GOOD MORNING

Animals are such agreeable friends they ask no questions, they pass no criticisms!!!

A dog is the only thing on earth that loves you more than he loves himself. -Josh Billings

CONTENTS
INTRODUCTION RATS MICE HAMSTERS DOGS FERRETS NON-HUMAN PRIMATES GENERAL CHARACTERISTICS OF NON-HUMAN PRIMATES USED IN PERIODONTAL RESEARCH CHARACTERISTICS OF THE ANIMALMODEL IN PERIODONTAL HEALTH. CHARACTERISTICS OF THE MODEL IN THE TRANSITION FROM HEALTH TO GINGIVITIS TO PERIODONTITIS IN SPONTANEOUS PERIODONTAL DISEASE. CHARACTERISTICS OF THE MODEL IN EXPERIMENTAL GINGIVITIS AND PERIODONTITIS .

NON-HUMAN PRIMATE MODELS FOR TESTING THE EFFICACY AND SAFETY OF PERIODONTAL .REGENERATION PROCEDURES PERIODONTAL REPAIR IN DOGS : SUPRRALVEOLAR DEFECT MODELS FOR EVALUATION OF SAFETY AND EFFICACY OF PERIODONTAL RECONSTRUCTIVE THERAPY. ANIMAL SELECTION AND MANAGEMENT . COMPARISON OF CANINE AND NON-HUMAN PRIMATE ANIMAL MODELS FOR PERIODONTAL REGENERATIVE THERAPY : RESULTS FOLLOWING A SINGLE ADMINISTRATION OF PDGF/IGF-I CONCLUDING REMARKS.

INTRODUCTION
In the 1920s, experimental epidemiology meant the study of epidemics among colonies of experimental animals such as rats and mice. In modern usage, experimental epidemiology is often equated with randomized controlled trials.

The aims of experimental studies may be stated as follows : a) to provide scientific proof of aetiological (or risk) factors which may permit the modification or control of those diseases and b) to provide a method of measuring the effectiveness and efficiency of health services for the prevention, control and treatment of disease and improve the health of the community.

EXPERIMENTAL STUDIES
1.In Vitro study-Lab specimens. 2.In Vivo study -Animal models -Human beings

Animal studies :
Throughout history animals have played an important role in mens quest for knowledge about himself and his environment. Animal studies have contributed to our knowledge of anatomy, physiology, pathology, microbiology, immunology, genetics, chemotherapy and so many others. At the beginning of this century, Webster in United States and Topley, Wilson and Greenwood in England had carried out classical animal experiments. Their studies centered round including epidemics in animals and in studies of herd immunity under laboratory conditions.

More important application of animal experiments have been in

a) experimental reproduction of human disease in animals to confirm aetiological hypotheses and to study the pathogenetic phenomena or mechanisms b) testing the efficacy of preventive and therapeutic measures such as vaccines and drugs, and c) completing the natural history of disease. For example,naturally occurring leprosy has been found in armadillos. Data obtained from studying these animals indicate that lepra bacilli might exist outside of humans.

 Animal models are needed to objectively evaluate the pathogenesis of human periodontal diseases and its various treatment modalities. Selection of the appropriate animal model depends on the similarity of the periodontium and the nature of the disease to that of humans. The more commonly used animal models for studying the pathogenesis of periodontal disease, use of implants and guided tissue regeneration have been dogs and nonhuman primates.

 Periodontal disease in rodents has not been found to be as closely related to the human varieties. Rats and hamsters are best suited for caries and calculus research. Ferrets may be a promising new model for studying periodontal disease and calculus formation. Variables unique to each animal species are manifested by a wide range of clinical and histopathological features. Different species have distinct diets, habits, life spans, tissue structures, host defense mechanisms and genetic traits.

HISTORICAL PERSPECTIVE
The modern era of use of animal models in field of periodontal research began in 1960s with documentation of the fact that gingivitis and periodontitis are caused by specific bacteria. Sir Frank Colyer (1947), first described the occurrence of periodontal disease in a variety of animals. Bleby and Festing, 1976 -The rats, mice, hamsters are often used in the preliminary screening experiments, but data on rabbits, dogs, monkeys and possibly primates may be necessary before testing the safety of experiments for its application in human.

