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Review Article

Rat as laboratory animal model in periodontology

Sowjanya Guvva, Mallanagouda B Patil1, Mehta DS2
Department of Periodontics and Implantology, Bapuji Dental College, Davangere, 1Department of Periodontics, College of Dental Sciences,
2
Department of Periodontics and Implantology, Bapuji Dental College and Hospital, Davengere, Karnataka, India

Abstract For ethical reason, initiation and progression of periodontal disease as well as certain types of periodontal
treatment cannot be studied in humans. Instead, numerous studies in this field have been carried out in
laboratory animals. Animal models are needed to objectively evaluate the pathogenesis of human periodontal
disease and its various treatment modalities. A number of animal models have been used in studying etiology
and pathology of periodontitis. The primate model is recommended because the pathogenesis in primate model
closely resembles that in humans. In addition, the dog model is used frequently because of ease of ligature
placement as well as the natural occurrence of periodontal disease. However, ease of handling, inexpensive,
short study time, low variation among strains and controlled microflora, relatively disease resistant, make
the rat model extremely versatile and suited for a wide range of research endeavors. Rats are used mainly for
research in toxicity, nutrition, behavior, and cancer. Normal oral structure and physiology and the pathogenesis
of periodontal diseases have been studied more extensively in the rat than in any other rodents. Rats are used
extensively to study the effects of drugs on the gingiva because their tissue overgrowth is similar to that of
humans. The purpose of this review is to evaluate rats as models for studying various aspects of periodontal
disease, including disease process and its treatment handling, advantages and limitations of these models.

Keywords: Animal models, periodontitis, rat, research

Address for correspondence: Dr. Mallanagouda B Patil, Department of Periodontics, College of Dental Sciences, Room No. 4, Davangere ‑ 577 004,
Karnataka, India.
E‑mail: mal_pati@rediffmail.com

INTRODUCTION tissues of a variety of mammals/animals have been

For very long time, periodontal diseases, in particular,
the severe forms of periodontitis, have been and still are
enigmatic both to those who suffer from them and to those
who try to understand and prevent them. Attempts to
understand these diseases through clinical and experimental
research have been made continuously and with varying
success for more than a 100 years.[1]
In addition to man, several other mammals, particularly those
kept in captivity or in the domestic domain, spontaneously
develop periodontitis with age. Therefore, the periodontal
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studied, some more extensively than others, in the
hope of finding a true analog to human gingivitis and
periodontitis. This search has been going on since
about the turn of the century. We now have a great
deal of reliable information about the histopathologic
and pathophysiology of the normal periodontium and
about the histopathologic and pathophysiologic aspects
of spontaneously and experimentally diseased periodontal
tissue of several animals.
In the process of searching for common denominators,
Page and Schroeder introduced us to some very interesting
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DOI:
10.4103/ijohs.ijohs_47_17 How to cite this article: Guvva S, Patil MB, Mehta DS. Rat as laboratory
animal model in periodontology. Int J Oral Health Sci 2017;7:68-75.

