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...
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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 |
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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> |
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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 |
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