Preservation of heparin-binding EGF-like growth factor activity on heparin-modified poly(-isopropylacrylamide)-grafted surfaces

A heparin-modified poly( N -isopropylacrylamide) (PIPAAm)-grafted surface bound with heparin-binding epidermal growth factor-like growth factor (HB-EGF) was able to culture hepatocytes maintaining high albumin secretion and high expression of hepatocyte-specific genes. However, the activity of HB-EG...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:RSC advances 2021-11, Vol.11 (59), p.37225-37232
Hauptverfasser: Kobayashi, Jun, Arisaka, Yoshinori, Yui, Nobuhiko, Yamato, Masayuki, Okano, Teruo
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 37232
container_issue 59
container_start_page 37225
container_title RSC advances
container_volume 11
creator Kobayashi, Jun
Arisaka, Yoshinori
Yui, Nobuhiko
Yamato, Masayuki
Okano, Teruo
description A heparin-modified poly( N -isopropylacrylamide) (PIPAAm)-grafted surface bound with heparin-binding epidermal growth factor-like growth factor (HB-EGF) was able to culture hepatocytes maintaining high albumin secretion and high expression of hepatocyte-specific genes. However, the activity of HB-EGF on the surface and its binding effects on hepatocytes remain unclear. In this study, we investigated the temperature-dependent interactions of HB-EGF and EGF receptor (EGFR) with heparin-modified PIPAAm to evaluate the activity of HB-EGF on the surface. Quartz crystal microbalance (QCM) measurements revealed that the amounts of adsorbed HB-EGF on either the heparin-modified PIPAAm-grafted surface (heparin-IC1) or PIPAAm-grafted surfaces were almost the same regardless of swelling/deswelling of grafted PIPAAm chains. The heparin-IC1 surface bound to HB-EGF at 37 °C had the ability to bind to hepatocytes through specific affinity interaction with EGFR, whose activation was confirmed by western blotting. However, the physisorbed HB-EGF on the PIPAAm surface greatly diminished its activity. Taken together, the introduction of heparin into grafted PIPAAm chains on the surface plays a pivotal role in holding HB-EGF while preserving its activity. Hydration and swelling of surface-grafted PIPAAm chains at 20 °C greatly diminished the attachment of hepatocytes with HB-EGF bound to heparin-IC1, whereas hepatocytes were able to bind to HB-EGF bound to heparin-IC1 at 37 °C. Thus, the equilibrated affinity interaction between EGFRs and surface-bound HB-EGF was considered to be attenuated by steric hindrance due to hydration and/or swelling of grafted PIPAAm chains. Activity of HB-EGF bound to a heparin-modified poly( N -isopropylacrylamide) (PIPAAm)-grafted surface was preserved through specific binding to heparin, whereas physisorbed HB-EGF on a PIPAAm-grafted surface greatly diminished its activity.