 In mid 1970s, there was a transition from animal based research to cell and tissue culture and other in vitro systems. By 1980s a sufficient understanding of the nature of periodontitis had been obtained to make application of the new knowledge directly to problems of diagnosis and treatment.

ADVANTAGES OF ANIMAL EXPEIRMENTS .The experimental animals can be bred in laboratories and manipulated easily according to the wishes of the investigator. A more important point is that they multiply rapidly and enable the investigators to carry out certain experiments (e.g., genetic experiments) which in human population would take several years and involve many generations.

LIMITATIONS OF ANIMAL EXPEIRMENTS Not all human diseases can be reproduced in animals. Secondly, all the conclusions derived from animal experiments may not be strictly applicable to human beings. An excellent example to illustrate this point is the WHO trial of typhoid vaccine in Yugoslavia in the mid-1950s. Laboratory tests in animals showed the alcohol-killed and preserved vaccine to be more effective than the traditional heatkilled phenol-preserved vaccine.

 But randomized controlled trails in human begins demonstrated that, contrary to laboratory evidence, the alcoholpreserved vaccine was found to be less than half as effective in preventing typhoid fever as the traditional phenolpreserved vaccine introduced by Almorth Wright. This highlights the difficulties encountered in extrapolating findings from animal experiments in man.

Criteria for selection of animal models :

High reproducibility. Genetically matching. Hierarchy of evolution. Not in threatened extinction Which can reproduce same disease. Histological, serological, biological parameters similar to humans. Should be possible to treat the defects. Ethical clearance from the person of animal house. There are over 4,500 known mammalian species and over 9,000 species of birds, but only few species of these classes have been subjected to experimentation. The choice is frequently dictated by convenience and economic factors rather than sound biologic principles (Hegarthy, 1981).

Animals used in biomedical research favor to

Uderstand how our bodies work. Understand the course of disease progression. Find cures and treatments for diseases. Test new drugs for safety. Evaluate medical and dental procedures before they are used in general population.

Classification of animal models

1. Experimental Model: one which the experimentally reproduced condition mimics a human disease (eg. Leprosy in armadillos). 2.Negative Model (Non-model): are animal species in which a particular disease cannot be produced.These are used to study why this animal is resistant to a particular disease (eg. Wood ratimmune to snake bite, Opossum-resistant to rabies.)

3.Spontaneous Model: is an animal species that has a disease which occurs naturally and mimicsa human disease at least in some way (eg. Stumptailed macaques-baldness, Doberman pinscher-von Willebrands disease-factor A haemophilia.

RATS
Physiological changes in the dentition occur throughout the life span of the rodent. Rats have 1 set of teeth consisting of 1 incisor that is rootless and 3 molars in each quadrant (Navia 1977). There is rapid wear of the occlusal surfaces with continuous eruption of the teeth and apposition of cementum and bone. This causes progressive changes in tooth position, especially the molars which continuously move in an occlusal-distal-buccal direction. Most histologic features of the epithelium and connective tissue in the rat are similar to humans except for the sulcular epithelium which is keratinized (Listgarten 1975).

 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 Spraque-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 (Socransky et al. 1970). Several gram-positive species of bacteria isolated from the human oral cavity were used as monocontaminants in rats, causing periodontal destruction in 84 days. These species included Actinomyces viscosus,Actinomyces israelii, Streptococcus mutans (Klausen et al. 1986, Crawford et al. 1978) and Actinomyces naeslundii (Garant 1976).

 When gnotobiotic rats were monoinfected with a gram-negative anaerobic rod(A. actinomycetemcomitans) isolated from a localized juvenile periodontitis patient (Irving et al.1975) or Eikenella corrodens (Listgarten et al. 1978), 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 connective tissue infiltrate contained primarily neutrophils, few lymphocytes, and no plasma cells. Thus, the destructive process in the response to gram negative bacteria can occur in the absence of a cell-mediated immune response (Irving & Socransky 1974), which is not similar to humans. Calculus formation can be studied in different strains of rats where diet seems to be the most consistent factor (Baer et al. 1961). Rats fed a sucrose-rich diet developed a rapid proliferation and overgrowth of bacteria plaque, mainly gram-positive, covering the molar fissures, the interdental spaces and the marginal gingiva. This ultimately resulted in rounded or crater like gingival pockets.

