Infrared hyperspectral imaging of 3Dprinting with prostate cancer cells for tissue engineering applications

Abstract

Prostate cancer is the most common cancer among men. Leading to approximately 10,000deaths in the UK each year. Different models have been used to study this disease suchas animal models, 2D and 3Din vitromodels. These modelsall have their own limitations.The limitations of these models strongly compromise our understanding of important cellularmechanisms underpinning prostate cancer initiation and progression. This study uses tissueengineering approaches to design a 3D model capable of mimicking the tumour environment.Peptide hydrogels (alpha1, alpha2 and alpha2*) were used to engineer the ExtraCellularMatrix (ECM) in which prostate cancer cell lines (PC3, PNT2 and LNCaP) are encapsulated.The culture media were found to have a significant effect on the mechanical properties of thehydrogel; however, they have no impact on their viscosity. A number of biological tests areused to assess the encapsulated cells in which the viability of the cells and their ability toproliferate are assessed. The constructs built with alpha1 hydrogel were printed accurately,but the cells were not viable on it. Alpha2 was found to work as a better ECM for all thecell lines but accurate constructs shapes were impossible to be printed successfully. Alpha2*,on the other hand, was printed in more precise shapes than alpha2. The viability of PC3 andPNT2 cells was high on alpha2*; however, LNCaP cells shows a variation of their viabilitywhich confirmed by the metabolic activity test. The cell-laden constructs generated usingalpha2* hydrogel were successfully embedded in paraffin and cut. The sections were stainedwith H&E, and five different antibodies were used to check the protein expression of theseeded and printed cells. Seeded and printed cells were found to express the same proteins.FTIR imaging is used to analyse the printed constructs. The hydrogel consists of amino acids,and hence it has the same absorption peaks as the cells which complicates the separation ofthe cells from the hydrogel. Multivariate Curve Resolution (MCR) was used to separate thesignals of the cell, but further development of this methodology is required

Date of Award	1 Aug 2021
Original language	English
Awarding Institution	The University of Manchester
Supervisor	Peter Gardner (Supervisor) & Marco Domingos (Supervisor)