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Scalable Production of Monodisperse Bioactive Spider Silk Nanowires
Elongated protein‐based micro‐ and nanostructures are of great interest for a wide range of biomedical applications, where they can serve as a backbone for surface functionalization and as vehicles for drug delivery. Current production methods for protein constructs lack precise control of either sh...
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| Published in: | Macromolecular bioscience 2023-04, Vol.23 (4), p.e2200450-n/a |
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| creator | Gustafsson, Linnea Kvick, Mathias Åstrand, Carolina Ponsteen, Nienke Dorka, Nicolai Hegrová, Veronika Svanberg, Sara Horák, Josef Jansson, Ronnie Hedhammar, My van der Wijngaart, Wouter |
| description | Elongated protein‐based micro‐ and nanostructures are of great interest for a wide range of biomedical applications, where they can serve as a backbone for surface functionalization and as vehicles for drug delivery. Current production methods for protein constructs lack precise control of either shape and dimensions or render structures fixed to substrates. This work demonstrates production of recombinant spider silk nanowires suspended in solution, starting with liquid bridge induced assembly (LBIA) on a substrate, followed by release using ultrasonication, and concentration by centrifugation. The significance of this method lies in that it provides i) reproducability (standard deviation of length |
| doi_str_mv | 10.1002/mabi.202200450 |
| format | article |
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The process for production and release of spider silk nanowires using liquid bridge induced assembly is successfully upscaled to produce over 12 million of 10 µm long nanowires suspended in solution within 20 min. The nanowires can easily be functionalized both pre‐and post‐formation and are successfully used as an ECM mimic to support cell adhesion and integrated into 3D cell aggregates.</description><identifier>ISSN: 1616-5187</identifier><identifier>ISSN: 1616-5195</identifier><identifier>EISSN: 1616-5195</identifier><identifier>DOI: 10.1002/mabi.202200450</identifier><identifier>PMID: 36662774</identifier><language>eng</language><publisher>Germany: Wiley Subscription Services, Inc</publisher><subject>Animals ; Biocompatibility ; Biological activity ; Biomedical materials ; Biotechnology ; Bioteknologi ; cell aggregates ; Cell culture ; Cell Culture Techniques ; Centrifugation ; Drug delivery ; Fabrication ; Humans ; liquid bridge induced assembly ; Liquid bridges ; Medical research ; Mesenchymal stem cells ; Nanostructures ; Nanotechnology ; Nanowires ; Production methods ; Proteins ; scalable production ; Silk ; Silk - chemistry ; spider silk ; Spiders ; Stem cells ; Substrates</subject><ispartof>Macromolecular bioscience, 2023-04, Vol.23 (4), p.e2200450-n/a</ispartof><rights>2023 The Authors. Macromolecular Bioscience published by Wiley‐VCH GmbH</rights><rights>2023 The Authors. Macromolecular Bioscience published by Wiley-VCH GmbH.</rights><rights>2023. This article is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4510-179d3726d890bf95e9c882f972eacffa5252e5f52d89c2a660d2e638acfa58a43</citedby><cites>FETCH-LOGICAL-c4510-179d3726d890bf95e9c882f972eacffa5252e5f52d89c2a660d2e638acfa58a43</cites><orcidid>0000-0003-0140-419X ; 0000-0002-8925-2815 ; 0000-0001-8248-6670</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36662774$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-323201$$DView record from Swedish Publication Index$$Hfree_for_read</backlink></links><search><creatorcontrib>Gustafsson, Linnea</creatorcontrib><creatorcontrib>Kvick, Mathias</creatorcontrib><creatorcontrib>Åstrand, Carolina</creatorcontrib><creatorcontrib>Ponsteen, Nienke</creatorcontrib><creatorcontrib>Dorka, Nicolai</creatorcontrib><creatorcontrib>Hegrová, Veronika</creatorcontrib><creatorcontrib>Svanberg, Sara</creatorcontrib><creatorcontrib>Horák, Josef</creatorcontrib><creatorcontrib>Jansson, Ronnie</creatorcontrib><creatorcontrib>Hedhammar, My</creatorcontrib><creatorcontrib>van der Wijngaart, Wouter</creatorcontrib><title>Scalable Production of Monodisperse Bioactive Spider Silk Nanowires</title><title>Macromolecular bioscience</title><addtitle>Macromol Biosci</addtitle><description>Elongated protein‐based micro‐ and nanostructures are of great interest for a wide range of biomedical applications, where they can serve as a backbone for surface functionalization and as vehicles for drug delivery. Current production methods for protein constructs lack precise control of either shape and dimensions or render structures fixed to substrates. This work demonstrates production of recombinant spider silk nanowires suspended in solution, starting with liquid bridge induced assembly (LBIA) on a substrate, followed by release using ultrasonication, and concentration by centrifugation. The significance of this method lies in that it provides i) reproducability (standard deviation of length <13% and of diameter <38%), ii) scalability of fabrication, iii) compatibility with autoclavation with retained shape and function, iv) retention of bioactivity, and v) easy functionalization both pre‐ and post‐formation. This work demonstrates how altering the function and nanotopography of a surface by nanowire coating supports the attachment and growth of human mesenchymal stem cells (hMSCs). Cell compatibility is further studied through integration of nanowires during aggregate formation of hMSCs and the breast cancer cell line MCF7. The herein‐presented industrial‐compatible process enables silk nanowires for use as functionalizing agents in a variety of cell culture applications and medical research.
