Abstract
Gingival re-epithelialization represents an essential phase of oral wound healing in which epithelial integrity is re-establish. We developed an automated high-throughput re-epithelialization kinetic model, using the gingival epithelial cell line Ca9–22. The model was employed to screen 39 lactic acid bacteria, predominantly including oral isolates, for their capacity to accelerate gingival re-epithelialization. This screen identified several strains of Streptococcus salivarius that stimulated re-epithelialization. Further analysis revealed that S. salivarius strain MS-oral-D6 significantly promoted re-epithelialization through a secreted proteinaceous compound and subsequent experiments identified a secreted serine protease as the most likely candidate to be involved in re-epithelialization stimulation. The identification of bacteria or their products that stimulate gingival wound repair may inspire novel strategies for the maintenance of oral health.
Highlights
IntroductionThe model was employed to screen 39 lactic acid bacteria, predominantly including oral isolates, for their capacity to accelerate gingival reepithelialization
The throughput and reproducibility of the scratch assay was improved in this study by using a 96-well system and the HTSScratcher, which consists of a 96 pin-array controlled by a counterforce mechanism that ensured that the downward movement of the array towards the 96-well plate was conducted smoothly and consistently
The data acquired through this pipeline was visualized using the dedicated software fetal calf serum (FCS) Express 4 Plus (De Novo Software, CA, USA) in which the number of cells infiltrating into the scratched area over time was determined for each well
Summary
The model was employed to screen 39 lactic acid bacteria, predominantly including oral isolates, for their capacity to accelerate gingival reepithelialization. This screen identified several strains of Streptococcus salivarius that stimulated reepithelialization. Probiotics have been reported to exert numerous beneficial effects on gut health, for example by antagonizing the growth of pathogenic bacteria[2], promoting host-microbe homeostasis through modulation of immunity[3, 4], alleviating symptoms of lactose intolerance[5], enhancing mucosal barrier function[6,7,8], and promoting intestinal epithelial survival and growth[9, 10]. Wounds that fail to proceed through the normal stages of wound repair in a timely and orchestrated manner result in chronic wounds, which are often colonized by bacteria that may contribute to the delayed or incomplete healing process www.nature.com/scientificreports/
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