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Synthetic Microbial Surrogates Consisting of Lipid Nanoparticles Encapsulating DNA for the Validation of Surface Disinfection Procedures.

ABSTRACT: Effective cleaning and disinfection procedures are an integral part of good manufacturing practice and in maintaining hygiene standards in health-care facilities. In this study, a method to validate such cleaning and disinfection procedures of surfaces was established employing lipid nanoparticles (LNPs) encapsulating DNA. It was possible to determine and distinguish between the physical cleaning effect (dilution) and the chemical cleaning effect (disintegration) on the LNPs during the cleaning and disinfection procedure (wiping). After treatment with 70 v % ethanol as a disinfectant and SDS solution as a cleaning agent, LNPs showed log₁₀ reductions of 4.5 and 4.0, respectively. These values are similar to the log₁₀ reductions exhibited by common bacteria, such as Escherichia coli and Serratia marcescens. Therefore, LNPs pose as useful tools for cleaning validation with advantages over the already existing tools and enable a separate detection of dilution and chemical disinfectant action.

SUBMITTER: Pfuderer L

PROVIDER: S-EPMC10031560 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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Synthetic Microbial Surrogates Consisting of Lipid Nanoparticles Encapsulating DNA for the Validation of Surface Disinfection Procedures.

Pfuderer Lara L Stark Wendelin J WJ Grass Robert N RN

ACS applied bio materials 20230228 3

Effective cleaning and disinfection procedures are an integral part of good manufacturing practice and in maintaining hygiene standards in health-care facilities. In this study, a method to validate such cleaning and disinfection procedures of surfaces was established employing lipid nanoparticles (LNPs) encapsulating DNA. It was possible to determine and distinguish between the physical cleaning effect (dilution) and the chemical cleaning effect (disintegration) on the LNPs during the cleaning ...[more]

PMID: 36854082

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Project description:BackgroundParo and other robot animals can improve wellbeing for older adults and people with dementia, through reducing depression, agitation and medication use. However, nursing and care staff we contacted expressed infection control concerns. Little related research has been published. We assessed (i) how microbiologically contaminated robot animals become during use by older people within a care home and (ii) efficacy of a cleaning procedure.MethodsThis study had two stages. In stage one we assessed microbial load on eight robot animals after interaction with four care home residents, and again following cleaning by a researcher. Robot animals provided a range of shell-types, including fur, soft plastic, and solid plastic. Stage two involved a similar process with two robot animals, but a care staff member conducted cleaning. The cleaning process involved spraying with anti-bacterial product, brushing fur-type shells, followed by vigorous top-to-tail cleaning with anti-bacterial wipes on all shell types. Two samples were taken from each of eight robots in stage one and two robots in stage two (20 samples total). Samples were collected using contact plate stamping and evaluated using aerobic colony count and identification (gram stain, colony morphology, coagulase agglutination). Colony counts were measured by colony forming units per square centimetre (CFU/cm2).ResultsMost robots acquired microbial loads well above an acceptable threshold of 2.5 CFU/cm2 following use. The bacteria identified were micrococcus species, coagulase negative staphylococcus, diptheriods, aerobic spore bearers, and staphylococcus aureus, all of which carry risk for human health. For all devices the CFU/cm2 reduced to well within accepted limits following cleaning by both researcher and care staff member.ConclusionsCompanion robots will acquire significant levels of bacteria during normal use. The simple cleaning procedure detailed in this study reduced microbial load to acceptable levels in controlled experiments. Further work is needed in the field and to check the impact on the transmission of viruses.

Project description:Messenger RNA (mRNA) has recently emerged as a promising class of nucleic acid therapy, with the potential to induce protein production to treat and prevent a range of diseases. However, the widespread use of mRNA as a therapeutic requires safe and effective in vivo delivery technologies. Libraries of ionizable lipid nanoparticles (LNPs) have been designed to encapsulate mRNA, prevent its degradation, and mediate intracellular delivery. However, these LNPs are typically characterized and screened in an in vitro setting, which may not fully replicate the biological barriers that they encounter in vivo. Here, we designed and evaluated a library of engineered LNPs containing barcoded mRNA (b-mRNA) to accelerate the screening of mRNA delivery platforms in vivo. These b-mRNA are similar in structure and function to regular mRNA, and contain barcodes that enable their delivery to be quantified via deep sequencing. Using a mini-library of b-mRNA LNPs formulated via microfluidic mixing, we show that these different formulations can be pooled together, administered intravenously into mice as a single pool, and their delivery to multiple organs (liver, spleen, brain, lung, heart, kidney, pancreas, and muscle) can be quantified simultaneously using deep sequencing. In the context of liver and spleen delivery, LNPs that exhibited high b-mRNA delivery also yielded high luciferase expression, indicating that this platform can identify lead LNP candidates as well as optimal formulation parameters for in vivo mRNA delivery. Interestingly, LNPs with identical formulation parameters that encapsulated different types of nucleic acid barcodes (b-mRNA versus a DNA barcode) altered in vivo delivery, suggesting that the structure of the barcoded nucleic acid affects LNP in vivo delivery. This platform, which enables direct barcoding and subsequent quantification of a functional mRNA, can accelerate the in vivo screening and design of LNPs for mRNA therapeutic applications such as CRISPR-Cas9 gene editing, mRNA vaccination, and other mRNA-based regenerative medicine and protein replacement therapies.

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Synthetic Microbial Surrogates Consisting of Lipid Nanoparticles Encapsulating DNA for the Validation of Surface Disinfection Procedures.

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Synthetic Microbial Surrogates Consisting of Lipid Nanoparticles Encapsulating DNA for the Validation of Surface Disinfection Procedures.

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