The process for production and release of spider silk nanowires using liquid bridge induced assembly is successfully upscaled to produce over 12 million of 10 µm long nanowires suspended in solution within 20 min. The nanowires can easily be functionalized both pre‐and post‐formation and are successfully used as an ECM mimic to support cell adhesion and integrated into 3D cell aggregates.</description><subject>Animals</subject><subject>Biocompatibility</subject><subject>Biological activity</subject><subject>Biomedical materials</subject><subject>Biotechnology</subject><subject>Bioteknologi</subject><subject>cell aggregates</subject><subject>Cell culture</subject><subject>Cell Culture Techniques</subject><subject>Centrifugation</subject><subject>Drug delivery</subject><subject>Fabrication</subject><subject>Humans</subject><subject>liquid bridge induced assembly</subject><subject>Liquid bridges</subject><subject>Medical research</subject><subject>Mesenchymal stem cells</subject><subject>Nanostructures</subject><subject>Nanotechnology</subject><subject>Nanowires</subject><subject>Production methods</subject><subject>Proteins</subject><subject>scalable production</subject><subject>Silk</subject><subject>Silk - chemistry</subject><subject>spider silk</subject><subject>Spiders</subject><subject>Stem cells</subject><subject>Substrates</subject><issn>1616-5187</issn><issn>1616-5195</issn><issn>1616-5195</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><recordid>eNqFkM9PwjAUxxujEUSvHs0Sz8P2be3aI-AvElAT1GvTbZ0WBp0tk_DfOwLOo6f3ku_nffLyReiS4D7BGG6WKjV9wAAYxxQfoS5hhIWUCHrc7jzpoDPv5xiThAs4RZ2IMQZJEnfRaJapUqWlDl6czetsbewqsEUwtSubG19p53UwNFY1ybcOZpXJtQtmplwET2plN8Zpf45OClV6fXGYPfR2f_c6egwnzw_j0WASZjElOCSJyKMEWM4FTgtBtcg4h0IkoFVWFIoCBU0LCg2QgWIM56BZxJtQUa7iqIfCvddvdFWnsnJmqdxWWmXkrXkfSOs-5GL9KSOIAJOGv97zlbNftfZrObe1WzUvSuCYcB4Jyhuqv6cyZ713umi9BMtdx3LXsWw7bg6uDto6Xeq8xX9LbQCxBzam1Nt_dHI6GI7_5D_CSYfG</recordid><startdate>202304</startdate><enddate>202304</enddate><creator>Gustafsson, Linnea</creator><creator>Kvick, Mathias</creator><creator>Åstrand, Carolina</creator><creator>Ponsteen, Nienke</creator><creator>Dorka, Nicolai</creator><creator>Hegrová, Veronika</creator><creator>Svanberg, Sara</creator><creator>Horák, Josef</creator><creator>Jansson, Ronnie</creator><creator>Hedhammar, My</creator><creator>van der Wijngaart, Wouter</creator><general>Wiley Subscription Services, Inc</general><scope>24P</scope><scope>WIN</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>ADTPV</scope><scope>AOWAS</scope><scope>D8V</scope><orcidid>https://orcid.org/0000-0003-0140-419X</orcidid><orcidid>https://orcid.org/0000-0002-8925-2815</orcidid><orcidid>https://orcid.org/0000-0001-8248-6670</orcidid></search><sort><creationdate>202304</creationdate><title>Scalable Production of Monodisperse Bioactive Spider Silk Nanowires</title><author>Gustafsson, Linnea ; Kvick, Mathias ; Åstrand, Carolina ; Ponsteen, Nienke ; Dorka, Nicolai ; Hegrová, Veronika ; Svanberg, Sara ; Horák, Josef ; Jansson, Ronnie ; Hedhammar, My ; van der Wijngaart, Wouter</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4510-179d3726d890bf95e9c882f972eacffa5252e5f52d89c2a660d2e638acfa58a43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Animals</topic><topic>Biocompatibility</topic><topic>Biological activity</topic><topic>Biomedical materials</topic><topic>Biotechnology</topic><topic>Bioteknologi</topic><topic>cell aggregates</topic><topic>Cell culture</topic><topic>Cell Culture Techniques</topic><topic>Centrifugation</topic><topic>Drug delivery</topic><topic>Fabrication</topic><topic>Humans</topic><topic>liquid bridge induced assembly</topic><topic>Liquid bridges</topic><topic>Medical research</topic><topic>Mesenchymal stem cells</topic><topic>Nanostructures</topic><topic>Nanotechnology</topic><topic>Nanowires</topic><topic>Production methods</topic><topic>Proteins</topic><topic>scalable production</topic><topic>Silk</topic><topic>Silk - chemistry</topic><topic>spider silk</topic><topic>Spiders</topic><topic>Stem