doi_str_mv 10.1039/d1ra07317f
format Article
fullrecord <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_proquest_miscellaneous_2658643685</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2658643685</sourcerecordid><originalsourceid>FETCH-LOGICAL-c494t-21902e8f6d23947d7e36e4adeabfa83da9248d6807cbcd724efbe48d5fb19c1e3</originalsourceid><addsrcrecordid>eNpdkk1v1DAQhi0EolXphTsoEpeCFPBX7PiCVJVuQaoEQnC2HHu865LEwU4W7Ym_jmHbpeDD2Jp55tVY7yD0lODXBDP1xpFksGRE-gfomGIuaoqFenjvfYROc77B5YiGUEEeoyPWcCU4Jsfo56cEGdLWzCGOVfTVBiaTwlh3YXRhXFeXV6u6D9-gWqf4Y95U3tg5pqrEsA3zripddy1DdMEHcNUU-91ZHXKcUpx2vbGphCE4eFmvk_FzQfKSihLkJ-iRN32G09v7BH1dXX65eF9ff7z6cHF-XVuu-FxTojCF1gtHmeLSSWACuHFgOm9a5oyivHWixdJ21knKwXdQMo3viLIE2Al6u9edlm4AZ2Gck-n1lMJg0k5HE_S_lTFs9DputcKcSUmKwNmtQIrfF8izHkK20PdmhLhkTUXTCs5E2xT0xX_oTVzSWL5XKEy5FK1oC_VqT9kUc07gD8MQrH9bq9-Rz-d_rF0V-Pn98Q_onZEFeLYHUraH6t_dYL8AjjusSA</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2602476868</pqid></control><display><type>article</type><title>Preservation of heparin-binding EGF-like growth factor activity on heparin-modified poly(-isopropylacrylamide)-grafted surfaces</title><source>DOAJ Directory of Open Access Journals</source><source>PubMed Central Open Access</source><source>EZB-FREE-00999 freely available EZB journals</source><source>PubMed Central</source><creator>Kobayashi, Jun ; Arisaka, Yoshinori ; Yui, Nobuhiko ; Yamato, Masayuki ; Okano, Teruo</creator><creatorcontrib>Kobayashi, Jun ; Arisaka, Yoshinori ; Yui, Nobuhiko ; Yamato, Masayuki ; Okano, Teruo</creatorcontrib><description>A heparin-modified poly( N -isopropylacrylamide) (PIPAAm)-grafted surface bound with heparin-binding epidermal growth factor-like growth factor (HB-EGF) was able to culture hepatocytes maintaining high albumin secretion and high expression of hepatocyte-specific genes. However, the activity of HB-EGF on the surface and its binding effects on hepatocytes remain unclear. In this study, we investigated the temperature-dependent interactions of HB-EGF and EGF receptor (EGFR) with heparin-modified PIPAAm to evaluate the activity of HB-EGF on the surface. Quartz crystal microbalance (QCM) measurements revealed that the amounts of adsorbed HB-EGF on either the heparin-modified PIPAAm-grafted surface (heparin-IC1) or PIPAAm-grafted surfaces were almost the same regardless of swelling/deswelling of grafted PIPAAm chains. The heparin-IC1 surface bound to HB-EGF at 37 °C had the ability to bind to hepatocytes through specific affinity interaction with EGFR, whose activation was confirmed by western blotting. However, the physisorbed HB-EGF on the PIPAAm surface greatly diminished its activity. Taken together, the introduction of heparin into grafted PIPAAm chains on the surface plays a pivotal role in holding HB-EGF while preserving its activity. Hydration and swelling of surface-grafted PIPAAm chains at 20 °C greatly diminished the attachment of hepatocytes with HB-EGF bound to heparin-IC1, whereas hepatocytes were able to bind to HB-EGF bound to heparin-IC1 at 37 °C. Thus, the equilibrated affinity interaction between EGFRs and surface-bound HB-EGF was considered to be attenuated by steric hindrance due to hydration and/or swelling of grafted PIPAAm chains. Activity of HB-EGF bound to a heparin-modified poly( N -isopropylacrylamide) (PIPAAm)-grafted surface was preserved through specific binding to heparin, whereas physisorbed HB-EGF on a PIPAAm-grafted surface greatly diminished its activity.