Novel glycidyl methacrylated dextran gelatin nanoparticles loaded with basic fibroblast growth factor: formulation and characteristics

Objective: To develop a novel efficient nanoparticulate carrier loaded with basic fibroblast growth factor (bFGF). Methods: Gelatin and glycidyl methacrylate-derivatized dextran (dex-GMA) were cross-linked and polymerized to form interpenetrating polymeric networks. The properties of the nanoparticl...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Drug development and industrial pharmacy 2009-12, Vol.35 (12), p.1419-1429
Hauptverfasser: Gu, Chunhu, Zheng, Renhong, Yang, Zhifu, Wen, Aidong, Wu, Hong, Zhang, Hui, Yi, Dinghua
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 1429
container_issue 12
container_start_page 1419
container_title Drug development and industrial pharmacy
container_volume 35
creator Gu, Chunhu
Zheng, Renhong
Yang, Zhifu
Wen, Aidong
Wu, Hong
Zhang, Hui
Yi, Dinghua
description Objective: To develop a novel efficient nanoparticulate carrier loaded with basic fibroblast growth factor (bFGF). Methods: Gelatin and glycidyl methacrylate-derivatized dextran (dex-GMA) were cross-linked and polymerized to form interpenetrating polymeric networks. The properties of the nanoparticles (NPs) were investigated as a function of the degree of dex-GMA substitution and the concentration of gelatin used in the preparation of the hydrogels. The morphology was observed with scanning eletromicroscopy and transmission eletromicroscopy. The swelling, degradation, and entrapment efficiency were also determined by dynamic evaluation methods in vitro. The protein release ratio and in vitro release kinetics were evaluated by routine procedure, and the biological activity of bFGF-loaded NPs was studied by cell proliferation assay, cell attachment, and cell function. Results: The NPs have a particle size of 320 ± 20 nm. bFGF was entrapped in the nanoparticles quantitatively (the encapsulation efficiency, 89.6 ± 0.9%). The bFGF in vitro release kinetics fitted to zero-order and Higuchi equations. Proliferation assay, attachment assay, and western blot showed that bFGF NPs had good biological effects on cultured bone marrow mesenchymal stem cells and could achieve a much longer action time than bFGF solution. Conclusion: These results suggested that a novel biodegradable dex-GMA gelatin hydrogel NPs loaded with bFGF could be successfully developed from both dextran- and gelatin-based biomaterials.
doi_str_mv 10.3109/03639040902988558
format Article
fullrecord <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_proquest_miscellaneous_733674812</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>733674812</sourcerecordid><originalsourceid>FETCH-LOGICAL-c365t-286a47b7bb7999ec2f7b79ac4610aafc60a433ca372c02e59cecefd255e57ba43</originalsourceid><addsrcrecordid>eNp9kMtu1DAUhi0EokPhAdggb1BXob7ESQxsqopLpQo2sI5OnJOJKycebIchL8Bz49EMIITUlXV8vv-X_RHynLNXkjN9yWQlNSuZZkI3jVLNA7LhSrBC1ZV4SDaHfZEBdUaexHjHGBdaqcfkjGsttGB8Q35-8t_R0a1bje1XRydMI5iwOkjY0x5_pAAz3WKe7UxnmP0OQrLGYaTOQ5-hvU0j7SBaQwfbBd85iIlug9_n-wFM8uE1HXyYlkOJnynMPTUjhLzCYGNui0_JowFcxGen85x8ff_uy_XH4vbzh5vrq9vCyEqlQjQVlHVXd12ttUYjhjxoMGXFGcBgKgallAZkLQwTqLRBg0MvlEJVd3l3Ti6Ovbvgvy0YUzvZaNA5mNEvsa2lrOqy4SKT_Eia4GMMOLS7YCcIa8tZe7Df_mc_Z16c2pduwv5v4qQ7Ay9PAEQDbshujY1_OCGk4nXJMvf2yNn5IA72Pri-TbA6H36H5H3vePNPfERwaTQQsL3zS5iz4Xt-8QtiLrYv</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>733674812</pqid></control><display><type>article</type><title>Novel glycidyl methacrylated dextran gelatin nanoparticles loaded with basic fibroblast growth factor: formulation and characteristics</title><source>Taylor &amp; Francis:Master (3349 titles)</source><source>MEDLINE</source><source>Taylor &amp; Francis Medical Library - CRKN</source><source>Business Source Complete</source><creator>Gu, Chunhu ; Zheng, Renhong ; Yang, Zhifu ; Wen, Aidong ; Wu, Hong ; Zhang, Hui ; Yi, Dinghua</creator><creatorcontrib>Gu, Chunhu ; Zheng, Renhong ; Yang, Zhifu ; Wen, Aidong ; Wu, Hong ; Zhang, Hui ; Yi, Dinghua</creatorcontrib><description>Objective: