Spectrophotometric Analysis of Streptococcus mutans Growth and Biofilm Formation in Saliva and Histatin-5 Relate to pH and Viscosity

ABSTRACT Objective: To analyze the ability of saliva in controlling the growth and the biofilm formation of Streptococcus mutans (S. mutans) as well as the effect of histatin-5 anti-biofilm relate to pH and saliva viscosity. Material and Methods: The S. mutans biofilm assayed by crystal violet 1% and its growth measured by spectrophotometer. The saliva viscosity was analyzed by viscometer, and pH of saliva was measured by pH meter. Results: Based on the optical density values, growth of S. mutans in saliva ranged <300 CFU/mL (0.1 nm) at concentrations of 25%, 12.5% and 6.25% for 24 hours. Whereas at the 48 h and 72 h period of incubation shown an increase in growth of S. mutans ranged 300-600 CFU/mL (0.2-0.36 nm). The inhibitory biofilm formation of S. mutans in saliva was significantly higher at concentrations of 12.5% and 6.25% at 24 h incubation times on a moderate scale, whereas the histatin-5 was effective to inhibit S. mutans biofilm on the 50 and 25 ppm. The saliva possessed a higher inhibitory of biofilm S. mutans than histatin-5 and good level viscosity (0.91-0.92 cP). Conclusion: The saliva was able to control the growth of S. mutans, and histatin-5 can inhibit the biofilm formation S. mutans. Furthermore, the saliva was also able to respond to the pH change with good viscosity of saliva.

A preliminary study on the effect of histatin 5 inhibiting Porphyromonas gingivalis and Fusobacterium nucleatum co-aggregation / 中华口腔医学杂志

Objective@#To detect the inhibitory ability of histatin 5 on the auto-aggregation of Porphyromonas gingivalis (Pg), and the co-aggregation of Pg with Fusobacterium nucleatum (Fn); and to provide a theoretical basis for the role of oral innate immunity played in the inhibition of chronic periodontitis.@*Methods@#Saliva and supragingival, subgingival plaque samples were collected from 49 chronic periodontitis patients in School of Stomatology, China Medical University and 27 periodontal healthy individuals. Enzyme linked immunosorbent assay was used to assess the amount of histatin 5 in saliva, absolute quantitative real-time PCR (qPCR) was applied to detect the DNA copies of Fn, Pg and total bacteria in supragingival and subgingival plaque samples. The effects of histatin 5 on auto- and co-aggregation were assessed by bacterial adhesion test and scanning electron microscopy. Hemagglutinin gene, arginine-gingipains gene in Pg and FomA gene in Fn were tested by relative qPCR. Independent samples t-test was used to calculate the significance between the experimental group and the control group. P-value<0.05 was considered statistically significant.@*Results@#For chronic periodontitis patients, there was an inverse correlation between the concentration of histatin 5 and Fn and Pg in supragingival plaque samples (r=-0.379, r=-0.624). Similarly, an inverse correlation was also observed between the concentration of histatin 5 and subgingival Fn and Pg, respectively (r=-0.404, r=-0.314). As for periodontally healthy individuals, there was an inverse correlation between the concentration of histatin 5 and supragingival and subgingival Pg (r=-0.572, r=-0.533). Bacterial adhesion test and scanning electron microscopy certified that 25 mg/L histatin 5 inhibited the auto-aggregation of Pg-Pg and the co-aggregation of Pg-Fn. Results of qPCR showed that 25 mg/L histatin 5 up-regulated hemagglutinin gene by (14.52±3.25) fold and down-regulated FomA gene to (0.22±0.10) fold.@*Conclusions@#Histatin 5 could inhibit the auto-aggregation of Pg-Pg and the co-aggregation of Pg-Fn by regulating hemagglutinin gene and FomA gene expression.

Profound Study for Functions of Antimicrobial Peptides in Prevention of Oral Disease.

Antimicrobial peptides (AMPs) have a widespread distribution in human body and have antimicrobial activity against microorganisms with wide-range class of host-defense molecules. These are small cationic peptides that play an important role in the development of innate immunity with activity against gram-positive and negative bacteria, parasites, fungi and some viruses. In the oral cavity, the AMPs are produced by the salivary glands and the oral epithelium and serve as defensive purposes. At least forty-five identifiable antimicrobial gene products found in saliva are secreted from oral epithelial cells, salivary glands and neutrophils. AMPs also serve as effective biological molecules in immune response activation, inflammation and wound healing The aim of this review was to discuss the types and functions of oral AMPs and their role in combating microorganisms and infections in the oral cavity. AMPs have a promising potential to be used against oral microbes in order to control their growth and biofilm formation. There are many challenges that need to be overcome in order to design and synthesize AMPs that have the ability to with stand the unique and harsh oral environment. AMPs are expected in the future to be used as models for developing effective oral microbial antibiotics.