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Animal models in periodontal research

Presented by:Dr. Shubhra Maheshwari II P. G Student Guided by:Dr.Ruchi Banthia Prof. & Guide
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Introduction Selection of animal models for periodontal research Animal models: 1. Non-human primates 2. Dogs 3. Rats 4. Hamsters 5. Ferrets 6. Other species Conclusion References

Index

Introduction
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 cells. Appropriate experimental animal models are required for testing and validating new regenerative therapies for damaged periodontal tissues before applying it to humans. 3

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.

Selection of animal models for periodontal research:


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. 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.
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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 choice of species for research should be guided by a number of factors:1. Available laboratory facilities. 2. Presence of a breeding colony, cost, ease of handling, and ease of housing. 3. Furthermore, relatedness to humans and limitations imposed by the size of oral structures, as well as the availability of appropriately-sized periodontal instruments, must be considered.
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4. Selection of species for investigations of surgical techniques may primarily consider anatomic factors such as size. 5. Whereas studies of periodontal disease etiology, wound healing, or therapeutic response must focus upon the anatomic and biologic relatedness to humans.
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Different animal species:


1. 2. 3. 4. 5. 6. Non-human primates Dogs Rats Hamsters Ferrets Other species

Non-human primates
Non-human primates are similar to humans, having comparable periodontal tissue structures and healthy and diseased periodontal states, as observed in humans. Monkeys have the advantage of probably being phylogenetically similar to humans. The anatomy of teeth and roots is close to that of humans, but the size is smaller.
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Histologically, the structure of the periodontium is also similar to that observed in humans. Microbiologically, in Macaca fascicularis (cynomolgus monkeys), the composition of the plaque is Gram positive rods and cocci for supragingival plaque and anaerobic Gram negative rods for subgingival plaque. The inflammatory response to periodontal disease is quite similar to that found in humans.

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In the last ten years, 25 articles have been published using monkeys for research relating to periodontal healing filling with biomaterials, guided tissue regeneration, enamel matrix derivatives, or implant surgery. These surgical approaches, are, for the most part, carried out on Macaca fascicularis. All the teeth can be used, which makes it possible to obtain an important number of test sites, with a limited number of animals.
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However, most non-human primates used for research purposes are large, expensive, and difficult to handle.

Furthermore, the genetic background of many of these animals has not been established, because animals used in research are often wild-captured animals, with heterogenecity in age, body weight, and oral and general health conditions.
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Clinically healthy gingiva can be established and maintained in non-human primates, and gingivitis as well as periodontitis occur in these animals.

It is possible to induce experimental periodontitis by placement of peri-dental silk ligatures or orthodontic elastics as well as by surgical removal of alveolar bone.

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In the original non-human primate model described by Caton & Kowalski (1976), experimentally-induced periodontal disease created with elastic ligatures that were removed after 3-6 months. Thereafter, Caton et al (1994) described 3 types of experimentally-induced periodontal lesions i.e acute defect, chronic defect, and acute/chronic defect model.

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Periodontal disease in some non-human primates is histologically very different from human periodontal disease. Therefore, results from studies in such species may be only slightly related to the human condition. In young cynomolgus monkeys A. a was frequently isolated from subgingival plaque of sites with localized gingivitis and no alveolar bone loss, indicating that the mere presence of these bacteria does not cause overt periodontal disease in this animal species or that the simian strains of A. a were less virulent.
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Small non-human primates like squirrel monkeys and marmosets, although less expensive and easier to cage than macaque species, develop a different periodontal disease histopathology, with few to no lymphocytes and plasma cells. These primates may also be difficult to manipulate because of their small size. Therefore, the use of squirrel monkeys and marmosets may not be appropriate in many studies of periodontal disease pathogenesis.
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Therefore, if the use of laboratory animals is requisite and lower species are not applicable, the apparent close anatomic and biologic similarities of many non-human primates to humans make them highly appropriate laboratory animals for studies of periodontal disease even though such studies generally involve small sample sizes.

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Dogs
According to the electronic search, it was shown that dogs were most used in periodontal research as biological models (31.16%). Their docile temperament and natural susceptibility to periodontal disease, dogs, particularly beagles, are used in dental research for the study of periodontal disease progression, guided tissue regeneration, tissue wound healing, and dental implants. 19

More than one hundred publications were found for periodontal research involving dogs for healing defects with various biomaterials, membranes, or with enamel matrix derivatives. The etiologic factors of periodontal disease seem to be identical in humans and dogs. Dogs may therefore be of value as a model for experimental gingivitis.

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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 they lack of lateral movements, no occlusal contacts for all the premolars and presence of open contacts between teeth.
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There is also frequent lack of gingival sulci and crevicular fluid, a different composition of periodontal plaque and calculus are other important differences between dogs and humans.

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Williams et al published their observation on the effects of the phenylpropionic acid derivative flurbiprofen, propionic acid derivative ibuprofen on periodontal disease on beagle dogs.

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Rats
Physiological changes occur throughout the lifespan of the rodents. There is rapid wear of the occlusal surfaces with the continuous eruption of the teeth and apposition of cementum and bone. This causes progressive changes in tooth position which moves in occlusal-buccal-distal direction which is unlike humans.
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Also the destructive process in the response to gram negative bacteria can occur in absence of cell-mediated immunity which is not similar to humans. (Irving & Socransky 1974).