To develop a novel efficient nanoparticulate carrier loaded with basic fibroblast growth factor (bFGF). Methods: Gelatin and glycidyl methacrylate-derivatized dextran (dex-GMA) were cross-linked and polymerized to form interpenetrating polymeric networks. The properties of the nanoparticles (NPs) were investigated as a function of the degree of dex-GMA substitution and the concentration of gelatin used in the preparation of the hydrogels. The morphology was observed with scanning eletromicroscopy and transmission eletromicroscopy. The swelling, degradation, and entrapment efficiency were also determined by dynamic evaluation methods in vitro. The protein release ratio and in vitro release kinetics were evaluated by routine procedure, and the biological activity of bFGF-loaded NPs was studied by cell proliferation assay, cell attachment, and cell function. Results: The NPs have a particle size of 320 ± 20 nm. bFGF was entrapped in the nanoparticles quantitatively (the encapsulation efficiency, 89.6 ± 0.9%). The bFGF in vitro release kinetics fitted to zero-order and Higuchi equations. Proliferation assay, attachment assay, and western blot showed that bFGF NPs had good biological effects on cultured bone marrow mesenchymal stem cells and could achieve a much longer action time than bFGF solution. Conclusion: These results suggested that a novel biodegradable dex-GMA gelatin hydrogel NPs loaded with bFGF could be successfully developed from both dextran- and gelatin-based biomaterials.</description><identifier>ISSN: 0363-9045</identifier><identifier>EISSN: 1520-5762</identifier><identifier>DOI: 10.3109/03639040902988558</identifier><identifier>PMID: 19929201</identifier><language>eng</language><publisher>Colchester: Informa UK Ltd</publisher><subject>Animals ; Basic fibrous growth factor ; Biological and medical sciences ; Bone Marrow Cells - drug effects ; Bone Marrow Cells - secretion ; Cell Adhesion - drug effects ; Cell Proliferation - drug effects ; Cells, Cultured ; Cross-Linking Reagents ; dextran ; Dextrans - chemical synthesis ; Dextrans - chemistry ; Dextrans - metabolism ; Drug Carriers - administration &amp; dosage ; Drug Carriers - chemistry ; Drug Carriers - pharmacology ; drug delivery system ; Epoxy Compounds - chemical synthesis ; Epoxy Compounds - chemistry ; Epoxy Compounds - metabolism ; Fibroblast Growth Factor 2 - administration &amp; dosage ; Fibroblast Growth Factor 2 - chemistry ; Fibroblast Growth Factor 2 - pharmacology ; gelatin ; Gelatin - chemistry ; General pharmacology ; Hydrogels - chemical synthesis ; Hydrogels - chemistry ; Kinetics ; Medical sciences ; Mesenchymal Stromal Cells - drug effects ; Mesenchymal Stromal Cells - secretion ; Methacrylates - chemical synthesis ; Methacrylates - chemistry ; Methacrylates - metabolism ; Mice ; nanoparticle ; Nanoparticles - chemistry ; Nanoparticles - ultrastructure ; Particle Size ; Pharmaceutical technology. Pharmaceutical industry ; Pharmacology. Drug treatments ; Water - analysis</subject><ispartof>Drug development and industrial pharmacy, 2009-12, Vol.35 (12), p.1419-1429</ispartof><rights>Informa UK, Ltd. 2009</rights><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c365t-286a47b7bb7999ec2f7b79ac4610aafc60a433ca372c02e59cecefd255e57ba43</citedby><cites>FETCH-LOGICAL-c365t-286a47b7bb7999ec2f7b79ac4610aafc60a433ca372c02e59cecefd255e57ba43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.tandfonline.com/doi/pdf/10.3109/03639040902988558$$EPDF$$P50$$Ginformahealthcare$$H</linktopdf><linktohtml>$$Uhttps://www.tandfonline.com/doi/full/10.3109/03639040902988558$$EHTML$$P50$$Ginformahealthcare$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,59620,59726,60409,60515,61194,61229,61375,61410</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=22351740$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19929201$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gu, Chunhu</creatorcontrib><creatorcontrib>Zheng, Renhong</creatorcontrib><creatorcontrib>Yang, Zhifu</creatorcontrib><creatorcontrib>Wen, Aidong</creatorcontrib><creatorcontrib>Wu, Hong</creatorcontrib><creatorcontrib>Zhang, Hui</creatorcontrib><creatorcontrib>Yi, Dinghua</creatorcontrib><title>Novel