Biomaterials functionalization by addition of biomolecules is an interesting approach for enhancing cell-biomaterial interactions and therefore improve their bioactivity. The global objective of this project is to enhance the bioactivity of biomaterials such as implantable devices and 3D scaffolds by adding extracellular matrix components and therefore facilitate the adhesion, growth and survival of human mesenchymal stem cells (hMSC) in biomaterials for tissue repair and tissue engineering applications.
In this project chondroitin sulfate (CS) and growth factors were studied for their effect on hMSC in biomaterials. First, the effect of these biomolecules was tested in solution. Then, two kinds of biomaterials were created: bioactive surfaces for enhancing bioactivity of implantable devices and bioactive hydrogels which can be used as 3D scaffolds for cell encapsulation and delivery.
A pro-survival effect of the growth factors studied in this project (epidermal growth factor, vascular endothelial growth factor and fibroblast growth factor) was not observed when tested in solution, therefore the project further focused on CS effect only. Interestingly, CS did not affect cell growth in media containing serum, while inducing cell detachment from substrate in serum free conditions.
For the bioactive surfaces construction, CS was grafted to either an amine-rich plasmapolymerized coating created on polyethylene terephthalate (PET) films (further referred as LP) or to commercial cell culture plates functionalized with amino groups. The bioactive surfaces were characterized by different techniques such as contact angle, atomic force microscopy, Orange II dye and Toluidine Blue O dye colorimetric assays (for amino group and CS quantification respectively) and finally, cell culture experiments (adhesion, growth and survival). Results confirmed the presence of CS grafted on both substrates. Commercial amine plates grafted almost five times more CS compared to LP. This rendered the surface antifouling for proteins and cells as confirmed by protein adsorption and cell culture assays. Cell culture assays on bioactive surfaces based on LP demonstrated improved cell adhesion and growth when compared to tissue culture plates or bare PET films in serum containing conditions.
Chitosan based hydrogels containing CS at a concentration of 500 μg/ml resulted in a cohesive hydrogel which supported hMSC viability up to 7 days. However increasing CS concentration to high level such as 10000 μg/ml led to decrease of cell viability after 4 or 7 days, probably due to lack of porosity and water since the hydrogel precipitates upon formation and expulses water.
In conclusion, this work demonstrated that CS immobilization can enhance the biological interactions of hMSC with biomaterials used for implantable devices such as PET. Further studies are needed to evaluate the possible effect of CS on hMSC differentiation and phenotype. Hydrogels with CS could be very interesting for tissue engineering applications such as cartilage formation.
| Date | 6 Jul 2016 |
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| Original language | American English |
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| Awarding Institution | - École de technologie supérieure
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| Supervisor | Sophie Lerouge (Supervisor) |
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De La Torre Torres, J. E. (Author),
Lerouge (Supervisor),
6 Jul 2016Student thesis: Master's thesis › Master in Engineering: Engineering