Influence of salivary conditioning and sucrose concentration on biofilm-mediated enamel demineralization

Abstract The acquired pellicle formation is the first step in dental biofilm formation. It distinguishes dental biofilms from other biofilm types. Objective To explore the influence of salivary pellicle formation before biofilm formation on enamel demineralization. Methodology Saliva collection was approved by Indiana University IRB. Three donors provided wax-stimulated saliva as the microcosm bacterial inoculum source. Acquired pellicle was formed on bovine enamel samples. Two groups (0.5% and 1% sucrose-supplemented growth media) with three subgroups (surface conditioning using filtered/pasteurized saliva; filtered saliva; and deionized water (DIW)) were included (n=9/subgroup). Biofilm was then allowed to grow for 48 h using Brain Heart Infusion media supplemented with 5 g/l yeast extract, 1 mM CaCl2.2H2O, 5% vitamin K and hemin (v/v), and sucrose. Enamel samples were analyzed for Vickers surface microhardness change (VHNchange), and transverse microradiography measuring lesion depth (L) and mineral loss (∆Z). Data were analyzed using two-way ANOVA. Results The two-way interaction of sucrose concentration × surface conditioning was not significant for VHNchange (p=0.872), ∆Z (p=0.662) or L (p=0.436). Surface conditioning affected VHNchange (p=0.0079), while sucrose concentration impacted ∆Z (p<0.0001) and L (p<0.0001). Surface conditioning with filtered/pasteurized saliva resulted in the lowest VHNchange values for both sucrose concentrations. The differences between filtered/pasteurized subgroups and the two other surface conditionings were significant (filtered saliva p=0.006; DIW p=0.0075). Growing the biofilm in 1% sucrose resulted in lesions with higher ∆Z and L values when compared with 0.5% sucrose. The differences in ∆Z and L between sucrose concentration subgroups was significant, regardless of surface conditioning (both p<0.0001). Conclusion Within the study limitations, surface conditioning using human saliva does not influence biofilm-mediated enamel caries lesion formation as measured by transverse microradiography, while differences were observed using surface microhardness, indicating a complex interaction between pellicle proteins and biofilm-mediated demineralization of the enamel surface.

Influence of substratum position and acquired pellicle on Candida albicans biofilm

The purpose of this study was to evaluate the influence of the substratum position and the saliva acquired pellicle (AP) on Candida albicans biofilm development. Poly(methylmethacrylate) (PMMA) disks were fabricated and randomly allocated to experimental groups: HNP (disks placed in a horizontal position and uncoated by pellicle), VNP (disks placed in a vertical position and uncoated by pellicle), HCP (disks placed in a horizontal position and coated by pellicle), and VCP (disks placed in a vertical position and coated by pellicle). Disks were placed in a 24-well plate and a suspension of 107 cells/mL of Candida albicans was added to each well for biofilm development. The plates were aerobically incubated at 35°C. The biofilms were evaluated at 1.5 (adhesion time point), 24, 48, 72, and 96 hours. The number of viable cells was quantified in terms of the colony-forming units per milliliter (CFU/mL). Metabolic activity was measured by the XTT assay. The biofilm structure was analyzed by scanning electron microscopy. The data were analyzed by three-way ANOVA followed by Tukey's test, with significance set at 5%. The vertical groups showed less biofilm formation and lower metabolic activity than the horizontal groups (p< 0.05). Significant differences in cell viability and metabolic activity were observed between the adhesion and other time points (p< 0.05), but these variables were not affected by the presence of the pellicle (p > 0.05). It can be concluded that the substratum position influenced biofilm development.

Oral streptococci growth on aging and non-aging esthetic restorations after radiotherapy

The aim of this study was to examine Streptococcus mutans biofilm growth on both aged and non-aged restorative dental resins, which were submitted to therapeutic irradiation. Sixty-four disks of an esthetic restorative material (Filtek Supreme) were divided into 2 groups: aged group (AG) and a non-aged group (NAG). Each group was subdivided into 4 subgroups: non-irradiated and irradiated with 10Gy, 35Gy, and 70Gy. The biofilms were produced by Streptococcus mutans UA159 growing on both AG and NAG surfaces. The colony-forming units per mL (CFU/mL) were evaluated by the ANOVA and the Tukey LSD tests (α=0.05). AG presented smaller amounts of CFU/mL than the NAG before irradiation and after 10Gy of irradiation (p<0.05). AG irradiated with 35 and 70Gy showed increased amount of bacterial biofilm when compared to non-irradiated and 10Gy-irradiated disks (p<0.05). The exposure to ionizing radiation at therapeutic doses promoted changes in bacterial adherence of aged dental restorative material.

O objetivo deste estudo foi avaliar a formação do biofilme de Streptococcus mutans crescido em resina restauradora envelhecida e não-envelhecida, submetidas à radiação terapêutica. Sessenta e quatro discos do material restaurador Filtek Supreme foram divididos em 2 grupos: grupo envelhecido (AG) e grupo não-envelhecido (NAG) e cada grupo foi dividido em 4 sub-grupos: não-irradiado e irradiado com 10Gy, 35Gy e 70Gy. O biofilme de S. mutans UA159 foi produzido na superfície de ambos os discos AG e NAG. As unidades formadoras de colônia/mL (UFC/mL) foram avaliadas por ANOVA e teste de Tukey (α=0,05). O grupo AG demonstrou menores quantidades de UFC/mL que o grupo NAG antes da radiação e após a radiação de 10Gy (p<0,05). Os sub-grupos AG irradiados com 35 e 70Gy demonstraram aumento na quantidade de biofilme quando comparado aos não irradiados e irradiados com 10Gy (p<0,05). A exposição à radiação ionizante nas doses terapêuticas promoveu mudanças na aderência bacteriana no material restaurador.

Película adquirida salival: revisión de la literatura / Acquired salivary pellicle: a review of the literature

En este artículo de revisión se presenta y analiza la información actualizada disponible sobre la composición química, mecanismo de formación y factores que afectan la producción de la película adquirida salival. Asimismo se discuten aspectos vinculados con la función que cumple dicho integumento, en especial la relacionada con su desempeño como antecesor de la placa bacteriana de la cual dependen las afecciones de mayor prevalencia e incidencia en odontología, como son la caries dental y la enfermedad periodontal.

In this article is presented and analyzes the information brought up to date on the chemical composition, mechanism of formation and factors that affect the production of salivary acquired pellicle. Likewise aspects related to the function are discussed that complies said integument, especially it related to its performance as the ancestor of the bacterial plaque of which the affections of grater prevalence and incidence in dentistry they depend, like are the dental decay and the periodontal illness.