Development of Acellular Hepatic Scaffolds Through a Low-Cost Gravity-Assisted Perfusion Decellularization Method

TL;DR

This study introduces a low-cost method for creating liver scaffolds using gravity instead of pumps, making it accessible for labs with limited resources.

Contribution

A novel gravity-assisted perfusion decellularization method that is cost-effective and suitable for low-resource settings.

Findings

Gravity-assisted perfusion achieved better cellular removal and ECM preservation than immersion methods.

Perfused samples had residual DNA levels below the accepted threshold for effective decellularization.

Histological analysis confirmed the absence of nuclei and preserved collagen architecture in perfused scaffolds.

Abstract

Background: Developing reliable and cost-effective decellularization methods is critical for advancing tissue engineering and regenerative medicine, particularly in regions with limited access to specialized perfusion systems. Methods: This study standardized a gravity-assisted perfusion protocol for rat liver decellularization, designed to operate without pumps or pressurized equipment. Adult Wistar rat livers were processed through a gravity-driven vascular flushing method and compared with a conventional immersion-based protocol. The resulting scaffolds were evaluated by macroscopic inspection, histological staining (Masson’s trichrome), and residual DNA quantification. Results: The gravity-assisted perfusion method achieved more efficient cellular removal and superior preservation of extracellular matrix (ECM) integrity compared with immersion. Residual DNA levels were 3.7 ng/mg in perfused samples, 209.47 ng/mg in immersed samples, and 331.97 ng/mg in controls, confirming a statistically significant reduction (p < 0.05). Only the perfused group met the accepted threshold for effective decellularization (<50 ng/mg dry tissue). Histological analysis corroborated these findings, showing the absence of nuclei and the preservation of collagen architecture characteristic of a structurally intact ECM. Conclusions: This low-cost, reproducible, and technically simple system enables the generation of high-quality acellular hepatic scaffolds without mechanical pumps. Its accessibility and scalability make it suitable for laboratories with limited infrastructure and educational settings. Moreover, this gravity-assisted approach provides a foundation for future recellularization and preclinical studies aimed at developing bioengineered liver constructs for regenerative and transplant applications.

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