5. Microbiology of Periodontal Disease Flashcards
Periodontal Diseases Have Microbial Etiology
* Stop brushing/flossing > gingivitis (\_\_\_\_) * 10% > periodontitis (\_\_\_\_) * Gumline > increase in volume of gingival margin (due to inflam cells/fuids) and then loss of atatchment, PDL and bone (in perio)
reversible
irreversible
Contemporary Model of Pathogenesis of Periodontitis
* Amount of plaque and content of plaque changes as you go to disease * \_\_\_\_ (low biomass, and content is of bacteria that is beneficial) in clinical health > protect the sites to prevent colonization * Gingivitis > high \_\_\_\_, and bc of changes you select for diff bacteria > lead to diff outcomes; host fighting the bacteria * \_\_\_\_ and \_\_\_\_ that affect bacteria levels > modulating the biofilm to affect composition and behavior
symbiosis
biomass
smoking
diabetes
Different Oral Habitats Harbor Different Biofilms
• Oral cavity - \_\_\_\_ sq/cm2 (palm of hand); soft and hard tissue • Bacterium > prefer to stay on teeth, \_\_\_\_ surface; close to gingival margin has a lot of "food" for you • \_\_\_\_ is how we subdivide the bacteria • Take samples form each area, they are different from each other and group differently > some bacteria are better at binding to some areas than others ○ The closer two of the names are the more similar > \_\_\_\_ and \_\_\_\_ more similar to each other • 1ml of saliva = \_\_\_\_ microbial cells; each areas has different profile due to physical characteristics of each area
220 non-shedding areas subgingival supragingival 10^8
• More advantageous to live in ____ than in a ____; more difficult to treat for clinicians
biofilm
planktonic state
Biofilm: a Bacterial Lifestyle
• Carpentier & Cerf (1993): “a community of microbes embedded in an organic polymer matrix, adhering to a surface”
• Elder et al (1995): “a functional consortium of microorganisms organised within an extensive exopolymer matrix”
• Costerton et al (1999): “a structured community of bacterial cells enclosed in a self-produced polymeric matrix and adherent to an inert or living surface
* \_\_\_\_ - father of biofilm biology in dentistry * Commonalities, necessary for biofilm formation > \_\_\_\_, \_\_\_\_ and an ideal \_\_\_\_ * Oral cavity > different areas with diff communities and diff surfaces
costerton
matrix
community
ideal surface
COMPONENTS OF A BIOFILM
• Require a \_\_\_\_ that bring bacteria in, and taking the waste out, bring nutrients and substrate ○ Tooth, soft tissue, implant, composite, a lot • Fluids > saliva and GCF ○ Saliva - \_\_\_\_, GCF - \_\_\_\_
bulk fluid
carbs
proteins
Advantages of Biofilm Lifestyle
• Protection from ____ system
• Protection from antibiotics and antimicrobials
• Bacteria: ____ of the dry weight; ____ is matrix of polysaccharides, salivary proteins and glycoproteins
• ____: maintains integrity of the community
• ____: 75-80% of volume, concentration of enzymes
• Enzymes produced by bacteria can be concentrated in the glycocalyx
• Cells at the bottom of a biofilm are alive but metabolically ____: metabolic activity in outer layers
• Tolerance to antibiotics > dose, and the target bacteria is down further in the biofilm; and, resistant bacteria on the outside, and can also conduct gene transfer of \_\_\_\_ genes • Bacteria ry to bind electrostatic and van der waals at first, and have appendages \_\_\_\_, fimbrarie > as soon bind, they change their \_\_\_\_ > secreting matrix (and exopolysacc's) ○ You have viruses, yeast and archaea as well • \_\_\_\_ > formation of food webs
immune 70% 30% exopolysaccharides glycocalyx inactive resistance pilliae behaviors cross-feeding
Biofilm Structure
* \_\_\_\_-like colonies * Within biofilm > \_\_\_\_ system (bring fluid, oxygen, nutrient, substrate) * \_\_\_\_ > outside of biofilm > clusters that are detaching that try to go colonize elsewhere
mushroom
primitive circulatory
streamer
The Biofilm Life Cycle
