Dental caries and periodontal disease occur in almost 95% of the population. Although the use of probiotics to improve oral health is still in its infancy, many clinical trials are ongoing in this field.The aim of this study is to select the optimal conditions for quantification in the oral cavity of Lactobacillus brevis CD2, a probiotic strain contained in the commercially available dietary supplement Inersan® and to assess its persistency after oral administration. Different sets of primers for L. brevis detection have been tested in real-time PCR reactions to find optimal amplification efficiency and target specificity. Primers ability in the identification of L. brevis DNA was assessed in a pilot study conducted on 12 healthy volunteers clinically free of oral pathologies. The subjects took 3 tablets/day for 3 days and one tablet the 4th day. Clinical samples (dorsal surface of tongue, first molar, vestibular fornix and saliva) were collected at baseline (before beginning the trial), at T0 (in the morning before the assumption of the last tablet) and 3, 6, and 9 hours after taking the last tablet. The quantities of L. brevis DNA were compared to the total number of bacteria present in the samples. The activity of the enzyme arginine deiminase, responsible for oral pH homeostasis (particularly abundant in the CD2 strain) was also assessed in the saliva samples before and after treatment by HPLC measurements. Enzymatic analyses revealed that arginine deiminase activity was higher at T0 and at 3 hours than at 6-9 hours.We observed absence of tooth colonization by L. brevis at each time point, whereas its amount in the other oral sites was inversely correlated with elapsed time from the assumption. Persistence was maximal 3 hours after administration of tablets, but decreased more than one hundred times at 6 hours. However, the overnight recovery was significantly better than that obtained during the day, suggesting that probiotics administration before night could result in a better effectiveness.
DEVELOPMENT OF A REAL-TIME PCR ASSAY FOR THE ASSESSMENT OF PROBIOTIC Lactobacillus brevis CD2 STRAIN PERSISTENCE IN ORAL CAVITY / Mastromarino, Paola; F., Cacciotti; F., Tammaro; C., Nardis; Masci, Alessandra; Mosca, Luciana. - STAMPA. - (2012), p. 174. (Intervento presentato al convegno Anaerobe 2012 tenutosi a San Francisco, USA nel June 27- July 1, 2012).
DEVELOPMENT OF A REAL-TIME PCR ASSAY FOR THE ASSESSMENT OF PROBIOTIC Lactobacillus brevis CD2 STRAIN PERSISTENCE IN ORAL CAVITY
MASTROMARINO, Paola;MASCI, ALESSANDRA;MOSCA, Luciana
2012
Abstract
Dental caries and periodontal disease occur in almost 95% of the population. Although the use of probiotics to improve oral health is still in its infancy, many clinical trials are ongoing in this field.The aim of this study is to select the optimal conditions for quantification in the oral cavity of Lactobacillus brevis CD2, a probiotic strain contained in the commercially available dietary supplement Inersan® and to assess its persistency after oral administration. Different sets of primers for L. brevis detection have been tested in real-time PCR reactions to find optimal amplification efficiency and target specificity. Primers ability in the identification of L. brevis DNA was assessed in a pilot study conducted on 12 healthy volunteers clinically free of oral pathologies. The subjects took 3 tablets/day for 3 days and one tablet the 4th day. Clinical samples (dorsal surface of tongue, first molar, vestibular fornix and saliva) were collected at baseline (before beginning the trial), at T0 (in the morning before the assumption of the last tablet) and 3, 6, and 9 hours after taking the last tablet. The quantities of L. brevis DNA were compared to the total number of bacteria present in the samples. The activity of the enzyme arginine deiminase, responsible for oral pH homeostasis (particularly abundant in the CD2 strain) was also assessed in the saliva samples before and after treatment by HPLC measurements. Enzymatic analyses revealed that arginine deiminase activity was higher at T0 and at 3 hours than at 6-9 hours.We observed absence of tooth colonization by L. brevis at each time point, whereas its amount in the other oral sites was inversely correlated with elapsed time from the assumption. Persistence was maximal 3 hours after administration of tablets, but decreased more than one hundred times at 6 hours. However, the overnight recovery was significantly better than that obtained during the day, suggesting that probiotics administration before night could result in a better effectiveness.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