 Therefore, it appears that the laboratory rat, although an acceptable model for studying calculus and caries, has limitations as a model for periodontal disease. Periodontal disease in rats is different from that of humans. After inoculation of micro-organisms 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 (Heijl et al.1980).

Fate of guided tissue regeneration with or without grafting of Bio-Oss or Bio-gran in rats.

Radiograph of a rat

MICE
The healthy and diseased periodontium of the mouse has been studied in a large number of strains, most of which were highly inbred. Out of the many strains populating the natural habitat, only two strains of the deep mouse (Peromyscus maniculatus and Peromyscus Oreas) have been examined, and used only briefly. Inbred strains which have attached attention are the RAP-albino.

 Mice present the typical rodent dentition with the formula: I 1/1, C 0/0, Pm 0/0, M 3/3. The periodontal tissues of the continuously growing incisors are rarely, if at all, affected by periodontitis.
Cementum is deposited on the apical third of the root. Also, the junctional epithelium shifts apically onto the root surface with age. Consequently, the distance between the crest of the alveolus and the CEJ increases, particularly at the lingual and palatal aspects of the mouse molars. Gilmore and Glickman (1959) suggested, probably rightly so, that with age, the rate of bone apposition at the alveolar crest does not keep pace with occlusal eruption of the tooth.

 Furthermore, the histological material published by Baer and Bernick (1957) and Baer and Lieberman (1959) demonstrates clearly the occlusal wear and the elongation of the maxillary molar roots, due to marked apposition of cellular cementum. Two additional strains of mice were subjected to a periodontal investigation. Tonna (1972) examined the BNL (Brookhaven National Laboratory, USA) mouse, which is an inbred Swiss albino whose mean life span is less than a year. The histological features of gingival and periapical tissues of animals 5-78 weeks of age were studied.

HAMSTERS
Hamsters have the same teeth formula as rats with a continuously erupting incisor and can open their mouths almost 180 degrees wide (Navia 1977). Hamsters have been used to demonstrate the transmissibility of periodontal disease with plaque bacteria (Jordan & Keyes 1964). The type of periodontal disease hamsters develop is similar to rats in that there is primarily gingival retraction with horizontal bone loss, the interdental septum being too narrow to induce infrabony defects. Inflammation is not a prominent feature, as it is in humans. Albino hamsters remain essentially disease-free while the golden and cream-colored hamsters develop spontaneous periodontal disease when fed a high carbohydrate diet (King & Rowles 1955).

 They naturally harbor an infectious agent capable of inducing the disease when experimental conditions are favorable. The disease can be induced in noninfected albinos by inoculating subgingival plaque from affected hamsters, and can be transmitted from generation to generation (Keyes & Jordan 1964). Subepithelial inflammatory response characteristic of human gingivitis, has not been identified for periodontal disease in the hamster.

When a cariogenic streptococci strain BA1 was inoculated with a plaque-producing filament (strain T6), the hamsters developed both caries and extensive periodontal disease. This is of special interest because the 2 diseases, caries and periodontal disease, could be evaluated in vivo at the same time (Jordan & Keyes 1964). However, it has been speculated that the tissue destruction may actually be due to pressure from the subgingival accumulations.

RODENTS
RATS

MICE

HAMSTERS

DOGS
Permanent teeth of the dog consists of 3 incisors, 1 canine, 4 premolars and 3 molars in the mandible and 2 molars in the maxilla (Navia 1977). In view of 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.

 Dogs maintained plaque free by repeated scalings and meticulous plaque control can develop clinically healthy gingiva. The etiologic factors of periodontal disease seem to be identical in humans and dogs (Attstrm et al. 1975, Ericsson et al. 1975, Gad 1968). Dogs may therefore be of value as a model for experimental gingivitis. The fact that it is possible to maintain periodontal health in sites where plaque accumulation was prevented confirms similar reports in humans (Lindhe & Nyman 1975, Theilade & Attstrm 1985).