68 © 2018 International Journal of Oral Health Sciences | Published by Wolters Kluwer - Medknow
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Guvva, et al.: Rat as laboratory animal model
forms of periodontitis in animals, such as “cara inchada”
(swollen face), a disease found in cattle in West Central
Brazil and the “broken mouth” periodontitis of sheep.
Because of these observations are then related to
periodontal diseases in rodents and nonhuman primates,
to ligature‑induced disease, to the nature of the disease in
germ‑free animals, to the incidence of periodontal disease
in nondominated versus dominated animals and to many
other aspects of comparative pathology.[1]
Animal models for testing periodontal regenerative
procedure are necessary because controlled quantitative
histological analysis is required to evaluate the quality and
extent of the rarely formed supporting tissues. Histometric
evaluation can determine the amount of new cementum,
periodontal ligament, and alveolar bone formed the amount
of new regenerative periodontal surgery. These studies are
not possible in man because of the need to retrieve the
teeth and their surrounding periodontium in large blocks
appropriate for histological analysis. Furthermore, proper
evaluation of a new therapy necessarily involves the use
of treated and untreated controls which are difficult to
obtain in the human. The testing of potentially harmful
new devices and pharmaceuticals may be unethical in man
before thorough evaluation in higher animals.[2]
Animal research and its value to human experience
remain controversial. Animal model data can provide
us with models of biologic trends before proceeding to
human application. Each animal species are manifested
by a wide range of clinical and histopathological features.
Different species have distinct traits, habits, life spans,
tissue structures, host defense mechanisms and genetic
microbiological immunological, and clinical features of
periodontal disease and its prevention and treatment.
Here, we have made an attempt to review to evaluate rats as
models for studying various periodontal diseases, including
disease process, its prevention and treatment, advantages,
and limitations of these models.
ORIGIN
The laboratory rat, Rattus norvegicus, is a rodent of the
family Muridae [Figure 1]. Wild rats’ apparently originated,
in recent times at least, in the temperate regions of central
Asia, from southern U. S. S. R. through northern China.
Through migration along trade and military routes, the
cosmopolitan rat has spread around the world.
Domesticated rats were raised by fanciers in the seventeenth
century and for combat with terriers in subsequent
International Journal of Oral Health Sciences | Volume 7 | Issue 2 | July-December 2017 69
centuries. By the mid‑1800s, rats were used in scientific
experimentation. Rats, such as mice, are now available
in various ecologic and genetic varieties, however, most
research rats are barrier‑raised in specific pathogens free
colonies.[3,4]
Rat is one of the most widely used species of laboratory
animals. Its popularity is next only to that of mice and
found all over the world, especially in association with
human habitation. R. norvegicus  (scientific name) adopts
readily in laboratories all over the world. Rats are used
mainly for research in toxicology, nutrition, behavior, and
cancer. Proportionately, a smaller number, are also used
in physiological and pharmacological investigations and
for teaching.
The most frequently used inbred strains of rats
include the Wister albino, Lewis, Norwegian grey, Rice
(Oryzomys palustris) (Leonard, 1979), Kyoto and Carworth
Wistar, Osborne‑Mendel, Holtzman, white Lobund, CD
and CDF‑Fisher 344, and several strains of the Sprague
Dawley rat (Forsyth strain, CR strain, and RIC strain).[1]
Sprague Dawley rat, an albino, has a narrow head and tail
longer than the body, whereas the Wistar rat has a wide
head and shorter tail. The Long‑Evans and other “hooded”
varieties are smaller than the albino strains and have darker
hair over portions of the head and anterior body.
Normal oral structure and physiology and the pathogenesis
of periodontal diseases have been studied more extensively
in the rat than in any other rodent. For this reason, the
rat serves as an example for other rodents such as mice
and hamsters. Although particular dental and periodontal
tissues differ in size among various rat strains, principles
of tissue structure and growth phenomena are identical
in all strains.
ANATOMIC PHYSIOLOGIC CHARACTERISTICS
The rat is the most extensively studied rodent for the
pathogenesis of periodontal disease. Typical rodent
dentition is Incisor 1/1, canine 0/0, premolar 0/0/ and
molars 3/3. The incisors are rootless with continuously
erupting [Figure 2-5]. The cheeks closes into the diastema,
separating incisors from the oral cavity and have an
articulated mandibular symphysis. The structure of the
dental gingival area in rat is quite similar to that observed
in humans. They have a tapered head with pinnae relatively
smaller than those of mouse, a long tail, and an anus usually
pressed on the ground. An adult rat weights about 530-900
grams and varies from species to species.
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Guvva, et al.: Rat as laboratory animal model
Figure 1: Rat
Figure 3: Mandibular molar teeth