</description><identifier>ISSN: 2046-2069</identifier><identifier>EISSN: 2046-2069</identifier><identifier>DOI: 10.1039/d1ra07317f</identifier><identifier>PMID: 35496401</identifier><language>eng</language><publisher>England: Royal Society of Chemistry</publisher><subject>Affinity ; Albumins ; Binding ; Chains ; Chemistry ; Epidermal growth factor ; Gene expression ; Grafting ; Growth factors ; Heparin ; Hydration ; Microbalances ; Polyisopropyl acrylamide ; Quartz crystals ; Steric hindrance ; Swelling ; Temperature dependence</subject><ispartof>RSC advances, 2021-11, Vol.11 (59), p.37225-37232</ispartof><rights>This journal is © The Royal Society of Chemistry.</rights><rights>Copyright Royal Society of Chemistry 2021</rights><rights>This journal is © The Royal Society of Chemistry 2021 The Royal Society of Chemistry</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c494t-21902e8f6d23947d7e36e4adeabfa83da9248d6807cbcd724efbe48d5fb19c1e3</citedby><cites>FETCH-LOGICAL-c494t-21902e8f6d23947d7e36e4adeabfa83da9248d6807cbcd724efbe48d5fb19c1e3</cites><orcidid>0000-0003-0493-6386 ; 0000-0002-3146-1024 ; 0000-0001-5212-1371 ; 0000-0002-4470-9842 ; 0000-0002-8133-4189</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9043771/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9043771/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35496401$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kobayashi, Jun</creatorcontrib><creatorcontrib>Arisaka, Yoshinori</creatorcontrib><creatorcontrib>Yui, Nobuhiko</creatorcontrib><creatorcontrib>Yamato, Masayuki</creatorcontrib><creatorcontrib>Okano, Teruo</creatorcontrib><title>Preservation of heparin-binding EGF-like growth factor activity on heparin-modified poly(-isopropylacrylamide)-grafted surfaces</title><title>RSC advances</title><addtitle>RSC Adv</addtitle><description>A heparin-modified poly( N -isopropylacrylamide) (PIPAAm)-grafted surface bound with heparin-binding epidermal growth factor-like growth factor (HB-EGF) was able to culture hepatocytes maintaining high albumin secretion and high expression of hepatocyte-specific genes. However, the activity of HB-EGF on the surface and its binding effects on hepatocytes remain unclear. In this study, we investigated the temperature-dependent interactions of HB-EGF and EGF receptor (EGFR) with heparin-modified PIPAAm to evaluate the activity of HB-EGF on the surface. Quartz crystal microbalance (QCM) measurements revealed that the amounts of adsorbed HB-EGF on either the heparin-modified PIPAAm-grafted surface (heparin-IC1) or PIPAAm-grafted surfaces were almost the same regardless of swelling/deswelling of grafted PIPAAm chains. The heparin-IC1 surface bound to HB-EGF at 37 °C had the ability to bind to hepatocytes through specific affinity interaction with EGFR, whose activation was confirmed by western blotting. However, the physisorbed HB-EGF on the PIPAAm surface greatly diminished its activity. Taken together, the introduction of heparin into grafted PIPAAm chains on the surface plays a pivotal role in holding HB-EGF while preserving its activity. Hydration and swelling of surface-grafted PIPAAm chains at 20 °C greatly diminished the attachment of hepatocytes with HB-EGF bound to heparin-IC1, whereas hepatocytes were able to bind to HB-EGF bound to heparin-IC1 at 37 °C. Thus, the equilibrated affinity interaction between EGFRs and surface-bound HB-EGF was considered to be attenuated by steric hindrance due to hydration and/or swelling of grafted PIPAAm chains. Activity of HB-EGF bound to a heparin-modified poly( N -isopropylacrylamide) (PIPAAm)-grafted surface was preserved through specific binding to heparin, whereas physisorbed HB-EGF on a PIPAAm-grafted surface greatly diminished its activity.