Efficacy of Diagnostic Treatments Flashcards

1
Q

What are the current diagnostic methods used for periodontal diseases?

A

Periodontal disease is currently diagnosed almost entirely on the basis of its clinical manifestations
- sign of gingival inflammation: redness and swelling
- periodontal probing: PD, BOP, CAL
- tooth mobility
- furcation involvement
- radiographs: bone changes - amount of bone loss
**(periodontal examination and radiographs are traditionally used diagnostic procedures for people over the age of 50)
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2
Q

What are some diagnostic techniques and methods for periodontal diseases that are not routinely used in clinical practice?

A
  • Microbiologic testing
  • Assessment of the Host response
  • genetic analysis (only for research)
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3
Q

How is microbiologic testing used as a diagnostic technique for periodontal diseases?

A

These are expensive and require well-trained personnel to conduct these tests (obtain saliva and plaque for testing)

  • bacterial culturing
  • direct microscopy
  • immunodiagnostic methods
  • enzymatic methods
  • molecular biology techniques
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4
Q

How do you assess the host response to diagnose periodontal diseases?

A
  • this is an biochemical analysis of host as part of periodontal diagnosis:
  • some sources of samples to test are are GCF (most commonly used), saliva, and serum (Blood)
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5
Q

How can genetic analysis be be used to diagnose periodontal disease?

A
  • there is a genetic susceptiloitilty to periodontitis

- gene polymorphism is a risk marker for periodontitis

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6
Q

What are some limitations of using probe penetration as a diagnostic method?

A
  • lack of sensitivity and reproducibility
  • probing depth depends on : gingival inflammation, insertion force, placement and angulation, size, probing technique, probe calibration, presence of sub gingival calculus, overhand restorations
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7
Q

what are some limitation of using CAL as a diagnostic method?

A
  • poor reliability and reproducibility

- limited practical value

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8
Q

what are some limitation of using radiograph examination as a diagnostic method?

A
  • limited sensitivity in small bone change
  • changes in bone can be identified by eye only after 30% to 50% of the bone mineral has been lost (subtraction radiography: detect bone density change as low as 5%)
  • no value in evaluating disease activity or progression
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9
Q

What is a ultrasonic periodontal probing?

A

ultrasonic periodontal probe uses a hollow tapered tip that is filled with water for coupling of the ultrasonic beam into the tissues (non-invasive)

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10
Q

What is cone-beam computed tomography?

A
  • conventional radiographs (PA; Pano) are very specific, but lack sensitivity
  • recently, CBCT has been introduced in periodontology for the detection of periodontal defects in in vivo settings
  • CBCT is promising for periodontal applications, especially for intrabony defects, dihiscence and fenestration defects, periodontal cysts, furcation defects and thickness of palatal masticatory mucosa
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11
Q

what is bacteria culturing for diagnostic testing?

A
  • is the gold standard (reference) method for microbe testing
  • assess for antibiotic susceptibility of microbes
  • can only grow live bacteria; strict sampling and transport conditions are essential
  • some putative pathogens are fastidious and difficult to culture
  • sensitivity is low: detection limits for selective and nonselective media average 10^4 to 10^ 5 bacteria
  • sophisticated equipment and experienced personnel required; relatively time- consuming and expensive
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12
Q

What’s direct microscopy for diagnostic testing?

A
  • alternative to bacterial culture methods
  • dark-field or phase-contrast microscopy
  • see the morphology and motility of bacteria in a plaque sample
  • most of the main putative perio pathogens are non-motile (so its is difficult to identify)
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13
Q

what are some immunodiagnostic methods for diagnostic testing?

A

use Ab that targets specific bacteria Ag

  • direct and indirect immunofluorescent microscopic assay (IFA)
  • able to identify pathogens using a plaque smear
  • used mainly to detect Aa and Pg
  • comparable to bacterial culture
  • does not require viable bacterial cells
  • cytoflurorography (flow cytometry)
    • complexity and cost prevent its wide use
  • enzyme-linked immunosorbent assay (ELISA)
    • used primarily to detect serum antibodies to periodontal pathogens
    • membrane immunoassay (Evalusite): chairside use to detect Aa, Pg, and Pi (detection limit of 10^5 for Aa and 10^6 for Pg)
  • latex agglutination:
    • based on the binding of protein to latex: latex beads are coated with species-specific antibody
    • currently these assays only for research purposes
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14
Q

what are some enzymatic methods for diagnostic testing?

A
  • several putative periodontal pathogens such as Pg, Tf, and Aa possess in common have a trypsin-like enzyme that hydrolyzes a subtract N-benzoyl-DL-arginine-2- naphthylamide (BANA)
  • chair-side kit (Perioscan) was available in the 1990’s
  • inability to distinguish between individual bacteria
  • it may be positive at clinically healthy sites
  • negative result doesn’t rule out the presence of other important periodontal pathogens
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15
Q

What are some molecule biology techniques for diagnostic testing?

A

Analysis of DNA, RNA, and structure or function of protein from target microorganisms

  • Nucleotide acid probes
    • synthesized and labeled DNA (20-30 nucleotides)
    • genomic DNA probe (whole DNA strand); significantly decreased in sensitivity and specific due to cross-reactivity to non-target microorganism
    • 16s rRNA - oligonucleotide probes (high sensitivity and specificity)
  • checkerboard DNA-DNA hybridization
    • whole genomic digoxigenin-labebled DNA
    • up to 40 oral bacterial species in a single test
    • not been generalized for diagnostic purpose
  • PCR
    • high sensitivity and specificity for the identification of target pathogens
    • PCR lower detection limit: 25-100 cells (culture: 10^4-10^5 cells)
      * unable to quantify pathogens accurately in clinical samples
  • real-time PCR
    * real-time PCR: good correlation between the fluorescent signal measured and the number of bacterial cells been used
    * expensive and sophisticated technology in real-time PCR
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16
Q

How do you collect GCF?

