MALDI MSI analysis of lipid changes in living skin equivalents in response to emollient creams containing palmitoylethanolamide

Mitchell, Christopher A.; Donaldson, Michael; Francese, Simona; Clench, Malcolm R.

doi:10.1016/j.ymeth.2016.02.001

Cited by 17 publications

(15 citation statements)

References 17 publications

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“…who examined the absorption of the drug imipramine into a commercial 3D tissue model of the epidermis “Straticel.” Other studies of a similar type have been reported by Francese et al . and Mitchell et al . In the work of Francese et al .…”

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confidence: 77%

Mass Spectrometry Imaging of 3D Tissue Models

et al. 2018

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A 3D cell culture is an artificially created environment in which cells are permitted to grow/interact with their surroundings in all three dimensions. Derived from 3D cell culture, organoids are generally small-scale constructs of cells that are fabricated in the laboratory to serve as 3D representations of in vivo tissues and organs. Due to regulatory, economic and societal issues concerning the use of animals in scientific research, it seems clear that the use of 3D cell culture and organoids in for example early stage studies of drug efficacy and toxicity will increase. The combination of such 3D tissue models with mass spectrometry imaging provides a label-free methodology for the study of drug absorption/penetration, drug efficacy/toxicity, and drug biotransformation. In this article, some of the successes achieved to date and challenges to be overcome before this methodology is more widely adopted are discussed.

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confidence: 77%

Mass Spectrometry Imaging of 3D Tissue Models

et al. 2018

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“…This allows the direct sampling of lipid ions from the skin tissue, without prior extraction, so that lipids can be analysed in situ . While direct ionisation techniques such as MALDI and DESI can be used for homogenised samples, they have also proved useful in providing spatial distribution information through skin imaging analysis . A full‐thickness skin biopsy can be sectioned onto a glass slide, and by placing the sample on a moving stage, the mass spectrometer samples ions as it performs spot‐to‐spot analysis, or rasters, across the skin tissue, allowing reconstruction of an image of the full‐thickness skin representing ion intensities .…”

Section: Mass Spectrometry Lipid Imagingmentioning

confidence: 99%

“…By selecting an ion of interest and extracting the data for each pixel, a chemical map of the tissue can be built‐up, showing the distribution and abundance of that lipid molecular ion across the sample . This provides information on the different skin compartments in one image, revealing anatomical lipid distribution, as well as allowing semiquantitative comparison of lipid intensities between samples, and the MSI data can be coregistered with the same or a consecutive section that has undergone histochemical staining . The size of the pixels and the ions sampled depends on the ionisation approach, and MALDI and DESI offer different advantages.…”

Section: Mass Spectrometry Lipid Imagingmentioning

confidence: 99%

“…MALDI‐MSI involves coating the tissue with a matrix that promotes ionisation of the lipid species and has been used to study lipid changes in skin and skin equivalents . MALDI‐MSI can provide very good spatial resolution depending on the laser spot size on the surface, with small pixel sizes of 15 μm obtainable using commercial systems, but analysis requires special consideration of sample preparation, including matrix application, which depends on factors such as the target ions (recently reviewed in ref.…”

Section: Mass Spectrometry Lipid Imagingmentioning

confidence: 99%

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Lipidomics for translational skin research: A primer for the uninitiated