</description><subject>Affinity</subject><subject>Albumins</subject><subject>Binding</subject><subject>Chains</subject><subject>Chemistry</subject><subject>Epidermal growth factor</subject><subject>Gene expression</subject><subject>Grafting</subject><subject>Growth factors</subject><subject>Heparin</subject><subject>Hydration</subject><subject>Microbalances</subject><subject>Polyisopropyl acrylamide</subject><subject>Quartz crystals</subject><subject>Steric hindrance</subject><subject>Swelling</subject><subject>Temperature dependence</subject><issn>2046-2069</issn><issn>2046-2069</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><recordid>eNpdkk1v1DAQhi0EolXphTsoEpeCFPBX7PiCVJVuQaoEQnC2HHu865LEwU4W7Ym_jmHbpeDD2Jp55tVY7yD0lODXBDP1xpFksGRE-gfomGIuaoqFenjvfYROc77B5YiGUEEeoyPWcCU4Jsfo56cEGdLWzCGOVfTVBiaTwlh3YXRhXFeXV6u6D9-gWqf4Y95U3tg5pqrEsA3zripddy1DdMEHcNUU-91ZHXKcUpx2vbGphCE4eFmvk_FzQfKSihLkJ-iRN32G09v7BH1dXX65eF9ff7z6cHF-XVuu-FxTojCF1gtHmeLSSWACuHFgOm9a5oyivHWixdJ21knKwXdQMo3viLIE2Al6u9edlm4AZ2Gck-n1lMJg0k5HE_S_lTFs9DputcKcSUmKwNmtQIrfF8izHkK20PdmhLhkTUXTCs5E2xT0xX_oTVzSWL5XKEy5FK1oC_VqT9kUc07gD8MQrH9bq9-Rz-d_rF0V-Pn98Q_onZEFeLYHUraH6t_dYL8AjjusSA</recordid><startdate>20211118</startdate><enddate>20211118</enddate><creator>Kobayashi, Jun</creator><creator>Arisaka, Yoshinori</creator><creator>Yui, Nobuhiko</creator><creator>Yamato, Masayuki</creator><creator>Okano, Teruo</creator><general>Royal Society of Chemistry</general><general>The Royal Society of Chemistry</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SR</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-0493-6386</orcidid><orcidid>https://orcid.org/0000-0002-3146-1024</orcidid><orcidid>https://orcid.org/0000-0001-5212-1371</orcidid><orcidid>https://orcid.org/0000-0002-4470-9842</orcidid><orcidid>https://orcid.org/0000-0002-8133-4189</orcidid></search><sort><creationdate>20211118</creationdate><title>Preservation of heparin-binding EGF-like growth factor activity on heparin-modified poly(-isopropylacrylamide)-grafted surfaces</title><author>Kobayashi, Jun ; Arisaka, Yoshinori ; Yui, Nobuhiko ; Yamato, Masayuki ; Okano, Teruo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c494t-21902e8f6d23947d7e36e4adeabfa83da9248d6807cbcd724efbe48d5fb19c1e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Affinity</topic><topic>Albumins</topic><topic>Binding</topic><topic>Chains</topic><topic>Chemistry</topic><topic>Epidermal growth factor</topic><topic>Gene expression</topic><topic>Grafting</topic><topic>Growth factors</topic><topic>Heparin</topic><topic>Hydration</topic><topic>Microbalances</topic><topic>Polyisopropyl acrylamide</topic><topic>Quartz crystals</topic><topic>Steric hindrance</topic><topic>Swelling</topic><topic>Temperature dependence</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kobayashi, Jun</creatorcontrib><creatorcontrib>Arisaka, Yoshinori</creatorcontrib><creatorcontrib>Yui, Nobuhiko</creatorcontrib><creatorcontrib>Yamato, Masayuki</creatorcontrib><creatorcontrib>Okano, Teruo</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Engineered Materials Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>RSC advances</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kobayashi, Jun</au><au>Arisaka, Yoshinori</au><au>Yui, Nobuhiko</au><au>Yamato, Masayuki</au><au>Okano, Teruo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Preservation of heparin-binding EGF-like growth factor activity on heparin-modified poly(-isopropylacrylamide)-grafted surfaces</atitle><jtitle>RSC advances</jtitle><addtitle>RSC Adv</addtitle><date>2021-11-18</date><risdate>2021</risdate><volume>11</volume><issue>59</issue><spage>37225</spage><epage>37232</epage><pages>37225-37232</pages><issn>2046-2069</issn><eissn>2046-2069</eissn><abstract>A