| Autor: |
Ong HT; 1 Ear Science Institute Australia , Nedlands, Australia .; 2 School of Veterinary and Life Sciences, Murdoch University , Murdoch, Australia ., Redmond SL; 1 Ear Science Institute Australia , Nedlands, Australia .; 3 Ear Sciences Centre, University of Western Australia , Perth, Australia ., Marano RJ; 1 Ear Science Institute Australia , Nedlands, Australia .; 3 Ear Sciences Centre, University of Western Australia , Perth, Australia ., Atlas MD; 1 Ear Science Institute Australia , Nedlands, Australia .; 3 Ear Sciences Centre, University of Western Australia , Perth, Australia ., von Unge M; 4 Division of Surgery, Akershus University Hospital and University of Oslo , Oslo, Norway .; 5 Centre for Clinical Research Västerås, University of Uppsala , Uppsala, Sweden ., Aabel P; 4 Division of Surgery, Akershus University Hospital and University of Oslo , Oslo, Norway ., Dilley RJ; 1 Ear Science Institute Australia , Nedlands, Australia .; 3 Ear Sciences Centre, University of Western Australia , Perth, Australia .; 6 Centre for Cell Therapy and Regenerative Medicine, University of Western Australia , Perth, Australia . |
| Abstrakt: |
Stem cell therapies for tympanic membrane repair have shown initial experimental success using mesenchymal stem cells in rat models to promote healing; however, the mechanisms providing this benefit are not known. We investigated in vitro the paracrine effects of human adipose-derived stem cells (ADSCs) on wound healing mechanisms for human tympanic membrane-derived keratinocytes (hTM) and immortalized human keratinocytes (HaCaT). ADSC conditioned media (CM ADSC ) were assessed for paracrine activity on keratinocyte proliferation and migration, with hypoxic conditions for ADSC culture used to generate contrasting effects on cytokine gene expression. Keratinocytes cultured in CM ADSC showed a significant increase in cell number compared to serum-free cultures and further significant increases in hypoxic CM ADSC . Assessment of ADSC gene expression on a cytokine array showed a range of wound healing cytokines expressed and under stringent hypoxic and serum-free conditions was upregulated (VEGF A, MMP9, Tissue Factor, PAI-1) or downregulated (CXCL5, CCL7, TNF-α). Several of these may contribute to the activity of conditioned media on the keratinocytes with potential applications in TM perforation repair. VEGFA protein was confirmed by immunoassay to be increased in conditioned media. Together with gene regulation associated with hypoxia in ADSCs, this study has provided several strong leads for a stem cell-derived approach to TM wound healing. |
