Components of Gingival Crevicular Fluid (GCF) and Matrix Metalloproteinases (MMPs)

Gingival crevicular fluid (GCF) is a serum-like fluid found in the gingival sulcus that plays a significant role in periodontal health and disease. Understanding its composition, particularly glucose and protein content, as well as the role of matrix metalloproteinases (MMPs) in tissue remodeling, is essential for dental professionals.

Composition of Gingival Crevicular Fluid (GCF)

1. Glucose and Hexosamines:

   • GCF contains compounds such as glucose, hexosamines, and hexuronic acid.
   • Glucose Levels:
     • Blood glucose levels do not correlate with GCF glucose levels; in fact, glucose concentration in GCF is three to four times greater than that in serum.
     • This elevated glucose level is interpreted as a result of the metabolic activity of adjacent tissues and the influence of local microbial flora.

2. Protein Content:

   • The total protein content of GCF is significantly less than that of serum.
   • This difference in protein concentration reflects the unique environment of the gingival sulcus and the specific functions of GCF in periodontal health.

Matrix Metalloproteinases (MMPs)

1. Definition and Function:

   • MMPs are a family of proteolytic enzymes that degrade extracellular matrix molecules, including collagen, gelatin, and elastin.
   • They are produced by various cell types, including:
     • Neutrophils
     • Macrophages
     • Fibroblasts
     • Epithelial cells
     • Osteoblasts and osteoclasts

2. Classification:

   • MMPs are classified based on their substrate specificity, although it is now recognized that many MMPs can degrade multiple substrates. The classification includes:
     • Collagenases: e.g., MMP-1 and MMP-8 (break down collagen)
     • Gelatinases: Type IV collagenases
     • Stromelysins
     • Matrilysins
     • Membrane-type metalloproteinases
     • Others

3. Activation and Inhibition:

   • MMPs are secreted in an inactive form (latent) and require proteolytic cleavage for activation. This activation is facilitated by proteases such as cathepsin G produced by neutrophils.
   • Inhibitors: MMPs are regulated by proteinase inhibitors, which possess anti-inflammatory properties. Key inhibitors include:
     • Serum Inhibitors:
       • α1-antitrypsin
       • α2-macroglobulin (produced by the liver, inactivates various proteinases)
     • Tissue Inhibitors:
       • Tissue inhibitors of metalloproteinases (TIMPs), with TIMP-1 being particularly important in periodontal disease.
   • Antibiotic Inhibition: MMPs can also be inhibited by tetracycline antibiotics, leading to the development of sub-antimicrobial formulations of doxycycline as a systemic adjunctive treatment for periodontitis, exploiting its anti-MMP properties.

Merkel Cells

1. Location and Function:
   • Merkel cells are located in the deeper layers of the epithelium and are associated with nerve endings.
   • They are connected to adjacent cells by desmosomes and are identified as tactile receptors.
   • These cells play a role in the sensation of touch and pressure, contributing to the sensory functions of the oral mucosa.

Clinical Implications

1. GCF Analysis:

   • The composition of GCF, including glucose and protein levels, can provide insights into the inflammatory status of the periodontal tissues and the presence of periodontal disease.

2. Role of MMPs in Periodontal Disease:

   • MMPs are involved in the remodeling of periodontal tissues during inflammation and disease progression. Understanding their regulation and activity is crucial for developing therapeutic strategies.

3. Therapeutic Applications:

   • The use of sub-antimicrobial doxycycline as an adjunctive treatment for periodontitis highlights the importance of MMP inhibition in managing periodontal disease.

4. Sensory Function:

   • The presence of Merkel cells in the gingival epithelium underscores the importance of sensory feedback in maintaining oral health and function.

Dimensions of Toothbrushes

Toothbrushes play a crucial role in maintaining oral hygiene, and their design can significantly impact their effectiveness. The American Dental Association (ADA) has established guidelines for the dimensions and characteristics of acceptable toothbrushes. This lecture will outline these specifications and discuss their implications for dental health.

