Characterization of Porcine Aortic Valvular Interstitial Cell ‘Calcified’ Nodules
Publication Date
October 26, 2012
Journal
PLOS ONE
Authors
Kristy L. Cloyd, Ismail El Hamamsy, Suwimon Boonrungsiman, Martin Hedegaard, et al
Volume
7
Issue
10
Pages
e48154
DOI
https://dx.plos.org/10.1371/journal.pone.0048154
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0048154
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/23110195
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3482191
Europe PMC
http://europepmc.org/abstract/MED/23110195
Web of Science
000310262500042
Scopus
84868152490
Mendeley
http://www.mendeley.com/research/characterization-porcine-aortic-valvular-interstitial-cell-calcified-nodules
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Mendeley | Further Information

{"title"=>"Characterization of Porcine Aortic Valvular Interstitial Cell 'Calcified' Nodules", "type"=>"journal", "authors"=>[{"first_name"=>"Kristy L.", "last_name"=>"Cloyd", "scopus_author_id"=>"37109353900"}, {"first_name"=>"Ismail", "last_name"=>"El-Hamamsy", "scopus_author_id"=>"6507972699"}, {"first_name"=>"Suwimon", "last_name"=>"Boonrungsiman", "scopus_author_id"=>"28067526500"}, {"first_name"=>"Martin", "last_name"=>"Hedegaard", "scopus_author_id"=>"24779481700"}, {"first_name"=>"Eileen", "last_name"=>"Gentleman", "scopus_author_id"=>"6506866889"}, {"first_name"=>"Padmini", "last_name"=>"Sarathchandra", "scopus_author_id"=>"6603419148"}, {"first_name"=>"Francesca", "last_name"=>"Colazzo", "scopus_author_id"=>"26038986200"}, {"first_name"=>"Molly M.", "last_name"=>"Gentleman", "scopus_author_id"=>"9843715100"}, {"first_name"=>"Magdi H.", "last_name"=>"Yacoub", "scopus_author_id"=>"36041927900"}, {"first_name"=>"Adrian H.", "last_name"=>"Chester", "scopus_author_id"=>"7101723560"}, {"first_name"=>"Molly M.", "last_name"=>"Stevens", "scopus_author_id"=>"7402243195"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"365952333", "sgr"=>"84868152490", "issn"=>"19326203", "pmid"=>"23110195", "scopus"=>"2-s2.0-84868152490", "doi"=>"10.1371/journal.pone.0048154", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)"}, "id"=>"f7fe3107-37b3-34d6-8ad7-e5133ec40848", "abstract"=>"Valve interstitial cells populate aortic valve cusps and have been implicated in aortic valve calcification. Here we investigate a common in vitro model for aortic valve calcification by characterizing nodule formation in porcine aortic valve interstitial cells (PAVICs) cultured in osteogenic (OST) medium supplemented with transforming growth factor beta 1 (TGF-β1). Using a combination of materials science and biological techniques, we investigate the relevance of PAVICs nodules in modeling the mineralised material produced in calcified aortic valve disease. PAVICs were grown in OST medium supplemented with TGF-β1 (OST+TGF-β1) or basal (CTL) medium for up to 21 days. Murine calvarial osteoblasts (MOBs) were grown in OST medium for 28 days as a known mineralizing model for comparison. PAVICs grown in OST+TGF-β1 produced nodular structures staining positive for calcium content; however, micro-Raman spectroscopy allowed live, noninvasive imaging that showed an absence of mineralized material, which was readily identified in nodules formed by MOBs and has been identified in human valves. Gene expression analysis, immunostaining, and transmission electron microscopy imaging revealed that PAVICs grown in OST+TGF-β1 medium produced abundant extracellular matrix via the upregulation of the gene for Type I Collagen. PAVICs, nevertheless, did not appear to further transdifferentiate to osteoblasts. Our results demonstrate that 'calcified' nodules formed from PAVICs grown in OST+TGF-β1 medium do not mineralize after 21 days in culture, but rather they express a myofibroblast-like phenotype and produce a collagen-rich extracellular matrix. This study clarifies further the role of PAVICs as a model of calcification of the human aortic valve.", "link"=>"http://www.mendeley.com/research/characterization-porcine-aortic-valvular-interstitial-cell-calcified-nodules", "reader_count"=>47, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>5, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>25, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>2, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Researcher"=>5, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>25, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>2, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>13, "Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>2, "Materials Science"=>7, "Medicine and Dentistry"=>5, "Agricultural and Biological Sciences"=>12, "Chemical Engineering"=>1, "Chemistry"=>4}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>13}, "Materials Science"=>{"Materials Science"=>7}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Chemistry"=>{"Chemistry"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>12}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>3}, "Chemical Engineering"=>{"Chemical Engineering"=>1}}, "reader_count_by_country"=>{"United States"=>2}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/552461"], "description"=>"<p><b>A,D,G -</b> Phase contrast images of cultured PAVICs grown in CTL medium for 21 days, PAVICs grown in OST+TGF-β1 medium for 21 days, and MOBs grown in OST medium for 21 days respectively (scale = 100 µm). <b>B,E,H -</b> Alizarin Red S staining negative for PAVICs grown in CTL medium for 21 days, positive for PAVICs grown in OST+TGF-β1 medium for 21 days, and positive for MOBs grown in OST medium respectively (scale = 100 µm). <b>C,F,I</b> - SEM images of cultured PAVICs grown in CTL medium for 21 days, PAVICs grown in OST+TGF-β1 medium for 21 days, and MOBs grown in OST medium for 21 days respectively (scale = 50 µm).