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(1) Background: Clear orthodontic aligners support the development of oral biofilms, which could lead to interferences with the oral microbiota already existing and the deterioration of oral health, with the development of dental caries, periodontal disease and even systemic infections. Therefore, preventive oral health care requires a cleaning and disinfection procedure for aligners. (2) Methods: A systematic review of the literature was conducted across four databases following the PRISMA guidelines up to May 2021, combining an electronic and a manual search. Prospective studies, including randomized controlled trials (RCTs), crossover studies (COSs) and controlled clinical trials (CCTs), published in the English language without time restrictions, evaluating the efficacy of cleaning and disinfection protocols for clear orthodontic aligners by comparing them with a placebo or a negative control, were included. The article selection, data extraction and risk of bias assessment were performed by two independent blinded review authors. In case of disagreement, a third author was solicited throughout the selection process. (3) Results: Among the 221 articles screened in the search process, 4 studies were included in the review, all designed as crossover studies (single arm without randomization with the same sequence of different cleaning and disinfection protocols for each participant). Different cleaning and disinfection methods were studied such as mechanical methods (brushing with toothpaste or vibration), chemical methods/pharmaceutical products (chlorhexidine antibacterial substance, anionic or cationic detergents or effervescent tablets) or combinations of both. (4) Conclusion: Although the determination of the most remarkable method of cleaning and disinfection was impossible because no direct comparison was conducted between all these methods, a multi-step protocol, including the combination of a mechanical and a chemical method, seems to be the most effective approach. Further research is needed to define the most preventive oral health care protocol. Registration: PROSPERO CRD 42021278498.

(1) Background: The use of removable orthodontic appliances, which is common in early ages, requires careful hygiene, as several different microorganisms are found on their surface during the orthodontic treatment. (2) Methods: Comprehensive electronic searches were conducted up to May 2021. Randomized controlled trials (RCTs) or controlled clinical trials (CCTs), prospective or retrospective, evaluating the efficacy of cleaning and disinfection methods for acrylic removable orthodontic appliances, redacted in the English language, were included. Three independent blinding review authors were involved in study selection, data extraction, and bias assessment. (3) Results: A total of 2491 records were screened and eight studies (six RCTs and two CCTs) fulfilled the inclusion criteria. Among the overall cleaning and disinfection methods described in the included studies, four categories could be defined: liquid antimicrobial agents, commercial tablet cleansers, natural plant extracts and incorporation of quaternary ammonium methacryloxy silicate, all of which demonstrated superior efficacy compared to the placebo/negative control. However, the different methods were not compared with each other. (4) Conclusions: Biofilm control on acrylic orthodontic removable appliances can be performed using the different cleaning and disinfection methods considered in the included studies. Further studies are needed to define the most effective technique. Registration: PROSPERO CRD 42021269297.