Course 8
PERIODONTAL PATHOGENIC BACTERIA
In gingivitis:
Experiments showed that the installation of gingival
inflammation occurs 10-21 days after poor oral hygiene, with the appearance of gingival hyperplasia.
However, other studies have shown that the entire mass of bacterial plaque, in contact with the gingiva, causes
the inflammation of the gingiva, so, a single specific factor in gingivitis cannot be considered an etiological
factor.
In marginal periodontitis:
Isolation and identification by cultivating pathogenic species in diseases of the marginal periodontium is difficult.
The bacteria involved are mostly anaerobic gram-negative bacilli. Data obtained by isolation in culture and
identification have recently been supplemented by DNA hybridization techniques, thus establishing the role of
bacterial associations in the production of periodontal diseases.
SOCRANSKY s.a. (1998) specify the organization of bacteria present in periodontal areas in "bacterial complexes"
that act by successive associations until the appearance of the lesions that are specific to periodontal lesions.
The complex compatible with periodontal health includes species of Oral Streptococci (S. gordonii, S. mittis, S.
oralis, S. sanguis, S. intermedius), having the role of "pioneers" in early bacterial colonization and was called the
yellow complex.
Other complexes considered compatible with periodontal health include Actinomyces odontolyticus and
Veillonella parvula species, the purple complex and Capnocytophaga, Aggregatibacter actinomycetemcomitans
serotype A, Eikenella corrodens: green complex.
These complexes are followed, by association, by those formed by species of Fusobacterium, Prevotella,
Campylobacter, Eubacterium nodatum - the orange complex, species that make connections and allow
colonization of bacteria with increased virulence:
The complex with accentuated virulence consists of Porphyromonas gingivalis, Tannerella forsithensia
(Bacteroides forsythus), Treponema denticola: the red complex correlated with the loss of the gingivo-
periodontal junction from periodontal diseases.
Three microbial species could not be described as integrated into either complex: Aggregatibacter
actinomycetemcomitans serotype b, Actinomyces naeslundii and Selenomonas noxia (HAFFAJEE s.a., 1999
quoted by CHARON, MOUTON, 2003).
All bacteria listed above, which do not belong to complexes compatible with periodontal health, are considered
periodontal pathogenic bacteria.
Aggregatibacter actinomycetemcomitans
The bacterium is a major periodontal pathogen in aggressive periodontitis: localized juvenile periodontitis and
fervently isolated from rapidly progressive periodontitis being associated with advanced forms of periodontal
destruction. Aggregatibacter actinomycetemcomitans is the bacterium that respects the
postulates of KOCH modified by SOCRANSKl, to be considered the pathogen in localized
juvenile periodontitis.
Bacteria are in the form of gram-negative cocobacilli, structurally showing all the
characteristics of gram-negative bacteria, periplasmic space, outer membrane,
lipopolysaccharide, microcapsule polysaccharide. It grows on cultures, it is not strictly
anaerobic (optional anaerobic), being favored by the CO2-enriched atmosphere 5-10%. The
colonies on agar environment have a characteristic aspect: they are circular, translucent.
convex, with irregular structure above, with a star-like appearance. The pathogenicity of the species is ensured
by structural factors, exotoxic factors and enzymatic factors.
Enzymatic factors are released by bacteria as in Porphyromonas gingivalis in the form of vesicles that diffuse into
the tissue from the outer membrane. Enzymes act as trypsin-Iike proteases and also Aggregatibacter
actinomycetemcomitans produce cytokines with harmful action on fibroblasts, as well as a collagenolytic
enzyme.
Leukotoxin is a protein in the RTX family (Repeats - in ToXin) which acts by forming pores in the membrane of
some immune cells. The pathogenic action of Aggregatibacter actinomycetemcomitans is thus manifested in a
complex way, by antiphagocytic effect and by cellular cytotoxicity effects, stimulating bone resorption and
significative tissue destruction.