Acceptable Dimensions of Toothbrushes

1. Brushing Surface Dimensions:

   • Length:
     • Acceptable brushing surfaces should measure between 1 to 1.25 inches (25.4 to 31.8 mm) long.
   • Width:
     • The width of the brushing surface should range from 5/16 to 3/8 inch (7.9 to 9.5 mm).
   • Rows of Bristles:
     • Toothbrushes should have 2 to 4 rows of bristles to effectively clean the teeth and gums.
   • Tufts per Row:
     • Each row should contain 5 to 12 tufts of bristles, allowing for adequate coverage and cleaning ability.

2. Filament Diameter:

   • The diameter of the bristles can vary, affecting the stiffness and cleaning effectiveness:
     • Soft Filaments:
       • Diameter of 0.2 mm (0.007 inches). Ideal for sensitive gums and children.
     • Medium Filaments:
       • Diameter of 0.3 mm (0.012 inches). Suitable for most adults.
     • Hard Filaments:
       • Diameter of 0.4 mm (0.014 inches). Generally not recommended for daily use as they can be abrasive to the gums and enamel.

3. Filament Stiffness:

   • The stiffness of the bristles is determined by the diameter relative to the length of the filament. Thicker filaments tend to be stiffer, which can affect the brushing technique and comfort.

Special Considerations for Children's Toothbrushes

• Size:
  • Children's toothbrushes are designed to be smaller to accommodate their smaller mouths and teeth.
• Bristle Thickness:
  • The bristles are thinner, measuring 0.005 inches (0.1 mm) in diameter, making them gentler on sensitive gums.
• Bristle Length:
  • The bristles are shorter, typically around 0.344 inches (8.7 mm), to ensure effective cleaning without causing discomfort.

Clinical Implications

1. Choosing the Right Toothbrush:

   • Dental professionals should guide patients in selecting toothbrushes that meet ADA specifications to ensure effective plaque removal and gum protection.
   • Emphasizing the importance of using soft or medium bristles can help prevent gum recession and enamel wear.

2. Education on Brushing Technique:

   • Proper brushing technique is as important as the toothbrush itself. Patients should be educated on how to use their toothbrush effectively, regardless of the type they choose.

3. Regular Replacement:

   • Patients should be advised to replace their toothbrush every 3 to 4 months or sooner if the bristles become frayed. This ensures optimal cleaning effectiveness.

4. Special Considerations for Children:

   • Parents should be encouraged to choose appropriately sized toothbrushes for their children and to supervise brushing to ensure proper technique and effectiveness.

Sutures for Periodontal Flaps

Suturing is a critical aspect of periodontal surgery, particularly when managing periodontal flaps. The choice of suture material can significantly influence healing, tissue adaptation, and overall surgical outcomes.

1. Nonabsorbable Sutures

Nonabsorbable sutures are designed to remain in the tissue until they are manually removed. They are often used in situations where long-term support is needed.

A. Types of Nonabsorbable Sutures

1. Silk (Braided)

   • Characteristics:
     • Excellent handling properties and knot security.
     • Provides good tissue approximation.
   • Applications: Commonly used in periodontal surgeries due to its ease of use and reliability.

2. Nylon (Monofilament) (Ethilon)

   • Characteristics:
     • Strong and resistant to stretching.
     • Less tissue reactivity compared to silk.
   • Applications: Ideal for delicate tissues and areas requiring minimal tissue trauma.

3. ePTFE (Monofilament) (Gore-Tex)

   • Characteristics:
     • Biocompatible and non-reactive.
     • Excellent tensile strength and flexibility.
   • Applications: Often used in guided tissue regeneration procedures and in areas where long-term support is needed.

4. Polyester (Braided) (Ethibond)

   • Characteristics:
     • High tensile strength and good knot security.
     • Less pliable than silk.
   • Applications: Used in situations requiring strong sutures, such as in flap stabilization.

2. Absorbable Sutures

Absorbable sutures are designed to be broken down by the body over time, eliminating the need for removal. They are often used in periodontal surgeries where temporary support is sufficient.

A. Types of Absorbable Sutures

1. Surgical Gut

   • Plain Gut (Monofilament)

     • Absorption Time: Approximately 30 days.
     • Characteristics: Made from sheep or cow intestines; provides good tensile strength initially but loses strength quickly.
     • Applications: Suitable for soft tissue approximation where rapid absorption is desired.

