
{"id":42675,"date":"2025-01-01T13:09:16","date_gmt":"2025-01-01T12:09:16","guid":{"rendered":"https:\/\/www.btitrainingcenter.com\/impact-of-increasingly-complex-cell-culture-conditions-on-the-proteome-of-human-periodontal-ligament-stem-cells\/"},"modified":"2025-08-01T13:46:05","modified_gmt":"2025-08-01T12:46:05","slug":"impact-of-increasingly-complex-cell-culture-conditions-on-the-proteome-of-human-periodontal-ligament-stem-cells","status":"publish","type":"post","link":"https:\/\/www.btitrainingcenter.com\/de\/impact-of-increasingly-complex-cell-culture-conditions-on-the-proteome-of-human-periodontal-ligament-stem-cells\/","title":{"rendered":"Impact of increasingly complex cell culture conditions on the proteome of human periodontal ligament stem cells"},"content":{"rendered":"<div class=\"wpb-content-wrapper\"><p>[vc_row content_text_aligment=&#8221;left&#8221; row_background_lines=&#8221;no&#8221;][vc_column][vc_column_text css=&#8221;&#8221;]<\/p>\n<h3 class=\"title\">Abstract<\/h3>\n<div id=\"enc-abstract\" class=\"abstract-content selected\">\n<div id=\"enc-abstract\" class=\"abstract-content selected\">\n<p><strong class=\"sub-title\">Aims:\u00a0<\/strong>Human periodontal ligament stem cells (hPDLSCs) exhibit an enormous potential to regenerate periodontal tissue. However, their translatability to the clinical setting is constrained by technical difficulties in standardizing culture conditions. The aim was to assess complex culture conditions using a proteomic-based protocol to standardize multi-layer hPDLSC cultivation methodology.<\/p>\n<p><strong class=\"sub-title\">Materials and methods:\u00a0<\/strong>hPDLSC-derived constructs were created with varying biological complexity. The simplest constructs were monolayer sheets of hPDLSCs cultured with fetal bovine serum (FBS) or Plasma Rich in Growth Factors supernatant (PRGFsn). The most complex constructs were triple-layered cell structures cultured with PRGFsn, with or without PRGF fibrin membrane (mPRGF). Ultrastructure and proteomic analyses were performed on these constructs.<\/p>\n<p><strong class=\"sub-title\">Results:\u00a0<\/strong>PRGF supernatant improved protein expression related to extracellular matrix, adhesion, proliferation, and migration in hPDLSCs. PRGF fibrin scaffold upregulates proteins for cell activation, respiration, and electron transport. hPDLSCs on fibrin membrane show robust osteogenic potential through differential protein expression (ossification, tissue remodeling, morphogenesis, or cell migration) and overall homeostasis relative to less complex structures.<\/p>\n<p><strong class=\"sub-title\">Conclusions:\u00a0<\/strong>Our data reveal the far-reaching potential of 3-dimensional constructs in combination with PRGF technology in periodontal regenerative applications.<\/div>\n<p><strong class=\"sub-title\">Keywords:\u00a0<\/strong>Human periodontal ligament stem cells; cell sheet technology; osteogenic differentiation; plasma rich in growth factors; proteome; triple-layered cell constructs.<\/div>\n<p>[\/vc_column_text][vc_empty_space][vc_column_text]<\/p>\n<h3>Figures<\/h3>\n<p>[\/vc_column_text][vc_empty_space][\/vc_column][\/vc_row][vc_row content_placement=&#8221;middle&#8221; content_text_aligment=&#8221;center&#8221;][vc_column]<div class=\"qodef-image-gallery qodef-grid-list qodef-disable-bottom-space  qodef-ig-grid-type qodef-four-columns qodef-tiny-space  qodef-image-behavior-lightbox\">\n\t<div class=\"qodef-ig-inner qodef-outer-space\">\n\t\t\t\t\t<div class=\"qodef-ig-image qodef-item-space\">\n\t\t\t\t<div class=\"qodef-ig-image-inner\">\n\t\t\t\t\t\t\t\t\t\t\t<a itemprop=\"image\" class=\"qodef-ig-lightbox\" href=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_UF0001_OC.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-141]\" title=\"&lt;strong&gt;Graphical abstract&lt;\/strong&gt;\">\n\t\t\t\t\t\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_UF0001_OC-200x200.jpg\" alt=\"\" width=\"200\" height=\"200\" \/>\t\t\t\t\t\t\t\t\t\t\t<\/a>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t<\/div>\n\t\t\t\t\t<div class=\"qodef-ig-image qodef-item-space\">\n\t\t\t\t<div class=\"qodef-ig-image-inner\">\n\t\t\t\t\t\t\t\t\t\t\t<a itemprop=\"image\" class=\"qodef-ig-lightbox\" href=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0001_OC.