</p>", "links"=>[], "tags"=>["morphology", "staining", "pavics", "mobs"], "article_id"=>222942, "categories"=>["Physics"], "users"=>["Kristy L. Cloyd", "Ismail El-Hamamsy", "Suwimon Boonrungsiman", "Martin Hedegaard", "Eileen Gentleman", "Padmini Sarathchandra", "Francesca Colazzo", "Molly M. Gentleman", "Magdi H. Yacoub", "Adrian H. Chester", "Molly M. Stevens"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048154.g001", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Micrographs_showing_the_morphology_and_staining_of_PAVICs_and_MOBs_in_culture_/222942", "title"=>"Micrographs showing the morphology and staining of PAVICs and MOBs in culture.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-26 00:49:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/552980"], "description"=>"<p><b>A -</b> Raman spectra comparing a representative MOBs mineralized nodule spectrum (black), PAVICs grown in CTL media mean spectrum (dark grey), and PAVICs cultured in OST+TGF-β1 medium mean spectrum (shaded box over entire spectrum). Vertical shaded areas highlight the 960 cm<sup>−1</sup> apatite peak and the 1070 cm<sup>−1</sup> carbonate peak spectral range. <b>B -</b> Higher magnification of the PAVICs mean spectrum grown in CTL (dark grey) and in OST+TGF-β1 medium (light grey). The red spectrum is the difference between the two PAVICs mean spectra. Shaded red bands highlight spectral ranges which discriminate between groups. <b>C -</b> Scatter plot showing group separation between PAVICs grown in CTL medium (red triangles) vs. PAVICs grown in OST+TGF-β1 medium (green circles). <b>D -</b> Latent variables loadings used in the interval partial least squares discriminant analysis (iPLS-DA) model, bands highlighted are spectral ranges which contributed to the distinction between groups.</p>", "links"=>[], "tags"=>["spectroscopy", "pavics", "mobs", "compared", "univariate", "multivariate"], "article_id"=>223465, "categories"=>["Physics"], "users"=>["Kristy L. Cloyd", "Ismail El-Hamamsy", "Suwimon Boonrungsiman", "Martin Hedegaard", "Eileen Gentleman", "Padmini Sarathchandra", "Francesca Colazzo", "Molly M. Gentleman", "Magdi H. Yacoub", "Adrian H. Chester", "Molly M. Stevens"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048154.g004", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Raman_spectroscopy_of_PAVICs_and_MOBs_in_culture_compared_through_univariate_and_multivariate_statistical_analysis_/223465", "title"=>"Raman spectroscopy of PAVICs and MOBs in culture compared through univariate and multivariate statistical analysis.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-26 00:57:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/553110"], "description"=>"<p>Inorganic peaks are italicized. * indicates collagen assignments.</p>", "links"=>[], "tags"=>["bands", "contributing", "pavics", "grown", "ctl", "mobs", "mineralised"], "article_id"=>223586, "categories"=>["Physics"], "users"=>["Kristy L. Cloyd", "Ismail El-Hamamsy", "Suwimon Boonrungsiman", "Martin Hedegaard", "Eileen Gentleman", "Padmini Sarathchandra", "Francesca Colazzo", "Molly M. Gentleman", "Magdi H. Yacoub", "Adrian H. Chester", "Molly M. Stevens"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048154.t001", "stats"=>{"downloads"=>7, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Raman_bands_contributing_to_the_distinction_between_PAVICs_grown_in_OST_TGF_946_1_media_CTL_media_and_MOBs_mineralised_nodules_/223586", "title"=>"Raman bands contributing to the distinction between PAVICs grown in OST+TGF-β1 media, CTL media and MOBs mineralised nodules.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-10-26 00:59:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/552602"], "description"=>"<p><b>A -</b> Gene expression of collagen I (* = p<0.05, ** = p<0.0001, *** = p<0.05;**** = p<0.001) and <b>B -</b> Osteocalcin (bone gamma-carboxyglutamate protein (BGLAP)) (* = p<0.05; ** = p<0.001; *** = p<0.0001) comparing PAVICs expression at day 7 (D7), day 14 (D14), and day 21 (D21) in both OST+TGF-β1 and CTL media. <b>C –</b> Cross sectional plane of cultured nodules used for the D,E,F,G,H, and I. <b>D,E -</b> Modified Verhoeff van Gieson stain [purple – cell nuclei, pink – collagen, black-elastin] of a cross section of PAVICs cultured in CTL media for 21 days and PAVICs cultured in OST+TGF-β1 media for 21 days respectively (scale = 50 µm). <b>F,G -</b> Peroxidase stain with Sirius red staining [brown - αSMA, red – collagen] for PAVICs grown in CTL medium for 21 days and PAVICs grown in OST+TGF-β1 medium for 21 days respectively (scale = 50 µm) <b>H,I -</b> Fluorescence staining [red - αSMA, blue - cell nuclei] of a PAVICs nodule cultured for 21 days in CTL medium and a PAVICs nodule cultured in OST+TGF-β1 medium for 21 days respectively (scale = 50 µm).