Kendall

Koszyczarek

Jones

et al. 2018

Experimental Dermatology

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Healthy skin depends on a unique lipid profile to form a barrier that confers protection and prevents excessive water loss, aids cell-cell communication and regulates cutaneous homoeostasis and inflammation. Alterations in the cutaneous lipid profile can have severe consequences for skin health and have been implicated in numerous inflammatory skin conditions. Thus, skin lipidomics is increasingly of interest, and recent developments in mass spectrometry-based analytical technologies can deliver in-depth investigation of cutaneous lipids, providing insight into their role and mechanism of action. The choice of tissue sampling technique and analytical approach depends on the location and chemistry of the lipid of interest. Lipidomics can be conducted by various mass spectrometry approaches, including different chromatography and ionisation techniques. Targeted mass spectrometry is a sensitive approach for measuring low-abundance signalling lipids, such as eicosanoids, endocannabinoids and ceramides. This approach requires specific extraction, chromatography and mass spectrometry protocols to quantitate the lipid targets. Untargeted mass spectrometry reveals global changes and allows analysis of hundreds of complex lipids across a range of lipid classes, including phospholipids, glycerophospholipids, cholesteryl esters and sphingolipids. Mass spectrometry lipid imaging, including matrix-assisted laser desorption ionisation mass spectrometry and desorption electrospray ionisation mass spectrometry, can reveal information about abundance and anatomical distribution of lipids within a single skin sample. Skin lipidomics can provide qualitative and quantitative data on hundreds of biologically relevant lipid species with different properties and activities, all found within a single skin sample, and support translational studies exploring the involvement of lipids in skin health and disease.

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“…Labskin comprises a dermal layer, consisting of primary fibroblasts embedded in a fibrin matrix, and an epidermal layer created by seeding keratinocytes on the dermis, and the ability to culture microbes on the model is proven 35 . Clench and co-workers have applied MALDI MS and MALDI MS imaging of transverse sections of Labskin for the analysis of lipids and small molecules [36][37][38][39][40][41] . Here, we demonstrate the analysis and identification of three ESKAPE pathogens, S. aureus, K. pneumoniae, and P. aeruginosa directly from wounded and infected Labskin by LESA MS. For S. aureus, two strains were considered, an MRSA reference strain and an MSSA clinical isolate.…”

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Direct identification of bacterial and human proteins from infected wounds in living 3D skin models

Havlíková

May

Styles

et al. 2020

Sci Rep

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Trauma is one of the leading causes of death in people under the age of 49 and complications due to wound infection are the primary cause of death in the first few days after injury. The ESKAPE pathogens are a group of bacteria that are a leading cause of hospital-acquired infections and a major concern in terms of antibiotic resistance. Here, we demonstrate a novel and highly accurate approach for the rapid identification of ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) directly from infected wounds in 3D in vitro skin models. Wounded skin models were inoculated with bacteria and left to incubate. Bacterial proteins were identified within minutes, directly from the wound, by liquid extraction surface analysis mass spectrometry. This approach was able to distinguish closely related strains and, unlike genomic approaches, can be modified to provide dynamic information about pathogen behaviour at the wound site. In addition, since human skin proteins were also identified, this method offers the opportunity to analyse both host and pathogen biomarkers during wound infection in near real-time. Trauma is one of the leading causes of death in people under the age of 49 1. Complications due to infection are the primary cause of death in patients who survive the first few days after a traumatic injury. In recent military conflicts, over half of the total injuries sustained were the result of trauma to the extremities 2. Over 25% of those patients suffered complications due to infection (rising to 50% for those requiring intensive care), either in the bones (osteomyelitis) or deep-wound infection 2,3. Similar rates of infection are associated with civilian trauma 2. Infectious complications result in significantly higher amputation rates, which are also of greater severity. The ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) are a group of opportunistic pathogens which account for most hospital-acquired infections, and their antibiotic resistance is rising 4,5. Rapid diagnosis and treatment of wound infection is therefore of utmost importance. Current diagnosis involves visual inspection of the wound for signs of infection (inflammation), followed by collection of swabs or tissues, microbial culture and identification in a clinical laboratory, which can take hours or even days. Mass spectrometry (MS) has emerged as a powerful technique for identification and classification of microorganisms. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS has been in development for that purpose since the late 1990s 6,7 and is now well-established in the clinic 8. In the analysis, patient-derived samples are cultured and individual colonies are smeared onto a MALDI target plate. A matrix is applied and the sample is subjected to MALDI-TOF MS. Identification is achieved by spectral fingerprinting...

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MALDI MSI analysis of lipid changes in living skin equivalents in response to emollient creams containing palmitoylethanolamide

Cited by 17 publications

References 17 publications

Mass Spectrometry Imaging of 3D Tissue Models

Mass Spectrometry Imaging of 3D Tissue Models

Lipidomics for translational skin research: A primer for the uninitiated

Direct identification of bacterial and human proteins from infected wounds in living 3D skin models

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