Sequential DLP Bioprinting of Dual-Layered Brain Organoid-Like Neural Microtissues Using Nanocomposite Bioinks

Area of Science:

• Biotechnology
• Neuroscience
• Materials Science

Background:

• Conventional methods for creating neural microtissues often lack structural control and reproducibility.
• Developing physiologically relevant models for neurodevelopment and disease is crucial.

Purpose of the Study:

• To present a detailed protocol for fabricating architecturally defined neural microtissues using digital light processing (DLP) bioprinting.
• To demonstrate a novel nanocomposite bioink for enhanced neural tissue engineering.

Main Methods:

• Utilized a sequential DLP bioprinting strategy with a layer-by-layer vat-switching technique.
• Formulated a nanocomposite bioink comprising gelatin methacryloyl (GelMA), reduced graphene oxide (rGO), and bacterial cellulose.
• Optimized DLP printing parameters and conducted long-term static culture and functional validation.

Main Results:

• Successfully fabricated complex, heterogeneous neural microtissues with defined architecture.
• The nanocomposite bioink exhibited enhanced electrical conductivity, printability, and mechanical stability.
• The fabricated microtissues supported neural network maturation and functional development.

Conclusions:

• The DLP bioprinting protocol offers improved reproducibility and structural control compared to conventional methods.
• This approach provides a more physiologically relevant platform for modeling neurodevelopment and neurological diseases.
• The developed technique advances the field of neural tissue engineering and brain organoid research.