 Originally, it was reported that periodontal disease began slowly in young dogs and increased with age (Gad 1968), progressing about 5x faster in dogs than humans (Ericsson et al. 1975). Later, it was documented that the range and severity of gingivitis and periodontitis varied in both young and older dogs and that gingivitis in younger dogs did not necessarily progress into periodontitis (Hull 1974). Gingival recession is an outstanding feature in dogs with periodontitis (Ericsson et al. 1975).

 Differences exist between dog and man in the location of the inflammatory infiltrate in early gingivitis. In the dog, the initial infiltrate located in the marginal part of the gingiva, proceeds along the junctional epithelium leaving the connective tissue in a relatively normal state. In healthy dogs kept plaque free for prolonged periods of time, a gingival sulcus is not evident, but a pocket develops with the onset of gingivitis (Lindhe & Rylander 1975). Gingivitis did not necessarily progress into periodontitis and a variety of factors may influence this conversion (Lindhe et al. 1975).

It has been demonstrated that induced gingivitis in young beagle dogs can progress into periodontitis, simply by allowing additional plaque to accumulate (Soames & Davies 1980). This process can be accelerated by the placement of ligatures (Lindhe & Ericsson 1978). Histologic features are characteristic of an advanced lesion, with the majority of tissue destruction occurring within the 1st 4 weeks following ligature placement (Schroeder & Lindhe 1975). Class II and III furcations may be found in the beagle dog. Wikesj et al. (1994) was able to create the same ligature-induced intrabony defects demonstrated in monkeys by Caton et al. 1976, 1994.

 This was done by surgically removing alveolar bone around the circumference of mandibular premolars Wikesj et al. 1994. The surgically created bony defect in the dog is different than that of a naturallyoccurring bony defect. Variations in attachment loss between adjacent surfaces make it difficult to document the presurgical size of the defect and to evaluate the postsurgical healing response (Selvig, 1994). A much faster bone turnover rate, and a different architecture and thickness of bone limit the suitability of dogs for regenerative studies (Giannobile et al. 1994).

 Early implant research utilized dogs, as well as rats, hamster, guinea pigs, rabbits and cats to explore the pathophysiology of tissue injury and repair. Osseointegration was demonstrated in various parts of the animal skeleton when titanium fixtures were placed Branemark 1983. Even though dogs have been used most frequently due to their size and the simple morphology of the roots, there are differences in the distances of the anterior and posterior teeth relative to the point of mandibular articulation. Furthermore, eating movements consisting of biting and grasping plays different functional demands on the bone (Navia 1977).

Although the features of gingivitis and periodontitis lesions in dogs closely resemble those in humans, there are still differences. Inflammatory lesions in dogs begin in the most coronal portion of the connective tissue at the gingival margin, rather than lateral and apical to the base of the gingival sulcus as in humans. With increasing severity of gingivitis, the entire thickness of the marginal gingiva is involved and not just the tissue lateral to the gingival pocket wall (Matsson & Attstrm 1979). Limitations of the use of a canine model includes great inter-animal variability, expense, limited number of bony defects available and faster bone formation (Giannobile et al. 1994).

Various species of dogs

Bone healing around implants placed in a jaw defect augmented with Bio-Oss in dogs.

FERRETS
Ferrets were thought to be first domesticated by the Egyptians to control rodents around 1300 B.C.
The dentition, wear patterns, calculus formation, salivary glands, and periodontal lesion of the ferret have been studied, although not to the extent of the previously-discussed species. The domestic ferret (Mustela putorius furo) is believed to have been derived from the wild (European) polecat (Fox 1988).

 Use of the domestic ferret as an animal study model in periodontics was originally described in the 1940s by King et al., who documented that the occurrence of periodontal disease in ferrets was similar to that occurring in humans (King 1954). The ferret has both a deciduous and a permanent dentition. The permanent dentition consists of incisors, canine, 2nd, 3rd and 4th premolar and first and 2nd molar (Berkovitz & Silverstone 1969).

