Semi-automated layer-by-layer biofabrication using rotational internal flow layer engineering technology

Area of Science:

• Biotechnology
• Tissue Engineering
• Regenerative Medicine

Background:

• Biofabrication aims to scale and improve reproducibility of human tissue production.
• Rotational Internal Flow Layer Engineering (RIFLE) creates microscale layered tubular constructs.
• Current RIFLE methods require manual application of cell-laden hydrogels.

Purpose of the Study:

• To automate key elements of the RIFLE biofabrication process, specifically liquid dispensing.
• To evaluate the performance of the automated RIFLE system using alginate and collagen hydrogels.
• To assess the impact of automation on cell viability and layer precision in engineered tissues.

Main Methods:

• Development and implementation of an automated liquid dispensing system for RIFLE.
• Utilizing alginate and collagen as cell-laden hydrogel materials.
• Comparing cell viability and layer formation between manual and semi-automated RIFLE processes.
• Characterizing the resolution of cell layering in engineered constructs.

Main Results:

• Semi-automated assembly achieved cell viabilities comparable to manual methods.
• Automated collagen hydrogel demonstrated high cell viability (>91%) over 10 days.
• The semi-automated RIFLE system precisely assembled cell populations into cell-width layers (≈14 µm).
• Automation reduced manual operations and potential for human error.

Conclusions:

• Automating the RIFLE process improves efficiency and reproducibility in biofabrication.
• Collagen hydrogels are advantageous for automated tissue engineering applications using RIFLE.
• Further automation, particularly in cell-hydrogel suspension preparation, is recommended for broader biofabrication advancement.