Nanoparticle Labeling of Bone Marrow-Derived Rat Mesenchymal Stem Cells: Their Use in Differentiation and Tracking
Mesenchymal stem cells (MSCs) are promising candidates for cellular therapies due to their ability to migrate to damaged tissue without inducing immune reaction. Many techniques have been developed to trace MSCs and their differentiation efficacy; however, all of these methods have limitations. Conj...
Gespeichert in:
| Veröffentlicht in: | BioMed research international 2015-01, Vol.2015 (2015), p.1-9 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 9 |
|---|---|
| container_issue | 2015 |
| container_start_page | 1 |
| container_title | BioMed research international |
| container_volume | 2015 |
| creator | Akhan, Ece Akcali, Kamil C. Tuncel, Donus |
| description | Mesenchymal stem cells (MSCs) are promising candidates for cellular therapies due to their ability to migrate to damaged tissue without inducing immune reaction. Many techniques have been developed to trace MSCs and their differentiation efficacy; however, all of these methods have limitations. Conjugated polymer based water-dispersible nanoparticles (CPN) represent a new class of probes because they offer high brightness, improved photostability, high fluorescent quantum yield, and noncytotoxicity comparing to conventional dyes and quantum dots. We aimed to use this tool for tracing MSCs’ fate in vitro and in vivo. MSC marker expression, survival, and differentiation capacity were assessed upon CPN treatment. Our results showed that after CPN labeling, MSC markers did not change and significant number of cells were found to be viable as revealed by MTT. Fluorescent signals were retained for 3 weeks after they were differentiated into osteocytes, adipocytes, and chondrocytes in vitro. We also showed that the labeled MSCs migrated to the site of injury and retained their labels in an in vivo liver regeneration model. The utilization of nanoparticle could be a promising tool for the tracking of MSCs in vivo and in vitro and therefore can be a useful tool to understand differentiation and homing mechanisms of MSCs. |
| doi_str_mv | 10.1155/2015/298430 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4310257</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A458160892</galeid><sourcerecordid>A458160892</sourcerecordid><originalsourceid>FETCH-LOGICAL-c561t-15499d7916371f5d15d518d6e2666032bab36ad6a5a564d179389ce2e6e303c63</originalsourceid><addsrcrecordid>eNqNks1vEzEQxVcIRKvSE3dkiQsCLXjstXfNAamkfEkpSJCercnubOKysYO9adX_HkcpoXCKD_ZI_ulpZt4riqfAXwMo9UZwyJdpKskfFMdCQlVqqODhvpbyqDhN6Yrn04DmRj8ujoTSquJGHBfxK_qwxji6diA2xTkNzi9Y6Nn74IldYIzhpjyn6K6pY99xZBeUyLfL2xUO7MdIKzahYUhv2WxJLrLLRMx5du76niL50eHogmfoOzaL2P7M4k-KRz0OiU7v3pPi8uOH2eRzOf326cvkbFq2SsNYgqqM6WoDWtbQqw5Up6DpNAmtNZdijnOpsdOoUOmqg9rIxrQkSJPkstXypHi3011v5ivq2txNxMGuo1thvLUBnf33x7ulXYRrW0ngQtVZ4MWdQAy_NpRGu3KpzdOip7BJFnRdmxqMMQeguWWjjYIDUCUqZYRSGX3-H3oVNtHnpWWqarKjGpq_1AIHss73IU_TbkXtWaW2njdGZOrVjmpjSClSv18EcLuNkt1Gye6ilOln93e3Z_8EJwMvd8DS-Q5v3GFqlBHq8R6cTc6D_gbkYNcG</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1648000618</pqid></control><display><type>article</type><title>Nanoparticle Labeling of Bone Marrow-Derived Rat Mesenchymal Stem Cells: Their Use in Differentiation and Tracking</title><source>MEDLINE</source><source>PubMed Central Open Access</source><source>Wiley Online Library Open Access</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><creator>Akhan, Ece ; Akcali, Kamil C. ; Tuncel, Donus</creator><contributor>Jones, Elena</contributor><creatorcontrib>Akhan, Ece ; Akcali, Kamil C. ; Tuncel, Donus ; Jones, Elena</creatorcontrib><description>Mesenchymal stem cells (MSCs) are promising candidates for cellular therapies due to their ability to migrate to damaged tissue without inducing immune reaction. Many techniques have been developed to trace MSCs and their differentiation efficacy; however, all of these methods have limitations. Conjugated polymer based water-dispersible nanoparticles (CPN) represent a new class of probes because they offer high brightness, improved photostability, high fluorescent quantum