Biological effects of polystyrene micro- and nano-plastics on human intestinal organoid-derived epithelial tissue models without and with M cells

Chen, Ying; Williams, Ashleigh  M.; Gordon, Edward; Rudolph, Sara; Longo, Brooke N.; Li, Gang; Kaplan, David L.

doi:10.1016/j.nano.2023.102680

Cited by 19 publications

(14 citation statements)

References 56 publications

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“…THP-1 cells were prepared for SEM imaging by fixation and dehydration. , THP-1 cells were seeded on transwells with a pore size of 8 μm at a density of 10 5 cells cm –2 and fixed in 1% glutaraldehyde in 1× PBS for 1 h (hr) at room temperature. After fixation, the sample was gently rinsed once with the same fixed solution and twice with DI water.…”

Section: Materials and Methodsmentioning

confidence: 99%

Impact of Silk-Ionomer Encapsulation on Immune Cell Mechanical Properties and Viability

Kumarasinghe,

Hasturk,

Wang

et al. 2024

ACS Biomater. Sci. Eng.

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Encapsulation of single cells is a powerful technique used in various fields, such as regenerative medicine, drug delivery, tissue regeneration, cell-based therapies, and biotechnology. It offers a method to protect cells by providing cytocompatible coatings to strengthen cells against mechanical and environmental perturbations. Silk fibroin, derived from the silkworm Bombyx mori, is a promising protein biomaterial for cell encapsulation due to the cytocompatibility and capacity to maintain cell functionality. Here, THP-1 cells, a human leukemia monocytic cell line, were encapsulated with chemically modified silk polyelectrolytes through electrostatic layer-by-layer deposition. The effectiveness of the silk nanocoating was assessed using scanning electron microscopy (SEM) and confocal microscopy and on cell viability and proliferation by Alamar Blue assay and live/dead staining. An analysis of the mechanical properties of the encapsulated cells was conducted using atomic force microscopy nanoindentation to measure elasticity maps and cellular stiffness. After the cells were encapsulated in silk, an increase in their stiffness was observed. Based on this observation, we developed a mechanical predictive model to estimate the variations in stiffness in relation to the thickness of the coating. By tuning the cellular assembly and biomechanics, these encapsulations promote systems that protect cells during biomaterial deposition or processing in general.

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Section: Materials and Methodsmentioning

confidence: 99%

Impact of Silk-Ionomer Encapsulation on Immune Cell Mechanical Properties and Viability

Kumarasinghe,

Hasturk,

Wang

et al. 2024

ACS Biomater. Sci. Eng.

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“…Finally, SEM is used to trace the localization and adherence of PS MPs/NPs in both MDM-and SDM-differentiated monolayers (Fig. 3B), helping to elucidate their interactions with the intestinal epithelial cells (Chen et al, 2023). This…”

Section: Scanning Electron Microscopy: Sample Preparation and Imagingmentioning

confidence: 99%

“…This protocol outlines the cultivation and maintenance of human intestinal organoids and their subsequent dissociation to form monolayers based on previously established methods (Chen et al, 2023). Organoids derived from patient biopsies of small intestine (ileum) are cultivated in a 3D Matrigel matrix, nourished by conditioned growth medium and various small molecules.…”

Section: Human Intestinal Organoid Culture and Generation Of Intestin...mentioning

confidence: 99%

“…In light of their stem cell origin, organoids can be differentiated into monolayers composed of various cell types like enterocytes, goblet cells, enteroendocrine cells, Paneth cells, and M cells, mimicking in vivo conditions. In this article, based on our previous study (Chen et al, 2023), we outline the culture and maintenance of human intestinal organoids along with their dissociation and differentiation into monolayer cultures, with and without M cells. We also detail essential protocols regarding imaging techniques, and the functional evaluation of the monolayer cultures after exposure to polystyrene (PS) MPs/NPs.…”

Section: Introductionmentioning

confidence: 99%

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Impact of Micro‐ and Nano‐Plastics on Human Intestinal Organoid‐Derived Epithelium