A
  • paper strips are placed within the crevice for 30 seconds
  • fluid volume can be quantified by Periotron
  • captured samples may not be present the entire periodontist
  • selection of the teeth and sites is often difficult
17
Q

What do you test when you obtain the GCF?

A
  • over 1166 proteins are present in GCF - 65 of them. We are gong to check for their:
    1) host-derived enzymes and their inhibitors
    2) by products of tissue breakdown
    3) inflammatory mediators and host-response modifiers
18
Q

host-derived enzymes - what are intracellular destruction enzymes?

A
  • possible markers of active periodontal destruction
  • released from dead or dying PMN/neutrophils from periodontium
  • aspartate amino-transferase: released during tissue destruction (cell death)
  • alkaline phosphatase: a membrane-bound glycoprotein involved in maintenance of alveolar bone
  • beta-glucuronidase: a lysosomal enzyme degrades proteoglycans and ground substance
  • elastase: a proteolytic enzyme found in lysosomal granules of neutrophils
19
Q

What are some intracellular destruction enzymes?

A

Aspartate aminotransferase (AST):
- periogard periodontal tissue monitors (chair side test kit)
- a marked elevation in AST levels in GCF from sites with severe gingival inflammation
- inability to discriminate between sites with sever inflammation with or without attachment loss
Alkaline phosphatase (ALP):
- ALP in GCF are higher in diseased then healthy sites
Beta-glucuronidase (betaG):
- elevated in betaG in GCF from sites with severe periodontal disease
- high sensitivity and specificity when related to occurrence of clinical attachment loss
- good predictor for future periodontal breakdown
Elastase:
- Periocheck (char side test kit)
- positive correlation of elastase in GCF with clinical attachment loss

20
Q

host-derived enzymes - what are extracellular enzymes?

A
  • associated with the activity of matrix metalloproteinases

- produced by inflammatory, epithelial and connective tissue cells

21
Q

what is an extracellular destruction enzyme?

A

Matrix Metalloproteinases (MMPs):

  • secreted by fibroblasts and macrophages
  • responsible for remodeling and degradation of ECM components
  • regulated by tissue inhibitors of MMPs (TIMPS)
  • high MMP levels in GCF are at significantly greater risk for progression of periodontitis
  • GCF MMPs level reduces in response to treatment
  • MMP-2 (gelatinase A), MMP -9 (gelatinase B), MMP-8 (collagenase 2), MMP -13 (collagenous 3), and MMP-3 (Stromelysin-1) involve in the initial destruction of periodontal ECM
22
Q

What are some tissue breakdown products from the destruction of collagen?

A
  • the ECM of the periodontist is composed of collagen (predominant), proteoglycans (version, decorin, biglycan, syndecan) and non-collagen proteins (elastin, fibronectin, laminin, osteocalcin, and tenascin)
  • elevated levels of hydroxyproline (breakdown from collage), glycosaminoglycans (From matrix degradation) and osteoclacin and type I collagen (from alveolar bone destruction) can be found note GCF from sites with periodontitis.
23
Q

what are some inflammatory mediators from GCF?

A

Cytokines:

  • TNFalpha
  • IL-1alpha
  • IL-1beta
  • IL-6
  • IL-8
  • PGE2
    1) traditional immunoassays analyze only a single cytokine at a time
    2) Bio-plex cytokine Assay: multiplex bead-based assays designed to quantitative multiple cytokines
24
Q

What’s optical spectroscopy - Infrared (IR) spectroscopy)?

A
  • vibrating covalent bonds of organic molecules absorb a characteristic wavelength of IR light
  • the wavelength of light absorbed depends on the nature of the covalent bond ( e.g., C=O and N-H), the type of vibration (e.g., bending and stretching), and the environment of the bond
  • the spectrum of absorbed light can be used to establish a molecule fingerprint of a tissue or fluid
25
Q

How do you diagnose periodontitis based on IR spectra of GCF?

A
  • measure the total contents of GCF
  • IR Spectroscopy can be used to characterize GCF from healthy, gingivitis, and periodontitis sites
  • vibrations of peptide groups: C=O stretching (amide I band) and N-H bending (amide II band)
  • IR spectroscopy of GCF is reagent free, requiring only small sample volumes, requiring minimal training for operator
  • high sensitivity and specificity
26
Q

what’s Near Infrared (NIR) spectroscopy?

A

-measure of oxygen saturation of the tissues
- the wavelength region 500 to 600 nm is dominated by the absorption from oxygenated hemoglobin (HbO2) and deoxygenated hemoglobin (Hb)
- tissue oxygenation at periodontitis sites significantly decrease as compared to gingivitis and healthy control sites
(tissue hypoxia reflects increased oxygen consumption that occurs with persistent inflammation)

27
Q

what are the current salivary diagnostic tests for periodontal diseases?

A

why salivary?

  • abundant fluid and easy to collect and store
  • highly enriches content of disease biomarkers
    • scubas Sjogren’s syndrome and oral cancer
      1) type and concentration of specific periodontal pathogens
    • apply DNA PCR - identify specific periodontal pathogens
      2) genetic susceptibility to periodontitis in individuals
    • test genetic variation: over-expression of IL-1alpha and IL-1beta
    • these tests identify general risk factors for the development of periodontal diseases, but fail to determine when periodontal destruction will occur
  • -> NOT being able to specifically predict periods of disease activity
28
Q

What’s the future diagnostic method for periodontal diseases?

A

Salivary proteome analysis

  • there are 2290 proteins which have been found in whole saliva (WS)
  • analysis of saliva may offer a cost-effective approach to screening periodontal disease in large populations