yield, and noncytotoxicity comparing to conventional dyes and quantum dots. We aimed to use this tool for tracing MSCs’ fate in vitro and in vivo. MSC marker expression, survival, and differentiation capacity were assessed upon CPN treatment. Our results showed that after CPN labeling, MSC markers did not change and significant number of cells were found to be viable as revealed by MTT. Fluorescent signals were retained for 3 weeks after they were differentiated into osteocytes, adipocytes, and chondrocytes in vitro. We also showed that the labeled MSCs migrated to the site of injury and retained their labels in an in vivo liver regeneration model. The utilization of nanoparticle could be a promising tool for the tracking of MSCs in vivo and in vitro and therefore can be a useful tool to understand differentiation and homing mechanisms of MSCs.</description><identifier>ISSN: 2314-6133</identifier><identifier>EISSN: 2314-6141</identifier><identifier>DOI: 10.1155/2015/298430</identifier><identifier>PMID: 25654092</identifier><language>eng</language><publisher>Cairo, Egypt: Hindawi Publishing Corporation</publisher><subject>Animals ; Biocompatibility ; Biology ; Biomedical materials ; Bone marrow ; Bone Marrow Cells - cytology ; Cell Differentiation ; Cell Lineage ; Cell Tracking - methods ; Differentiation ; Dyes ; Female ; Gene expression ; Health aspects ; In vitro testing ; In vivo testing ; In vivo tests ; Labeling ; Mesenchymal Stromal Cells - cytology ; Mesenchymal Stromal Cells - metabolism ; Microscopy ; Nanoparticles - chemistry ; Quantum dots ; Rats ; Rats, Sprague-Dawley ; Staining and Labeling ; Stem cells ; Surgical implants ; Water - chemistry</subject><ispartof>BioMed research international, 2015-01, Vol.2015 (2015), p.1-9</ispartof><rights>Copyright © 2015 Ece Akhan et al.</rights><rights>COPYRIGHT 2015 John Wiley & Sons, Inc.</rights><rights>Copyright © 2015 Ece Akhan et al. Ece Akhan et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</rights><rights>Copyright © 2015 Ece Akhan et al. 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c561t-15499d7916371f5d15d518d6e2666032bab36ad6a5a564d179389ce2e6e303c63</citedby><cites>FETCH-LOGICAL-c561t-15499d7916371f5d15d518d6e2666032bab36ad6a5a564d179389ce2e6e303c63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4310257/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4310257/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,724,777,781,882,27905,27906,53772,53774</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25654092$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Jones, Elena</contributor><creatorcontrib>Akhan, Ece</creatorcontrib><creatorcontrib>Akcali, Kamil C.</creatorcontrib><creatorcontrib>Tuncel, Donus</creatorcontrib><title>Nanoparticle Labeling of Bone Marrow-Derived Rat Mesenchymal Stem Cells: Their Use in Differentiation and Tracking</title><title>BioMed research international</title><addtitle>Biomed Res Int</addtitle><description>Mesenchymal stem cells (MSCs) are promising candidates for cellular therapies due to their ability to migrate to damaged tissue without inducing immune reaction. Many techniques have been developed to trace MSCs and their differentiation efficacy; however, all of these methods have limitations. Conjugated polymer based water-dispersible nanoparticles (CPN) represent a new class of probes because they offer high brightness, improved photostability, high fluorescent quantum yield, and noncytotoxicity comparing to conventional dyes and quantum dots. We aimed to use this tool for tracing MSCs’ fate in vitro and in vivo. MSC marker expression, survival, and differentiation capacity were assessed upon CPN treatment. Our results showed that after CPN labeling, MSC markers did not change and significant number of cells were found to be viable as revealed by MTT. Fluorescent signals were retained for 3 weeks after they were differentiated into osteocytes, adipocytes, and chondrocytes in vitro. We also showed that the labeled MSCs migrated to the site of injury and retained their labels in an in vivo liver regeneration model. The utilization of nanoparticle could be a promising tool for the tracking of MSCs in vivo and in vitro and therefore can be a useful tool to understand differentiation and homing mechanisms of MSCs.