heparin-modified poly( N -isopropylacrylamide) (PIPAAm)-grafted surface bound with heparin-binding epidermal growth factor-like growth factor (HB-EGF) was able to culture hepatocytes maintaining high albumin secretion and high expression of hepatocyte-specific genes. However, the activity of HB-EGF on the surface and its binding effects on hepatocytes remain unclear. In this study, we investigated the temperature-dependent interactions of HB-EGF and EGF receptor (EGFR) with heparin-modified PIPAAm to evaluate the activity of HB-EGF on the surface. Quartz crystal microbalance (QCM) measurements revealed that the amounts of adsorbed HB-EGF on either the heparin-modified PIPAAm-grafted surface (heparin-IC1) or PIPAAm-grafted surfaces were almost the same regardless of swelling/deswelling of grafted PIPAAm chains. The heparin-IC1 surface bound to HB-EGF at 37 °C had the ability to bind to hepatocytes through specific affinity interaction with EGFR, whose activation was confirmed by western blotting. However, the physisorbed HB-EGF on the PIPAAm surface greatly diminished its activity. Taken together, the introduction of heparin into grafted PIPAAm chains on the surface plays a pivotal role in holding HB-EGF while preserving its activity. Hydration and swelling of surface-grafted PIPAAm chains at 20 °C greatly diminished the attachment of hepatocytes with HB-EGF bound to heparin-IC1, whereas hepatocytes were able to bind to HB-EGF bound to heparin-IC1 at 37 °C. Thus, the equilibrated affinity interaction between EGFRs and surface-bound HB-EGF was considered to be attenuated by steric hindrance due to hydration and/or swelling of grafted PIPAAm chains. Activity of HB-EGF bound to a heparin-modified poly( N -isopropylacrylamide) (PIPAAm)-grafted surface was preserved through specific binding to heparin, whereas physisorbed HB-EGF on a PIPAAm-grafted surface greatly diminished its activity.</abstract><cop>England</cop><pub>Royal Society of Chemistry</pub><pmid>35496401</pmid><doi>10.1039/d1ra07317f</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0003-0493-6386</orcidid><orcidid>https://orcid.org/0000-0002-3146-1024</orcidid><orcidid>https://orcid.org/0000-0001-5212-1371</orcidid><orcidid>https://orcid.org/0000-0002-4470-9842</orcidid><orcidid>https://orcid.org/0000-0002-8133-4189</orcidid><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 2046-2069
ispartof RSC advances, 2021-11, Vol.11 (59), p.37225-37232
issn 2046-2069
2046-2069
language eng
recordid cdi_proquest_miscellaneous_2658643685
source DOAJ Directory of Open Access Journals; PubMed Central Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central
subjects Affinity
Albumins
Binding
Chains
Chemistry
Epidermal growth factor
Gene expression
Grafting
Growth factors
Heparin
Hydration
Microbalances
Polyisopropyl acrylamide
Quartz crystals
Steric hindrance
Swelling
Temperature dependence
title Preservation of heparin-binding EGF-like growth factor activity on heparin-modified poly(-isopropylacrylamide)-grafted surfaces
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-01T05%3A28%3A32IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Preservation%20of%20heparin-binding%20EGF-like%20growth%20factor%20activity%20on%20heparin-modified%20poly(-isopropylacrylamide)-grafted%20surfaces&rft.jtitle=RSC%20advances&rft.au=Kobayashi,%20Jun&rft.date=2021-11-18&rft.volume=11&rft.issue=59&rft.spage=37225&rft.epage=37232&rft.pages=37225-37232&rft.issn=2046-2069&rft.eissn=2046-2069&rft_id=info:doi/10.1039/d1ra07317f&rft_dat=%3Cproquest_pubme%3E2658643685%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2602476868&rft_id=info:pmid/35496401&rfr_iscdi=true