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Innovative Parabiotics for oral health

Periodic Reporting for period 1 - INPAROL (Innovative Parabiotics for oral health)

Periodo di rendicontazione: 2021-05-10 al 2023-05-09

Oral diseases constitute an increased major public health burden. Alternative dental prevention and therapeutic approaches such as probiotics are gaining popularity, but partly due to these products being live bacteria their use has not gained significant popularity in dentistry. Parabiotics, or inactivated probiotics, in appropriate doses have been shown to induce health benefits in other medical fields but have only as yet been tested sparingly for dental use.

Parabiotics have been previously produced through the use of heat technologies, which can lead to significant breakdown of protein components of probiotics’ membrane components. INPAROL aimed to produce parabiotics through innovative ultrasound technologies from strains of probiotics aimed at improving oral health. Different protocols were tested to assess inactivation of probiotics and therefore parabiotic production. The produced products were tested to assess physical changes following treatment.

Testing of live or partially inactivated products comes with its own complex challenges. In light of this, apart from using traditionally used antimicrobial assays INPAROL also developed protocols adapted from food microbiology modelling techniques to test the produced altered probiotics and parabiotics’ antimicrobial effects against oral microorganisms.
INPAROL tested probiotic production from 2 types of probiotics, Lactobacillus rhamnosus and Lactobacillus retueri as well as their mixes. Viability after treatment was tested and all probiotic groups tested lost viability after 15 minutes of treatment confirming parabiotic formation to the bacterial cells after treatment. Imaging through the use of scanning electron microscopy was carried out to assess structural changes to the probiotic cells. Imaging showed progressive collapse of the bacterial membrane with increased treatment times. Lactobacillus rhamnosus products tested resulted in the greatest inhibition towards oral microorganisms, although inhibition decreased as treatment time increased. This led to investigations of liquid portion, known as the supernatant, as opposed to the cell portion post-treatment. The supernatant was then analysed and this investigation highlighted that the supernatant contained portions of the bacterial cells and therefore a parabiotic in itself. Modelling techniques were then adapted to test the supernatant-derived parabiotic effects on oral microorganisms growth.

Dissemination of the project was carried at different levels.
Disseminating the information to the public was carried out through:
-Facebook (https://www.facebook.com/INPAROL
) and Linkedin (https://www.linkedin.com/company/80963509/admin/) INPAROL project pages aiming to share any videos or articles generated from dissemination activities as well as general information on probiotics.
-INPAROL participated in a video highlighting research carried out at the Faculty of Health Sciences at the University of Malta. This video was screened during Science in the City 2021, a local European researchers night which was held as a hybrid event.
-INPAROL was invited to a radio interview (https://www.facebook.com/watch/?v=627429381630433) on Radju Malta , Malta's Public Service Broadcaster to talk about the project and the role of probiotics and their derivates as part of RadioMocha Malta, a weekly science program.
-An article on the Sunday Times of Malta was written on the role of the oral microbiome. This was printed, as well as published online (https://timesofmalta.com/articles/view/the-human-microbiome-the-good-the-bad-and-the-ugly.933538?fbclid=IwAR2RDT-_o1l4kcuOahh0fGjWsznTmr2vVuyBIvRGr99L1rqDAwXavA4wESA).

INPAROL’s researcher participated in online workshops held by another European consortium, Combating and Evaluation of Mixed Biofilms (CEMBO; cembo.eu) to raise awareness on MSCA Individual fellowships and the project itself through an invited presentation, while also updating the project and researcher’s current knowledge on current techniques that are being developed to test mixed biofilms. Dissemination to the scientific community was also carried out through participation at the Continental European Division of the International Association of Dental Research (IADR) meeting in Brussels (https://ced-iadr2021.com/) which had a session dedicated to probiotics; participation in the British Society of Periodontology meeting held in London (https://www.bsperio.org.uk/events/bsp-event-calendar/bsp-conference-2020) and submission of an abstract for the upcoming Pan European Region IADR Meeting in Marseille in September 2022 (https://per-iadr2020.com/). INPAROL also hosted a trainee from National and Kapodistrian University of Athens (Greece) through an ERASMUS undergraduate internship scheme to further train other scientists in probiotic testing and parabiotic generation.
INPAROL also reached out to industry leaders in the field of probiotics to assess their current views and potential collaboration for development of innovative probiotics.
The project’s results provided important information on such a niche field to other bodies involved in probiotics and their derivatives, including researchers, industry and the end user, the general public. To our knowledge this is the first time that ultrasound technology has been proposed as a method of producing parabiotics. This not only has potential impact in the field of oral health through the production of a wider range of treatments but also adaptability and translation to other fields in medicine, food science and beyond.
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