The gnawing apparatus, shared with other rodents, is
remarkable. The continuously erupting (hypsodontic),
chisel‑like incisors, powerful jaw muscles, articulated
symphysis, and long diastema with loose cheek skin all
contribute to the rat’s omnivorous habits and gnawing
ability. The 12 molars, on the other hand, are permanently
rooted (brachiodontic), located far back in the mouth, and
are used for grinding.[3,4]
LIFE SPAN
Rats may live longer than 3 years and have a productive
breeding life of approximately 9 or more months or until
they are 12 or more months of age, during which time
they may bear 7–10 L with 6–14 offspring per litter. After
12 months, litter size decreases and litter interval increases
until sexual senescence at 450–500 days.
HANDLING
Rats gently handled become tame and will rarely bite unless
startled or hurt. Because the tail may tear, the tail should
be grasped only at the base and for short periods. Rats are
picked up by placing the hand firmly over the back and rib
70 International Journal of Oral Health Sciences | Volume 7 | Issue 2 | July-December 2017
Figure 2: Maxilla and mandible of rat
Figure 4: Lower incisors
cage and restraining the head with thumb and forefinger
immediately behind the mandibles. Rats held upside down
are more concerned with righting themselves than with
biting [Figure 6].
HOUSING
Rats are housed either in metal cages [Figure 7] with mesh or
in plastic cages with solid floors. “Rats mesh” should have
openings of 2 wires per inch. A homemade, shoebox‑shaped
cage with a hardware cloth top is satisfactory for pet rats,
Terraria are commonly used to house pet rodents, but care
must be taken to assure accessibility to water and feed. Care
should be taken to prevent escape, loss of neonates through
wire floors, and fractured limbs from too close a mesh.
Young rats housed on the wire and deprived of the nest
may become dehydrated, especially during months when
ambient humidity is low. Rat cages should provide at least
259 cm square floor space per adult (300 g) rat. A female
with litter requires 1000 cm square floor space. Rat cages
should be at least 18 cm high. Bedding for rats, which
may be paper, wood chips or shavings, ground corn cob,
or sawdust, should be nonallergenic, dust free, absorptive,
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Guvva, et al.: Rat as laboratory animal model
Figure 5: Upper incisor
Figure 6: Rat holding
Figure 7: Rat cage
nontoxic, and clean. Soft shavings are more easily formed
into nests.[3,4]
Temperatures in rat rooms should be maintained between
20°C and 25°C relative humidity should be between
40% and 70%. Light intensity is reduced within translucent
or filtered cages. A 12:12 dark light cycle is usually used in
rat colonies; continuous light will depress cycling. Rodents
in general especially nocturnal rodents, do no very well
in dimly lighted rooms. Apparently, the bright light used
International Journal of Oral Health Sciences | Volume 7 | Issue 2 | July-December 2017 71
in most benefit than for the rodent’s wellbeing. Bedding
should be charged as often as necessary to keep odor
minimal and the rats dry and clean, one to three bedding
changes per day week are usually sufficient. Cages are
washed once or twice weekly.
FEEDING AND WATERING
Rats are fed a clean, wholesome, fresh, and nutritious
pelleted rodent diet free choice. The commercially
available diets are complete that is they do not require
supplementation. Rats should be watered from water
bottles with sipper tubes or through an automatic watering
system. Several varieties of special purpose rodent chows
are available. Rats are cautious feeders and will avoid
strange foods.
An adult rat will consume approximately 5 g food and 10 ml
water/100 g body weight per day. Consumption varies with
the ambient temperature, humidity, health status, breeding
stage, diet, and the time of day. Rats are nocturnal and feed
primarily at night.
PUBLIC HEALTH CONCERNS
Allergies to animal dander and urinary proteins (serum proteins)
occur commonly in humans, and cutaneous and upper and
lower respiratory allergies to rats (large crowded, poor air
circulation, infrequent cage changing, and cleaning lead to
accumulation of ammonia gas) are very common.
Zoonotic diseases carried or transmitted by rats include
leptospirosis, streptococcal infections, salmonellosis,
cestodiasis, Korean hemorrhagic fever and rat bite, or
Haverhill fevers. Sylvatic plague is carried by rat fleas.
The mite liponyssus sylviarum can transmit the St, Louis
encephalitis virus, and L bacoti attacks man directly.
Rat‑bite fever is caused by Streptobacillus moniliformis,
which is carried in the nasopharynx of asymptomatic rats.
People experiencing rat bite fever may develop recurrent
fever with petechial hemorrhages, endocarditits, and
polyarthritis. Although these diseases and infections may
occur in wild rats brought into a laboratory, they are rare
or unknown in domestic rats.[3,4]
USES IN RESEARCH
Laboratory rats include studies of aging, neoplasia, drug
effects and toxicity, gnotobiology, dental caries, lipid
metabolism, vitamin effects, behavior, alcoholism and
cirrhosis, arthritis, phenylketonuria, jaundice, fructose
intolerance, hypertension, embryology, teratology, diabetes
insipidus, and infectious disease.
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Guvva, et al.: Rat as laboratory animal model

