Experimental Gingivitis in

Man
Harald Löe, Else Theilade, S. Börglum Jensen, 1965
Aim
• Researchers sought to induce gingivitis in subjects with healthy
gingiva by ceasing oral hygiene practices
• Study change in oral microflora
• How long it would take for gingivitis to occur
• Which sites are more affected
• How long would it take for gingivitis to resolve after resuming oral
hygiene practices
Method
• 12 healthy subjects (mean age 23) with healthy gingiva
• Gingiva assessed for health and plaque accumulation at the start
• Subjects instructed to cease oral hygiene routine
• No-hygiene period ended when gingivitis was diagnosed at
subsequent examination
• Detailed hygiene instructions given
• Experiment ended when gingivitis resolved
Gingival index
• From Löe & Silness, 1963
• Each tooth given a score for 3 sites (buccal,
lingual, mesial)
• Average score of 3 sites gave GI for the tooth
• Teeth grouped (incisors, premolars, molars)
and score for group derived from average
score of teeth
• Average score of all teeth became GI score
for patient
Plaque index
• From Silness & Löe, 1964
• Each tooth given a score for 3 sites (buccal,
lingual, mesial)
• Average score of 3 sites gave GI for the
tooth
• Teeth grouped (incisors, premolars, molars)
and score for group derived from average
score of teeth
• Average score of all teeth became GI score
for patient
• Presence/absence of calculus was not
recorded
Bacteria examination
• Two Methods
• Impression technique, stained with gentian violet
• Conventional Smears made from plaque samples at the same sites. Samples
collected with scaling instruments right after impression and stained with gram stain.
• Observed under microscope and classified by morphological characteristics
• Microflora at buccal gingival margin 24, 25, 26, and papillary area mesial 26
and mesial to 25
• Levels recorded
• Examined at intervals from start of experiment until end of no-hygiene
period, final examination done when healthy gingiva is established
Key findings – Plaque
• All subjects accumulated a large amount of plaque by the end of the
no-hygiene period. Mean PI increased from .43 to 1.67
• No major difference in plaque accumulation between maxillary and
mandibular arches, nor between different groups of teeth
• Lingual surfaces accumulated less than buccal and interproximal
surfaces
• PI dropped soon after oral hygiene measures resumed
Key findings – Gingivitis
• All subjects developed gingivitis
• Mean GI increased from .27 to 1.05
• 3 subjects developed gingivitis in 10 days
• 9 others between 15 and 21 days
• No major difference in gingival inflammation between maxillary and
mandibular arches, nor between different groups of teeth
• Interproximal surfaces scored higher and lingual surfaces scored lower
• Interproximal areas of maxillary molars most affected, lingual of mandibular
premolars least affected
• Gingivitis resolved a week after oral hygiene resumed
Key findings – Plaque composition

• Derived from both conventional smears and impression technique

• Preparations from healthy gingiva had small groups of bacteria, being mainly Gram-
positive cocci and short rods (90%), as well as desquamated epithelial cells and some
leukocytes
• During no-hygiene period, change in the microflora occurred in 3 stages
• Immediately after, amount of cocci increased, and leukocytes observed along gingival
margin
• 2 to 4 days after, filamentous bacteria (identified as Leptotrichia and Fusobacteria) began
to appear. Accumulations of leukocytes increased in size
• 6 to 10 days, Vibrio and Spirochetes gradually began to appear. This was more difficult to
observe. At first, localized to single papilla or gingival margin of a tooth, then spread to
cover entire gingiva
• Once gingiva returned to normal cocci and short rods dominated the flora again,
filamentous bacteria remained in 2 subjects. No Vibrio or Spirochetes.
Changes in plaque composition
Healthy Gingiva Gingivitis

>Gram positive rods + cocci >Gram positive rods + cocci

90-100% 45-60%
>Gram negative cocci + short
rods 22%
>Gram positive filaments
>Fusobacteria 10%
>Spirochetes + Vibrios 7%
General increase in amount of bacteria
Discussion
• Shortcomings
• Clinical and bacteriological examination was not made on the same
day for all 12 patients, with varying intervals in between
• Tables show broad trends rather than precise data
• Undermines the correlations that can be inferred
Discussion
• Withdrawing all form of oral hygiene leads to rapid increase of oral
debris, and gingivitis
• Shift in microbial composition precedes onset of gingivitis
‘There is no doubt that microorganisms play a role in the initiation of
gingival inflammation’
• Correlation between specific bacterial populations and gingivitis
unknown

• Removal of bacterial plaque causes resolution of gingival inflammation