Alginate-Based 3D A549 Cell Culture Model to Study Paracoccidioides Infection

Abstract

A three-dimensional (3D) lung aggregate model based on sodium alginate scaffolds was developed to study the interactions between Paracoccidioides brasiliensis (Pb) and lung epithelial cells. The suitability of the 3D aggregate as an infection model was examined using cell viability (cytotoxicity), metabolic activity, and proliferation assays. Several studies exemplify the similarity between 3D cell cultures and living organisms, which can generate complementary data due to the greater complexity observed in these designed models, compared to 2D cell cultures. A 3D cell culture system of human A549 lung cell line plus sodium alginate was used to create the scaffolds that were infected with Pb18. Our results showed low cytotoxicity, evidence of increased cell density (indicative of cell proliferation), and the maintenance of cell viability for seven days. The confocal analysis revealed viable yeast within the 3D scaffold, as demonstrated in the solid BHI Agar medium cultivation. Moreover, when ECM proteins were added to the alginate scaffolds, the number of retrieved fungi was significantly higher. Our results highlight that this 3D model may be promising for in vitro studies of host–pathogen interactions.

Silver(i) complexes containing antifungal azoles: significant improvement of the anti-Candida potential of the azole drug after its coordination to the silver(i) ion

“…These cells were shown to be susceptible and permissive for infection, since they exhibit mucin overexpression and were used as promising models for studying in vitro infections using both bacteria and fungi. 56–58 The adherence of pathogenic fungi to human cells is considered as the initial step in the invasion of host cells. First, it was shown that the lowest MIC against fluorescently labelled C. albicans SC5314-RFP, used in the adherence assay, was determined for 3 (0.31 μM, data not shown), so this complex and the corresponding ctz ligand were selected for further analysis.…”

Alternative Non-Mammalian Animal and Cellular Methods for the Study of Host–Fungal Interactions

“…Thus, three-dimensional (3D) cell cultures are extensively employed for novel drug screening of antifungals. Several advantages are tied to obtaining more precise toxicity and efficacy results than in vivo models, along with host–pathogen interactions closely mimicking reality [ 25 , 26 ]. Recently, pulmonary spheroids have been employed to screen broad-spectrum antifungal drugs, yielding outcomes that indicate three-dimensional cell cultures are less susceptible to the toxic effects of drug treatment compared to monolayer cultures.…”