COLLECTION OF BLOOD

In long‑term experiments, it is essential that the same

method of blood collection should be used thought
for both control and experimental animals because the
composition of a blood sample will depend on the method
of sampling.

Small samples can be obtained from a tail vein by snipping

the tip of the tail [Figure 8]. The tip of the tail is cleaned
with alcohol and when this has dried the tip is snipped
with a clean scalpel and the blood collected in a pipette or
directly on to a slide or tube, after discarding the first few
drops. The tail must not be massaged, or the sample will
not be diluted with tissue fluids. Figure 8: Blood collected from tail

Large blood samples should be removed by cardiac puncture

and rats orbital sinus with a hematocrit tube [Figure 9].

The anesthetized rat is placed on its back and the heart

located under the 5th or 6th rib by palpation with the
index finger of the left hand the left thumb being held
on the rats’ right side. A 2–5 ml capacity syringe and
a 19–25 mm, 24–26 gauge needle is suitable for the
purpose. The needle is inserted at 45° to the long axis
of the body into thoracic cavity up to the point where
the heart is felt throbbing against the needle. The needle
is then pushed into the ventricle and blood sample
is withdrawn. The syringe and needle can contain an
anticoagulant if necessary.
Figure 9: Blood collection from eye
In rat the total blood can be collected at one bleeding is
5.5 ml/kg body weight.

DRUG ADMINISTRATION

Drug can be administered through oral, subcutaneous,

intramuscular, intraperiotoneal and intravenous routes.
Convenient volumes for injections are 0.2–0.3 ml/100 g
body weight concentration adjusted accordingly. The
dorsal tarsal vein is useful for injection as also the
saphenous vein where it crosses the leg above the hock.
Intravenous injection is best given in the tail vein using a
12.7 mm, 24 gauge needle. Tail veins can be made visible
by diping the tail in a saturated solution of sodium sulfide
for 2 min.

RATS AS MODEL IN PERIODONTAL RESEARCH Figure 10: Radiograph of molars

Rats have the typical rodent dentition, and the molars are after birth, whereas in the Sprague Dawley rat this occurs at
fully erupted between about the 15th and the 40th day of about 15–17 days and in the osbeorne Mendel rat at about
postnatal life. Eruption time differs from strain to strain. day 17. On the other hand, the first molars begin to emerge
In the Cotton rat, the first molars begin to erupt 4–7 days in the rice rat within 9–10 days after birth.[5]

