Transcriptome Analysis of Porphyromonas Gingivalis in Response to Inhibition with Lactobacillus Rhamnosus Atcc 7469 Cell Free Supernatant
Periodontal disease is a chronic disease that is majorly caused by polymicrobial biofilm formation at the periodontal pocket. The important pathogen of this disease is P. gingivalis where it is involved in biofilm formation and virulent activity on the gingival tissue. Probiotics are suggested as...
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| Summary: | Periodontal disease is a chronic disease that is majorly caused by polymicrobial
biofilm formation at the periodontal pocket. The important pathogen of this disease is P.
gingivalis where it is involved in biofilm formation and virulent activity on the gingival
tissue. Probiotics are suggested as an adjunct method to overcome this pathogen's
colonization and biofilm formation. The main objective of this study is to investigate the
molecular mechanism of antimicrobial and anti-biofilm properties of L. rhamnosus
ATCC 7469 cell-free supernatant (CFS) against periodontal pathogen, P. gingivalis. To
achieve the objective, disc diffusion assay, minimum inhibitory concentration (MIC) and
minimum bactericidal concentration (MBC) were carried out to determine the
antimicrobial properties while anti-biofilm assay were carried out with various
concentration of L. rhamnosus ATCC 7469 CFS to assess the anti-biofilm activity.To
identify the effect of L. rhamnosus ATCC 7469 on the gene expression of P. gingivalis,
Next Generation Sequencing (NGS) were employed followed by RT-qPCR to validate
the gene expressions.The antimicrobial screening revealed that L. rhamnosus ATCC
7469 CFS was able to inhibit P. gingivalis with the inhibition zone of 10.64 ± 0.44 mm
with the MIC and MBC value of 25% CFS concentration.. In addition, L. rhamnosus
ATCC 7469 CFS able to reduce biofilm formation by 84.94% at the highest
concentration of CFS as observed by the biofilm assay. The molecular investigation by
NGS showed that various genes and pathways related to cell survival, metabolism, and
virulent activity were differentially expressed. Then, the affected gene mfa1, kgp, and rgp
with the addition of one more biofilm related gene, FimA were validated by RT-qPCR
technique. The genes of interest expression were in agreement with the NGS outcomes.
In conclusion, probiotics L. rhamnosus ATCC 7469 CFS possess antimicrobial and antibiofilm
properties against P. gingivalis and the effects of exposure to L. rhamnosus
ATCC 7469 CFS are reflected in the gene expression of P. gingivalis |
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