</p>", "links"=>[], "tags"=>["sectional", "staining", "cultured"], "article_id"=>223083, "categories"=>["Physics"], "users"=>["Kristy L. Cloyd", "Ismail El-Hamamsy", "Suwimon Boonrungsiman", "Martin Hedegaard", "Eileen Gentleman", "Padmini Sarathchandra", "Francesca Colazzo", "Molly M. Gentleman", "Magdi H. Yacoub", "Adrian H. Chester", "Molly M. Stevens"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048154.g002", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Gene_expression_and_cross_sectional_staining_of_cultured_PAVICs_/223083", "title"=>"Gene expression and cross sectional staining of cultured PAVICs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-26 00:51:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/552782"], "description"=>"<p>PAVICs were grown in <b>A -</b> CTL medium and <b>B -</b> OST+TGF-β1 medium (VIC – valvular interstitial cell, ECM – extracellular matrix) (scale = 2 µm).</p>", "links"=>[], "tags"=>["electron", "micrographs", "cultured"], "article_id"=>223266, "categories"=>["Physics"], "users"=>["Kristy L. Cloyd", "Ismail El-Hamamsy", "Suwimon Boonrungsiman", "Martin Hedegaard", "Eileen Gentleman", "Padmini Sarathchandra", "Francesca Colazzo", "Molly M. Gentleman", "Magdi H. Yacoub", "Adrian H. Chester", "Molly M. Stevens"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048154.g003", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Transmission_Electron_Micrographs_of_cultured_PAVICs_/223266", "title"=>"Transmission Electron Micrographs of cultured PAVICs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-26 00:54:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/294918", "https://ndownloader.figshare.com/files/295117", "https://ndownloader.figshare.com/files/295193"], "description"=>"<div><p>Valve interstitial cells populate aortic valve cusps and have been implicated in aortic valve calcification. Here we investigate a common <em>in vitro</em> model for aortic valve calcification by characterizing nodule formation in porcine aortic valve interstitial cells (PAVICs) cultured in osteogenic (OST) medium supplemented with transforming growth factor beta 1 (TGF-β1). Using a combination of materials science and biological techniques, we investigate the relevance of PAVICs nodules in modeling the mineralised material produced in calcified aortic valve disease. PAVICs were grown in OST medium supplemented with TGF-β1 (OST+TGF-β1) or basal (CTL) medium for up to 21 days. Murine calvarial osteoblasts (MOBs) were grown in OST medium for 28 days as a known mineralizing model for comparison. PAVICs grown in OST+TGF-β1 produced nodular structures staining positive for calcium content; however, micro-Raman spectroscopy allowed live, noninvasive imaging that showed an absence of mineralized material, which was readily identified in nodules formed by MOBs and has been identified in human valves. Gene expression analysis, immunostaining, and transmission electron microscopy imaging revealed that PAVICs grown in OST+TGF-β1 medium produced abundant extracellular matrix via the upregulation of the gene for Type I Collagen. PAVICs, nevertheless, did not appear to further transdifferentiate to osteoblasts. Our results demonstrate that <b>‘</b>calcified’ nodules formed from PAVICs grown in OST+TGF-β1 medium do not mineralize after 21 days in culture, but rather they express a myofibroblast-like phenotype and produce a collagen-rich extracellular matrix. This study clarifies further the role of PAVICs as a model of calcification of the human aortic valve.</p> </div>", "links"=>[], "tags"=>["characterization", "porcine", "aortic", "valvular", "interstitial", "nodules"], "article_id"=>118071, "categories"=>["Physics"], "users"=>["Kristy L. Cloyd", "Ismail El-Hamamsy", "Suwimon Boonrungsiman", "Martin Hedegaard", "Eileen Gentleman", "Padmini Sarathchandra", "Francesca Colazzo", "Molly M. Gentleman", "Magdi H. Yacoub", "Adrian H. Chester", "Molly M. Stevens"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0048154.s001", "https://dx.doi.org/10.1371/journal.pone.0048154.s002", "https://dx.doi.org/10.1371/journal.pone.0048154.s003"], "stats"=>{"downloads"=>6, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Characterization_of_Porcine_Aortic_Valvular_Interstitial_Cell_Calcified_Nodules__/118071", "title"=>"Characterization of Porcine Aortic Valvular Interstitial Cell ‘Calcified’ Nodules", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-10-26 02:14:31"}

PMC Usage Stats | Further Information

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Relative Metric

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