1: ____ attachment
2: ____ attachment
3: Maturation I
4: Maturation II
5: ____
* Saliva > glycoproteins; antibodies (\_\_\_\_) cover the surface > bacteria come to adhere, but only specific bacteria for glyco's > multiply and secrete \_\_\_\_ > spread \_\_\_\_ and then \_\_\_\_ > form mushroom-like colonies > dispersion (release of planktonic) * Behave differently in biofilm vs planktonic; as soon as bacteria attaches and change \_\_\_\_ expression (up to \_\_\_\_ of genetic changes)
reversible irreversible dispersion IgA exopoly lateral vertical gene 30%
Dental Plaque Development Pellicle formation Initial colonization Horizontal spread and co-aggregation Vertical growth Climax community (niches)
* Pellicle formation immediately after porphy > initial colonization (good at binding glycoproteins, \_\_\_\_, \_\_\_\_ [early]) > horizontal spread > secrete matrix and coaggregate > bacteria tthat bind these bugs (coagg) > vertical growth > channels formed > more layers, change the types of bacteria that are here, and they are ammenable to binding more bacteria; each layer has diff \_\_\_\_, etc. * \_\_\_\_ > no more microbial change; * Cell-cell communication > \_\_\_\_ > way for bacteria to sense population and do not need to proliferate anyway
streptococcus actinomyces metabolisms climax community quorum sensing
Dental Biofilm Development
* As it accumulates, occurs at \_\_\_\_, and it is \_\_\_\_ * Grows first laterally, and then vertically * In rougher areas of tooth, why gingival margin? GCF, high protein food
gingival margin
self-limiting
Patients Present Different Rates of Plaque Formation
• Difference in plaque \_\_\_\_ rates • \_\_\_\_ that promote plaque growth ○ Properties of tissue, microbiome, etc.
formation
factors
Microbial Ecology
Study of relationships of microorganisms with one another and with the environment
* Relationships that affect plaque * Conditioning film/pellicle > only bugs that can bind will stay * Resistance > some bacteria may be resistant > can xfer \_\_\_\_ genes, and they also have \_\_\_\_ of resistance (protect surrounding bac) * Quorum signaling * Gene transfer * Coadhesion/coagg > some bugs that bind bug who bind bugs to surface * Antagonims > \_\_\_\_ so they establisht hemsvles * \_\_\_\_ > xhcnaged of metabolic activity
resitant
radius
bacteriocins
cross-feeding
Bacterial Interactions
Early and late colonizers do not ____, but they all aggregate with ____
A more ____ state is conducive to shift towards gram negative anaerobes
Streptococci are less sensitive to ____. Modify environment to more ____ state
Receptors on non shedding surface; recognized primarily by ____
• Streptococci, best at binding pellicle (v specific, receptor to mucins, etc.) • 2nd wave > bugs bind strepto > actinocyes > veinoella > propenma > early colonizers > change environment > consume oxygen and sugar > create new environment for late colonizers • Pathogenic bacteria • F. nucleatum > bridges early and late colonizers ○ \_\_\_\_; p gingi is small \_\_\_\_
co-agg f. nucleatum reduced air reduced streptococci
filamentous
rod
HUMAN SUPRAGINGIVAL
PLAQUE
* Crown with epoxy > observed accumulation * Different \_\_\_\_, and some mushrooms and channels
shapes
Experimental Gingivitis
• Plaque accumulate for \_\_\_\_ days • Most changed in \_\_\_\_ days > gingivitis ○ Change in color, contour, texture and consistency • \_\_\_\_ migration of epithelium due to inflam cells and edema
21
10-11
coronal
Early vs. Mature Supragingival Plaque
* Gram and shapes only * Day 0 > most \_\_\_\_ cocci and rods * Progression of time > losing streptococcus, in expesnse of \_\_\_\_ cocci rods and then \_\_\_\_
gram+
gram-
spirochetes
HUMAN SUBGINGIVAL PLAQUE