glycidyl methacrylated dextran gelatin nanoparticles loaded with basic fibroblast growth factor: formulation and characteristics</title><title>Drug development and industrial pharmacy</title><addtitle>Drug Dev Ind Pharm</addtitle><description>Objective: To develop a novel efficient nanoparticulate carrier loaded with basic fibroblast growth factor (bFGF). Methods: Gelatin and glycidyl methacrylate-derivatized dextran (dex-GMA) were cross-linked and polymerized to form interpenetrating polymeric networks. The properties of the nanoparticles (NPs) were investigated as a function of the degree of dex-GMA substitution and the concentration of gelatin used in the preparation of the hydrogels. The morphology was observed with scanning eletromicroscopy and transmission eletromicroscopy. The swelling, degradation, and entrapment efficiency were also determined by dynamic evaluation methods in vitro. The protein release ratio and in vitro release kinetics were evaluated by routine procedure, and the biological activity of bFGF-loaded NPs was studied by cell proliferation assay, cell attachment, and cell function. Results: The NPs have a particle size of 320 ± 20 nm. bFGF was entrapped in the nanoparticles quantitatively (the encapsulation efficiency, 89.6 ± 0.9%). The bFGF in vitro release kinetics fitted to zero-order and Higuchi equations. Proliferation assay, attachment assay, and western blot showed that bFGF NPs had good biological effects on cultured bone marrow mesenchymal stem cells and could achieve a much longer action time than bFGF solution. Conclusion: These results suggested that a novel biodegradable dex-GMA gelatin hydrogel NPs loaded with bFGF could be successfully developed from both dextran- and gelatin-based biomaterials.</description><subject>Animals</subject><subject>Basic fibrous growth factor</subject><subject>Biological and medical sciences</subject><subject>Bone Marrow Cells - drug effects</subject><subject>Bone Marrow Cells - secretion</subject><subject>Cell Adhesion - drug effects</subject><subject>Cell Proliferation - drug effects</subject><subject>Cells, Cultured</subject><subject>Cross-Linking Reagents</subject><subject>dextran</subject><subject>Dextrans - chemical synthesis</subject><subject>Dextrans - chemistry</subject><subject>Dextrans - metabolism</subject><subject>Drug Carriers - administration &amp; dosage</subject><subject>Drug Carriers - chemistry</subject><subject>Drug Carriers - pharmacology</subject><subject>drug delivery system</subject><subject>Epoxy Compounds - chemical synthesis</subject><subject>Epoxy Compounds - chemistry</subject><subject>Epoxy Compounds - metabolism</subject><subject>Fibroblast Growth Factor 2 - administration &amp; dosage</subject><subject>Fibroblast Growth Factor 2 - chemistry</subject><subject>Fibroblast Growth Factor 2 - pharmacology</subject><subject>gelatin</subject><subject>Gelatin - chemistry</subject><subject>General pharmacology</subject><subject>Hydrogels - chemical synthesis</subject><subject>Hydrogels - chemistry</subject><subject>Kinetics</subject><subject>Medical sciences</subject><subject>Mesenchymal Stromal Cells - drug effects</subject><subject>Mesenchymal Stromal Cells - secretion</subject><subject>Methacrylates - chemical synthesis</subject><subject>Methacrylates - chemistry</subject><subject>Methacrylates - metabolism</subject><subject>Mice</subject><subject>nanoparticle</subject><subject>Nanoparticles - chemistry</subject><subject>Nanoparticles - ultrastructure</subject><subject>Particle Size</subject><subject>Pharmaceutical technology. Pharmaceutical industry</subject><subject>Pharmacology. Drug treatments</subject><subject>Water - analysis</subject><issn>0363-9045</issn><issn>1520-5762</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMtu1DAUhi0EokPhAdggb1BXob7ESQxsqopLpQo2sI5OnJOJKycebIchL8Bz49EMIITUlXV8vv-X_RHynLNXkjN9yWQlNSuZZkI3jVLNA7LhSrBC1ZV4SDaHfZEBdUaexHjHGBdaqcfkjGsttGB8Q35-8t_R0a1bje1XRydMI5iwOkjY0x5_pAAz3WKe7UxnmP0OQrLGYaTOQ5-hvU0j7SBaQwfbBd85iIlug9_n-wFM8uE1HXyYlkOJnynMPTUjhLzCYGNui0_JowFcxGen85x8ff_uy_XH4vbzh5vrq9vCyEqlQjQVlHVXd12ttUYjhjxoMGXFGcBgKgallAZkLQwTqLRBg0MvlEJVd3l3Ti6Ovbvgvy0YUzvZaNA5mNEvsa2lrOqy4SKT_Eia4GMMOLS7YCcIa8tZe7Df_mc_Z16c2pduwv5v4qQ7Ay9PAEQDbshujY1_OCGk4nXJMvf2yNn5IA72Pri-TbA6H36H5H3vePNPfERwaTQQsL3zS5iz4Xt-8QtiLrYv</recordid><startdate>200912</startdate><enddate>200912</enddate><creator>Gu, Chunhu</creator><creator>Zheng, Renhong</creator><creator>Yang, Zhifu</creator><creator>Wen, Aidong</creator><creator>Wu, Hong</creator><creator>Zhang, Hui</creator><creator>Yi, Dinghua</creator><general>Informa UK Ltd</general><general>Taylor &amp; Francis</general><scope>IQODW</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>7X8</scope></search><sort><creationdate>200912</creationdate><title>Novel glycidyl methacrylated dextran gelatin nanoparticles loaded with basic fibroblast growth factor: formulation and characteristics</title><author>Gu, Chunhu ; Zheng, Renhong ; Yang, Zhifu ; Wen, Aidong ; Wu, Hong ; Zhang, Hui ; Yi, Dinghua</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c365t-286a47b7bb7999ec2f7b79ac4610aafc60a433ca372c02e59cecefd255e57ba43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Animals</topic><topic>Basic fibrous growth factor</topic><topic>Biological and medical sciences</topic><topic>Bone Marrow Cells - drug effects</topic><topic>Bone Marrow Cells - secretion</topic><topic>Cell Adhesion - drug effects</topic><topic>Cell Proliferation - drug effects</topic><topic>Cells, Cultured</topic><topic>Cross-Linking Reagents</topic><topic>dextran</topic><topic>Dextrans - chemical synthesis</topic><topic>Dextrans - chemistry</topic><topic>Dextrans - metabolism</topic><topic>Drug Carriers - administration &amp; dosage</topic><topic>Drug Carriers - chemistry</topic><topic>Drug Carriers - pharmacology</topic><topic>drug delivery system</topic><topic>Epoxy Compounds - chemical synthesis</topic><topic>Epoxy Compounds - chemistry</topic><topic>Epoxy Compounds - metabolism</topic><topic>Fibroblast Growth Factor 2 - administration &amp; dosage</topic><topic>Fibroblast Growth Factor 2 - chemistry</topic><topic>Fibroblast Growth Factor 2 - pharmacology</topic><topic>gelatin</topic><topic>Gelatin - chemistry</topic><topic>General pharmacology</topic><topic>Hydrogels - chemical synthesis</topic><topic>Hydrogels - chemistry</topic><topic>Kinetics</topic><topic>Medical sciences</topic><topic>Mesenchymal Stromal Cells - drug effects</topic><topic>Mesenchymal Stromal Cells - secretion</topic><topic>Methacrylates - chemical synthesis</topic><topic>Methacrylates - chemistry</topic><topic>Methacrylates - metabolism</topic><topic>Mice</topic><topic>nanoparticle</topic><topic>Nanoparticles - chemistry</topic><topic>Nanoparticles - ultrastructure</topic><topic>Particle Size</topic><topic>Pharmaceutical technology. Pharmaceutical industry</topic><topic>Pharmacology. Drug treatments</topic><topic>Water - analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gu, Chunhu</creatorcontrib><creatorcontrib>Zheng, Renhong</creatorcontrib><creatorcontrib>Yang, Zhifu</creatorcontrib><creatorcontrib>Wen, Aidong</creatorcontrib><creatorcontrib>Wu, Hong</creatorcontrib><creatorcontrib>Zhang, Hui</creatorcontrib><creatorcontrib>Yi, Dinghua</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Drug development and industrial pharmacy</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gu, Chunhu</au><au>Zheng, Renhong</au><au>Yang, Zhifu</au><au>Wen, Aidong</au><au>Wu, Hong</au><au>Zhang, Hui</au><au>Yi, Dinghua</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Novel glycidyl methacrylated dextran gelatin nanoparticles loaded with basic fibroblast growth factor: formulation and characteristics</atitle><jtitle>Drug development and industrial pharmacy</jtitle><addtitle>Drug Dev Ind Pharm</addtitle><date>2009-12</date><risdate>2009</risdate><volume>35</volume><issue>12</issue><spage>1419</spage><epage>1429</epage><pages>1419-1429</pages><issn>0363-9045</issn><eissn>1520-5762</eissn><abstract>Objective: