Rat liver ECM incorporated into electrospun polycaprolactone scaffolds as a platform for hepatocyte culture
Thomas S R Bate1, William Shanahan1, Joseph P Casillo2
1School of Engineering, Institute for Bioengineering, University of Edinburgh, Edinburgh, UK.
Abstract:
Liver disease is expanding across the globe; however, health-care systems still lack approved pharmaceutical treatment strategies to mitigate potential liver failures. Organ transplantation is the only treatment for liver failure and with increasing cases of liver disease, transplant programs increasingly cannot provide timely transplant availability for all patients. The development of pharmaceutical mitigation strategies is clearly necessary and methods to improve drug development processes are considered vital for this purpose. Herein, we present a methodology for incorporating whole organ decellularised rat liver ECM (rLECM) into polycaprolactone (PCL) electrospun scaffolds with the aim of producing biologically relevant liver tissue models. rLECM PCL scaffolds have been produced with 5 w/w% and 10 w/w% rLECM:PCL and were analyzed by SEM imaging, tensile mechanical analyses and FTIR spectroscopy. The hepatocellular carcinoma cell line, HepG2, was cultured upon the scaffolds for 14 days and were analyzed through cell viability assay, DNA quantification, albumin quantification, immunohistochemistry, and RT-qPCR gene expression analysis. Results showed significant increases in proliferative activity of HepG2 on rLECM containing scaffolds alongside maintained key gene expression. This study confirms that rLECM can be utilized to modulate the bioactivity of electrospun PCL scaffolds and has the potential to produce electrospun scaffolds suitable for enhanced hepatocyte cultures and in-vitro liver tissue models.
Insights
Researchers developed new liver tissue models using decellularized rat liver ECM and PCL scaffolds. These enhanced scaffolds support hepatocyte growth, offering a promising alternative for liver disease research and drug development.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Regenerative Medicine
Background:
- Global rise in liver disease necessitates novel therapeutic strategies beyond transplantation.
- Current organ transplant programs face limitations in meeting patient demand.
- Development of pharmaceutical mitigation strategies and improved drug development processes are crucial.
Purpose of the Study:
- To develop biologically relevant liver tissue models using decellularized rat liver extracellular matrix (rLECM).
- To incorporate rLECM into polycaprolactone (PCL) electrospun scaffolds.
- To assess the suitability of these scaffolds for enhanced hepatocyte cultures and in-vitro liver models.
Main Methods:
- Fabrication of rLECM:PCL electrospun scaffolds at 5% and 10% rLECM concentrations.
- Characterization of scaffolds using SEM imaging, tensile mechanical analysis, and FTIR spectroscopy.
- Culture of HepG2 cells on scaffolds for 14 days, followed by analysis of cell viability, DNA, albumin, gene expression, and immunohistochemistry.
Main Results:
- Scaffolds incorporating rLECM demonstrated significantly increased HepG2 cell proliferative activity.
- Key gene expression related to liver function was maintained in HepG2 cells cultured on rLECM scaffolds.
- rLECM effectively modulated the bioactivity of PCL electrospun scaffolds.
Conclusions:
- Decellularized rat liver ECM can enhance the bioactivity of electrospun PCL scaffolds.
- These novel scaffolds show potential for creating advanced in-vitro liver tissue models.
- This approach could facilitate improved hepatocyte cultures and aid in liver disease research and drug development.