 Ferrets have been used as a medical and dental model. Harper et al. (1990) and Mann et al. (1990) have found ferrets to be a suitable model for the study of calculus. Calculus in ferrets has a physical structure similar to hydroxyapatite. The main difference between the ferret and human calculus is a lesser degree of calcification in the ferret deposits. Diet did influence the rate of formation, but not as much as in rats. Calculus in ferrets can be scored while the animal is alive, whereas this is not possible in rats (Harper et al. 1990).

 The course of the periodontal lesion follows a similar path as in humans. The tissues responded by characteristic inflammatory reactions, which are identical in all respects to those found in human gingivitis (King & Gimson 1947). Ferrets can develop ligature-induced periodontitis within 28 days (Harper et al. 1989, Fischer & Kinge 1994) where the ligated sites lost 50-75% of attachment, and lesions exhibited large populations of PMNs adjacent to the ligatures.

 Plasma cells and lymphocytes were observed apical to the lesions. The ferret is a suitable model for the study of calculus because of its resemblance to human calculus and the fact that formation of calculus is not dietdependent as in the rat and hamster. Further research is still needed to ascertain the role of ferrets as a model in the pathogenesis of periodontal disease.

FERRETS

NON-HUMAN PRIMATES
In designing any medical or dental animal study, it is often advantageous to select an animal that is phylogenetically similar to humans. The wide range of non-human primate species allows appropriate selection for different investigations. Each species has unique similarities and dissimilarities to humans.

 Non-human primates have similar oral structures to humans and have naturally occurring dental plaque, calculus and gingivitis, but small increase in pocket depths. The majority of non-human primates have similar deciduous and permanent dental formulas as man with closely related dental anatomy, although the size of teeth are dramatically smaller. The organization of collagen fibers in gingival and periodontal connective tissue are also similar to that of humans.

 Clinically, healthy monkey gingiva is histologically indistinguishable from human gingiva. A shift in the composition of plaque flora from an early gingivitis to a later stage is also comparable to humans (Krygier et al. 1973,Schou et al. 1993). The inflammatory infiltrate associated with periodontal disease is microscopically similar to humans in some species such as the cynomolgus monkeys (Macaca fascicularis), but the squirrel monkeys (Saimiri sciureus) and marmosets have limited numbers of lymphocytes and plasma cells, making them inappropriate models for studying pathogenesis of periodontitis. (Shou et al. 1993, Brecx et al. 1985, Kornman et al. 1981a,Listgarten & Ellegaard 1973, Page & Schroeder 1982, Page et al. 1972).

 It was found that the incidence and degree of angular defects was not affected by the presence or absence of traumatic injury (Polson 1974). Monkeys have been used widely as an animal model for studying periodontal surgical procedures. Smaller non-human primates such as marmosets have small oral cavities which may preclude their use for certain periodontal procedures. Large non-human primates have a naturally occurring periodontitis, but it occurs later in life and the lesions are asymmetrical (Caton et al. 1994). Therefore, if osseous lesions are needed for clinical studies, they are usually experimentally induced.

 Similar dental anatomy, periodontal wound healing (Caton & Kowalski 1976), suitability of furcation sites (Giannoble et al. 1994) and experimentally induced defects that do not spontaneously regenerate, make mature adult cynomolgus and rhesus (Macaca mulatta) species good models for studying ligature-induced periodontitis. Periodontal lesions in these animals are also suitable for evaluating periodontal regenerative procedures (Shou et al.1993), especially since histometric analysis needed to quantify the amount of new cementum, periodontal ligament and alveolar bone formed as the result of regenerative periodontal surgery (Caton et al. 1994), can only be done with animals, usually monkeys or dogs.

 In the original non-human primate model described by Caton & Kowalski (1976), experimentally-induced periodontal disease was created with elastic ligatures that were removed after 36 months. Thereafter, Caton et al.(1994) described 3 types of experimentally-induced periodontal lesions. The acute defect model required surgical removal of bone, cementum and periodontal ligament to create the defect where spontaneous regeneration of the defects occurs. The chronic defect model, required the placement of orthodontic elastics around the circumference of teeth to create defects which may take up to 6 months to produce, with deep defects found more likely in proximal sites than on the facial and lingual surfaces.

