Abstract
Early childhood caries (ECC) is a severe manifestation of carious pathology with rapid and disruptive progression. The ECC microbiota includes a wide variety of bacterial species, among which is an anaerobic newly named species, Scardovia wiggsiae, a previously unidentified Bifidobacterium. Our aim was to provide the first ultrastructural characterization of S. wiggsiae and its biofilm by scanning electron microscopy (SEM) using a protocol that faithfully preserved the biofilm architecture and allowed an investigation at very high magnifications (order of nanometers) and with the appropriate resolution. To accomplish this task, we analyzed Streptococcus mutans’ biofilm by conventional SEM and VP-SEM protocols, in addition, we developed an original procedure, named OsO4-RR-TA-IL, which avoids dehydration, drying and sputter coating. This innovative protocol allowed high-resolution and high-magnification imaging (from 10000× to 35000×) in high-vacuum and high-voltage conditions. After comparing three methods, we chose OsO4-RR-TA-IL to investigate S. wiggsiae. It appeared as a fusiform elongated bacterium, without surface specialization, arranged in clusters and submerged in a rich biofilm matrix, which showed a well-developed micro-canalicular system. Our results provide the basis for the development of innovative strategies to quantify the effects of different treatments, in order to establish the best option to counteract ECC in pediatric patients.
Highlights
Childhood caries (ECC) or baby bottle syndrome [1] is a severe manifestation of carious pathology with rapid and disruptive progression, that affects children between birth and 71 months of age, as defined by the American Dental Association
S. mutans Prepared by Conventional scanning electron microscopy (SEM) Procedure
Biofilm has a typical structure b5aosfed16 on a delicate three-dimensional network; in order to avoid alteration of this network, we modified the d3r.yRinegsuplrtostocol by replacing critical point drying with drying in an ascending hexamethyldisilazane series, due to the extracellular polymeric substance (EPS) loss effect of the conventional critical point drying procedure [53]. 3.1
Summary
Childhood caries (ECC) or baby bottle syndrome [1] is a severe manifestation of carious pathology with rapid and disruptive progression, that affects children between birth and 71 months of age, as defined by the American Dental Association. The ECC main cause is the prolonged use of sugary substances (taken mainly at night, when the salivary flow is considerably reduced) combined with the presence of an oral complex microbiota. This latter is rich in acidogenic bacterial species [4], and they metabolize sugars producing lactic acid, which cause dental hard tissues demineralization. Present are only a few preliminary studies on S. wiggsiae, but no information from a microbiological and morphological point of view For this reason, our aim is to investigate the ability of S. wiggsiae to form a biofilm, and to provide, for the first time, an ultrastructural and morphological characterization by scanning electron microscopy
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