Biomimetics, Vol. 10, Pages 777: Development of Acellular Hepatic Scaffolds Through a Low-Cost Gravity-Assisted Perfusion Decellularization Method

Biomimetics doi: 10.3390/biomimetics10110777

Authors:
María Fernanda Duarte-Ortega
Luis Bernardo Enríquez-Sánchez
Manuel David Pérez-Ruiz
Alfredo Nevárez-Rascón
María Alejandra Favila-Pérez
Alva Rocío Castillo-González
Celia María Quiñonez-Flores
Luis Carlos Hinojos-Gallardo
Víctor Adolfo Ríos-Barrera
Carlos Arzate-Quintana

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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