 The acute/chronic defect model also creates defects surgically. These defects enter a chronic inflammatory state by placement of stainless steel bands into the defects and plaque retentive ligatures.
Due to the possibility of obtaining block biopsies, the rhesus monkey, cynomolgus monkey, and baboons have been used to study osseointegrated oral implants. A study (Fritz et al.1997) suggested that ligature-induced periodontitis around teeth and ligature-induced peri-implantitis follow similar destructive patterns, namely alteration of microbiological flora.

 Although various species of non-human primates are adequate for studying different aspects of periodontal diseases, monkeys are expensive to purchase and maintain and are ferocious. Wild captured monkeys can be disease carriers. Mycobacterium tuberculosis.Herpes Simplex Type B, Shigella species and Simian B virus are infectious to man (Shou et al. 1993). Monkeys are also prone to systemic infections and diseases, and pose difficulties in controlling postsurgical infections and trauma.

Galago senegalensis bratticus :

Grant et al (1973) studied the bush baby, a prosimian residing Central and East Africa. A breeding pair and its offspring were used. By 14 months of age gingival inflammation was apparent. In some specimens from animals 2 years old the junctional epithelium was located on the root surface, some transseptal fibers had been destroyed, and bone resorption was apparent. In one 6 year old animal, an abscess was noted without the furcation region of one molar. Cemental resorption and reposition often resulted in ankylosis of the teeth to the bone.

Galago senegalensis bratticus

Macaca :
Johnson and his co-workers have assessed the nature of the inflammatory cell infiltrate in connective tissues at various stages of spontaneously developing gingivitis in M.fascicularis as well as in animals whose teeth were cleaned and chlorhexidine applied for 3 weeks and then left to accumulate plaque. The composition of the inflammatory infiltrate in the gingival tissues of young M.fascicularis allowed to accumulate plaque was assessed in a variety of unrelated specimens (Johnson and Hopps, 1975). Biopsy specimens were harvested at various times for upto 243 days after the beginning of plaque accumulation and from older animals allowed to collect plaque for 3 years.

 The clinical and histopathologic features of plaque induced gingivitis in M.mulatta were studied by Listgarten and Ellegaard (1973). In the animals on the tooth cleaning regimen, inflammatory cells continued to be present in most of the biopsy specimens, although their numbers fluctuated. Leukocytes in the area of the infiltrate and neutrophils in the junctional epithelium both increase in number. In addition to lymphoid cells and neutrophils, macrophages, mast cells and large clear cells considered to be sick fibroblasts were observed.

Maccaca

Maccaca-Skulls

Marmosets :
Gingivitis and periodontitis have been studied extensively in wild caught and colony maintained marmosets. Generally, cotton tops (s.Oedipus) and cotton ears (C.Jacchus) have been used. Early reports indicated that marmosets do get chronic periodontitis. That the prevalence is high and that the lesions closely resemble those seen in humans. More extensive studies were later done on wild caught and colony maintained animals (Page et al, 1971, 1972; Levy 1971).

Marmossets

Baboons :
Hodosh et al (1971) assessed the periodontal status of 40 African baboons, P. anubis and Theropithecus gelada, approximately 7-10 years old. Clinical manifestations of gingivitis were observed in 33 of the 40 animals, and in 3, periodontal pockets were found but without radiographic evidence for bone loss. Gingivitis is characterized by the presence of subgingival plaque and calculus and apical displacement and proliferation of the junctional epithelium but without ulceration of the epithelium or suppuration. A dense inflammatory infiltrate consisting mostly of plasma cells and lymphocytes is located in the connective tissues lateral to the pocket.

 The most obvious clinical feature was soft tissue hyperplasia. The gingival index and oral hygiene index scores are quite high. The junctional epithelium is converted to pocket epithelium which is by neutrophils and lymphocytes, including T blasts. Periodontal disease in the baboon is thus strikingly similar to the disease in humans, both clinically and microscopically.