Wang,

Iglesias‐Ledon,

Bishop

et al. 2024

Current Protocols

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The development of patient‐derived intestinal organoids represents an invaluable model for simulating the native human intestinal epithelium. These stem cell‐rich cultures outperform commonly used cell lines like Caco‐2 and HT29‐MTX in reflecting the cellular diversity of the native intestinal epithelium after differentiation. In our recent study examining the effects of polystyrene (PS), microplastics (MPs), and nanoplastics (NPs), widespread pollutants in our environment and food chain, on the human intestinal epithelium, these organoids have been instrumental in elucidating the absorption mechanisms and potential biological impacts of plastic particles. Building on previously established protocols in human intestinal organoid culture, we herein detail a streamlined protocol for the cultivation, differentiation, and generation of organoid‐derived monolayers. This protocol is tailored to generate monolayers incorporating microfold cells (M cells), key for intestinal particle uptake but often absent in current in vitro models. We provide validated protocols for the characterization of MPs/NPs via scanning electron microscopy (SEM) for detailed imaging and their introduction to intestinal epithelial monolayer cells via confocal immunostaining. Additionally, protocols to test the impacts of MP/NP exposure on the functions of the intestinal barrier using transendothelial electrical resistance (TEER) measurements and assessing inflammatory responses using cytokine profiling are detailed. Overall, our protocols enable the generation of human intestinal organoid monolayers, complete with the option of including or excluding M cells, offering crucial techniques for observing particle uptake and identifying inflammatory responses in intestinal epithelial cells to advance our knowledge of the potential effects of plastic pollution on human gut health. These approaches are also amendable to the study of other gut‐related chemical and biological exposures and physiological responses due to the robust nature of the systems. © 2024 Wiley Periodicals LLC.Basic Protocol 1: Human intestinal organoid culture and generation of monolayers with and without M cellsSupport Protocol 1: Culture of L‐WRN and production of WRN‐conditioned mediumSupport Protocol 2: Neuronal cell culture and integration into intestinal epitheliumSupport Protocol 3: Immune cell culture and integration into intestinal epitheliumBasic Protocol 2: Scanning electron microscopy: sample preparation and imagingBasic Protocol 3: Immunostaining and confocal imaging of MP/NP uptake in organoid‐derived monolayersBasic Protocol 4: Assessment of intestinal barrier function via TEER measurementsBasic Protocol 5: Cytokine profiling using ELISA post‐MP/NP exposure.

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“…Animals may get entangled in plastic objects dispersed in the environment and die. Microplastic fragments (small pieces of plastics, less than 5 mm in size) and nanoplastic particles (plastic particles with sizes in the nanometer range and, therefore, invisible) are already found everywhere in soil and in water bodies; nanoplastics are also found in drinking water, in food, and within living organisms. , They have harmful effects on many animal species, and the findings concerning animals suggest harmful effects also on human health . Figure S2 in the Supporting Information offers a basic outline of the major sources of plastic pollution and their fate and effects in the environment, thus also highlighting the variety of fates and effects and the resulting complexity of the ensuing overall problem.…”

Section: Green Chemistry Education and Students’ Outreach Initiativesmentioning

confidence: 99%

How Green Chemistry Education Can Empower Chemistry Students to Be Promoters of Sustainable Substance-Handling Practices in Their Communities

Mammino

2023

ACS Sustainable Chem. Eng.

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Pursuing sustainability entails a huge number of components. Some of them are related to everyday practices, i.e., to the behaviors of individual persons. The handling of substances and materials is one of the major aspects having a great impact on people's health and the environment and, thus, on sustainability. It becomes crucial that citizens in all communities acquire adequate awareness about behavior patterns that enhance safety, protect health, and maximize pollution prevention. Human beings tend to adopt certain behavior patterns more willingly if they are convinced of their importance than if they are simply instructed to implement them. Students taking chemistry courses and getting acquainted with green chemistry are in an ideal position to design the most suitable ways to disseminate knowledge about the appropriate uses and final disposal of substances and materials to their communities. The paper considers selected classes of substances and materials used in everyday life and requiring shifts to more sustainable uses and disposal, analyzes the main messages that need to be conveyed, and proposes explorations of chemistry students' active engagement as a valuable resource to promote sustainability in a convincing and considerate way.

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Biological effects of polystyrene micro- and nano-plastics on human intestinal organoid-derived epithelial tissue models without and with M cells

Cited by 19 publications

References 56 publications

Impact of Silk-Ionomer Encapsulation on Immune Cell Mechanical Properties and Viability

Impact of Silk-Ionomer Encapsulation on Immune Cell Mechanical Properties and Viability

Impact of Micro‐ and Nano‐Plastics on Human Intestinal Organoid‐Derived Epithelium

How Green Chemistry Education Can Empower Chemistry Students to Be Promoters of Sustainable Substance-Handling Practices in Their Communities

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