In vitro model of bacterial biofilm mineralization in complex humid environments: a proof of concept study.

IF 4.3 3区 工程技术 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Frontiers in Bioengineering and Biotechnology Pub Date : 2025-01-10 eCollection Date: 2024-01-01 DOI:10.3389/fbioe.2024.1496117
L Zorzetto, S Hammer, S Paris, C M Bidan
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Abstract

Background: Bacteria in physiological environments can generate mineralizing biofilms, which are associated with diseases like periodontitis or kidney stones. Modelling complex environments presents a challenge for the study of mineralization in biofilms. Here, we developed an experimental setup which could be applied to study the fundamental principles behind biofilm mineralization on rigid substrates, using a model organism and in a tailored bioreactor that mimics a humid environment. We developed a simple yet effective method to produce rigid specimens with the desired shape.

Materials and methods: To simulate humid growth conditions, rigid specimens were conditioned with human saliva, inoculated with the chosen model bacterial strain and placed in a chamber with continuous drop-wise supply of nutritious media. The preconditioning stage did not affect significantly the bacteria proliferation, but considering this option was instrumental to future evolutions of the model, where saliva could be substituted with other substances (e.g., urine, plasma or antimicrobial solutions). Two different growth media were used: a control medium with nutritious substances and a mineralizing medium consisting in control medium supplemented with mineral precursors. Both the specimen shape and the bioreactor designs resulted from an optimization process thoroughly documented in this work. As a proof of concept, we showed that it is possible to locate the bacteria and minerals using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM).

Results: We achieved an in vitro model representative of the conditions of growth and mineralization of biofilms in humid environments on a rigid substrate: something between the traditional solid-air and solid-liquid interface models.

Conclusion: Such model will be useful to understand fundamental mechanisms happening in complex environments.

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来源期刊
Frontiers in Bioengineering and Biotechnology
Frontiers in Bioengineering and Biotechnology Chemical Engineering-Bioengineering
CiteScore
8.30
自引率
5.30%
发文量
2270
审稿时长
12 weeks
期刊介绍: The translation of new discoveries in medicine to clinical routine has never been easy. During the second half of the last century, thanks to the progress in chemistry, biochemistry and pharmacology, we have seen the development and the application of a large number of drugs and devices aimed at the treatment of symptoms, blocking unwanted pathways and, in the case of infectious diseases, fighting the micro-organisms responsible. However, we are facing, today, a dramatic change in the therapeutic approach to pathologies and diseases. Indeed, the challenge of the present and the next decade is to fully restore the physiological status of the diseased organism and to completely regenerate tissue and organs when they are so seriously affected that treatments cannot be limited to the repression of symptoms or to the repair of damage. This is being made possible thanks to the major developments made in basic cell and molecular biology, including stem cell science, growth factor delivery, gene isolation and transfection, the advances in bioengineering and nanotechnology, including development of new biomaterials, biofabrication technologies and use of bioreactors, and the big improvements in diagnostic tools and imaging of cells, tissues and organs. In today`s world, an enhancement of communication between multidisciplinary experts, together with the promotion of joint projects and close collaborations among scientists, engineers, industry people, regulatory agencies and physicians are absolute requirements for the success of any attempt to develop and clinically apply a new biological therapy or an innovative device involving the collective use of biomaterials, cells and/or bioactive molecules. “Frontiers in Bioengineering and Biotechnology” aspires to be a forum for all people involved in the process by bridging the gap too often existing between a discovery in the basic sciences and its clinical application.
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