Baboons

Chimpanzees :
Gingivitis closely resembled that seen in humans, with neutrophils present in the junctional epithelium and the gingival vessels, along with an infiltrate comprising mostly plasma cells and lymphocytes (Arnold and Baram, 1973). Pathologically altered firbroblasts were present within the infiltrated connective tissue. In an animal of 8 years of age, severe periodontitis resulting in tooth exfoliation and confined to the first molars and incisors was observed (Page et al 1975). The disease in this animal appeared to be a counterpart of juvenile periodontitis in humans.

CHIMPANZEES

 For ethical reasons, initiation and progression of periodontal disease as well as certain types of periodontal treatment cannot be studied in humans. The inability to examine initiation and progression of periodontal disease and to assess certain therapies in humans has led to a great interest in the use of animal models in periodontal research. Some of the most prominent animals used are non-human primates in periodontal health, in the transition from health to gingivitis to periodontitis, and in experimental gingivitis and periodontitis.

Non-Human Primates Used in Studies of Periodontal Disease Pathogenesis:

 Where possible, the results of these studies are compared with results from human studies. Only a few studies have compared in detail the anatomy, physiology, immunology, and tissue interactions in monkeys with those of humans. With the exceptions of differences and variations in size of the dentition, the number of each tooth type as well as larger canines, presence of diastemata between anterior teeth, and an edge-to-edge relationship of the incisors, the dental and periodontal anatomy of non-human primates seem quite similar to that of humans.

GENERAL CHARACTERISTICS OF NONHUMAN PRIMATES USED IN PERIODONTAL RESEARCH :

Sources of Animal Subjects. Considerations for Species Selection.

a.Sources of Animal Subjects : Most of the non-human primates previously used in research were wild animals captured in their natural habitats. Such animals used in laboratory studies present investigators with the problem of pronounced heterogeneity in age, body weight, and dental health status. In addition, the unknown medical history of captured animals may introduce risks to animal handlers and great variability in studies that, due to cost, generally involve small sample sizes.

 The high incidence of latent and subclinical diseases especially Marburg disease, tuberculosis, rabies, and Salmonella/Shigella infections among wild-captured non-human primates necessitates specialized handling procedures for such animals. Furthermore, some viruses, such as Simian B virus, are fatal when transmitted to humans.

 In order to prevent transfer of these diseases to other non-human primates and other animals and humans, and to ensure that investigations do not include sick primates, the animals must be screened for disease and maintained in quarantine both before delivery to the laboratory and again after receipt at the animal care facility at which the investigation will be conducted.
Wild captured animals may be transferred among several holding facilities between the time of capture and their use in a study, and each facility may use various anti-infective agents to optimize the health and survival of the animals.

Classification and name

Distribu tion WildLiving Species

Africa

Average Weight
200-250 g

Number of Each Tooth Type

I2/2, CI/1, Pm3/3, M3/3

Main Periodontal Characteristics

Prosimians : Bushbaby (Galago senegalensis) Simians Cebidae : Howler monkey (Allouta caraya) Squirrel monkey (Saimiri sciureus)

Spontaneously occurring periodontal disease characterized histologically by lymphocytes and plasma celsl Spontaneously occurring periodontal disease characterized histologically by lymphocytes and plasma cells Experimental periodontitis characterized histologically by lymphocytes and plasma cells Spontaneously occurring periodontal disease characterized clinically by marked gingival telangiectsia and histologically by predominance of macrophages Spontaneously occurring periodontal disease characterized clinically by marked gingival telangiectasia and histologically by predominance of macrophages Spontaneously occurring periodontal disease characterized histologically by lymphocytes and plasma cells

S. America

Male: maximum 10 kg; female: maximum 6.5 kg

12/2, CI/1, Pm3/3, M3/3

S. America

750-1100 g

12/2, Cl/1, Pm3/3, M3/3

Callithricidae Cotton-ear marmoset (Callithrix jacchus)

S. America

250-500 g

12/2, Cl/1, Pm3/3, M2/2

Cotton-top marmoset (Saguinus Oedipus)

