@article{047d478b20a147b7993117182a7b4841,
title = "Immobilization of vitronectin-binding heparan sulfates onto surfaces to support human pluripotent stem cells",
abstract = "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.",
keywords = "allylamine, biomaterial, extracellular matrix, glycosaminoglycan, implant, stem cells, surface coating",
author = "Lynn Yap and Sadasivam Murali and Gajadhar Bhakta and Titmarsh, {Drew M.} and Chen, {Allen Kuan Liang} and {Chiin Sim}, Lyn and Muriel Bardor and Lim, {Yu Ming} and Goh, {James C.H.} and Oh, {Steve K.W.} and Choo, {Andre B.H.} and {van Wijnen}, {Andre J.} and Robinson, {David E.} and Whittle, {Jason D.} and Birch, {William R.} and Short, {Robert D.} and Victor Nurcombe and Cool, {Simon M.}",
note = "Funding Information: Special thanks to Dr. N.R. Dunn, Principal Investigator, Institute of Medical Biology (IMB), Agency for Science, Technology and Research (A*STAR), Singapore, for assistance in the preparation of the manuscript, particularly for data concerning teratoma assays. We also thank Dr. J. Trott, IMB, A*STAR for insightful comments during the preparation of the manuscript. We also acknowledge grant support from the Joint Council Office, A*STAR and the Institute of Medical Biology, A*STAR. Thanks also to the National University of Singapore (NUS) Graduate School for Integrative Sciences and Engineering (NGS) for PhD scholarship support. The authors indicate no potential conflicts of interest. Funding Information: Special thanks to Dr. N.R. Dunn, Principal Investigator, Institute of Medical Biology (IMB), Agency for Science, Technology and Research (A*STAR), Singapore, for assistance in the preparation of the manuscript, particularly for data concerning teratoma assays. We also thank Dr. J. Trott, IMB, A*STAR for insightful comments during the preparation of the manuscript. We also acknowledge grant support from the Joint Council Office, A*STAR and the Institute of Medical Biology, A*STAR. Thanks also to the National University of Singapore (NUS) Graduate School for Integrative Sciences and Engineering (NGS) for PhD scholarship support. Publisher Copyright: {\textcopyright} 2017 Wiley Periodicals, Inc.",
year = "2018",
month = jul,
doi = "10.1002/jbm.b.33999",
language = "English (US)",
volume = "106",
pages = "1887--1896",
journal = "Journal of Biomedical Materials Research - Part B Applied Biomaterials",
issn = "1552-4973",
number = "5",
}