3D Bioprinted Neural‐Like Tissue as a Platform to Study Neurotropism of Mouse‐Adapted SARS‐CoV‐2

Abstract

The effects of neuroinvasion by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) become clinically relevant due to the numerous neurological symptoms observed in Corona Virus Disease 2019 (COVID‐19) patients during infection and post‐COVID syndrome or long COVID. This study reports the biofabrication of a 3D bioprinted neural‐like tissue as a proof‐of‐concept platform for a more representative study of SARS‐CoV‐2 brain infection. Bioink is optimized regarding its biophysical properties and is mixed with murine neural cells to construct a 3D model of COVID‐19 infection. Aiming to increase the specificity to murine cells, SARS‐CoV‐2 is mouse‐adapted (MA‐SARS‐CoV‐2) in vitro, in a protocol first reported here. MA‐SARS‐CoV‐2 reveals mutations located at the Orf1a and Orf3a domains and is evolutionarily closer to the original Wuhan SARS‐CoV‐2 strain than SARS‐CoV‐2 used for adaptation. Remarkably, MA‐SARS‐CoV‐2 shows high specificity to murine cells, which present distinct responses when cultured in 2D and 3D systems, regarding cell morphology, neuroinflammation, and virus titration. MA‐SARS‐CoV‐2 represents a valuable tool in studies using animal models, and the 3D neural‐like tissue serves as a powerful in vitro platform for modeling brain infection, contributing to the development of antivirals and new treatments for COVID‐19.

The Relevance of Astrocytic Cell Culture Models for Neuroinflammation in Neurodegeneration Research

“…Although 3D culture is promising for modeling neuroinflammation and neurodegeneration, there is still a lack of systematic studies that compare neuroinflammatory readouts in 3D cultures versus canonical 2D. For instance, SARS-CoV-2 infection and AD has been investigated in 3D cultures [214,215], but neuroinflammatory readouts are not explored in general.…”

Using nanomaterials to address SARS-CoV-2 variants through development of vaccines and therapeutics

“…Researchers have applied 3D bioprinting techniques in combination with tunable scaffold based, cell-laden, bioinks to produce 3D bioprinted neural tissue models to model neurotropism to mimic the in vivo extracellular matrix of neural tissues for enhanced infectious modelling of SARS-CoV-2.3D Bioprinting allows for precise placement of cells alongside components of its extracellular matrix to support the cells leading to better modelling of cellular behaviours in vitro. A key example of this development is exemplified by researchers de Melo et al, 2022, where they infected a 3D bioprinted neural-tissue like model consisting of murine astrocytes in a gelatin, GelMa, and fibrinogen-based bioink with mouse-adapted SARS-CoV-2. In observing their 3D bioprinted model, this paper validates the potential of 3D bioprinting techniques for disease modelling of infectious diseases especially in tissue areas that are challenging to study (de Melo et al, 2022).…”

“…A key example of this development is exemplified by researchers de Melo et al, 2022, where they infected a 3D bioprinted neural-tissue like model consisting of murine astrocytes in a gelatin, GelMa, and fibrinogen-based bioink with mouse-adapted SARS-CoV-2. In observing their 3D bioprinted model, this paper validates the potential of 3D bioprinting techniques for disease modelling of infectious diseases especially in tissue areas that are challenging to study (de Melo et al, 2022).…”

Disruptive 3D in vitro models for respiratory disease investigation: A state-of-the-art approach focused on SARS-CoV-2 infection

“…Although a few research studies have developed 3D models for SARS-CoV-2 infection investigation [20] , [21] , [22] , [23] , the tridimensional cultures focused on COVID-19 are still insufficient for thorough disease investigation and drug development. Besides that, no current 3D culture has ever correlated ACE2 expression with viral kinetics investigation nor precisely represented viral hematogenous dissemination through a pulmonary air-liquid interface [ALI] associated with endothelial cells.…”