</description><subject>Animals</subject><subject>Biocompatibility</subject><subject>Biology</subject><subject>Biomedical materials</subject><subject>Bone marrow</subject><subject>Bone Marrow Cells - cytology</subject><subject>Cell Differentiation</subject><subject>Cell Lineage</subject><subject>Cell Tracking - methods</subject><subject>Differentiation</subject><subject>Dyes</subject><subject>Female</subject><subject>Gene expression</subject><subject>Health aspects</subject><subject>In vitro testing</subject><subject>In vivo testing</subject><subject>In vivo tests</subject><subject>Labeling</subject><subject>Mesenchymal Stromal Cells - cytology</subject><subject>Mesenchymal Stromal Cells - metabolism</subject><subject>Microscopy</subject><subject>Nanoparticles - chemistry</subject><subject>Quantum dots</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>Staining and Labeling</subject><subject>Stem cells</subject><subject>Surgical implants</subject><subject>Water - chemistry</subject><issn>2314-6133</issn><issn>2314-6141</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>RHX</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNks1vEzEQxVcIRKvSE3dkiQsCLXjstXfNAamkfEkpSJCercnubOKysYO9adX_HkcpoXCKD_ZI_ulpZt4riqfAXwMo9UZwyJdpKskfFMdCQlVqqODhvpbyqDhN6Yrn04DmRj8ujoTSquJGHBfxK_qwxji6diA2xTkNzi9Y6Nn74IldYIzhpjyn6K6pY99xZBeUyLfL2xUO7MdIKzahYUhv2WxJLrLLRMx5du76niL50eHogmfoOzaL2P7M4k-KRz0OiU7v3pPi8uOH2eRzOf326cvkbFq2SsNYgqqM6WoDWtbQqw5Up6DpNAmtNZdijnOpsdOoUOmqg9rIxrQkSJPkstXypHi3011v5ivq2txNxMGuo1thvLUBnf33x7ulXYRrW0ngQtVZ4MWdQAy_NpRGu3KpzdOip7BJFnRdmxqMMQeguWWjjYIDUCUqZYRSGX3-H3oVNtHnpWWqarKjGpq_1AIHss73IU_TbkXtWaW2njdGZOrVjmpjSClSv18EcLuNkt1Gye6ilOln93e3Z_8EJwMvd8DS-Q5v3GFqlBHq8R6cTc6D_gbkYNcG</recordid><startdate>20150101</startdate><enddate>20150101</enddate><creator>Akhan, Ece</creator><creator>Akcali, Kamil C.</creator><creator>Tuncel, Donus</creator><general>Hindawi Publishing Corporation</general><general>John Wiley & Sons, Inc</general><general>Hindawi Limited</general><scope>ADJCN</scope><scope>AHFXO</scope><scope>RHU</scope><scope>RHW</scope><scope>RHX</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>3V.</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7TK</scope><scope>7U7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>CWDGH</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>7U5</scope><scope>F28</scope><scope>L7M</scope><scope>5PM</scope></search><sort><creationdate>20150101</creationdate><title>Nanoparticle Labeling of Bone Marrow-Derived Rat Mesenchymal Stem Cells: Their Use in Differentiation and Tracking</title><author>Akhan, Ece ; Akcali, Kamil C. ; Tuncel, Donus</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c561t-15499d7916371f5d15d518d6e2666032bab36ad6a5a564d179389ce2e6e303c63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Animals</topic><topic>Biocompatibility</topic><topic>Biology</topic><topic>Biomedical materials</topic><topic>Bone marrow</topic><topic>Bone Marrow Cells - cytology</topic><topic>Cell Differentiation</topic><topic>Cell Lineage</topic><topic>Cell Tracking - methods</topic><topic>Differentiation</topic><topic>Dyes</topic><topic>Female</topic><topic>Gene expression</topic><topic>Health aspects</topic><topic>In vitro testing</topic><topic>In vivo testing</topic><topic>In vivo tests</topic><topic>Labeling</topic><topic>Mesenchymal Stromal Cells - cytology</topic><topic>Mesenchymal Stromal Cells - metabolism</topic><topic>Microscopy</topic><topic>Nanoparticles - chemistry</topic><topic>Quantum dots</topic><topic>Rats</topic><topic>Rats, Sprague-Dawley</topic><topic>Staining and Labeling</topic><topic>Stem cells</topic><topic>Surgical implants</topic><topic>Water - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Akhan, Ece</creatorcontrib><creatorcontrib>Akcali, Kamil C.