On the way back from 3D to 2D: Chitosan promotes adhesion and development of neuronal networks onto culture supports

Lisa, Donatella Di; Muzzi, Lorenzo; Dellacasa, Elena; Frega, Monica; Martinoia, Sergio; Pastorino, Laura

doi:10.1016/j.carbpol.2022.120049

Cited by 6 publications

(4 citation statements)

References 49 publications

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“…In the field of neural engineering, natural hydrogels, having a low modulus of elasticity and high-water content, are ideal 3D matrices mimicking human brain ECM [56,57]. Based on previous work [17,35,38], the natural polysaccharide CHITO has demonstrated its bioactivity toward nervous cells, including both primary neurons and iNeurons, emerging thus as a suitable candidate for the development of 2D and 3D in vitro neuronal networks. Among all the properties of CHITO, one of the most interesting is its ability to form thermogels at physiological temperature and pH in the presence of a weak base such as β-GP.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, we have demonstrated that the polysaccharide chitosan (CHITO) by itself is able to support neuronal adhesion, growth and functional maturation in 2D cultures derived from both primary neurons and iNeurons co-cultured with astrocytes [35,36]. We also demonstrated that the same cell phenotypes seeded onto a granular CHITO scaffold can develop 3D highly interconnected and functional networks, up to 21 and 42 d in vitro respectively [17,37,38].…”

Section: Introductionmentioning

confidence: 95%

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Long-term in vitro culture of 3D brain tissue model based on chitosan thermogel

Lisa,

Muzzi,

Lagazzo

et al. 2023

Biofabrication

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Methods for studying brain function and disease heavily rely on in vivo animal models, ex-vivo tissue slices, and 2D cell culture platforms. These methods all have limitations that significantly impact the clinical translatability of results. Consequently, models able to better recapitulate some aspects of in vivo human brain are needed as additional preclinical tools. In this context, 3D hydrogel-based in vitro models of the brain are considered promising tools. To create a 3D brain-on-a-chip model, a hydrogel capable of sustaining neuronal maturation over extended culture periods is required. Among biopolymeric hydrogels, chitosan-β-glycerophosphate (CHITO-β-GP) thermogels have demonstrated their versatility and applicability in the biomedical field over the years. In this study, we investigated the ability of this thermogel to encapsulate neuronal cells and support the functional maturation of a 3D neuronal network in long-term cultures. To the best of our knowledge, we demonstrated for the first time that CHITO-β-GP thermogel possesses optimal characteristics for promoting neuronal growth and the development of an electrophysiologically functional neuronal network derived from both primary rat neurons and neurons differentiated from human induced pluripotent stem cells (h-iPSCs) co-cultured with astrocytes. Specifically, two different formulations were firstly characterized by rheological, mechanical and injectability tests. Primary nervous cells and neurons differentiated from h-iPSCs were embedded into the two thermogel formulations. The 3D cultures were then deeply characterized by immunocytochemistry, confocal microscopy, and electrophysiological recordings, employing both 2D and 3D micro-electrode arrays. The thermogels supported the long-term culture of neuronal networks for up to 100 d. In conclusion, CHITO-β-GP thermogels exhibit excellent mechanical properties, stability over time under culture conditions, and bioactivity toward nervous cells. Therefore, they are excellent candidates as artificial extracellular matrices in brain-on-a-chip models, with applications in neurodegenerative disease modeling, drug screening, and neurotoxicity evaluation.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Long-term in vitro culture of 3D brain tissue model based on chitosan thermogel

Lisa,

Muzzi,

Lagazzo

et al. 2023

Biofabrication

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“…The day before seeding, MEAs were assembled with donuts-shaped Poly-dimethyl-siloxane (PDMS) structures (internal and external diameters: 5 and 22 mm respectively) to confine the active electrodes area. MEAs were sterilized in oven at 120° for 2 h. The active area of MEAs were functionalized with 1% chitosan solution pre-sterilized by autoclaving it at 120°C and left in the incubator overnight at 37°C, ( Di Lisa et al, 2022 ). The coating solution was removed from the MEA which was then washed twice with water and left to dry under the laminar hood until seeding.…”

Section: Methodsmentioning

confidence: 99%

Electrophysiological and morphological modulation of neuronal-glial network by breast cancer and nontumorigenic mammary cell conditioned medium

Di Lisa,

Cortese,

Chiappalone

et al. 2024

Front. Bioeng. Biotechnol.

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Breast cancer is a significant global health concern, with the overexpression of human epidermal growth factor receptor 2 (HER2/ERBB2) being a driver oncogene in 20%–30% of cases. Indeed, HER2/ERBB2 plays a crucial role in regulating cell growth, differentiation, and survival via a complex signaling network. Overexpression of HER2/ERBB2 is associated with more aggressive behavior and increased risk of brain metastases, which remains a significant clinical challenge for treatment. Recent research has highlighted the role of breast cancer secretomes in promoting tumor progression, including excessive proliferation, immune invasion, and resistance to anti-cancer therapy, and their potential as cancer biomarkers. In this study, we investigated the impact of ERBB2+ breast cancer SKBR-3 cell line compared with MCF10-A mammary non-tumorigenic cell conditioned medium on the electrophysiological activity and morphology of neural networks derived from neurons differentiated from human induced pluripotent stem cells. Our findings provide evidence of active modulation of neuronal-glial networks by SKBR-3 and MCF10-A conditioned medium. These results provide insights into the complex interactions between breast cancer cells and the surrounding microenvironment. Further research is necessary to identify the specific factors within breast cancer conditioned medium that mediate these effects and to develop targeted therapies that disrupt this interaction.

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“…Recently, we demonstrated that the cationic polysaccharide chitosan is by itself able to sustain neuronal adhesion and the maturation of both 2D and 3D functional neuronal networks [23][24][25][26] . Moreover, we demonstrated its printability [27] .…”

Section: Introductionmentioning

confidence: 99%