Immobilization of vitronectin-binding heparan sulfates onto surfaces to support human pluripotent stem cells
Autor: | Drew M. Titmarsh, James C.H. Goh, Victor Nurcombe, Sadasivam Murali, Steve Oh, Andre J. van Wijnen, David Robinson, Jason D. Whittle, Gajadhar Bhakta, Lynn Yap, Yu Ming Lim, Robert D. Short, William R. Birch, Muriel Bardor, Lyn Chiin Sim, Allen Chen, Simon M. Cool, Andre Choo |
---|---|
Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Materials science biology Biomedical Engineering 02 engineering and technology Heparan sulfate 021001 nanoscience & nanotechnology Biomaterials Glycosaminoglycan Extracellular matrix 03 medical and health sciences chemistry.chemical_compound Surface coating 030104 developmental biology chemistry Biochemistry Biophysics biology.protein Vitronectin Stem cell Surface plasmon resonance 0210 nano-technology Induced pluripotent stem cell |
Zdroj: | Journal of Biomedical Materials Research Part B: Applied Biomaterials. 106:1887-1896 |
ISSN: | 1552-4973 |
Popis: | Functionalizing medical devices with polypeptides to enhance their performance has become important for improved clinical success. The extracellular matrix (ECM) adhesion protein vitronectin (VN) is an effective coating, although the chemistry used to attach VN often reduces its bioactivity. In vivo, VN binds the ECM in a sequence-dependent manner with heparan sulfate (HS) glycosaminoglycans. We reasoned therefore that sequence-based affinity chromatography could be used to isolate a VN-binding HS fraction (HS9) for use as a coating material to capture VN onto implant surfaces. Binding avidity and specificity of HS9 were confirmed by enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR)-based assays. Plasma polymerization of allylamine (AA) to tissue culture-treated polystyrene (TCPS) was then used to capture and present HS9 as determined by radiolabeling and ELISA. HS9-coated TCPS avidly bound VN, and this layered surface supported the robust attachment, expansion, and maintenance of human pluripotent stem cells. Compositional analysis demonstrated that 6-O- and N-sulfation, as well as lengths greater than three disaccharide units (dp6) are critical for VN binding to HS-coated surfaces. Importantly, HS9 coating reduced the threshold concentration of VN required to create an optimally bioactive surface for pluripotent stem cells. We conclude that affinity-purified heparan sugars are able to coat materials to efficiently bind adhesive factors for biomedical applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1887-1896, 2018. |
Databáze: | OpenAIRE |
Externí odkaz: |