Human Platelet Lysate-Derived Nanofibrils as Building Blocks to Produce Free-Standing Membranes for Cell...

Area of Science:

• Biotechnology and Biomaterials Science
• Tissue Engineering and Regenerative Medicine
• Supramolecular Chemistry

Background:

• Amyloid-like fibrils are valuable in biotechnology as biomimetic platforms for controlling cell behavior.
• Fibrils are crucial for tissue structure, mechanical properties, and cell adhesion, necessitating scalable synthesis methods.

Purpose of the Study:

• To develop a method for instantaneous and bulk formation of amyloid-like nanofibrils from human platelet lysate (PL).
• To create free-standing nanofibril membranes for supporting 3D cell microtissue formation.
• To evaluate the potential of these membranes as bioactive platforms for emulating native tissues.

Main Methods:

• Instantaneous fibrillation of PL proteins using cholinium tosylate, an ionic liquid.
• Confirmation of protein conformational transition to cross-β-sheet-rich structures.
• Solvent casting of nanofibrils into thin, flexible, free-standing membranes for cell culture.

Main Results:

• PL proteins rapidly formed amyloid-like nanofibrils with cross-β-sheet structures.
• PL-derived nanofibril membranes exhibited nanotopographical roughness and stability for 14 days.
• Mesenchymal stem cells and tumor cells formed 3D spheroid-like microtissues on the membranes, showing improved viability and metabolic activity.

Conclusions:

• PL-derived nanofibril membranes serve as effective bioactive platforms for generating 3D cell-guided microtissues.
• These membranes can improve cell viability and metabolic activity within engineered microtissues.
• The developed strategy offers a bottom-up approach to emulate native tissue organization in a human microenvironment.