smith, anti-SSA and anti-SSB. On the other hand, the third and the fourth control normal saline groups were overall autoantibody-negative. The above selected groups were sacrificed after fourteen weeks of exposure to nickel chloride or normal saline and then thy-roidectomized. The tissues taken from them were bisected and fixed in 10% buffered formalin for 24 hours. The tissues were then processed in the Tissue-Tek vacuum infiltration processor and stained using hematoxylin and eosin stain. The slides were examined blindly by histopathologist using light microscope.


The maximum serum antibody titers in the selected rats from our previous study are shown in Table 1.

The histopathological results are shown in Table 2. The changes in the thyroid gland were accepted as negative if all of the parameters shown in Table 2 were negative, otherwise, positive. Positive changes in the thyroid gland depicted in Figure 1 were observed in 3/3 (p=0.05) of subcutaneous nickel chloride group, on the other hand, p value was 0.49 in the oral nickel group.


There are a limited number of publications concerning the structure of thyroid gland following the exposure to some drugs and toxins (7,10,12-14). To our knowledge, this is the first experimental animal study showing the correlation between the histopathological changes in the thyroid gland secondary to challenge with nickel and the serum autoimmune response. One study showed that in rats, injection of nickel sulfate (NISO4) induced the epithelial proliferation in the thyroid glands (16).

In our previous study, almost all serum autoantibodies in Brown Norway rats were undetectable prior to intervention. After challenge with nickel chloride, serum ANA appeared early and in a significant number of rats (p<0.05) in both subcutaneous and orally treated rats. The same response (p<0.05) was seen with anti-SSA in the subcutaneous nickel chloride group. Other serum autoanti-bodies including antidsDNA, anti-Smith and anti-SSB were insignificant (17). The main histopathological finding of the present study was increased epithelial follicular proliferation in the subcutaneous nickel group (p=0.05). In the oral nickel group, increased epithelial follicular changes were observed only in one rat, which was probably due to the processing of nickel chloride substance by gastrointestinal M-cells (18).

The mechanism causing these effects could be due to direct toxic injury, autoimmunity or due to disruption of the hypothalamic-pitu-itary-thyroid (HPT) axis, a disturbance in the calcium homeostasis or in the energy metabolism (7).


With the above significant response of Serum ANA and anti-SSA, the autoimmunity is probably playing a role in the epithelial follicular proliferation in the thyroid gland.

In conclusion, the findings from this study indicate that nickel chloride might induce autoimmune thyroid disease and we recommend, in the future, a immunohistochemical tissue study to be conducted on murine antithyroid antibodies, if available.


The study was supported by a grant from Research Center, College of Medicine, King Saud University. The author is grateful to Dr. Sufia Husain, FRC Path for reporting histology. The author is also grateful for the following: Dr. B. Al-Mohaimeed, Dr. N. Khalil, Mr. M. Marzook, and Mr. S. Abu-al-Ghaith, S. Seno, Mitzi Guinto, Joann Octubre for their cooperation, and the Research Ethic Committee, College of Medicine for the approval of this study.

Recevied: 18.01.2010 Accepted: 26.01.2010


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(17.) Al-Mogairen SM,Al-Arfaj AS,Meo SA,Adam M,Al-Hammad A,Gad El Rab MO. Induction of autoimmunity in Brown Norway rats by oral and par-enteral administration of sodium silicater. Lupus 2009; 18: 413-417.

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Address for Correspondence: Sultan Al-Mogairen, MD, Department of Medicine, Faculty of Medicine, King Saud University, Saudi Arabia Email:

Sultan Al-Mogairen

King Saud University, Medicine, Riyadh, Saudi Arabia
Table 1. Maximum serum antibody titres of the rast in which
histopathology was performed

RATS           ANA ([less than        Anti-dsDNA
              or equal to] 1/10)    ([greater than
                 significant       or equal to] 0.08)

Nickel SC1           1/20                0.088
Nickel SC2           1/80                0.080
Nickel SC3           1/80                0.194
Nickel PO1          1/160                0.211
Nickel PO2           1/20                0.231
Nickel PO3           1/80                0.231
Control SC1          -ve                  -ve
Control SC2          -ve                  -ve
Control SC3          -ve                  -ve
Control PO1          -ve                  -ve
Control PO2          -ve                  0.88
Control PO3          -ve                  -ve

RATS             Ant-Smith            Anti-SSA
               ([greater than      ([greater than
              or equal to] 0.06)  or equal to] 0.12)
                 significant         significant

Nickel SC1           -ve                0.274
Nickel SC2          0.122                0.14
Nickel SC3          0.147                -ve
Nickel PO1          0.109               0.253
Nickel PO2          0.23                 0.17
Nickel PO3          0.149                1.8
Control SC1         0.06                 -ve
Control SC2          -ve                 -ve
Control SC3          -ve                 -ve
Control PO1          -ve                 -ve
Control PO2          -ve                 -ve
Control PO3          -ve                 -ve

RATS              Anti-SSB
               ([greater than
              or equal to] 0.05)

Nickel SC1           0.09
Nickel SC2           -ve
Nickel SC3           -ve
Nickel PO1           -ve
Nickel PO2          0.204
Nickel PO3           0.16
Control SC1          -ve
Control SC2          -ve
Control SC3          -ve
Control PO1          -ve
Control PO2          -ve
Control PO3          -ve

SC: Subcutaneous
PO: Per oral
-ve: Negative

Table 2. Histopathological features of silicate-tested&control groups

Thyroid of the   Inflammation   Autoimmune    Granuloma
following rast                  thyroiditis

Nickel SC1           -ve            -ve          -ve
Nickel SC2           -ve            -ve          -ve
Nickel SC3           -ve            -ve          -ve
Nickel PO1           -ve            -ve          -ve
Nickel PO2           -ve            -ve          -ve
Nickel PO3           -ve            -ve          -ve
Control SC1          -ve            -ve          -ve
Control SC2          -ve            -ve          -ve
Control SC3          -ve            -ve          -ve
Control PO1          -ve            -ve          -ve
Control PO2          -ve            -ve          -ve
Control PO3          -ve            -ve          -ve

Thyroid of the   Epithelial follicular   Hurthle cell   Neoplastic
following rast       proliferation          change        change

Nickel SC1                ++                 -ve           -ve
Nickel SC2                 +                 -ve           -ve
Nickel SC3                ++                 -ve           -ve
Nickel PO1                -ve                -ve           -ve
Nickel PO2                -ve                -ve           -ve
Nickel PO3                 +                 -ve           -ve
Control SC1               -ve                -ve           -ve
Control SC2               -ve                -ve           -ve
Control SC3               -ve                -ve           -ve
Control PO1               -ve                -ve           -ve
Control PO2                +                 -ve           -ve
Control PO3                +                 -ve           -ve

Thyroid of the   Fibrosis
following rast    atrophy

Nickel SC1          -ve
Nickel SC2          -ve
Nickel SC3          -ve
Nickel PO1          -ve
Nickel PO2          -ve
Nickel PO3          -ve
Control SC1         -ve
Control SC2         -ve
Control SC3         -ve
Control PO1         -ve
Control PO2         -ve
Control PO3         -ve

-ve = 0 = negative

+ = 1 = mild

++ = 2 = moderate

+++ = 3= severe

SC: subcutaneous

PO: per oral
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