Patient-derived organoids (PDOs) as a novel in vitro model for neuroblastoma tumours.

BACKGROUND: Neuroblastoma (NB) is a paediatric tumour of the sympathetic nervous system. Half of all cases are defined high-risk with an overall survival less than 40% at 5 years from diagnosis. The lack of in vitro models able to recapitulate the intrinsic heterogeneity of primary NB tumours has hindered progress in understanding disease pathogenesis and therapy response. METHODS: Here we describe the establishment of 6 patient-derived organoids (PDOs) from cells of NB tumour biopsies capable of self-organising in a structure resembling the tissue of origin. RESULTS: PDOs recapitulate the histological architecture typical of the NB tumour. Moreover, PDOs expressed NB specific markers such as neural cell adhesion molecules, NB84 antigen, synaptophysin (SYP), chromogranin A (CHGA) and neural cell adhesion molecule NCAM (CD56). Analyses of whole genome genotyping array revealed that PDOs maintained patient-specific chromosomal aberrations such as MYCN amplification, deletion of 1p and gain of chromosome 17q. Furthermore, the PDOs showed stemness features and retained cellular heterogeneity reflecting the high heterogeneity of NB tumours. CONCLUSIONS: We were able to create a novel preclinical model for NB exhibiting self-renewal property and allowing to obtain a reservoir of NB patients' biological material useful for the study of NB molecular pathogenesis and to test drugs for personalised treatments.

Salivary pH level and bacterial plaque evaluation in orthodontic patients treated with Recaldent products.

Dental caries and resulting tooth decay can produce a multifactorial destructive process with a very high incidence. Cariogenic bacteria attack enamel with acids that produce subsurface lesions, thereby weakening the enamel and allowing bacterial progression into the dentin. The formation of dental decay, because of demineralization of the tooth structure, can be prevented or delayed by increasing the rate of the tooth's remineralization and replacement relative to the tooth's rate of demineralization. This rebuilding of enamel may be accelerated by the addition of amorphous calcium phosphate (ACP) with the aid of casein phosphopeptide (CPP) (Recaldent molecule). In this study, the role of CPP in stabilizing and releasing ACP on the tooth surface has been investigated to better understand its efficacy in the prevention of tooth demineralization in orthodontic patients. Twenty-five patients who wore fixed orthodontic appliances were enrolled in this clinical trial. It was explained to the patients that CPP-ACP would be used for 3 weeks and then suspended for an additional 3 weeks. Salivary pH evaluation, plaque pH evaluation and oral hygiene index (OHI) were performed at T0, T1 and T2. Results showed an increase in OHI level and an increase of the salivary pH (76% of the patients). Instead of plaque pH level that showed trivial results, only 48% of the patients showed a bacterial plaque pH increase. In conclusion, this study has not provided unequivocal evidence for the protective properties of Recaldent molecule. Long-term studies are necessary to better understand the role of this molecule.