   • Chromic Gut (Monofilament)

     • Absorption Time: Approximately 45 to 60 days.
     • Characteristics: Treated with chromium salts to delay absorption; retains strength longer than plain gut.
     • Applications: Used in areas where a longer healing time is expected.

2. Synthetic Absorbable Sutures

   • Polyglycolic Acid (Braided) (Vicryl, Ethicon)

     • Absorption Time: Approximately 16 to 20 days.
     • Characteristics: Provides good tensile strength and is absorbed predictably.
     • Applications: Commonly used in periodontal and oral surgeries due to its handling properties.

   • Dexon (Davis & Geck)

     • Characteristics: Similar to Vicryl; made from polyglycolic acid.
     • Applications: Used in soft tissue approximation and ligation.

   • Polyglycaprone (Monofilament) (Maxon)

     • Absorption Time: Similar to Vicryl.
     • Characteristics: Offers excellent tensile strength and is absorbed more slowly than other synthetic options.
     • Applications: Ideal for areas requiring longer support during healing.

Junctional Epithelium

The junctional epithelium (JE) is a critical component of the periodontal tissue, playing a vital role in the attachment of the gingiva to the tooth surface. Understanding its structure, function, and development is essential for comprehending periodontal health and disease.

Structure of the Junctional Epithelium

1. Composition:

   • The junctional epithelium consists of a collar-like band of stratified squamous non-keratinized epithelium.
   • This type of epithelium is designed to provide a barrier while allowing for some flexibility and permeability.

2. Layer Thickness:

   • In early life, the junctional epithelium is approximately 3-4 layers thick.
   • As a person ages, the number of epithelial layers can increase significantly, reaching 10 to 20 layers in older individuals.
   • This increase in thickness may be a response to various factors, including mechanical stress and inflammation.

3. Length:

   • The length of the junctional epithelium typically ranges from 0.25 mm to 1.35 mm.
   • This length can vary based on individual anatomy and periodontal health.

Development of the Junctional Epithelium

• The junctional epithelium is formed by the confluence of the oral epithelium and the reduced enamel epithelium during the process of tooth eruption.
• This fusion is crucial for establishing the attachment of the gingiva to the tooth surface, creating a seal that helps protect the underlying periodontal tissues from microbial invasion.

Function of the Junctional Epithelium

• Barrier Function: The junctional epithelium serves as a barrier between the oral cavity and the underlying periodontal tissues, helping to prevent the entry of pathogens.
• Attachment: It provides a strong attachment to the tooth surface, which is essential for maintaining periodontal health.
• Regenerative Capacity: The junctional epithelium has a high turnover rate, allowing it to regenerate quickly in response to injury or inflammation.

Clinical Relevance

• Periodontal Disease: Changes in the structure and function of the junctional epithelium can be indicative of periodontal disease. For example, inflammation can lead to increased permeability and loss of attachment.
• Healing and Repair: Understanding the properties of the junctional epithelium is important for developing effective treatments for periodontal disease and for managing healing after periodontal surgery.

Automated Probing Systems

Automated probing systems have become increasingly important in periodontal assessments, providing enhanced accuracy and efficiency in measuring pocket depths and clinical attachment levels. This lecture will focus on the Florida Probe System, the Foster-Miller Probe, and the Toronto Automated Probe, discussing their features, advantages, and limitations.

1. Florida Probe System

• Overview: The Florida Probe System is an automated probing system designed to facilitate accurate periodontal assessments. It consists of several components:

  • Probe Handpiece: The instrument used to measure pocket depths.
  • Digital Readout: Displays measurements in real-time.
  • Foot Switch: Allows for hands-free operation.
  • Computer Interface: Connects the probe to a computer for data management.

• Specifications:

  • Probe Diameter: The end of the probe is 0.4 mm in diameter, allowing for precise measurements in periodontal pockets.

• Advantages:

  • Constant Probing Force: The system applies a consistent force during probing, reducing variability in measurements.
  • Precise Electronic Measurement: Provides accurate and reproducible measurements of pocket depths.
  • Computer Storage of Data: Enables easy storage, retrieval, and analysis of patient data, facilitating better record-keeping and tracking of periodontal health over time.