Periodontal pathogens of the red complex
Porphyromonas gingivalis
Porphyromonas gingivalis is the dominant species in aggressive periodontitis, rapidly progressive ones, in
generalized juvenile periodontitis, mostly chronic periodontitis (adult periodontal lesions) and in ulcero-necrotic
gingivostomatitis.
The species is cultivable, being predominantly isolated from periodontal pockets and has been intensively
studied. They are immobile bacteria, have a microcapsule and adhesion factors: fimbriae. On the smear it is in
the form of gram-negative cocobacilli.
They grow on cultures that require the addition of hemin and vitamin K under
conditions of strict anaerobiosis. The colonies may be black pigmented.
The pathogenic potential of the species is accentuated by the synergism with other
periodontopathogenic species. The pathogenicity of the bacteria is determined by its
factors of structure and elaboration of enzymes with the role of aggression in the
periodontal tissue.
Another mechanism of bacterial pathogenicity occurs through the production of
protease-like enzymes. They are released from the outer membrane in the form of vesicles which, through their
structure and the enzymes they contain, are both defense factors of the bacterium and invasion factors in the
periodontal tissue.
The proteases released by Porphyromonas gingivalis have multiple actions in the periodontal tissues and are of
three types according to their action:
- cysteine proteases (or trypsin-like enzymes, pseudotrypsins);
- aminopepetidases and
- collagenase.
Their act in a destructive way on the complement, immunoglobulins (especially igA) and on collagen with
important consequences in tissue penetration, diffusion and destruction. The pathogenic effect of the species
thus appears as a result of multiple antiphagocytic, leukopenic, toxicolytic actions on epithelial cells, fibroblasts,
macrophages.
Tannerella forsithensia (Bacteroides forsythus)
Along with Porphyromonas gingivalis, the species is considered a major periodontopathogen. The appearance
on the smear is gram-negative bacillus, like small spindles, it has spuned heads, sometimes
even filamentous aspects. It is immobile, unsporulated. Their cultivation is difficult, they
require environments supplemented with muramic acid or they are favored by the
concomitant cultivation with bacteria of the species Fusobacterium nucleatum, in conditions
of strict anaerobiosis. The colonies are small, appearing after 7-14 days of incubation.
Pathogenicity is ensured by structural and enzymatic factors. It develops a protease enzyme
(trypsin-like), which can be highlighted as in Porphyromonas gingivalis by the BANA test, due
to the action on the substrate benzoyl-DL-arginine- β -naphtylamide.
The bacteriua has been isolated from gingivitis and periodontitis, especially in aggressive forms. It increases
significantly in the depth of the periodontal pockets, being isolated right from the wall of the periodontal
pockets, which means penetration into the tissues.
Treponema denticola
Treponema comprises spiral bacteria present in the oral cavity, having a preferential location in the gingival
sulcus. Treponemes are gram-negative bacteria with a special structure. It has a sarcolemma arranged in spirals
with tufts of flagella at the ends and covered by a sheath. Due to the flagella, they show varied
and complex movements, they are mobile even in viscous environments, as in the conditions of
the oral biofilm. From cultivation point of view, many treponemes currently uncultivable and
identified by molecular techniques have been described.
Other treponemes are difficult to grow under strict anaerobic conditions.
Pathogenicity is due to the development of enzymatic factors: proteases, hemolysin,
hemagglutinin. With the help of proteases it attaches to eukaryotic cells with cytotoxic effects to fibroblasts and
epithelial cells, and they also attack collagen and fibrinogen.
Hemolysins, by their action on erythrocytes, produce hemolysis, and the resulting hemoglobin provides bacteria
with the iron essential for growth.A special factor of invasion in the tissues is the mobility of the treponema
even in the conditions of a viscous environment
Treponemes are present in large quantities in severe forms of periodontitis. In the evolution of the disease, as
well as in the appreciation of the efficiency treatments, variation in the number of treponemes can be an
important criterion.