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Guvva, et al.: Rat as laboratory animal model
The structure of the dentogingival area in the rat, including
the junctional epithelium and its attachment to the tooth
surface, the configuration, and topography of the epithelial
tissues at the gingival margin, and the very shallow gingival
sulcus with the free surface of the junctional epithelium
at its bottom or at the level of the gingival margin, is very
similar to that of man.[6]
Although there are occasional statements to the contrary
such as, the crevicular epithelium in rats is keratinized,[7]
the only significant difference in gingival tissue structure
between rats is in the relationship between the oral
gingival epithelium and the junctional epithelium. In the
rat as the gingival epithelium bends apically and joins the
coronal portion of junctional epithelium at buccal or
lingual aspects of the molars, it often produces a stratum
granulosum and a stratum corneum, the most superficial
cells are in desmosomal contact with the nondifferentiating
and nonkeratinizing cells of the junctional epithelium.[7]
Nevertheless, the junctional epithelium is the pathway
for the influx of foreign substances and the pathway for
inflammatory cell exudation in the rat as it is in the human.
AGE CHANGES IN RAT
Age‑dependent phenomena in rat include a series of finely
tuned and interrelated tissue changes including growth of
the jaws, wear of the occlusal surfaces, continuous eruption
of the teeth, and apposition of cementum and bone,
etc. These changes concomitantly result with increasing
postnatal age in a shifting of the molars in three‑dimensional
space, i.e., in the vertical (apical‑occlusal) as well as in the
horizontal (mesial‑distal and buccal‑lingual) planes and thus
in a continues change of tooth position as it relates to the
bony socket, the alveolar process and the entire jaw.
Hoffman and Schour[8] were the first to document many
of these phenomena and reported that first mandibular
molar,[1] the height of the anatomic crown decreases as a
result of attrition from 1.3 to 0.7 mm with the most rapid
attrition taking place within the first 100 days of postnatal
life,[2] the length of the roots increases from 0.4 to about
3.7 mm, with most of this elongation resulting from
apposition of cellular cementum,[3] the cementoenamel
junction (CEJ), which is located at the alveolar bone crest
at about 35 days of age, continues to move in an occlusal
direction more rapidly than bone is deposited at the alveolar
crest, resulting in an increasing distance of up to 0.8 mm.[4]
The distance between the root apices and the bone in which
the roots are embedded increases with age as well, reflecting
an occlusal drift of the entire tooth. As this drift occurs,
bone is deposited at the fundus of the alveolus [Figure 10].
International Journal of Oral Health Sciences | Volume 7 | Issue 2 | July-December 2017 73
ERUPTION RATE
The eruption of rat molars, accompanies by rapid occlusal
wear and hard tissue apposition, continues through life.
The rate of eruption, at least in young animals, is faster
than the concomitant increase in bone height. During the
period from 21 to 35 days, the first mandibular molar erupts
at a rate of 590 μm/week, its root elongates at a rate of
540 μm/week and alveolar bone apposition at the fundus
occurs at a rate of 54 μm/week and at the alveolar crest,
at 160 μm/week.[8]
Sicher and Weinmann [9] calculated the rate of distal
molar drift to be 60–80 μm/week and concluded that the
continuous occlusal distal buccal movement of rat molars
is the physiological expression of adaptive changes required
by growth changes of the jaws and by rapid occlusal wear.
This physiological, age‑related drifting of erupting rat
molars in the buccal‑distal direction has, at present, become
a fascinating rat model for studying the dynamics of bone
remodeling. With the use of stereological methods, we
determine the exact remodeling cycle and also the bone
balance. A period of 1.5 days is required for resorption,
3.5 days for reversal, and 1 day for bone deposition: the
exact volume of bone resorbed on the mesial side of the
tooth is deposited on the distal side. The molars erupt early
in life in the growing jaws; continuous vertical eruption
is not only compensating for attrition but correlated to
the growth of the jaws in height. The molars drift distally
through life in correlation with the growth of the maxilla
and the mandible in length at their posterior end.[10]
Belting et al. 1953[10] using sections cut in the mesiodistal
plane found that the distance between the CEJ and the
alveolar bone crest increases with age, more so at lingual
and distal than at buccal and mesial surfaces, but the
width of the supraalveolar interdental space and of the
entire interdental bony septum becomes greatly reduced
with age. While continuing (Belting et al.) to erupt in an
occlusal direction throughout the life of the animal, the
molar teeth tilt mesially while migrating distally and also
tilt lingually while shifting slightly in a buccal direction.
Accompanying these movements is a shift of the junctional
epithelium in the apical direction, which in the normal
course of aging covers coronal portions of root cementum
in germ‑free/conventional rats.[10]
PERIODONTAL DISEASE IN RATS
One of the most successful approaches to studying
oral disease in rats appears to be the utilization of the
gnotobiotic or germ‑free rat. Gnotobiotic rats of the
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Guvva, et al.: Rat as laboratory animal model
Sprague Dawley strain have been used to demonstrate
