1. Perio Protect Method™ Scanning Electron Microscope Bacterial Count Results D.C. Keller, Perio Protect LLC, St. Louis, MO, USA; J.W. Costerton, USC School of Dentistry, Los Angeles, CA, USA; C. Schaudinn, USC School of Dentistry, Los Angeles, CA, USA; P. Sedghizadeh, USC School of Dentistry, Los Angeles, CA, USA; G.S. Keller, Keller Professional Group P.C., St. Louis, MO, USA

2. Perio Protect Method™ The Perio Protect Method™ uses an appropriately formed, customized dental tray fabricated in a licensed dental lab to place prescribed solutions into periodontal pockets. Localized Medication Delivery System Next slide

3. © Perio Protect Perio Protect Method™ Localized Medication Delivery System Flexible polycarbonate carriers were inserted into periodontal pockets and worn in vivo for two day intervals during treatment with the Perio Protect Method™. The pictures in this presentation show beginning bacterial biofilm and subsequent SEM pictures taken after 2 days, 7 days, 12 days, and 17 days of treatment. Flexible Polycarbonate Carriers

4. © Perio Protect 19 mb, before Perio Protect ® Treatment

5. © Perio Protect 19 mb, before Perio Protect ® Treatment The magnified biofilm has formed on the carrier and consists of those bacteria present in the 6mm periodontal pocket. These can be virulent strains, such as Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Treponema, etc. Next slide

6. © Perio Protect 19 mb, 2 days after Perio Protect ® Treatment

7. © Perio Protect 19 mb, 2 days after Perio Protect ® Treatment

8. © Perio Protect 19 mb, 2 days after Perio Protect ® Treatment This slide demonstrates a much modified oral bacterial environment after 2 days of treatment. The colonization of fusiform-like bacteria and long rods has been affected by the patient delivered medications and some of the cocci-like bacteria also demonstrate modifications in form. Next slide

9. © Perio Protect 19 mb 7 days after Perio Protect ® Treatment

10. © Perio Protect These results after using the medications for 7 days demonstrate a dramatic change in the bacterial colonization. The bacterial population is decreased in this study by 98% and there are no rods observed in these scans. The remaining biofilm bacteria demonstrate significant modifications. Next slide 19 mb, 7 days after Perio Protect® Treatment

11. © Perio Protect 19 mb, 12 days after Perio Protect ® Treatment

12. © Perio Protect The periodontal pocket is significantly changed by 12 days. There are almost no bacteria observed and the pellicle is observed to be incompletely formed on the carrier through the action of the medications where hyaluronic acid is cleaved and histadine is converted to alanine and asparagine is converted to aspartate and the protein chains are disrupted. Next slide 19 mb, 12 days after Perio Protect® Treatment

13. © Perio Protect 19 mb, 17 days after Perio Protect ® Treatment

14. © Perio Protect 99.98% of the bacteria are missing from the carriers at 17 days. Those having the form of Actinomyces-type of bacteria and a few cocci- like bacteria are all that can be found and we know these from the normal aerobic microflora of healthy gums. Next slide 19 mb, 17 days after Perio Protect® Treatment

15. 10 4 10 5 10 6 2 0 12 7 days 17 ~2x10E6 ~4x10E3~5x10E3 ~2x10E6 ~4x10E2 bacteria cells short rods Fusiform-like Selenomonas-like Cocci-like Actinomyces-like 100% 0.2% 0.25% 0.02% 100% 1 2 2 3 23 3 4 4 4 3 5 Probing Changes after 12 days 10 3 Perio Protect Method™ effect on subgingival biofilms (graph) tooth 19 mesial buccal © Perio Protect

16. 100% 0.2%0.25% 0.02% 2 2 0 0 12 7 7 17 ~2x10E6 ~4x10E3 ~5x10E3 100% days 19mb ~6x10E5 ~7x10E2 ~3x10E7 carrier lost 2% 1% 0.05% 0% 2x10E5 1x10E4 2x10E7 0x10E0 ~2x10E6 ~4x10E2 0.002% 14mb 14db short rods Fusiform-like Selenomonas-like Cocci-like Actinomyces-like © Perio Protect Charts

17. © Perio Protect End of Presentation Cell analysis before treatment with the Perio Protect Method™ determined the pocket ecosystem consisted of large numbers of fusiform-like bacteria, cocci, short rods and in one pocket, treponema. After two days of using the Perio Tray™ for minutes each day according to the Perio Protect™ protocol most of the fusiform bacteria and treponema had disappeared. After 17 days there were less than 0.02% (99.98% Kill) of the periopathogenic bacteria and what remained were some cocci and pleomorphic rods that were visible next to a large number of eukaryotic cells. After 17 days the few remaining bacteria, cocci- and actinomyces-like, are known as normal aerobic microflora of healthy gums.