• Disadvantages:

  • Lack of Tactile Sensitivity: The automated nature of the probe means that clinicians do not receive tactile feedback, which can be important for assessing tissue health.
  • Fixed Force Setting: The use of a fixed force setting throughout the mouth may not account for variations in tissue condition, potentially leading to inaccurate measurements or patient discomfort.

2. Foster-Miller Probe

• Overview: The Foster-Miller Probe is another automated probing system that offers unique features for periodontal assessment.

• Capabilities:

  • Pocket Depth Measurement: This probe can measure pocket depths effectively.
  • Detection of the Cemento-Enamel Junction (CEJ): It is capable of coupling pocket depth measurements with the detection of the CEJ, providing valuable information about clinical attachment levels.

3. Toronto Automated Probe

• Overview: The Toronto Automated Probe is designed to enhance the accuracy of probing in periodontal assessments.

• Specifications:

  • Probing Mechanism: The sulcus is probed with a 0.5 mm nickel titanium wire that is extended under air pressure, allowing for gentle probing.
  • Angular Control: The system controls angular discrepancies using a mercury tilt sensor, which limits angulation within ±30 degrees. This feature helps maintain consistent probing angles.

• Limitations:

  • Reproducible Positioning: The probe requires reproducible positioning of the patient’s head, which can be challenging in some clinical settings.
  • Limited Access: The design may not easily accommodate measurements of second or third molars, potentially limiting its use in comprehensive periodontal assessments.

Anatomy and Histology of the Periodontium

Gingiva (normal clinical appearance): no muscles, no glands; keratinized

• Color: coral pink but does vary with individuals and races due to cutaneous pigmentation
• Papillary contour: pyramidal shape with one F and one L papilla and the col filling interproximal space to the contact area (col the starting place gingivitis)
• Marginal contour: knife-edged and scalloped
• Texture: stippled (orange-peel texture); blow air to dry out and see where stippling ends to see end of gingiva
• Consistency: firm and resilient (push against it and won’t move); bound to underlying bone
• Sulcus depth: 0-3mm
• Exudate: no exudates (blood, pus, water)

  Anatomic and histological structures

Gingival unit: includes periodontium above alveolar crest of bone

a. Alveolar mucosa: histology- non-keratinized, stratified, squamous epithelium, submucosa with glands, loose connective tissue with collagen and elastin, muscles.  No epithelial ridges, no stratum granulosum (flattened cells below keratin layer)

b. Mucogingival junction: clinical demarcation between alveolar mucosa and attached gingiva

c. Attached gingiva: histology- keratinized, stratified, squamous epithelium with epithelial ridges (basal cell layer, prickle cell layer, granular cell layer (stratum granulosum), keratin layer); no submucosa

• Dense connective tissue: predominantly collagen, bound to periosteum of bone by Sharpey fibers
• Reticular fibers between collagen fibers and are continuous with reticulin in blood vessels

d. Free gingival groove: demarcation between attached and free gingiva; denotes base of gingival sulcus in normal gingiva; not always seen

e. Free gingival margin: area from free gingival groove to epithelial attachment (up and over ® inside)

• Oral surface: stratified, squamous epithelium with epithelial ridges
• Tooth side surface (sulcular epithelium): non-keratinized, stratified, squamous epithelium with no epithelial ridges (basal cell and prickle cell layers)

f. Gingival sulcus: space bounded by tooth surface, sulcular epithelium, and junctional epithelium; 0-3mm depth; space between epithelium and tooth

g. Dento-gingival junction: combination of epithelial and fibrous attachment

• Junctional epithelium (epithelial attachment): attachment of epithelial cells by hemi-desmosomes and sticky substances (basal lamina- 800-1200 A, DAS-acid mucopolysaccharides, hyaluronic acid, chondroitin sulfate A, C, and B), to enamel, enamel and cementum, or cementum depending on stage of passive eruption.  Length ranges from 0.25-1.35mm.
• Fibrous attachment: attachment of collagen fibers (Sharpey’s fibers) into cementum just beneath epithelial attachment; ~ 1mm thick

h. Nerve fibers: myelinated and non-myelinated (for pain) in connective tissue.  Both free and specialized endings for pain, touch pressure, and temperature -> proprioception.  If dentures, rely on TMJ.

