Correlation between relative bacterial activity and lactate dehydrogenase gene expression of co-cultures in vitro

Abstract

Objectives: The present study aims at correlating the relative bacterial activity with the H⁺ concentration and the ldh expression of caries-associated bacteria in co-cultures.

Materials and methods: Well plates were prepared with BHI medium and cultures of Lactobacillus paracasei and Fusobacterium nucleatum. Bacterial growth at 37 °C was measured using a microplate-photometer before and after adding sucrose to the samples. Samples of co-cultures (n = 12) and single-species cultures (n = 3) were taken and pH was assessed. Real-time quantitative PCRs were applied targeting the 16S-gene, the 16S-rRNA, the ldh-gene, and the ldh-mRNA.

Results: For L. paracasei with sucrose, an increase in relative bacterial activity (62.8% ± 23.5% [mean, SE]) was observed, while F. nucleatum showed a clear decrease in relative bacterial activity (- 35.0% ± 9.6%). Simultaneously, the H⁺ concentration increased (1.15E-05 mol*l-1 ± 4.61E-07 mol*l-1). Consequently, a significant positive correlation was found between L. paracasei's relative bacterial activity and H⁺ concentration (Spearman rank correlation, r = 0.638; p = 0.002), while F. nucleatum exhibited a negative correlation (r = - 0.741; p ≤ 0.001). Furthermore L. paracasei with sucrose showed a moderate, but significant positive correlation between relative bacterial activity and ldh-expression (r = 0.307; p = 0.024).

Conclusions and clinical relevance: The relative bacterial activity after sucrose pulse showed a significant correlation not only to the acid production (H⁺ concentration) but also to ldh expression of L. paracasei. However, further research is required to confirm these findings in a mature biofilm in vivo.

Keywords: 16S rRNA; Biofilm; Dental caries/etiology; Fusobacteria; Lactobacilli; Relative bacterial activity; ldh-expression.