Mansoura Journal of Dentistry 2014;1(3):142-145.

Impact of Systemically Induced Rheumatoid

Arthritis on Tooth Development in Rats
Offsprings

Ahmed R. Zaher1, Mohamad E. Helal2, Asmaa S. Elmahdy3

1
Professor of Oral Biology, Faculty of Dentistry, Mansoura University, Egypt.
2
Professor of Oral Biology, Faculty of Dentistry, Mansoura University, Egypt.
3
Teaching assistant, Department of Oral Biology, Faculty of Dentistry, Mansoura University, Egypt.

Abstract:
Objectives: In this study, we evaluated the impact of maternal rheumatoid arthritis (RA) in Albino rats on tooth development in their
offspring.
Methods: 30 female rats were divided into 2 groups control group (CG), and rheumatoid non-treated group (RhG). Induction of RA was
performed in (RhG) by injection of collagen II, followed by induction of pregnancy in both groups.
Results: Fetuses of (RhG) showed slower rate of development both histologically and immunohistochemically at first few days
followed by rapid normal growth at 10th day.
Conclusion: Maternal RA, temporarily, slow down the rate of tooth development in their fetuses, then rapid catch up of normal rate of
growth occurs.
Keywords: Rheumatoid arthritis, collagen, tooth development.

Introduction
ooth development starts at 5th week of human
gestation and at 10th embryonic day of mouse
development. Each tooth passes through four
morphological stages: initiation, bud, cap, and bell stages,
named according to the shape of enamel organ [1]. At late
bell stage, odontoblasts secrete first layer of dentin starting
dentinogenesis, dentin formation, and amelogenesis,
enamel formation, through reciprocal induction.
Collagen is the major insoluble fibrous protein in the
extracellular matrix and in connective tissue [2]. Type I
collagen forms a fibrous three-dimensional network which
builds up the dentin matrix. Compared to bone, the collagen
matrix in dentin is more interwoven with numerous
crossings of fibrils [3]. Excessive degradation of type I
collagen is associated with a variety of human diseases such
as RA, tumor metastasis, and atherosclerosis [4].
Rheumatoid arthritis (RA) is a chronic inflammatory
disease characterized by joint swelling, joint tenderness,
and destruction of synovial joints, leading to severe
disability and premature mortality [5]. RA is considered an
autoimmune disease due to presence of auto-antibodies,
such as rheumatoid factor (RF) and anticitrullinated
protein antibody (ACPA) (tested as anticyclic citrullinated
peptide [anti-CCP]), which can precede the clinical
manifestation of RA by many years [6].
Many women with RA are of childbearing age, which
highlights the importance to study if there is any effect on
tooth development of their children. To our best
knowledge, this is the first study that evaluates the effect of
rheumatoid arthritis rat mother on tooth development of
their offspring.
Materials and methods
Animals
20 female Albino rats weighting (180-200 g) were used in
the study and divided into 2groups: control group (CG), and
rheumatoid non treated group (RhG).

Collagen preparation
Arthritis was induced successfully according to the protocol
recommended by Chondrex Inc. Type II collagen (CII),
isolated and puried from bovine articular cartilage
(Chondrex, Inc), dissolved overnight at
C in 0.01 M
acetic acid at a concentration of 2 mg/ml. The solution was
then emulsied in an equal volume of incomplete Freunds
adjuvant (IFA) (Sigma) in a drop-wise fashion with
continuous stirring with electric homogenizer. The stability
of the emulsion was tested by adding one drop of emulsion
into a beaker of water. A stable emulsion remained, as a
solid clump in water without dispersing, while the
spreading onto the water surface indicated an unstable
emulsion [7].
Histological and immunohistochemical analysis
Tongue specimens were fixed in 10% neutral buffered
formalin for 24 hours then were trimmed and processed by
standard paraffin-embedding methods. Sections were cut at
m, deparaffinized, and then stained with: H&E
&Immunohistochemical staining using monoclonal
antibodies to collagen I (COL-1).
Results
Histological analysis
H&E stained coronal sections were examined of fetuses of
CG and RhG for developing tooth germ of the 1st molar. In
CG, at 1st day after birth tooth germ of developing 1st molar
was at late bell stage with definite layer of dentin and at
10th day dentin and enamel matrix formation were almost
completed. In RhG, at 1st day after birth tooth germ of
developing 1st molar was at early bell stage; without dentin
formation and at 10th day normal thickness of enamel
matrix and dentin were formed.(Fig. 1 A&B) and (Fig. 2
A&B).
Immunohistochemical analysis
COL-1 was more brown intense in developing dentin and
bone in CG than in bone in RhG at 1st day after birth. On
the other hand, the colour intensity of COL-1 at 10th day in
Ahmed R. Zaheret al.

