REVIEW ARTICLE

Bhardwaj and Bhardwaj, IJAVMS, Vol. 6, Issue 3, 2012: 150-157

doi: 10.5455/ijavms.138

Contribution of Animal Models in Periodontal Research

Amit Bhardwaj, Shalu V. Bhardwaj
SGT Dental College, Gurgaon, Haryana; Private dental practitioner, Gurgaon
Corresponding author email:amitmds1980@rediffmail.com
Rec.Date: Apr 03, 2012 10:05
Accept Date: Apr 21, 2012 00:24
Abstract
Aim The aim of this review is to define the most pertinent animal models for periodontal
research.
Background In periodontal research, animal studies are complementary to in vitro experiments
prior to testing new treatments. Animal models and cell cultures have contributed new knowledge
in biological sciences, including periodontology. Although cultured cells can be used to study
physiological processes that occur during the pathogenesis of periodontitis, the complex host
response fundamentally responsible for this disease cannot be reproduced in vitro. Animal models
should make possible the validation of hypotheses and prove the safety and efficacy of new
regenerating approaches using biomaterials, growth factors or stem cells.
Review results Among the animal kingdom, rodents, rabbits, pigs, dogs, and nonhuman primates
have been used to model human periodontitis, each with advantages and disadvantages.
Experimental models have been developed in order to reproduce major periodontal diseases
(gingivitis, periodontitis), their pathogenesis and to investigate new surgical techniques.
Conclusion A practical and highly reproducible model that truly mimics the natural pathogenesis
of human periodontal disease has yet to be developed.
Clinical significance The anatomy, physiology and pathogenicity of experimental models should
relate as much as possible to those of patients in order to demonstrate the safety and efficacy of
new biomaterials or treatments in periodontal regeneration.
Key words: Biomaterials, experimental animal, periodontal disease.
Background
Periodontitis is an inflammatory disease of the gums, which affects all the dental supporting
tissues. This immune-inflammatory disease of the periodontium results in progressive loss of

ANIMAL MODELS IN PERIODONTAL RESEARCH

gingival tissue, the periodontal ligament, and adjacent supporting alveolar bone1. The objective of
periodontal treatment should ultimately be to regenerate the periodontal tissue by using non
surgical or surgical techniques, biomaterials for guided tissue regeneration, bone substitutes (e.g.
Calcium phosphates or others), growth factors (e.g. enamel matrix derivatives) or, as more
recently proposed, mesenchymal stem cells2. Research in periodontology involves different
strategies, principally the etiology of periodontal diseases using experimental models of
periodontitis, and the regeneration of damaged periodontal tissues by surgically creating bone
defects in combination, or not, with experimental periodontitis. Animal models have contributed
to the generation of new knowledge in biological sciences, including periodontology3.
Periodontal disease can occur naturally or be experimentally induced in animals. Various species
have been used to study the pathogenesis of periodontitis and to assess therapeutic modalities
against the disease. Appropriate experimental animal models are required for testing and
validating new regenerative therapies for damaged periodontal tissues. Animal studies are
effectively complementary to in vitro experiments prior to testing new clinical treatments where
biopsy harvesting for histology is generally not accepted in human. Different animal species
could be used for modeling periodontitis and treatments, but primates, dogs, rats, rabbits, pigs,
hamsters and ferrets are the most commonly employed. Experimental animal models should be
used with care, particularly primates and canines, for ethical reasons4,5. Concerning
etiopathologic studies, non-human primates are preferred but large series are not ethically and
economically acceptable. Small animal models (e.g. rats or hamsters) have been developed for
periodontal research but these studies mainly focus on bacteriology and immune response.
Depending on species, periodontal diseases could be induced spontaneously, experimentally or
both. In the context of regenerative medicine using biomaterials, large animal models have been
preferred due to the reproducibility and surgical accessibility of experimental defects. Apart from
monkeys, which are the ideal model in pre-clinical studies, dogs have been widely used for
modeling the regeneration of periodontal defects with biomaterials. Some studies have also used
rats, mini-pigs, sheep, rabbits and cats. The aim of this review is to summarize the experimental
animal models used for periodontal research.
Review results
The selection of an experimental model is determined by research objectives, as well as
laboratory constraints such as housing of large or non standard animals. In most cases, small
animal models such as rats or hamsters will be sufficient to assess the role of bacteria, diet or
other factors in periodontal inflammation at the histological level, providing sufficient statistical
significance and pre-clinical relevance. The use of large animals with ethical and social issues
such as monkeys and dogs should be reserved for last phase validation of new treatments prior to
use in human clinical practice. Experimental studies conducted in monkeys are highly relevant for
human clinical practice as they present comparable anatomy and develop similar periodontal
diseases with similar clinical symptoms. However, experimental research in monkeys requires a
strong ethical justification of their care and use and should take into account the purchasing,
transportation and housing expenses of these animals over long periods. The selection of these
animals was based on similar pathologies and the ease of surgically created clinically relevant
defects. Experimental periodontal defects may be obtained in three different ways6:
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ANIMAL MODELS IN PERIODONTAL RESEARCH