To develop a novel efficient nanoparticulate carrier loaded with basic fibroblast growth factor (bFGF). Methods: Gelatin and glycidyl methacrylate-derivatized dextran (dex-GMA) were cross-linked and polymerized to form interpenetrating polymeric networks. The properties of the nanoparticles (NPs) were investigated as a function of the degree of dex-GMA substitution and the concentration of gelatin used in the preparation of the hydrogels. The morphology was observed with scanning eletromicroscopy and transmission eletromicroscopy. The swelling, degradation, and entrapment efficiency were also determined by dynamic evaluation methods in vitro. The protein release ratio and in vitro release kinetics were evaluated by routine procedure, and the biological activity of bFGF-loaded NPs was studied by cell proliferation assay, cell attachment, and cell function. Results: The NPs have a particle size of 320 ± 20 nm. bFGF was entrapped in the nanoparticles quantitatively (the encapsulation efficiency, 89.6 ± 0.9%). The bFGF in vitro release kinetics fitted to zero-order and Higuchi equations. Proliferation assay, attachment assay, and western blot showed that bFGF NPs had good biological effects on cultured bone marrow mesenchymal stem cells and could achieve a much longer action time than bFGF solution. Conclusion: These results suggested that a novel biodegradable dex-GMA gelatin hydrogel NPs loaded with bFGF could be successfully developed from both dextran- and gelatin-based biomaterials.</abstract><cop>Colchester</cop><pub>Informa UK Ltd</pub><pmid>19929201</pmid><doi>10.3109/03639040902988558</doi><tpages>11</tpages></addata></record>
fulltext fulltext
identifier ISSN: 0363-9045
ispartof Drug development and industrial pharmacy, 2009-12, Vol.35 (12), p.1419-1429
issn 0363-9045
1520-5762
language eng
recordid cdi_proquest_miscellaneous_733674812
source Taylor & Francis:Master (3349 titles); MEDLINE; Taylor & Francis Medical Library - CRKN; Business Source Complete
subjects Animals
Basic fibrous growth factor
Biological and medical sciences
Bone Marrow Cells - drug effects
Bone Marrow Cells - secretion
Cell Adhesion - drug effects
Cell Proliferation - drug effects
Cells, Cultured
Cross-Linking Reagents
dextran
Dextrans - chemical synthesis
Dextrans - chemistry
Dextrans - metabolism
Drug Carriers - administration & dosage
Drug Carriers - chemistry
Drug Carriers - pharmacology
drug delivery system
Epoxy Compounds - chemical synthesis
Epoxy Compounds - chemistry
Epoxy Compounds - metabolism
Fibroblast Growth Factor 2 - administration & dosage
Fibroblast Growth Factor 2 - chemistry
Fibroblast Growth Factor 2 - pharmacology
gelatin
Gelatin - chemistry
General pharmacology
Hydrogels - chemical synthesis
Hydrogels - chemistry
Kinetics
Medical sciences
Mesenchymal Stromal Cells - drug effects
Mesenchymal Stromal Cells - secretion
Methacrylates - chemical synthesis
Methacrylates - chemistry
Methacrylates - metabolism
Mice
nanoparticle
Nanoparticles - chemistry
Nanoparticles - ultrastructure
Particle Size
Pharmaceutical technology. Pharmaceutical industry
Pharmacology. Drug treatments
Water - analysis
title Novel glycidyl methacrylated dextran gelatin nanoparticles loaded with basic fibroblast growth factor: formulation and characteristics
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-13T18%3A47%3A56IST&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=Novel%20glycidyl%20methacrylated%20dextran%20gelatin%20nanoparticles%20loaded%20with%20basic%20fibroblast%20growth%20factor:%20formulation%20and%20characteristics&rft.jtitle=Drug%20development%20and%20industrial%20pharmacy&rft.au=Gu,%20Chunhu&rft.date=2009-12&rft.volume=35&rft.issue=12&rft.spage=1419&rft.epage=1429&rft.pages=1419-1429&rft.issn=0363-9045&rft.eissn=1520-5762&rft_id=info:doi/10.3109/03639040902988558&rft_dat=%3Cproquest_pubme%3E733674812%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=733674812&rft_id=info:pmid/19929201&rfr_iscdi=true