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-141]\" title=\"&lt;strong&gt;Figure 1.&lt;\/strong&gt; PRGF fibrin membrane (mPRGF) preparation. (a) PRGF obtained after blood centrifugation (b) is ativated with calcium chloride and afterward the clot is formed. (c) This clot is then transferred into a fibrin membrane shaper and (d) a weigth is placed on top of it. (e) Finally, the mPRGF is obtained by pressing for 5\u2009minutes. Scale bar: 2\u2009cm.\">\n\t\t\t\t\t\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0001_OC-200x200.jpg\" alt=\"\" width=\"200\" height=\"200\" \/>\t\t\t\t\t\t\t\t\t\t\t<\/a>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t<\/div>\n\t\t\t\t\t<div class=\"qodef-ig-image qodef-item-space\">\n\t\t\t\t<div class=\"qodef-ig-image-inner\">\n\t\t\t\t\t\t\t\t\t\t\t<a itemprop=\"image\" class=\"qodef-ig-lightbox\" href=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0002_OC.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-141]\" title=\"&lt;strong&gt;Figure 2.&lt;\/strong&gt; Ultrastructural analysis of the triple-layered cell constructs with mPRGF. SEM microphotographs showing (a, b) The fibrin mesh of mPRGF at the bottom of the construct at different magnifications and (c) A uniform cell coverage on the other side of this structure. (d) The complexity of this composite is shown in a SEM image of a transversal section where the red arrow indicates hPDLSCs and the black one points at the fibrin net of mPRGF. Scale is the distance between the first and last points above the scale value in \u03bcm. On the bottom of the figure, a sequence of confocal microscopy captions highlights (e) The fibrin mesh, (f) Actin microfilaments of hPDLSCs and platelet aggregates and (G) hPDLSCs nuclei. (H) Image obtained by overlaying the fluorescence signal from the 3 channels, confirming the cell integration with the mPRGF fibrin membrane. Fibrin mesh, actin microfilaments, and hPDLSC nuclei are fluorescently labeled in green, red, and blue, respectively. Scale bar: 50\u2009\u03bcm.\">\n\t\t\t\t\t\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0002_OC-200x200.jpg\" alt=\"\" width=\"200\" height=\"200\" \/>\t\t\t\t\t\t\t\t\t\t\t<\/a>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t<\/div>\n\t\t\t\t\t<div class=\"qodef-ig-image qodef-item-space\">\n\t\t\t\t<div class=\"qodef-ig-image-inner\">\n\t\t\t\t\t\t\t\t\t\t\t<a itemprop=\"image\" class=\"qodef-ig-lightbox\" href=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0003_OC.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-141]\" title=\"&lt;strong&gt;Figure 3.&lt;\/strong&gt; Experimental design and proteomic analysis of hPdlscs. (a) Schematic representation of the experimental setup. hPDLSCs were grown under five different conditions (condition 1\u20135) and two replicas of each condition were prepared. Upon protein extraction and digestion, peptides were labeled with TMT 10-plex reagent, combined and subjected to high pH fractionation before LC-MS\/MS analysis. (b) Principal component analysis (PCA) of proteomics data.\">\n\t\t\t\t\t\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0003_OC-200x200.jpg\" alt=\"\" width=\"200\" height=\"200\" \/>\t\t\t\t\t\t\t\t\t\t\t<\/a>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t<\/div>\n\t\t\t\t\t<div class=\"qodef-ig-image qodef-item-space\">\n\t\t\t\t<div class=\"qodef-ig-image-inner\">\n\t\t\t\t\t\t\t\t\t\t\t<a itemprop=\"image\" class=\"qodef-ig-lightbox\" href=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0004_OC.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-141]\" title=\"&lt;strong&gt;Figure 4.&lt;\/strong&gt; GO analysis of differentially expressed proteins. (a) Enriched GO terms in hPDLSC monolayer cultures grown on PRGFsn or on FBS. (b) Enriched GO terms in hPDLSC triple-layer cultures grown on PRGFsn compared to hPDLSC monolayer cultures grown on PRGFsn. The length of the line indicates the fold enrichment of the GO term, whereas the size of the dot at the end of the line represents the number of genes included in the term. Only GOs of the upregulated subsets are shown, as the downregulated subsets did not exhibit any enriched GO categories. As a background, the total number of proteins identified in each of the comparisons were used.