Dentist Mark Bonner from Canada, assisted by Solange Dunoyé from France, proposes a healing technique of periodontal disease based on the original works of Kofoid (1929), Keyes (1981), Lyons (1989), Bonner (1998 -2014) and Trim (2011).

According to the literature, for over a century these researchers have identified with phase contrast microscopy the protozoan
Entamoeba gingivalis in the teeth crevices affected by periodontal disease. The recent use of polymerase chain reaction (PCR) has detected the amoeba in more than 76% of diseased sites (Trim 2011, Santi-Rocca 2012, Bonner 2014) and has demonstrated its absence in all situation of healthy gingiva. All these studies originally initiated from clinical microscopy allow the practicing dentists, on the one hand, to visualize either the pathogen biofilm in periodontal disease situation or healthy commensal biofilm according to the gingival condition. This technic, on the other hand, allows exceptional clinical results. This protocol is also pertinent and efficient for any
dental practice involved in parasitic component of periodontal diseases and implantology.

Healthy Biofilm

Under microscopy, healthy biofilm typically present as non-motile dots and dashes bacteria from the collected gingival crevice. Those are called cocci and filaments of different lengths and thicknesses. Note inflammatory cells are absent.

Gingivitis Biofilm

Gingivitis features coil-shaped bacteria, along with bacilli and mobile vibrio, very dense motile rod-like small cigars. Very restless, they are working around the remains of the white blood cells, clustered like vultures on a carcass of a zebu. This is bacterial gingivitis. Some inflammation is present.

Periodontitis biofilms

With a perfect microscopic correlation in the cases of active periodontitis, the amoeba Entamoeba gingivalis is caught in swallowing a white blood cell nucleus (critical activity we call “exonucleophagy”, annihilating normal immune defense response of the patient). The remains of leukocytes hollow of their substance – the ghost’s white blood cells – lay scattered around. Ravenous ghoul leaves scattered proteolytic enzymes spread on the fragile periodontal attachment and cause damage that we know so well. The NETs activity of PMN is necessarily disturbed. To avoid of course to obtain periodontal healing.

Microscopic Parasites Searching Method

Search primarily by scanning (100x) and follow by spotting (1000x) to find parasites. Once infected location within the biofilm is visually defined, spotting at 1000x confirms parasite diagnosis, thanks in part to the typical amoeba nucleus (central karyosome and peripheral
chromatin).

Spotting amoeba pseudopod and typical nucleus confirm the presence of the parasite surrounded by PMN activity at 1000x magnification.

11 Steps Protocol

1. Measure periodontal pockets: note the 3 teeth with the deepest pockets and their location
2. Take the sublingual saliva and spread 2 cm on a frosted glass slide: write patient name, 3 sites
3. Scrape the bottom of the three periodontal pockets with a periodontal probe and drop these 3 points of plaque
   quickly into the saliva
4. Avoid shaking or disturbing the arrangement of this biofilm
5. Deposit a large surface thin cover glass 22 x 40 mm and squeeze very strongly without sliding the coverglass (for maximum contrast) to obtain the thinnest possible biofilm
6. Avoid using a medium other than the saliva of the patient. The use of tap water, physiological saline or Ringer solution will deform the amoebae to the point of not recognizing it
7. Avoid any substance such as alcohol or ether gum to seal the coverslip
8. Make the first investigation at 100x of the first sample. Facing a suspicious area (high motility, white blood cells, nest of parasites), go to 1000x under immersion oil to confirm the diagnosis
9. Repeat the same procedure for the other two samples
10. To find amoebae, look for small black-black donuts (100x dark background) in a field of live neutrophils (exonucleophagy) or an arrangement of moving Actinomyces brush pattern (nesting)
11. Write down the most relevant or most pathogenic results as your main diagnosis