(i) The acute defect model
(ii) The chronic defect model
(iii) The acute/chronic defect model.
In the acute model, all defects are surgically-induced by removing surgically all the periodontal
components (bone, cementum and periodontal ligament). Reproducible defects in experimental
and control sites are created. In the chronic model, lesions are obtained by placing orthodontic
elastics, silk sutures or ligatures around teeth during 12 to 20 weeks, depending on the type of
animal studied. These defects are deeper in the interproximal spaces than in the buccal or lingual
surfaces. In the combined acute/chronic model, the defects are surgically-created and ligatures are
placed to ensure calculus accumulation and to prevent spontaneous regeneration of the defects.
Different animal models used in periodontal research based on pathogenesis of periodontal
disease are non-human primates, dogs, minipigs, ferrets and hamsters. They all have showed
excellent results during experimental studies. These animal models are also used in periodontal
research based on periodontal treatment modalities.
Discussion
Various experimental animal models in periodontal research
1. Non-Human Primates
Nonhuman primates have oral structures and teeth similar to those of humans and have naturally
occurring dental plaque, calculus, oral microbial pathogens (e.g., P. gingivalis), and periodontal
disease. In particular, rhesus monkeys (Macaca mulatta), cynomolgus monkeys (Macaca
fascicularis), and baboons (Papio anubis) are susceptible to naturally occurring periodontal
disease7. All these species are Diphyodont and have the same dental formula as human: I 2/2, C
1/1, Pm 2/2 and M 3/3. Although periodontitis in primates most closely resembles the human
disease, the expense of and special husbandry requirements for these animals limit their use in
periodontal studies.
2. Miniature Pigs
Miniature pigs have oral and maxillofacial structures similar to those of humans in terms of
anatomy, physiology, and disease development8. The Minnesota miniature pig (minipig) was
developed about 60 years ago9 and has been used extensively in biomedical research10. After the
age of 6 months, minipigs usually develop gingivitis, manifested by inflamed gingival tissue,
accumulated plaque and calculus, and bleeding when probed8. There is infiltration of
inflammatory cells in the gingival tissue that results in progression to severe periodontal
inflammation at 16 months of age with identical histopathology to that seen in humans.
Periodontitis in minipigs is promoted in about 48 weeks using ligatures, and in association with
bacterial inoculations of P. gingivalis, S. mutans, and A. actinomycetemcomitans8. Minipigs can
be suitable for periodontal as well as orofacial investigations. However, minipigs are relatively
expensive, with husbandry issues and few studies to support their use.
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ANIMAL MODELS IN PERIODONTAL RESEARCH

3. Dogs
Many experimental studies on gingival and periodontal diseases have been conducted in dogs.
The beagle is one of the most commonly used due to its size and its extremely cooperative
temperament. Globally, all periodontal tissues and the size of the teeth are quite similar to those
observed in humans. However, some major differences exist between dogs and humans as the
lack of lateral movements, no occlusal contacts for all the premolars and presence of open
contacts between teeth. The frequent lack of gingival sulci and crevicular fluid, a different
composition of periodontal plaque and calculus are other important differences between dogs and
humans11. All dogs are diphyodont with deciduous and permanent dentition. The formula for
permanent dentition is I 3/3, C1/1, Pm 4/4, M 2/3.
4. Mice
The Baker mouse model of periodontitis has been used to measure alveolar bone resorption
caused by oral bacterial inoculums as an outcome for the clinical presentation of periodontitis in
humans12. The dental formula of mice is typical rodent dentition: I 1/1, C 0/0, Pm 0/0, M 3/3.
Mice naturally develop periodontitis starting at about 9 months of age with further increases as a
function of age, similar to human periodontitis. This model, however, may not reproduce all
aspects of human periodontitis initiation and progression; the bacteria used are one or two of at
least 150 microbial types present in any dental plaque biofilm. However mice can be utilized to
understand the host-parasite interaction13.
5. Rats
The rat is the most extensively-studied rodent for the pathogenesis of periodontal diseases.
Typical rodent dentition is I 1/1, C 0/0, Pm 0/0, M 3/3. The incisor is rootless.The structure of the
dental gingival area in rats is quite similar to that observed in humans14, with a shallow gingival
sulcus and attachment of the junctional epithelium to the tooth surface. The most commonly-used
strains are Wistar or the Spraque-Dawley. In rats, periodontitis appears to be an infectious
process. Inoculations or injections of various periodontal pathogens such as Prophyromonas
gingivalis,
Aggregatibacter
actinomycetemcomitans,
Fusobacterium
nucleatum,
Capnocytophaga, Eikenella corrodens, Actinomyces viscosus, and Streptococus sobrinus can
induce periodontal lesions15. Furthermore, rats are easy to handle.
6. Hamsters
In hamsters, periodontal disease does not occur spontaneously but may be obtained
experimentally. The golden Syrian hamster is the most commonly used. The dentition formula is
identical to rodents: I 1/1, C 0/0, Pm 0/0, M 3/3. Histologically, the structure of the periodontal
tissue is very similar to that of rats16 but due to the small size of this kind of animal, the
interdental septum is narrower than in rats. Spontaneous periodontal disease was obtained using
an appropriate diet containing high concentrations of carbohydrates, particularly sucrose17. In
summary, as in rats, the inflammatory response is very limited in hamsters and is very different
from that observed in humans. The mechanisms of alveolar bone resorption in hamsters with dietIJAVMS, Vol. 6, Issue 3, 2012: 150-157

