Isolation and characterization of pepsin solubilized type II collagen from porcine cartilage and cartilagious fish for intervertebral disc (IVD) regeneration

Abstract

Event Abstract Back to Event Isolation and characterization of pepsin solubilized type II collagen from porcine cartilage and cartilagious fish for intervertebral disc (IVD) regeneration Zhuning Wu1, 2*, Anne Maria Mullen3, Abhay Pandit2 and Dimitrios Zeugolis1, 2 1 National University of Ireland Galway (NUI Galway), Regenerative, Modular & Developmental Engineering Laboratory (REMODEL), Ireland 2 National University of Ireland Galway (NUI Galway), Centre for Research in Medical Devices (CÚRAM), Ireland 3 Food Research Centre, Teagasc, Ireland Introduction: Collagen is the main component of ECM with 29 types identified. Type II collagen presents supportive function in cartilaginous tissues. Since type II collagen is the major component of nucleus pulposus in intervertebral disc, the potential of injectable type II collagen hydrogel as a cell carrier for intervertebral disc regeneration is being investigated in this project. At this stage, we isolated and characterised type II collagen from porcine cartilage nad cartilaginous fish. Type II collagen was cross-linked with 4arm and 8arm StarPEG. We hypothesise that pure type II collagen can be obtained from cartilaginous tissue and 8arm StarPEG crosslinking will increase the mechanical, biological and biomedical properties of collagen hydrogel. Materials and Methods: Tyep II collagen was extracted from porcine trachea, ear, articular cartilage and cartilaginous fish through acid-pepsin digestion at 4ºC. SDS-PAGE was used to reveal the number and size of collagen chains. Thermal stability was tested by differential scanning calorimetry (DSC). Enzymatic degradation was assessed by collagenase assay. TNBSA assay will be carried out to test the free amine group in cross-linked collagen hydrogel. Human chondrocyte cell phenotype maintenance will be assessed with proteina and gene assays. Results: Conclusions: Pure type II collagen can be extracted from porcine cartilage and cartilaginous fish. Further biological assays will be carried out to characterise collagen enzymatic degradation and thermal stability. The authors would like to thank Teagasc, Irish Agriculture and Food Development Authority, Funding agency for providing financial support for this project.