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RESEARCH PRODUCT

Establishment of a pulmonary epithelial barrier on biodegradable poly-L-lactic-acid membranes

Vincenzo La CarrubbaSalvatore MontesantoSalvatore MontesantoNatalie P. SmithersFabio BucchieriFranco ConfortiValerio BrucatoDonna E. Davies

subject

PhysiologyCell MembranesCell Culture TechniquesBiocompatible Materials02 engineering and technologyEpitheliumTissue engineeringAnimal CellsAbsorbable ImplantsMaterials TestingElectric ImpedanceMedicine and Health SciencesLungTissue homeostasisBarrier functionStaining0303 health sciencesMultidisciplinaryTissue ScaffoldsTight junctionPolyethylene TerephthalatesChemistryQRCell Staining021001 nanoscience & nanotechnologyMembrane StainingElectrophysiologyMembranePhysical SciencesMedicineCytokinesBiological CulturesCellular Structures and OrganellesJunctional ComplexesCellular TypesAnatomy0210 nano-technologyResearch ArticleCell PhysiologySciencePolyestersMaterials ScienceMaterial PropertiesResearch and Analysis MethodsMembrane PotentialPermeabilityCell LineTight Junctions03 medical and health sciencesCell AdhesionHumans030304 developmental biologyBiochemistry Genetics and Molecular Biology (all)Tissue EngineeringBiology and Life SciencesEpithelial CellsMembranes ArtificialCell BiologyCell CulturesBiological TissueAgricultural and Biological Sciences (all)Specimen Preparation and TreatmentCell culturePermeability (electromagnetism)BiophysicsCytokine secretion

description

Development of biocompatible and functional scaffolds for tissue engineering is a major challenge, especially for development of polarised epithelia that are critical structures in tissue homeostasis. Different in vitro models of the lung epithelial barrier have been characterized using non-degradable polyethylene terephthalate membranes which limits their uses for tissue engineering. Although poly-L-lactic acid (PLLA) membranes are biodegradable, those prepared via conventional Diffusion Induced Phase Separation (DIPS) lack open-porous geometry and show limited permeability compromising their use for epithelial barrier studies. Here we used PLLA membranes prepared via a modification of the standard DIPS protocol to control the membrane surface morphology and permeability. These were bonded to cell culture inserts for use in barrier function studies. Pulmonary epithelial cells (H441) readily attached to the PLLA membranes and formed a confluent cell layer within two days. This was accompanied by a significant increase in trans-epithelial electrical resistance and correlated with the formation of tight junctions and vectorial cytokine secretion in response to TNFα. Our data suggest that a structurally polarized and functional epithelial barrier can be established on PLLA membranes produced via a non-standard DIPS protocol. Therefore, PLLA membranes have potential utility in lung tissue engineering applications requiring bio-absorbable membranes.

https://doi.org/10.1371/journal.pone.0210830