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ANIMAL MODELS IN PERIODONTAL RESEARCH

dependent periodontal lesions are quite similar to those observed in rats infected with Grampositive bacteria.
7. Ferrets
Ferrets have a deciduous and permanent dentition. The formula is I 2/2, C 1/1, Pm 4/4, M 2/2.
Ferrets (Mustela putorius) naturally develop calculus and periodontal disease similar to
humans18,19. Unlike rodents, calculus formation in ferrets does not depend on the diet and can be
scored in live ferrets. Ferrets are a suitable model to study calculus; however, they can easily
escape from standard cages and they need special maintenance.
8. Minks
The dentition formula of the adult mink is I 3/3, C 1/1, Pm 3/3 and M 1/2. In minks, spontaneous
periodontitis, which is age- and plaque-dependent, is observed. Nevertheless, the extent of this
periodontal disease appears to be severe only in very old animals. In minks, neutrophils play a
key role in periodontal destruction due to deficiencies in the chemotactic response and massive
release of lysosomal enzymes and proteases into periodontal tissue. Minks are therefore
interesting experimental models in research on the etiology of periodontal diseases. Nevertheless,
housing these animals may be difficult or require specific authorizations that may explain the
absence of recent publications in the literature
9. Rabbits
Characterization of the oral microorganisms in rabbits showed numerous pathogenic bacteria,
including F. nucleatum, P. heparinolytica, Prevotella spp., P. micros, S. milleri group, A. israelii,
and A. haemolyticum, which is somehow consistent with the flora related to periodontal disease in
humans20. Rabbits have been used for creation of surgically induced periodontal defect and to
study periodontal regeneration, but they have been found less suitable for regeneration of
periodontal ligament21.
10. Other Animals
Other animal models have been investigated for modeling periodontal diseases.
Horses- Common naturally occurring oral diseases in horses include buccal abrasions, calculus,
gingival recession,and periodontal pockets. According a recent equine survey, the prevalence of
periodontal pockets and gingival recession is highest in older horses and mostly associated with
other dental disorders and tooth loss22. Because of their size and husbandry considerations, horses
are not a practical model for basic science studies of periodontitis or for testing of potential
therapies.
Sheep- They have also been studied in the context of periodontal diseases. The permanent
dentition of sheep consists of 32 teeth with a formula: I 0/3, C 0/1, Pm 3/3, M 3/3.

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ANIMAL MODELS IN PERIODONTAL RESEARCH

One publication23 used the cat as animal model. In this research, class III furcation defects were
surgically created at the level of the premolars in order to study ankylosis at root level during
periodontal healing.
Few studies have reported mini-pigs for research, mainly related to dental implant surgery and
periodontal regeneration by enamel matrix derivatives24,25, as well as the effects of dental lasers
on periodontal healing26.
Table 1. Advantages and disadvantages of select animal models for studying
periodontal disease development
Animal Model
Advantages
Disadvantages
Similar
dental
structure,
microflora,
Very expensive, with ethical
Non Human Primates
and disease to humans. Natural or and husbandry issues
experimentally
induced
periodontitis.
Develop natural or experimental Relatively expensive, need
Dogs
periodontitis similar to humans
special daily care, husbandry
issues. Dentition different
from humans.
Dental structure and periodontitis Relatively
expensive,
Minipig
have some similarity to humans. husbandry issues; relatively
Natural or experimentally induced few studies
periodontitis
Naturally
or
experimentally Some husbandry issues
Ferrets
induced disease with similarity to
humans
Experimentally induced disease. Naturally
resistant
to
Rodents
Similar molar structure to humans. periodontitis. Different
Inexpensive model
microbiota from humans.
Small size and therefore
amount of tissue for analysis.
Large number of animals
needed

Conclusion: Experimental models for periodontal diseases are essential for understanding the
origin and evolution of the pathology in humans. The use of animal models in periodontal
research is a necessary step prior to entering into clinical trials with new biomaterials and
treatments. Each animal model for periodontal disease has advantages and disadvantages
(summarized in Table 1). A more systematic use of these small animal models appears evident for
future research, especially from a surgical point of view.
Clinical significance: The anatomy, physiology and pathogenicity of experimental models
should relate as much as possible to those of patients in order to demonstrate the safety and
efficacy of new biomaterials or treatments in periodontal regeneration.
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doi: 10.5455/ijavms.138