TRIOLOGY™ In-Office Hygiene Protocol

U.S. Patent # 8,961,939
Abstract*: Exposure to Common Oral Products Causes Cell Death to Human Oral Fibroblasts
King, J.1, Winetroub, D.1, Bowen, J.2, Cole, C3
1 NOWsystem, Inc, Kansas City, KS
2 Dental Arts, Independence, MO 3 Likarda, LLC, Kansas City, KS
Antibacterial oral rinses or gels are designed to dampen bacterial growth or kill microorganisms in the mouth. However, when these
compounds are used chronically, they may also affect the healthy gum cells, namely the keratinocytes and the oral fibroblasts. In this study,
human oral fibroblasts were grown in culture for 1 week until they had reached 70% confluency. At that time, wells in triplicate were
exposed to one of the following common oral rinses at 100% concentration for 30 seconds: Rincinol P.R.N., chlorhexidine gluconate (0.12%),
TRIOLOGY™ Debriding Rinse, Listerine alcohol free, and Listerine original. Control wells had their media changed twice in a 30 second
period, to mimic the physical forces of the media changes in the other product wells. At the end of 30 seconds each compound was
thoroughly removed and media restored to the cells. Images of the fibroblasts were captured and cell counts completed. 24 hours later,
images were again captured for cell counts, and in addition, the cells were exposed to PrestoBlue cell viability reagents (InVitrogen) for cell
counts via fluorescent readings using an Enspire microplate reader (Perkin Elmer).
Cells exposed to Rincinol P.R.N., chlorhexidine gluconate (0.12%), Listerine alcohol free, and Listerine original all caused nearly 100% cell
death with a single 30 sec exposure. Only the wells exposed to TRIOLOGY™ Debriding Rinse had any remaining cells at 24 hours following
exposure or 3 days later with only 27% cell death. In summary, only TRIOLOGY™ Debriding Rinse maintained cell health after a 30 second
exposure.
*Complete abstract available upon request.
Abstract*: A Single Cleansing with Botanical Compounds Reduces Oral Bacteria
Bowen, J.1, Stehno-Bittel, L.2
1 Dental Arts, Independence, MO 2 Likarda, LLC, Kansas City, KS
Fifteen subjects, ages 24-58, were consented and produced a sputum sample, which served as the pre-treatment control. Next, each
subject rinsed with the TRIOLOGY™ Debriding Rinse or chlorhexidine gluconate rinse (CHX) for 30-60 seconds, followed by extensive
rinsing with water. A subsequent sputum sample was collected as the post-treatment sample.
The samples were tested for detection of bacterial DNA using polymerase chain reaction (PCR) conducted by OralDNA, Inc. Each sample
was normalized to an internal DNA control used to calculate a genome equivalent. The bacteria tested encompassed some of the most
predominant microflora of the mouth including:
Aggregatibacter actinomycetemomitans (Aa), Porphyromonas gingivalis (Pg), Treponema denticola (Td), Eubacterium nodatum (En),
Fusobacterium nucleatum/periodonticum (FN), Prevotella intermedia (Pi), Campylobacter rectus (Cr), Peptostreptococcus micros (Pm),
Eikenella corrodens (Ec), Capnocytophaga species (Cs)).
82% of all subjects that rinsed with TRIOLOGY™ had a reduction in their total bacterial load. A reduction in the bacterial level was achieved
with the single TRIOLOGY™ rinse treatment in 10 of 11 of the bacteria tested.
*Complete abstract available upon request.
US Patent No. 8,961,939