* Subgingival plaque > root surface, and GCF, and you have multiple surfaces > protected area * Content of microbiome > 3 types of biofilm > \_\_\_\_-attached, \_\_\_\_-attached, \_\_\_\_-attached biofilm * Type of bugs in eachaarea are \_\_\_\_; in coronal portion > actinomyces, (yellow = streptocuss), purple = veionella, these all like oxygen and metabolize sugar; in apical portion > orange (connect early and late colonizers), and then at bottom > red complex (p gingiv, trepmone, tener forsythia) > need heme to grow * these areas are \_\_\_\_ > \_\_\_\_ and \_\_\_\_ comes to feed them
tooth epithelium loosely different ulcerated GCF blood
Non-Specific Plaque Hypothesis
Periodontal Diseases result from plaque accumulation, ____ of the species present within the dental plaque
irrespective
Acute Necrotizing Ulcerative Gingivitis (ANUG)
* This brough back \_\_\_\_ theory * Trench mouth, from WWII * Areas of necrosis and papillae > \_\_\_\_ and smells, all gingival and no loss of \_\_\_\_ * Was not about amount of plaque, and bugs > \_\_\_\_ and \_\_\_\_ mostly > immune response
specific paifnul bone fuso spirochetes
Aggressive Periodontitis (Localized) - LJP
* \_\_\_\_ * \_\_\_\_ and \_\_\_\_ * Limited amount of \_\_\_\_
plaque
first molars
incisors
plaque
Specific Plaque Hypothesis
____ species of bacteria cause different forms of gingival and periodontal diseases
specific
- Studied using culture > take samples, petri and grow and identify
- Comparing: dx has more ____ and more ____(prevotella, and p gingivalis); normal has compatible healty colonies
- Use ____ > numbers and shapes of bacteria; right is dx (filaments and spirochetes, and more of them)
colonies
black
phase contrast
Checkerboard DNA-DNA Hybridization
* Before microarrays, think of this as a \_\_\_\_ * Some bugs more \_\_\_\_ than others (40) * DNA probes targeting bacteria; attached to \_\_\_\_ molecule; when probe is denatured (half of DNA strand with reporter) finds complement on bacteria > light/signal * Can compare to \_\_\_\_, so you can figure out how many bugs you have in each sample
macroarray
common
reporter
standards
Microbial complexes
Blue: \_\_\_\_ Yellow: \_\_\_\_ Purple: \_\_\_\_ and \_\_\_\_ Green Orange: \_\_\_\_ Red: \_\_\_\_, \_\_\_\_ and \_\_\_\_
actinomyces streptococcus v. parvula a. odontolyticus f. nucleatum p. gingi, t. forsythia, t. denticola
Early Biofilm Formation
Pellicle
* 0, 2, 4, 6 hours and analyze for bugs * At 0 > you have bacteria, and you had more and more as time goes on * Best at growing at this stage > \_\_\_\_ and \_\_\_\_ * Specificity > copmoisiton of saliva is different from the plaque (saliva is red)
yellow
actinomyces (blue
Experimental Gingivitis
* Over 21 days * 0 - 21 : increases in bacteria, most significant in \_\_\_\_ * Also confirms that you can restore \_\_\_\_ even better than before
orange
health
Ecological Plaque Hypothesis
* Focuses on \_\_\_\_ * Takes into consideration, mass, content, behvaior and influence of host and evniroenmtn * A little biofilm > a little inflam > not a lot of CGF > gram+, \_\_\_\_ compatbiily * A lot of biofilm > inflam > a lot of GCF, bleeding, higher \_\_\_\_, higher \_\_\_\_ > anaerobic, like GCF * \_\_\_\_ increases GCF levels * Factors that tip balance
dysbiosis health pH temp smoking
- Smokers > red; non-smokers > green
- Visualize how similar and different samples are; closer together in space are more similar in microbial composition, etc
- Reds are ____, greens are ____; different from one another
- Smoking chages ____
clumped
clumped
microbiome
Smoking Favors a Pathogenic Microbiome
* Used a checkerboard * All sites > orange, red; also seen in deep and \_\_\_\_ sites * Sites that look clinically healthy, but they have a more \_\_\_\_ microbiome bc of the environemental pressure that \_\_\_\_ represents
shallow
pathogenic
smoking
Ethnicity Modulates the Oral Microbiome
* Ethnicity, AA, chinese, latinos, whites * Some level of \_\_\_\_, but they had core \_\_\_\_ that distinguish them from each other * \_\_\_\_ can influence, not just ethinicity
commonality
bacteriotype
diet
What Does Microbiome Mean?
