A miniaturized hydrogel-based in vitro model for dynamic culturing of human cells overexpressing beta-amyloid precursor protein

Abstract

Recent findings have highlighted an interconnection between intestinal microbiota and the brain, referred to as microbiota–gut–brain axis, and suggested that alterations in microbiota composition might affect brain functioning, also in Alzheimer’s disease. To investigate microbiota–gut–brain axis biochemical pathways, in this work we developed an innovative device to be used as modular unit in an engineered multi-organ-on-a-chip platform recapitulating in vitro the main players of the microbiota–gut–brain axis, and an innovative three-dimensional model of brain cells based on collagen/hyaluronic acid or collagen/poly(ethylene glycol) semi-interpenetrating polymer networks and β-amyloid precursor protein-Swedish mutant-expressing H4 cells, to simulate the pathological scenario of Alzheimer’s disease. We set up the culturing conditions, assessed cell response, scaled down the three-dimensional models to be hosted in the organ-on-a-chip device, and cultured them both in static and in dynamic conditions. The results suggest that the device and three-dimensional models are exploitable for advanced engineered models representing brain features also in Alzheimer’s disease scenario.

Self-assembling peptide gels promote angiogenesis and functional recovery after spinal cord injury in rats

“…53,54 In order for hydrogels to be useful for constructing in vitro 3D culture systems in various fields, their stiffness, adhesion properties, and porosity need to be controllable to allow the growth and interactions of various types of cells. 55,56 The stiffness of RADA16 can be sufficiently modified by the addition functional groups, which might be useful for the development of ideal 3D culture systems for the field of tissue regeneration.…”

3D brain tissue physiological model with co-cultured primary neurons and glial cells in hydrogels

“…In this perspective, developing an OOC-based 3D system modeling brain tissue is particularly challenging. 4 Starting from the scalability of the thickness of COLLbased semi-IPNs to the millimeter scale and their suitability for dynamic culture in a microfluidic, optically accessible OOC device, 5 we set up a novel 3D brain-like tissue model potentially suitable for dynamic culture by coupling 1.5 mm-thick COLL-based semi-IPNs to the coculture of cortical neurons and glial cells. Semi-IPNs have the advantage of easy tunability of their final properties, 35 a key point when aiming at co-culturing different cell types, including neural cells.…”

“…For the development of 3D models, we cultured brain cells (as single cultures or co-cultures) in semi-IPNs based on COLL and HA (M w =1・10 5 Da, Altergon Italia, Morra De Sanctis, Italy) or COLL and PEG. We used PEG with two molecular weights (M w ): 1.945 kDa (referred to as PEG 2000 ) and 3.270 kDa (referred to as PEG 3350 ).…”

“… 4 In the present work, we focused on the setting up of a millimeter-thick brain-like tissue model potentially suitable for dynamic culture in a recently developed optically accessible organ-on-a-chip (OOC) device. 5 …”

“…4 In the present work, we focused on the setting up of a millimeter-thick brain-like tissue model potentially suitable for dynamic culture in a recently developed optically accessible organ-on-a-chip (OOC) device. 5 In the last decade, OOC technology has seen considerable improvements for its potential to reduce animal studies for drug development and toxicity testing. 6 Its reliability and reproducibility offer substantial advantages for tissue 3D brain tissue physiological model with co-cultured primary neurons and glial cells in hydrogels and disease modeling, 7,8 overcoming the limitations of 2D static cultures.…”