Mansoura Journal of Dentistry 2014;1(3):142-145.

both groups was nearly the same. (Fig. 3 A&B) and (Fig. 4
A&B).
Discussion
Systemic auto-immune diseases have higher pervelance in
woman and mainly at child bearing age [8]. Many studies
observed that autoimmune disease in women during
pregnancy may be associated with an increased risk for
learning disabilities in their sons. In this study, RA is one of
systemic autoimmune disease and is strongly associated
with oral health [9]. This is to say because immunological
and pathological processes occurring in periodontitis and
RA are nearly identical. Prospective studies on pregnant
mothers suggested that maternal periodontal disease may
cause preterm birth, low birth weight and may increase
their offspring's risk of developing early and severe dental
caries [10]. So this study was conducted to answer the
question, whether RA diseased albino rat mothers has any
effect on tooth development in their offsprings.
In the present study, histological examination of
developing 1st molar tooth germ at 1st day in RhG showed
stage of early bell rather than late bell stage that was found
in CG. This delay in tooth development of group B,
compared with that in group A, can be explained by
findings of Scott [11] who demonstrated that pregnant
women with connective tissue diseases, such as RA, are at
risk for preterm birth and intra-uterine fetal growth
restriction. As a contradict, many clinical experience and
the reports of over 500 patients in clinical studies
demonstrated that RA activity decreases for many women
during pregnancy and their baby born healthy [12].
However, this may be due to either low disease activity or
good choice of medication taken by mothers during
pregnancy.

Histolofical findings of RhG at 10th day, in this study,

showed normal development as in CG. This was in
agreement with findings of a cohort study of Dutch women
with RA and was accomplished by de Steenwinkel et al.
[13] who demonstrated that active rheumatoid arthritis
(RA) during pregnancy, without medication, and the
presence of RF or anticitrullinated protein antibodies
(ACPAs) are associated with lower birth weight of the child
followed by rapid postnatal catch-up in weight.
In the present study, Immunohistochemical analysis
showed that brown colour intensity of COL-1 in RhG at 1st
day was less than that of CG indicating that secreted
collagen I was decreased in RhG. This result could be
explained ad the following; auto-antibodies against
collagen type I that circulating in blood of high disease
activity RA pregnant mothers were able to cross placenta
and attack collagen type I in developing bone and dentin
resulting in transient slow rate of formation and decreased
amount of collagen in these tissues [14,15]. However, this
was not the case at 10th day, where the colour intensity of
COL-1 was the same in RhG and CG which may be also
due to rapid catch up mechanism.
Conclusion
With the limitation of our study as it was the first study that
investigate the effect of one of maternal autoimmune
disease, RA, on tooth development, we can conclude that
RA, under certain circumstances as absence of medication
or high disease activity, leads to slow rate of development
and decreased amount of collagen I production at first few
days after birth and compensated by rapid compensatory
mechanism and restoration of normal rate of development
and growth at the following days.

st

st

Figure 1: Photomicrograph of developing 1 molar at 1 day, CG (A) and RhG (B). (H&E stain, x 100)

Ahmed R. Zaheret al.

Mansoura Journal of Dentistry 2014;1(3):142-145.

Figure 2: Photomicrograph of developing 1st molar at 10th day, CG (A) and RhG (B). (H&E stain, x 400)

Figure 3: Photomicrograph of developing 1st molar at 1st day ,showing positive brown intense reaction of COL-1 in dentin and
bone of CG (A) and less intense reaction in bone of RhG (B). (COL-1 stain, x 100)

Ahmed R. Zaheret al.

Mansoura Journal of Dentistry 2014;1(3):142-145.

Figure 4: Photomicrograph of developing 1st molar at 10th day ,showing positive brown intense reaction of COL-1 in dentin
and bone of both CG (A) and RhG (B). (COL-1 stain, x 100)
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