Protein Deposition on Sport Mouthguards and the Effectiveness of Two Different Cleaning Protocols

Abstract

Objective: To determine which salivary proteins adhere onto sport mouthguards, and to evaluate the effectiveness of different cleaning strategies in removing deposited protein. Methods: Fifteen healthy volunteers used a mouthguard for 1 h. The deposited salivary proteins were analyzed using gel electrophoresis and Western blotting techniques and compared with the protein composition of unstimulated saliva. In addition, the effectiveness of two different cleaning strategies to remove proteins from the mouthguards were compared: rinsing the mouthguards after use with cold tap water and cleaning the mouthguard with a soluble effervescent tablet. Results: Gel electrophoresis showed deposition of proteins of 50-60 kDa and 14 kDa on the mouthguards used in the mouth for 1 h. Western blotting identified these bands as amylase and lysozyme, respectively. Rinsing the mouthguard with cold tap water after use removed 91% of the total amount of deposited proteins, while cleaning with an effervescent tablet removed 99%. Conclusions: During the use of mouthguards, salivary proteins are deposited on their surface. Because salivary proteins can potentially affect bacterial adhesion to mouthguards, proper cleaning after use is recommended. Cleaning the mouthguard with cold tap water or using an effervescent tablet both seem to be effective strategies to remove proteins deposited on sport mouthguards.

Keywords: adhesion; cleaning strategy; mouthguard; salivary proteins.

Figure 1

(A) SDS-PAGE results of unstimulated whole saliva (S) and used mouthguards without cleaning (1) from 5 different participants (A t/m E). Lane 1 (L) contains the molecular weight markers. (B) Western blot with antibodies against amylase (53 kDa) and lysozyme (14 kDa).

Figure 2

(A) SDS-PAGE of unused mouthguards and unstimulated saliva (S), used mouthguards without cleaning (1), mouthguards rinsed with cold tap water after use (2), and mouthguards cleaned with an effervescent tablet after use (3) from two different participants (A,B). The left lane (L) contains the molecular weight markers. (B) Western blot with antibody against amylase (53 kDa).

Figure 3

Peak Pixel Intensity of protein eluted from mouthguards without cleaning (1), mouthguards cleaned by rinsing with cold water (2), and mouthguards cleaned using an effervescent tablet (3), showing the peak volume of all proteins present (A), of amylase (B), and of lysozyme (C). Data are mean ± SD (n = 15). Level of significance: * ≤0.05, *** ≤0.001.

Figure 4

Amylase activity (in AU/mL) retrieved from mouthguards without cleaning (1), mouthguards rinsed with cold water (2), and mouthguards cleaned using an effervescent tablet (3). ns = not significant.