The aim of the study was to establish a live/dead qPCR with propidium monoazide (PMA) that can quantitatively differentiate between viable/non-viable microorganisms in dental hard tissues. Human premolars (n = 88) were prepared with nickel–titanium instruments and incubated with E. faecalis (21 d). Subsequently, the bacteria in half of the teeth were devitalized by heat inactivation (100 °C, 2 h). The following parameters were tested: PMA concentrations at 0 µmol (control), 50 µmol, 100 µmol, and 200 µmol; PMA incubation times of 30 min and 60 min, and blue light treatment for 30 min and 60 min. The teeth were ground using a cryomill and the bacterial DNA was quantified using qPCR, ANOVA, and p = 0.05. The qPCR of the control group detected a similar number of avital 9.94 × 106 and vital 1.61 × 107 bacterial cells. The use of PMA inhibited the amplification of DNA from non-viable cells during qPCR. As a result, the best detection of avital bacteria was achieved with the following PMA parameters: (concentration, incubation time, blue light treatment) 200-30-30; 5.53 × 104 (avital) and 1.21 × 100.7 (vital). The live/dead qPCR method using PMA treatment is suitable for the differentiation and quantification of viable/non-viable microorganisms in dentin, as well as to evaluate the effectiveness of different preparation procedures and antimicrobial irrigants in other biological hard substances.
Models of artificial root canals are used in several fields of endodontic investigations and pre-clinical endodontic training. They allow the physical testing of dental treatments, the operating of instruments used and the interaction between these instruments and the tissues. Currently, a large number of different artificial root canal models exist whose geometry is created either on the basis of selected natural root canal systems or to represent individual geometrical properties. Currently, only a few geometric properties such as the root canal curvature or the endodontic working width are taken into consideration when generating these models. To improve the representational capability of the artificial root canal models, the aim of the current study is therefore to generate an artificial root canal based on the statistical evaluation of selected natural root canals. Here, the approach introduced by Kucher for determining the geometry of a root canal model is used, which is based on the measurement and statistical evaluation of the root canal center line’s curvatures and their cross-sectional dimensions. Using the example of unbranched distal root canals of mandibular molars ( = 29), an artificial root canal model representing the mean length, curvature, torsion and cross-sectional dimensions of these teeth could be derived.
Microplastics - defined as tiny polymer fragments less than 5 mm in size - pose a clear threat to a wide range of small organisms through ingestion, leading to starvation. In addition, there are many reports of their occurrence both in remote geographical areas and in the human body. Small and made of mostly inert materials, they pose a significant analytical challenge. While primary microplastics i.e., those manufactured intentionally, have defined characteristics, secondary microplastics resulting from the degradation of larger plastic debris are much less understood. Cryogenic milling has emerged as one of the best ways to generate realistic samples with potential for use as references in studies. Here, we use this technique to generate and study the shapes and properties of a wide range of consumer plastics, providing an overview of optical and analytical properties.