Perfusion enhanced polydimethylsiloxane based scaffold cell culturing system for multi-well drug screening platform

Area of Science:

• Biomedical Engineering
• Hepatology
• Tissue Engineering

Background:

• Conventional 2D liver cell cultures lack in vivo architecture, compromising cell function and morphology.
• Limitations in nutrient/gas exchange and shear stress impact long-term hepatocyte viability in traditional models.

Purpose of the Study:

• To develop and evaluate a 3D polydimethylsiloxane (PDMS) scaffold integrated with a perfusion bioreactor for enhanced rat hepatocyte culture.
• To assess the scaffold-bioreactor system's ability to maintain liver-specific functions and cell morphology.
• To investigate the system's utility for preclinical drug testing, using acetaminophen as a model.

Main Methods:

• Fabrication of a 3D PDMS scaffold with interconnected macropores.
• Integration of the scaffold into a perfusion-enhanced bioreactor system.
• Culturing of rat hepatocytes for 9 days, assessing albumin/urea production and morphology via immunofluorescence.
• Drug response testing using N-Acetyl-Para-Amino-Phenol (acetaminophen).
• Numerical simulations to analyze oxygen distribution and fluid shear stress.

Main Results:

• The 3D PDMS scaffold in the perfusion bioreactor created a favorable microenvironment for hepatocytes.
• Enhanced cell-to-cell contact and maintenance of cuboidal morphology and liver-specific functions (albumin/urea production) were observed.
• Improved oxygen distribution and shielding from shear stress were confirmed by numerical simulations.
• The system effectively modeled acetaminophen's effects on liver cells.

Conclusions:

• The perfusion-enhanced PDMS scaffold-bioreactor system significantly improves hepatocyte culture conditions.
• This system offers a more physiologically relevant platform for long-term liver cell culture and preclinical drug toxicity testing.
• It holds promise for advancing hepatotoxicity testing and reducing reliance on animal models.