</creatorcontrib><creatorcontrib>Tuncel, Donus</creatorcontrib><collection>الدوريات العلمية والإحصائية - e-Marefa Academic and Statistical Periodicals</collection><collection>معرفة - المحتوى العربي الأكاديمي المتكامل - e-Marefa Academic Complete</collection><collection>Hindawi Publishing Complete</collection><collection>Hindawi Publishing Subscription Journals</collection><collection>Hindawi Publishing 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>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>Middle East & Africa Database</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>BioMed research international</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Akhan, Ece</au><au>Akcali, Kamil C.</au><au>Tuncel, Donus</au><au>Jones, Elena</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Nanoparticle Labeling of Bone Marrow-Derived Rat Mesenchymal Stem Cells: Their Use in Differentiation and Tracking</atitle><jtitle>BioMed research international</jtitle><addtitle>Biomed Res Int</addtitle><date>2015-01-01</date><risdate>2015</risdate><volume>2015</volume><issue>2015</issue><spage>1</spage><epage>9</epage><pages>1-9</pages><issn>2314-6133</issn><eissn>2314-6141</eissn><abstract>Mesenchymal stem cells (MSCs) are promising candidates for cellular therapies due to their ability to migrate to damaged tissue without inducing immune reaction. Many techniques have been developed to trace MSCs and their differentiation efficacy; however, all of these methods have limitations. Conjugated polymer based water-dispersible nanoparticles (CPN) represent a new class of probes because they offer high brightness, improved photostability, high fluorescent quantum yield, and noncytotoxicity comparing to conventional dyes and quantum dots. We aimed to use this tool for tracing MSCs’ fate in vitro and in vivo. MSC marker expression, survival, and differentiation capacity were assessed upon CPN treatment. Our results showed that after CPN labeling, MSC markers did not change and significant number of cells were found to be viable as revealed by MTT. Fluorescent signals were retained for 3 weeks after they were differentiated into osteocytes, adipocytes, and chondrocytes in vitro. We also showed that the labeled MSCs migrated to the site of injury and retained their labels in an in vivo liver regeneration model. The utilization of nanoparticle could be a promising tool for the tracking of MSCs in vivo and in vitro and therefore can be a useful tool to understand differentiation and homing mechanisms of MSCs.</abstract><cop>Cairo, Egypt</cop><pub>Hindawi Publishing Corporation</pub><pmid>25654092</pmid><doi>10.1155/2015/298430</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 2314-6133 |
| ispartof | BioMed research international, 2015-01, Vol.2015 (2015), p.1-9 |
| issn | 2314-6133 2314-6141 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4310257 |
| source | MEDLINE; PubMed Central Open Access; Wiley Online Library Open Access; PubMed Central; Alma/SFX Local Collection |
| subjects | Animals Biocompatibility Biology Biomedical materials Bone marrow Bone Marrow Cells - cytology Cell Differentiation Cell Lineage Cell Tracking - methods Differentiation Dyes Female Gene expression Health aspects In vitro testing In vivo testing In vivo tests Labeling Mesenchymal Stromal Cells - cytology Mesenchymal Stromal Cells - metabolism Microscopy Nanoparticles - chemistry Quantum dots Rats Rats, Sprague-Dawley Staining and Labeling Stem cells Surgical implants Water - chemistry |
| title | Nanoparticle Labeling of Bone Marrow-Derived Rat Mesenchymal Stem Cells: Their Use in Differentiation and Tracking |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-20T07%3A39%3A41IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Nanoparticle%20Labeling%20of%20Bone%20Marrow-Derived%20Rat%20Mesenchymal%20Stem%20Cells:%20Their%20Use%20in%20Differentiation%20and%20Tracking&rft.jtitle=BioMed%20research%20international&rft.au=Akhan,%20Ece&rft.date=2015-01-01&rft.volume=2015&rft.issue=2015&rft.spage=1&rft.epage=9&rft.pages=1-9&rft.issn=2314-6133&rft.eissn=2314-6141&rft_id=info:doi/10.1155/2015/298430&rft_dat=%3Cgale_pubme%3EA458160892%3C/gale_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1648000618&rft_id=info:pmid/25654092&rft_galeid=A458160892&rfr_iscdi=true |