the ability of various filamentous bacteria to form plaque
and induce periodontal disease in the absence of other
bacteria.[11]
Several Gram‑positive species of bacteria isolated from
the human oral cavity were used as monocontaminations
in rats, causing periodontal destruction in 84 days.
When gnotobiotic rats were monoinfected with a
Gram‑negative anaerobic rod isolated from a localized JP
patient[12] or Eikenella corrodents, plaque adhering to the
tooth surface was not formed. Once initiated, bone resorption
occurred continuously rather than sporadically as in humans.
In germ‑free rats, however, there is a considerable amount of
impacted hair and bedding material between the teeth whose
role in the disease process remains unclear. Lesions induced
by Gram‑negative bacteria showed minimal inflammation.
The computed tomography infiltrate contained primarily
neutrophils, few lymphocytes, and no plasma cells. Thus,
the destructive process in response to Gram‑negative
bacteria can occur in the absence of a cell‑mediated immune
response which is not similar to humans.[12]
Periodontal disease in rats is different from that of humans.
After inoculation of microorganisms into germ‑free rats,
periodontal destruction occurs very rapidly, so there is no
need for inducing disease with ligatures. The rat is relatively
resistant to periodontal disease and is therefore used mostly
for oral microflora research. Another difference between
the rat and human periodontal disease is that, instead of
the lesion extending along the root surface as in man, the
most apical extent of the lesion is located along the central
part of the interdental tissues. Bone loss could occur
without apical migration of the junctional epithelium.[13]
The gingival response involved is an acute, not a chronic,
immune infiltrate.
Periodontitis occurs spontaneously in selected strains of
inbred animals such as a particular disease susceptible
strains of rice rat. However, disease is diet dependent.[1]
The disease prone rats develop rampant periodontal lesions
within their 1st year of postnatal life. Gupta and Shaw 1956[5]
stated that the earliest evidence of abnormalities in the
periodontium was noted in 16‑day‑old rats whose second
molars were only partially erupted. Marginal gingivitis was
accompanied by edema and ulceration, with the formation
of deep pockets that often were filled with food debris and
hair, a higher frequency and greater severity of abnormalities
was noted in the mandible than in the maxilla. With varying
74 International Journal of Oral Health Sciences | Volume 7 | Issue 2 | July-December 2017
degrees of severity, these lesions affect both interdental
and inter‑radicular spaces. They are characterized by deep
crater‑like pockets filled with large bacterial masses which
are often attached to the root surfaces, varying amounts of
impacted hair, extensive alveolar bone resorption, and 50%
or more denudation of the molar roots. In final stages, the
teeth become extremely loose and eventually exfoliate. In
few animals, the entire alveolar process carrying the molars
may become completely degraded.
In spite of a statement to the contrary, periodontitis does
not occur in germ‑free Sprague Dawley, White Lobound
the CDF Fisher rats, all of which, even under conventional
conditions, are periodontal disease‑resistant animals.
When silk ligature is tied around the mandibular molars of
germfree rats, the gingival tissue becomes displaced but do
not react in any way except for an enhanced transmigration
of polymorphonuclear leukocytes a similar transmigration
through the junctional epithelium also occurs in untreated
germ‑free control animals.[14]
A similar response is obtained when stainless steel wire
ligatures are applied to the maxillary molars of conventional
rats systemically treated with either tetracycline or
cortisone. Experimentally, induced chronic traumatic injury
of the gingival tissues does not lead to periodontitis in
germ‑free animals.[15]
CONCLUSION
The modern era of periodontal research began in the mid
and late 1960s with documentation of the fact that gingivitis
and periodontitis in humans are caused by bacteria. Since
that time, information has been accumulating at an
astounding rate.
The diversity among animal species in susceptibility,
progression, and features of periodntitis provides rich
research possibilities and opportunities. On the other
hand, this diversity necessitates that the questions
asked be defined with precision and the animal to be
used selected with great care. Human periodontitis can
be dissected into several components with regard to
research. Excellent animal models do exist for some
of these components and in many cases, the unusual
features of the disease in a particular species make it
especially useful.
Because rat has been the most widely used rodent in
periodontal research, features of its normal periodontium,
in particular, the dentogingival area and the physiological
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Guvva, et al.: Rat as laboratory animal model
age‑dependent changes in the molar regions are especially
well documented and understood.
Animal research and its value to human experience remain
controversial. Regardless of how much data can be
presented, it is impossible to expect different species to
respond identically or even similarly to the same challenge
except within very narrow limits. Animal data can provide
us with models of biologic trends before proceeding to
human application.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
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