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A designer cell culture insert with a nanofibrous membrane toward engineering an epithelial tissue model validated by cellular nanomechanics

Kumar, P and Kedaria, D and Mahapatra, C and Mohandas, M and Chatterjee, K (2021) A designer cell culture insert with a nanofibrous membrane toward engineering an epithelial tissue model validated by cellular nanomechanics. In: Nanoscale Advances, 3 (16). pp. 4714-4725.

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Official URL: https://doi.org/10.1039/d1na00280e


Engineered platforms for culturing cells of the skin and other epithelial tissues are useful for the regeneration and development ofin vitrotissue models used in drug screening. Recapitulating the biomechanical behavior of the cells is one of the important hallmarks of successful tissue generation on these platforms. The biomechanical behavior of cells profoundly affects the physiological functions of the generated tissue. In this work, a designer nanofibrous cell culture insert (NCCI) device was developed, consisting of a free-hanging polymeric nanofibrous membrane. The free-hanging nanofibrous membrane has a well-tailored architecture, stiffness, and topography to better mimic the extracellular matrix of any soft tissue than conventional, flat tissue culture polystyrene (TCPS) surfaces. Human keratinocytes (HaCaT cells) cultured on the designer NCCIs exhibited a 3D tissue-like phenotype compared to the cells cultured on TCPS. Furthermore, the biomechanical characterization by bio-atomic force microscopy (Bio-AFM) revealed a markedly altered cellular morphology and stiffness of the cellular cytoplasm, nucleus, and cell-cell junctions. The nuclear and cytoplasmic moduli were reduced, while the stiffness of the cellular junctions was enhanced on the NCCI compared to cells on TCPS, which are indicative of the fluidic state and migratory phenotype on the NCCI. These observations were corroborated by immunostaining, which revealed enhanced cell-cell contact along with a higher expression of junction proteins and enhanced migration in a wound-healing assay. Taken together, these results underscore the role of the novel designer NCCI device as anin vitroplatform for epithelial cells with several potential applications, including drug testing, disease modeling, and tissue regeneration. © The Royal Society of Chemistry 2021.

Item Type: Journal Article
Publication: Nanoscale Advances
Publisher: Royal Society of Chemistry
Additional Information: The copyright for this article belongs to Authors
Keywords: Atomic force microscopy; Biomechanics; Cell membranes; Cell proliferation; Diagnosis; Nanofibers; Stiffness; Stiffness matrix; Tissue; Tissue culture; Tissue regeneration; Topography, Biomechanical behavior; Cell-cell junctions; Cellular morphology; Extracellular matrices; Human keratinocytes; Nanofibrous membranes; Physiological functions; Wound healing assays, Cell engineering
Department/Centre: Division of Interdisciplinary Sciences > Centre for Biosystems Science and Engineering
Division of Mechanical Sciences > Materials Engineering (formerly Metallurgy)
Date Deposited: 25 Sep 2021 12:39
Last Modified: 25 Sep 2021 12:39
URI: http://eprints.iisc.ac.in/id/eprint/69794

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