* Collection of microorgs in oral cavity; including \_\_\_\_, yeast, but usually just the bacteria * Couldn't culture some bugs back then; but then 2000's that allowed \_\_\_\_ to culture bacteria > went to knowing we had \_\_\_\_ bacteria that could colonize (not 700 all at once, etc.) * Develop database > look into 700 > \_\_\_\_ cultivated, \_\_\_\_% that cannot be cultivated and we don’t know much about that may be pathogens (\_\_\_\_ unnamed)
viruses DNA 700 50% 35% 15%
• Firstly Looked at ____, sizes and colnines [only few ____ at a time] > many bacteria at once > complexes how we go from health to disease > in 2000’s+ ____ techniques that didn’t need cultivation, only need the ____ (can look at ____, fungi, etc.)
shapes samples molecular DNA viruses
Periodontal Microbiome in Health and Disease
• Good to explore, and to confirm that these bugs were pathogens actually • People healthy, and perido people, and within that , some from sahllow and some from deep • Plotted in \_\_\_\_ order of relative abundance • Red - deep, green is shallow, blue is healthy • In disease > p gingiv, trepnoma, tanneroma forsythia; also see other bugs; and \_\_\_\_ organisms ○ All bugs are high in disease, and low in health and \_\_\_\_ in shallow • \_\_\_\_ in dx patients are diff from shallow in healthy (have more pathogenic bacteria) > these shallow sites will p\_\_\_\_
decreasing uncultivated intermediates shallow progress to disease
Periodontal Diseases Result from Dysbiotic Communities
• ____: shifts in the resident microbiota of certain body sites that lead to disease
• Biofilms as a whole cause and perpetuate chronic infections
– “Pathogenic ____”,
– “Pathogenic ____ community”,
– “The community as ____”
* Pathogen here, the bugs we already have them in our mouth (diff from other diseases) part of resident microbiota > pathogen \_\_\_\_ * Just presence of pathogens do affect community as whole > introduce p gingi > the community behaves \_\_\_\_ (not just pathogen being bad and influencing the community)
dybiosis
synergy
microbial
pathogen
synergy
differently
Classical vs. Opportunistic Infection
• \_\_\_\_, can be pathogenic but they can live in harmony (the case here in periodntitis)
pathobionts
Identification of Pathogens • Over \_\_\_\_ species • \_\_\_\_ species • \_\_\_\_ sample • Identification of \_\_\_\_ sites • Periodontal disease activity resulting from different species
• Need numbers of active sites in order to observe
700
uncultivable
representative
active
Identification of Pathogens
- Clinically similar Diseases
- Bacteria that grow as a result of the disease process
- ____ Infections
- ____ state
- ____ strains
- ____ state > people with perio, but had pathogens for a number of years, but something happened so they develo
- Different strains > strains may not be as ____ (not as much LPS); not taken into consideration
mixed
carrier
virulent
carrier
virulent
Identification of Periodontal Pathogens: Koch’s Postulates Modified
by Socransky
* How to tell whether pathogen > \_\_\_\_ postulates * \_\_\_\_ > preence/numbers in disease cased * \_\_\_\_ > * Host response > immunogenic * \_\_\_\_ factors > intent to do harm * Animal sutides > reproduce here * Done in \_\_\_\_, \_\_\_\_, \_\_\_\_ and \_\_\_\_ (hard to study), this why they're classical pathogens
koch's association elimination virulence AA PG TF spirochetes
Virulence Factors
- ____; coagg, multiplication
- ____ (LPS) on surface and in vesicles
• Interbacterial relationships - together they’re ____, more potent
• Bacterial enzymes: collagenases, proteases - i.e., some proteases that degrade ____
• Overcome host defense mechanisms - when go from health–> gingivitis–> periodontitis, either ____ bacterial action ex. LPS but a lot of damage (loss of fibers, bone) is due to the host response to that bacterial insult- ____.