Periodontal bacteria of the orange complex
Fusobacterium nucleatum
The bacteria are constantly present in the subgingival flora, they are frequently isolated from progressive forms
of periodontal disease, as well as from ulceronecrotic gingivitis.
The appearance of the bacteria is gram-negative bacillus, spindle-shaped with sharp extremities
and ballooned center, arranged in isolation, in pairs or in bundles. It has granular inclusions.
It grows on anaerobic media in the form of colonies compared to aspect with the occlusal face
of a molar.
Its pathogenicity is due to the lipopolysaccharide in the structure of the gram-negative wall, the significant
excretion of butyric acid as a toxic metabolite, but also to the pronounced synergism with the species
Porphyromonas gingivalis, causing significant tissue damage.
The special importance of this species is due to the presence of adhesive structures of a great variety. Bacteria
of the species Fusobacterium nucleatum may have adhesions that recognize the receptors of more than 10 oral
bacterial species.
The bacterium is thus considered to have a "key" role in the formation of bacterial plaque, implicitly in its
evolution towards pathogenicity.
Prevotella intermedia (Bacteroides intermedius)
They are isolated from chronic periodontitis with advanced lesions from adult, but also from ulcer-necrotic
gingivostomatitis.They are gram-negative coccobacillus. They grow in anaerobic environments and form small,
round colonies, wet, and pigmented in black.They ferment carbohydrates with the formation of
acetic and succinic acid.
Prevotella species have been shown to have the ability to acquire resistance to lactam β antibiotics
by an adaptive biochemical mechanism with the production of β lactamases (more commonly
nigrescens).
Micromonas micros (Peptostreptococcus micros)
Species from the group of peptostreptococci isolated from marginal periodontitis are Micromonas micros and
Peptostreptococcus anaerobius.
The Micromonas micros species has bacteria in the form of gram-positive cocci, placed in isolation or in chains.
Cultivation is performed on anaerobic medium.In periodontal lesions it is associated with other periodontal
pathogens; the number of these bacteria is very high in severe lesions, advanced form of disease.
The species Peptostreptococus anaerobius is correlated with aggressive periodontitis with early onset.
Campilobscter rectus (Wollnella recta)
Gram-negative bacteria with a helical appearance, very mobile due to a polar cilia. It grows under anaerobic
conditions and forms colonies that look different depending on the composition of the culture. The species can
be frequently isolated from subgingival bacterial plaque in patients with periodontitis accompanied by advanced
destruction. In AIDS patients with marginal ulcero necrotic periodontitis it is the dominant bacteria.
Eubacterium nodstum
The species belongs to the Eubacterium group, which includes many anaerobic species, difficult to cultivate.
Bacteria are gram-positive with a coco-bacillary appearance. According to some authors, the species
Eubacterium nodatum is associated with severe forms of marginal periodontitis.
Other bacterial species isolated in marginal periodontitis
Other cultivable species, isolated from periodontitis belong to the genus Capnocytophaga.
Bacteria are gram-negative and sometimes have a bacillary form, sometimes they are filamentous, are
capnophilic but favored by cultivation in anaerobiosis. All species included in this genus are resistant to
metronidazole.
The species Eikenella corrodens comprises gram-negative, facultative anaerobic bacilli and develops
characteristic colonies that erode agar. Bacteria have been isolated from patients with aggressive forms and
advanced lesions of periodontitis, but the direct pathogenic role of these species has not been established.
Capnocytophaga and Eikenella bacteria belong, according to SOCRANSKI, to the green complex and can be
isolated from periodontitis, but they are also present in healthy sites.
Other microorganisms, protozoan parasites such as Entamoeba gingivalis, Trichomonas tenax, have been
identified in advanced lesions of marginal periodontal disease. However, these parasites can also be present in
healthy people, especially in the elderly.
Herpes viruses have also been found in the gingivo-periodontal tissues, but they are also present in a latent
state in most adult patients. They could not be directly incriminated in the production of periodontal disease but
can cause a decrease in resistance and thus facilitate periodontal pathogenic bacterial infection.