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Periodontal disease in rats is different from that in humans. After inoculation of micro-organisms into germfree rats, periodontal destruction occurs very rapidly. Few rats is relatively resistant to periodontal disease and is therefore used mostly for oral microflora research.
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Instead of 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 tissue. Therefore, bone loss could occur without apical migration of the junctional epithelium (Heijl et al 1980) The gingival response is acute not a chronic, immune infiltrate unlike humans.
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Calculus formation can be studied in different strains of rats where diet seems to be the most consistent factor (Baer et al 1961). Therefore, laboratory rat, although acceptable model for studying calculus and caries, has limitations as a model for periodontal disease.

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One of the most successful approaches to studying oral disease in rats appears to be utilization of the gnotobiotic or germ-free rats. Gnotobiotic rats of Spraque-Dawley strain have been used to demonstrate the ability of various filamentous bacteria to form plaque and induce periodontal disease in absence of other bacteria (Socransky et al 1970)

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Hamsters
Hamsters have the same teeth formula as rats with a continuously erupting incisor and can open their mouth almost 180 degrees wide (Navia 1977). They have been used to demonstrate the transmissibility of periodontal disease with plaque bacteria (Jordan & Keyes 1964).

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The disease can be inoculated in noninfected rats by inoculating subgingival plaque from affected hamsters and can be transmitted from generation to generation (Keyes & Jordan 1964) They develop a periodontal disease similar to rats, in that there is primarily gingival retraction with horizontal bone loss, the interdental septum being too narrow to induce infrabony defects.
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Inflammation is not a prominent feature, as it is in humans. Subepithelial inflammatory response characteristic of human gingivitis, has not been identified for periodontal disease in hamsters.

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When a cariogenic streptococci strain was inoculated with a plaque producing filament, the hamsters developed both caries and extensive periodontal disease. (Jordan & Keyes 1964) Hamsters have buccal pouches lined with stratified squamous epithelium on each side of the head that have been utilized in oral carcinoma research (Navia 1977).

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Ferrets
Use of domestic ferret as an animal study model in periodontics was originally described in the 1940s by King et al, who documented that the occurence of periodontal disease in ferrets was similar to that occuring in humans (King et al 1954). Ferrets have been used as a medical and dental model.

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Harper et al 1996 & Mann et al 1990 have found ferrets to be a suitable model for the study of calculus. Calculus in ferrets have similar physical structure than in humans but the main difference is in ferrets it is lesser calcified than in humans. The course of the periodontal lesion follows a similar path as in humans.

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As the disease progressed, calculus increased both in quantity and extent. The gingiva showed signs of inflammations (King & Gimson 1947). Secondly, after the junctional epithelium split, gingival pockets are formed similar to humans.

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Therefore, ferret is a suitable model for the study of calculus because of its resemblance to human calculus. Further research is still needed to ascertain the role of ferrets as a model in the pathogenesis of periodontal disease.

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Other species
Minks: 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. 38

Mice: The extensive physiological alterations in molar position in the alveolar socket over time mean that mice are not the best model for studying natural or induced periodontal disease.

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Sheep: Periodontitis may affect these anterior teeth and is quite rapidly accompanied by deep periodontal pockets and severe bone loss. The inflammation of the gingiva appears to be moderate. However there are no recent publications available using sheep as the animalmodel in periodontology.

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Rabbits: Rabbits have mainly been used for testing biomaterials or for treatment of periimplantitis and appear as a very interesting model for testing the bone healing.

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Cats: One publication by Takahashi et al in 2005 used 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.

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Pigs: Five studies have reported mini-pigs for research, mainly related to dental implant surgery and periodontal regeneration by enamel matrix derivatives (Schliephake H et al 1998; Craig et al 2006), as well as the effects of dental lasers on periodontal healing (Romanos et al 2004).

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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.

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Wilensky & collegues concluded at pointing out that both the bacterial challenge as well as host response to that challenge need to be considered in a quantitative, time-dependant, site & tissue-specific manner so that conclusion drawn from the data obtained can be extrapolated to humans.

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Monkeys and dogs, as their anatomy and physiopathology are comparable to those of humans, should be restricted to pre-clinical studies for validating new treatments. Apart from these large animal models, smaller, easier to maintain and less expensive species have been proposed. Rats and hamsters develop experimental periodontal diseases.
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The use of gnobiotic or germ-free rats is a major model for all research on the microbiology of periodontal diseases. In the same way, the hamster remains an interesting model for any immunological research. A more systematic use of these small animal models appears evident for future research, especially from a surgical point of view.

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References
1. Struillou X, Boutigny H, Soueiden A, Layrolle P.Experimental animal models in periodontology: A Review. The Open Dentistry Journal 2010; 4: 3747. 2. Schou S, Holmstrup P, Kornman KS: Non-human primates used in studies of periodontal disease pathogenesis: a riview of the literature. Journal of Periodontology 1993.
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3. A. Dannan & F. Alkattan. Animal Models in Periodontal Research: A Mini-Review of the Literature . The Internet Journal of Veterinary Medicine. 2008 Volume 5 Number 1. 4. Weinberg Ma, Bral M: Laboratory animal models in periodontology. Journal of Clinical Periodontology 1999; 26: 335-340.
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5. Drugs, diseases, and the periodontium. Seymour RA, Heasman PA. 6. Fine DH. Of men and mice:animal models of human periodontal disease. Journal of Clinical Periodontology; 36: 913-914.

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THANK YOU

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