scholarly journals Real Time In Vivo Confocal Microscopic Analysis of the Enamel Remineralization by Casein Phosphopeptide-Amorphous Calcium Phosphate (CPP-ACP): A Clinical Proof-of-Concept Study

2020 ◽  
Vol 10 (12) ◽  
pp. 4155
Author(s):  
Maria Contaldo ◽  
Dario Di Stasio ◽  
Fedora della Vella ◽  
Dorina Lauritano ◽  
Rosario Serpico ◽  
...  

Enamel defects (EDs) are qualitative and/or quantitative disturbances of the dental surface. To date, the responsiveness to remineralizing treatments has been studied ex vivo, on dental sections from extracted teeth. The present research aims to establish if in vivo reflectance confocal laser scanning microscopy is able to visualize the changes in the enamel architecture on living teeth, before, during and after remineralizing treatments by casein phosphopeptide-amorphous calcium phosphate (CPP-ACP). As proof-of-concept study, 17 consecutive children affected by EDs were enrolled and 38 EDs were considered. A CPP-ACP mousse was applied twice a week for 6 weeks and clinical and microscopic images were collected before, during and after the treatment for evaluating the changes occurred. For in vivo microscopic imaging, a reflectance confocal laser scanning microscope (RCM) for in vivo use was adopted. In this study RCM was proven to be able to visualize in vivo and at microscopic resolution the changes occurred during the remineralizing processes without needing for dental extractions and histopathological procedures. This in vivo RCM capability could encourage its clinical application in monitoring responsiveness to enamel therapies.

2001 ◽  
Vol 10 (3) ◽  
pp. 329-342 ◽  
Author(s):  
Emmanouhl S. Tzanakakis ◽  
Chang-Chun Hsiao ◽  
Taku Matsushita ◽  
Rory P. Remmel ◽  
Wei-Shou Hu

Cytochrome P450 (CYP450) enzymes are essential for xenobiotic metabolism. Although CYP450s are found in many tissues, CYP2B1/2 are primarily expressed in the rat liver. The constitutive expression in vivo of CYP2B1/2 is low but it is induced in the presence of various drugs such as phenobarbital (PB). In this study, CYP2B1/2 activity in cultured hepatocytes was assessed in situ with the introduction of a fluorogenic sub-strate, pentoxyresorufin. The product of 7-pentoxyresorufin-O-dealkylation (PROD), which is catalyzed specifically by CYP2B1/2, was detected using confocal laser scanning microscopy (CLSM). Primary hepatocytes cultured as monolayers on collagen-coated surfaces exhibited background PROD activity and minimal PB inducibility after 4 days in culture. In contrast, rat hepatocytes organized in compacted aggregates, or spheroids, exhibited higher levels of PROD activity and retained their ability for PB induction. The results from the CLSM analysis were verified by RT-PCR and Western immunoblotting analysis. Furthermore, CLSM in conjunction with image processing techniques and three-dimensional reconstruction revealed the localization of enhanced PROD activity in the center of spheroids. The results support the use of CLSM as a powerful tool for investigating CYP2B1/2 activity in cultured rat hepatocytes.


1993 ◽  
Vol 104 (4) ◽  
pp. 1175-1185 ◽  
Author(s):  
P. Buchenau ◽  
H. Saumweber ◽  
D.J. Arndt-Jovin

The regulation of DNA topology by topoisomerase II from Drosophila melanogaster has been studied extensively by biochemical methods but little is known about its roles in vivo. We have performed experiments on the inhibition of topoisomerase II in living Drosophila blastoderm embryos. We show that the enzymatic activity can be specifically disrupted by microinjection of antitopoisomerase II antibodies as well as the epipodophyllotoxin VM26, a known inhibitor of topoisomerase II in vitro. By labeling the chromatin of live embryos with tetramethylrhodamine-coupled histones, the effects of inhibition on nuclear morphology and behaviour was followed in vivo using confocal laser scanning microscopy. Both the antibodies and the drug prevented or hindered the segregation of chromatin daughter sets at the anaphase stage of mitosis. In addition, high concentrations of inhibitor interfered with the condensation of chromatin and its proper arrangement into the metaphase plate. The observed effects yielded non-functional nuclei, which were drawn into the inner yolk mass of the embryo. Concurrently, undamaged nuclei surrounding the affected region underwent compensatory division, leading to the restoration of the nuclear population, and thereby demonstrating the regulative capacity of Drosophila blastoderm embryos.


PLoS ONE ◽  
2012 ◽  
Vol 7 (4) ◽  
pp. e34869 ◽  
Author(s):  
Carlos Fritzsche ◽  
Oliver Stachs ◽  
Martha Charlotte Holtfreter ◽  
Constanze Nohr-Łuczak ◽  
Rudolf Friedrich Guthoff ◽  
...  

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