cells</topic><topic>Substrates</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gustafsson, Linnea</creatorcontrib><creatorcontrib>Kvick, Mathias</creatorcontrib><creatorcontrib>Åstrand, Carolina</creatorcontrib><creatorcontrib>Ponsteen, Nienke</creatorcontrib><creatorcontrib>Dorka, Nicolai</creatorcontrib><creatorcontrib>Hegrová, Veronika</creatorcontrib><creatorcontrib>Svanberg, Sara</creatorcontrib><creatorcontrib>Horák, Josef</creatorcontrib><creatorcontrib>Jansson, Ronnie</creatorcontrib><creatorcontrib>Hedhammar, My</creatorcontrib><creatorcontrib>van der Wijngaart, Wouter</creatorcontrib><collection>Wiley Online Library Open Access</collection><collection>Wiley-Blackwell Open Access Backfiles (Open Access)</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>SwePub</collection><collection>SwePub Articles</collection><collection>SWEPUB Kungliga Tekniska Högskolan</collection><jtitle>Macromolecular bioscience</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gustafsson, Linnea</au><au>Kvick, Mathias</au><au>Åstrand, Carolina</au><au>Ponsteen, Nienke</au><au>Dorka, Nicolai</au><au>Hegrová, Veronika</au><au>Svanberg, Sara</au><au>Horák, Josef</au><au>Jansson, Ronnie</au><au>Hedhammar, My</au><au>van der Wijngaart, Wouter</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Scalable Production of Monodisperse Bioactive Spider Silk Nanowires</atitle><jtitle>Macromolecular bioscience</jtitle><addtitle>Macromol Biosci</addtitle><date>2023-04</date><risdate>2023</risdate><volume>23</volume><issue>4</issue><spage>e2200450</spage><epage>n/a</epage><pages>e2200450-n/a</pages><issn>1616-5187</issn><issn>1616-5195</issn><eissn>1616-5195</eissn><abstract>Elongated protein‐based micro‐ and nanostructures are of great interest for a wide range of biomedical applications, where they can serve as a backbone for surface functionalization and as vehicles for drug delivery. Current production methods for protein constructs lack precise control of either shape and dimensions or render structures fixed to substrates. This work demonstrates production of recombinant spider silk nanowires suspended in solution, starting with liquid bridge induced assembly (LBIA) on a substrate, followed by release using ultrasonication, and concentration by centrifugation. The significance of this method lies in that it provides i) reproducability (standard deviation of length <13% and of diameter <38%), ii) scalability of fabrication, iii) compatibility with autoclavation with retained shape and function, iv) retention of bioactivity, and v) easy functionalization both pre‐ and post‐formation. This work demonstrates how altering the function and nanotopography of a surface by nanowire coating supports the attachment and growth of human mesenchymal stem cells (hMSCs). Cell compatibility is further studied through integration of nanowires during aggregate formation of hMSCs and the breast cancer cell line MCF7. The herein‐presented industrial‐compatible process enables silk nanowires for use as functionalizing agents in a variety of cell culture applications and medical research.
The process for production and release of spider silk nanowires using liquid bridge induced assembly is successfully upscaled to produce over 12 million of 10 µm long nanowires suspended in solution within 20 min. The nanowires can easily be functionalized both pre‐and post‐formation and are successfully used as an ECM mimic to support cell adhesion and integrated into 3D cell aggregates.</abstract><cop>Germany</cop><pub>Wiley Subscription Services, Inc</pub><pmid>36662774</pmid><doi>10.1002/mabi.202200450</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0003-0140-419X</orcidid><orcidid>https://orcid.org/0000-0002-8925-2815</orcidid><orcidid>https://orcid.org/0000-0001-8248-6670</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Animals Biocompatibility Biological activity Biomedical materials Biotechnology Bioteknologi cell aggregates Cell culture Cell Culture Techniques Centrifugation Drug delivery Fabrication Humans liquid bridge induced assembly Liquid bridges Medical research Mesenchymal stem cells Nanostructures Nanotechnology Nanowires Production methods Proteins scalable production Silk Silk - chemistry spider silk Spiders Stem cells Substrates |
| title | Scalable Production of Monodisperse Bioactive Spider Silk Nanowires |
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