Optical Sensing Applied to Bioengineering of Decellularized Livers.
Name: HELLYEZER VILELA DE MORAIS
Publication date: 17/04/2026
Examining board:
Name |
Role |
|---|---|
| BRENO VALENTIM NOGUEIRA | Presidente |
| MARIA JOSE PONTES | Examinador Externo |
| TEODIANO FREIRE BASTOS FILHO | Examinador Interno |
Summary: The global prevalence of liver diseases and the shortage of donor organs for transplantation have driven the search for alternative therapeutic strategies, highlighting tissue engineering as a promising solution. In this context, decellularization
and recellularization of hepatic Extracellular Matrix (ECM) have emerged as viable approaches for the development of functional bioartificial organs. However, ensuring the quality and integrity of the decellularized ECM is critical for clinical success,
requiring efficient and non-invasive monitoring methods. This study aimed to adapt and validate an optical sensing system for real-time monitoring of the decellularization process of murine livers, seeking to correlate optical patterns with process efficiency
and ECM preservation. To this end, mouse livers were subjected to a perfusion-based decellularization protocol, approved by the Animal Ethics Committee (CEUA) under protocol no. 57/2019. The optical sensing system, originally developed for cardiac
monitoring, was redesigned through the incorporation of a converging lens (derived from an optical microscope condenser) and the optimization of the dark chamber. The results demonstrated that the insertion of the condenser significantly increased the
sensitivity of the system, enabling the detection of variations in light transmission and the identification of macroscopic perfusion-related alterations, such as clot formation. Quantitative analysis demonstrated that the decellularization protocol using SDS (Sodium Dodecyl Sulfate) significantly reduced DNA content; however, it was insufficient to achieve the levels recommended in the literature. The introduction of an additional DNase step resulted in a marked reduction of residual DNA, approaching
the threshold of 50 ng/mg. These findings were corroborated by histological analyses, in which hematoxylin and eosin (HE) staining demonstrated the absence of nuclei in the DNase-treated group, while Picrosirius Red staining under polarized light revealed improved organization of collagen fibers in this group. Together, these results indicate that effective hepatic decellularization depends on the integration of chemical and enzymatic steps, with DNase playing a critical role in the removal of residual genetic material. The optical sensing system proved to be a promising tool for real-time monitoring of structural changes in the tissue; however, it should be combined with quantitative and histological methods for a comprehensive evaluation of the process. Future perspectives include the development of multimodal systems integrating optical monitoring with techniques capable of directly assessing molecular components, such as residual DNA, as well as the application of predictive models based on optical data. This study contributes to the optimization and standardization of hepatic decellularization protocols, with potential impact on the advancement of tissue engineering and the clinical feasibility of bioartificial organs.