• Factors resulting in tissue damage:
– Indirect : caused by the immune response, upon bacterial stimulus
– Direct: caused by the bacteria
adhesins endotoxins stronger IgA direct indirect vesicles
TABLE OF ACTIONS OF LPS!
YA
This is a number of examples of these virulence factors in some of the most common pathogens: P. Gingivalis and AA. So both of them have ____.
P. Gingivalis has ____ that releases blebs with LPS, ____ to attach, ____ to penetrate tissues, ____ to degrade CT, trypsin like proteases.
AA. primary virulence factor is ____ which kill leukocytes but also has LPS, ____, bacteriocins, ____.
LPS capsules fimbrae invasins collageneases
leukotoxin
adhesins
invasins
Kissing???!
This study ruined kissing for everyone..looked into French kissing and to see what happened to microbiome.
Couples said how many times and how kiss and had them kiss.
Control group gave probiotics- ____. Then kissed again. For priobiotics group, knew what bacteria they gave, saw after kiss how much bact. the person gave the other: ____ M bacteria
But these bugs ____. Kissing could influence the microbiome but was the ____ of kissing. Basically its not because you were exposed to some new bug that is now part of our microbiome. But if you are exposed multiple times–> part of ____.
lactobacillus bravis 80 transient frequency microbiome
Take Home Messages
Oral Cavity is Unique in the Human Body:
• ____ Environment
• ____ Host-Microbial Interaction
• Etiologic agents reside within a biofilm that is located outside the body (double protection)
• Pathogens are part of the ____ microbiota: found in health for long periods prior to disease initiation
• Provides ____ surfaces: natural microbial colonization
• Multiple ____ for pathogens beyond the periodontal pocket
open dynamic indigenous non-shedding reservoirs
- There are clear differences in the development and composition of ____ and ____ biofilms (plaque).
- ____ (saliva/GCF), ____ (tooth, soft tissue, restorations) and ____ are key determinants of biofilm formation and composition.
- Bacterial colonization is initiated by colonization by ____cocci and rods, followed by ____ cocci and rods. Then ____ and ____ and finally ____ are detected.
supragingival subgingival bulk fluid surface bacteria gram+ gram- fusobacteria filaments spirochetes
- There are clear clinical differences between periodontal health, gingivitis and periodontitis
- Gingivitis and periodontitis result from microscopic changes in the underlying periodontal structures
- These changes result from interactions between the ____ and the local ____ and can be modulated by certain ____ and ____ factors
- Gingivitis is widespread in populations, while periodontitis is somewhat less common
- Periodontitis is preceded by gingivitis, but not all cases of ____ lead to periodontitis
host microbiota local systemic gingivitis
- Factors such as ____, diet, ____ and ethnicity can shape the oral microbiome
- The ____ plaque hypothesis provide the framework for the current understanding of the etiology of periodontal diseases
- Certain microbial complexes are associated with periodontal health and others to disease: their colonization follows a ____ pattern
- The modified ____ are the basis for the classification of pathogens
- The health-associated microbiome is composed primarily of ____ and ____ (yellow and blue complexes) whereas the disease-associated microbiome is enriched for ____, ____ (orange complex), ____ and ____ (red complex)
- Adherence capability, ____ and ____ are some of the most important virulence factors of periodontal pathogens
smokiing genetics specific specifc koch's postulates streptococci actinomyces prevotella fusobacterium porphyromonas gingivalis treponema denticola
LPS
enzymes