\">\n\t\t\t\t\t\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0004_OC-200x200.jpg\" alt=\"\" width=\"200\" height=\"200\" \/>\t\t\t\t\t\t\t\t\t\t\t<\/a>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t<\/div>\n\t\t\t\t\t<div class=\"qodef-ig-image qodef-item-space\">\n\t\t\t\t<div class=\"qodef-ig-image-inner\">\n\t\t\t\t\t\t\t\t\t\t\t<a itemprop=\"image\" class=\"qodef-ig-lightbox\" href=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0005_OC.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-141]\" title=\"&lt;strong&gt;Figure 5.&lt;\/strong&gt; GO analysis of differentially expressed proteins in hPDLSCs grown as triple-layered constructs. GO annotations of significantly enriched terms among (a) Downregulated and (b) Upregulated proteins are shown in the comparison of condition 4 (in the presence of the fibrin membrane) vs condition 3 (in the absence of the fibrin membrane). As a background, the total number of proteins identified in each of the comparisons were used.\">\n\t\t\t\t\t\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0005_OC-200x200.jpg\" alt=\"\" width=\"200\" height=\"200\" \/>\t\t\t\t\t\t\t\t\t\t\t<\/a>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t<\/div>\n\t\t\t\t\t<div class=\"qodef-ig-image qodef-item-space\">\n\t\t\t\t<div class=\"qodef-ig-image-inner\">\n\t\t\t\t\t\t\t\t\t\t\t<a itemprop=\"image\" class=\"qodef-ig-lightbox\" href=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0006_OC.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-141]\" title=\"&lt;strong&gt;Figure 6.&lt;\/strong&gt; GO analysis of differentially expressed proteins in hPDLSCs grown as triple-layered constructs on differentiation medium. GO annotations of significantly enriched terms among (a) Downregulated and (b) Upregulated proteins are shown in the comparison of condition 5 (in the presence osteogenic inducers) vs condition 3 (in the absence of osteogenic inducers). As a background, the total number of proteins identified in each of the comparisons were used.\">\n\t\t\t\t\t\t\t\t\t\t\t<img loading=\"lazy\" decoding=\"async\" src=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/IRME_A_2445931_F0006_OC-200x200.jpg\" alt=\"\" width=\"200\" height=\"200\" \/>\t\t\t\t\t\t\t\t\t\t\t<\/a>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t<\/div>\n\t\t\t<\/div>\n<\/div>[vc_empty_space height=&#8221;50px&#8221;][\/vc_column][\/vc_row][vc_row][vc_column]<a itemprop=\"url\" href=\"https:\/\/www.btitrainingcenter.com\/wp-content\/uploads\/2021\/08\/15-IRME_20_2445931.pdf.pdf\" target=\"_blank\"  class=\"qodef-btn qodef-btn-medium qodef-btn-solid btnWebRosa2\"  >\n    <span class=\"qodef-btn-text\">Artikel herunterladen<\/span>\n    <\/a>[\/vc_column][\/vc_row]<\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"<p><strong>Regen Med. 2025 Jan;20(1):21-34.<\/strong><br \/>\nHuman periodontal ligament stem cells (hPDLSCs) exhibit an enormous potential to regenerate periodontal tissue. However, their translatability to the clinical setting is constrained by technical difficulties in standardizing culture conditions. The aim was to assess complex culture conditions using a proteomic-based protocol to standardize multi-layer hPDLSC cultivation methodology.<\/p>\n","protected":false},"author":2002,"featured_media":42619,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[156],"tags":[],"class_list":["post-42675","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-regenerative-medizin"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/posts\/42675","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/users\/2002"}],"replies":[{"embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/comments?post=42675"}],"version-history":[{"count":1,"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/posts\/42675\/revisions"}],"predecessor-version":[{"id":42676,"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/posts\/42675\/revisions\/42676"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/media\/42619"}],"wp:attachment":[{"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/media?parent=42675"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/categories?post=42675"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/de\/wp-json\/wp\/v2\/tags?post=42675"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}