i.Mesh of terminal argyophilic fibers (stain silver), some extending into epithelium

ii  Meissner-type corpuscles: pressure sensitive sensory nerve encased in CT

iii.Krause-type corpuscles: temperature receptors

iv. Encapsulated spindles

i. Gingival fibers:

i.  Gingivodental group:

• Group I (A): from cementum to free gingival margin
• Group II (B): from cementum to attached gingiva
• Group III (C): from cementum over alveolar crest to periosteum on buccal and lingual plates

ii.  Circular (ligamentum circularis): encircles tooth in free gingiva

iii. Transeptal fibers: connects cementum of adjacent teeth, runs over interdental septum of alveolar bone.  Separates gingival unit from attachment apparatus.

Transeptal and Group III fibers the major defense against stuff getting into bone and ligament.

2.  Attachment apparatus: periodontium below alveolar crest of bone

Periodontal ligament: Sharpey’s fibers (collagen) connecting cementum to bone (bundle bone).  Few elastic and oxytalan fibers associated with blood vessels and embedded in cementum in cervical third of tooth.  Components divided as follows:

i. Alveolar crest fibers: from cementum just below CEJ apical to alveolar crest of bone

ii.Horizontal fibers: just apical to alveolar crest group, run at right angles to long axis of tooth from cementum horizontally to alveolar bone proper

iii.Oblique fibers: most numerous, from cementum run coronally to alveolar bone proper

iv. Apical fibers: radiate from cementum around apex of root apically to alveolar bone proper, form socket base

v. Interradicular fibers: found only between roots of multi-rooted teeth from cementum to alveolar bone proper

vi. Intermediate plexus: fibers which splice Sharpey’s fibers from bone and cementum

vii. Epithelial Rests of Malassez: cluster and individual epithelial cells close to cementum which are remnants of Hertwig’s epithelial root sheath; potential source of periodontal cysts.

viii. Nerve fibers: myelinated and non-myelinated; abundant supply of sensory free nerve endings capable of transmitting tactile pressure and pain sensation by trigeminal pathway and elongated spindle-like nerve fiber for proprioceptive impulses

Cementum: 45-50% inorganic; 50-55% organic (enamel is 97% inorganic; dentin 70% inorganic)

i.  Acellular cementum: no cementocytes; covers dentin (older) in coronal ½ to 2/3 of root, 16-60 mm thick

ii. Cellular cementum: cementocytes; covers dentin in apical ½ to 1/3 of root; also may cover acellular cementum areas in repair areas, 15-200 mm thick

iii. Precementum (cementoid): meshwork of irregularly arranged collagen in surface of cementum where formation starts

iv. Cemento-enamel junction (CEJ): 60-65% of time cementum overlaps enamel; 30% meet end-to-end; 5-10% space between

v. Cementum slower healing than bone or PDL.  If expose dentinotubules ® root sensitivity.

Alveolar bone: 65% inorganic, 35% organic

i. Alveolar bone proper (cribriform plate): lamina dura on x-ray; bundle bone receive Sharpey fibers from PDL

ii. Supporting bone: cancellous, trabecular (vascularized) and F and L plates of compact bone

Blood supply to periodontium

i. Alveolar blood vessels (inferior and superior)

A) Interalveolar: actually runs through bone then exits, main supply to alveolar bone and PDL

B) Supraperiosteal: just outside bone, to gingiva and alveolar bone

C) Dental (pulpal): to pulp and periapical area

D) Terminal vessels (supracrestal): anastomose of A and B above beneath the sulcular epithelium

E) PDL gets blood from: most from branches of interalveolar blood vessels from alveolar bone marrow spaces, supraperiosteal vessels when interalveolar vessels not present, pulpal (apical) vessels, supracrestal gingival vessels

ii. Lymphatic drainage: accompany blood vessels to regional lymph nodes (esp. submaxillary group)