C. and S. America

250-500 g

12/2, Cl/1, Pm3/3, M2/2

Cercopithecidae (Papio anubis)

Baboon

Africa

Male: 22-30 kg; female: 11-15 kg

12/2, Cl/1, Pm2/2, M3/3

Rhesus monkey (Macaca mulatta) Cynomolgus monkey (Macaca fascicularis)

C. Asia

Male: 5.6-12 kg; Female: 4.4-10.7 kg

12/2, Cl/1, Pm2/2, M3/3

S.E. Asia

Male: 3.5-8.3 kg; Female: 2.5-5.7 kg

12/2, Cl/1, Pm2/2, M3/3

Spontaneously occurring and experimental periodontal disease characterized histologically by lymphocytes and plasma cells. high carrier rate of A. actinomycetemcomitans and low carrier rate of Actinomyces species compared to humans A catalase-producign Prevotella melaninogenica strain has been identified during experimental periodontitis

Stump-tailed monkey (Macaca actoides)

S. E. Asia

Male: < 13 kg

12/2, Cl/1, Pm2/2, M3/3

Pig-tailed monkey (Macaca nemestrina)

S.E. Asia

Male: < 13 kg

12/2, Cl/1, Pm2/2, M3/3

Hominidae Chimpanzee (Pantroglodytes)

Africa

Male: 45-60 kg; Feamel: 35-45 kg

12/2, Cl/1 Pm2/2, M3/3

Spontaneously occurring periodontal disease characterized histologically by lymphocytes and plasma cells

Mountain gorilla (Gorilla gorilla beringei)

Africa

Male: 135-275 kg; Female: 70-140 kg

12/2, Cl/1, Pm2/2, M3/3

 The taxonomic order primate is extremely heterogeneous. Some species are terrestrial; some arboreal. Some are predominantly herbivorous, whereas others are predominantly carnivorous. According to weight, size of dentition, number or each tooth type, and food, the characteristics range from very similar to human to vastly different. It is therefore of great importance of select an animal species that is appropriate for the specific study. The choice of species for research should be guided by a number of factors : available laboratory facilities, presence of a breeding colony, cost, ease of handling, and ease of housing.

b.Considerations for Species Selection:

 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. Selection of species for investigations of surgical techniques may primarily consider anatomic factors such as size, whereas studies of periodontal disease etiology, wound healing, or therapeutic response must focus upon the anatomic and biologic relatedness to humans.

 Age is another important factor: young adults may be preferable for certain studies because they do not display the severe attrition observed in older animals. The age of young animals may be determined by tooth eruption, sexual maturation, or bone development in hand-wrist radiographs. It should be noted that the age at which permanent tooth eruption begins and is completed, varies greatly between species. However, there are currently no reliable means for determining the age of adult, sexually mature wild-captured primates.

Ages of Permanent Tooth Eruption in NonHuman Primates (Age in months)

First permanent tooth eruption 18 33 to 35 Last permanent tooth eruption

Species

Cynomolgus monkey (Macaca fascicularis)

Baboon (Papio anubis) Cotto-ear marmoset (Callithrix jacchus)

80 55

3 to 4

11 to 12

 The small oral cavities of several non-human primates necessitate selection of special examination methods. Direct observation of marginal inflammation has been performed in squirrel monkeys, but in marmosets, magnified photographs, magnifying glass, or stereomicroscopy must be used. Radiographs have routinely been used in macaque species and squirrel monkeys, but custom film packs may be necessary in the maxilla of the macaque species due the flat palatal vault and they are a necessity in both the maxilla and mandible of very small primates like squirrel monkeys.

 As an alternative to clinical and radiological methods, histometric methods enable measurement of the dimensions of junctional and pocket epithelium, connective tissue attachment level, and alveolar bone level and are especially useful for the evaluation of different periodontal treatment procedures. These methods have been used in squirrel monkeys and also in larger primates.

Clinical and radiographic pictures

Animal-rights advocates remind us of this admonition:

The ways in which people treat animals will be reflected in how people relate to one another. -William Greider