
{"id":40322,"date":"2023-07-18T10:52:11","date_gmt":"2023-07-18T09:52:11","guid":{"rendered":"https:\/\/www.btitrainingcenter.com\/the-dual-responsive-interaction-of-particulated-hyaline-cartilage-and-plasma-rich-in-growth-factors-prgf-in-the-repair-of-cartilage-defects-an-in-vitro-study\/"},"modified":"2025-07-11T11:42:45","modified_gmt":"2025-07-11T10:42:45","slug":"the-dual-responsive-interaction-of-particulated-hyaline-cartilage-and-plasma-rich-in-growth-factors-prgf-in-the-repair-of-cartilage-defects-an-in-vitro-study","status":"publish","type":"post","link":"https:\/\/www.btitrainingcenter.com\/en\/the-dual-responsive-interaction-of-particulated-hyaline-cartilage-and-plasma-rich-in-growth-factors-prgf-in-the-repair-of-cartilage-defects-an-in-vitro-study\/","title":{"rendered":"The Dual-Responsive Interaction of Particulated Hyaline Cartilage and Plasma Rich in Growth Factors (PRGF) in the Repair of Cartilage Defects: An In Vitro Study"},"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=\"eng-abstract\" class=\"abstract-content selected\">\n<div id=\"eng-abstract\" class=\"abstract-content selected\">\n<div id=\"eng-abstract\" class=\"abstract-content selected\">\n<p>The treatment of chondral and osteochondral defects is challenging. These types of lesions are painful and progress to osteoarthritis over time. Tissue engineering offers tools to address this unmet medical need. The use of an autologous cartilage construct consisting of hyaline cartilage chips embedded in plasma rich in growth factors (PRGF) has been proposed as a therapeutic alternative. The purpose of this study was to dig into the potential mechanisms behind the in vitro remodelling process that might explain the clinical success of this technique and facilitate its optimisation. Chondrocyte viability and cellular behaviour over eight weeks of in vitro culture, type II collagen synthesis, the dual delivery of growth factors by hyaline cartilage and PRGF matrix, and the ultrastructure of the construct and its remodelling were characterised. The main finding of this research is that the cartilage fragments embedded in the three-dimensional PRGF scaffold contain viable chondrocytes that are able to migrate into the fibrin network, proliferate and synthesise extracellular matrix after the second week of in vitro culture. The characterization of this three-dimensional matrix is key to unravelling the molecular kinetics responsible for its efficacy.<\/p><\/div>\n<\/div>\n<\/div>\n<p><strong class=\"sub-title\">Keywords:\u00a0<\/strong>PACI; PRGF; chondral defects; chondrocyte; fibrin; growth factors; particulated cartilage; platelet-rich plasma.[\/vc_column_text][vc_empty_space][vc_column_text]<\/p>\n<h3>Figures<\/h3>\n<div id=\"eng-abstract\" class=\"abstract-content selected\">\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\/2023\/07\/ijms-24-11581-g001.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-855]\" title=\"&lt;strong&gt;Figure 1&lt;\/strong&gt; Distribution of median weight and size of cartilage particles. The graphs show the median, mean, and interquartile range of the mean weight of cartilage fragments mixed with PRGF in the different experiments, and the mean size of these fragments. n = 66 wells for weight determination; n = 121 fragments for area calculation.\">\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\/2023\/07\/ijms-24-11581-g001-200x200.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\/2023\/07\/ijms-24-11581-g002.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-855]\" title=\"&lt;strong&gt;Figure 2&lt;\/strong&gt; Cell culture images (A\u2013D) showing the three-dimensional matrix at time zero (A,B) and after four weeks of in vitro culture (C,D). Safranin O staining (E\u2013L) showing representative images at t0 (E,F) and t4 (G,H). Detail at \u00d740 magnification of the matrices at t4 (I\u2013L). Immunohistochemistry of collagen type II (M,N) showing deposition of collagen II between fibrin fibres. Ki-67 immunohistochemistry (O,P) marking the nuclei of the proliferating cells. Original magnifications: \u00d72.5 (A,C,E,G); \u00d710 (F,H); \u00d740 (B,D,I\u2013P). Scale bars: (A,C,E,G) 800 \u00b5m; (F,H) 200 \u00b5m; (B,D,I\u2013P) 50 \u00b5m.\">\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\/2023\/07\/ijms-24-11581-g002-200x200.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\/2023\/07\/ijms-24-11581-g003.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-855]\" title=\"&lt;strong&gt;Figure 3&lt;\/strong&gt; Microphotographs illustrating (A) the background staining of the cartilage fragments with Hoechst 33,342 at low magnification. (B) At higher magnification, the nuclei of the chondrocytes stained with Hoechst 33,342 can be seen dispersed within a fragment of cartilage. (C) Detail of the chondrocytes at higher magnification (Hoechst 33,342). Sequence of microphotographs from two different fields, at low (D,E) and high magnification (F\u2013I), showing the viability of chondrocytes in a sample of 4 weeks in culture. The phase-contrast images (D,G) show the cartilage fragments. In the fluorescence channels, the green colour shows live cells (E,H), while no dead cells were visible in the red channel (F,I). Original magnifications: \u00d72.5 (A); \u00d75 (D\u2013F); \u00d720 (B,C,G\u2013I). Scale bars: (A) 800 \u00b5m; (D\u2013F) 400 \u00b5m; (B,C,G\u2013I) 100 \u00b5m.\">\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\/2023\/07\/ijms-24-11581-g003-200x200.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\/2023\/07\/ijms-24-11581-g004.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-855]\" title=\"&lt;strong&gt;Figure 4&lt;\/strong&gt; Scanning electron microscopy (SEM) images showing (A) cartilage fragments uniformly embedded within the fibrin network of PRGF. (B) Photomicrograph showing an area where the fibrin matrix has broken down and cartilage fragments are visible. (C) Chondrocyte lacunae exposed on the surface of a cartilage fragment. (D) High magnification image showing several empty lacunae and other lacunae with chondrocytes inside. (E) PRGF fibrin network over 4 weeks of incubation (t0\u2013t4) showing that its three-dimensional structure has been retained. Original magnifications: \u00d750 (A); \u00d7200 (C); \u00d7500 (B); \u00d71000 (D); \u00d75000 (top row of E); \u00d710000 (bottom row of E). Scale bars: (A) 500 \u00b5m; (B) 50 \u00b5m; (C) 125 \u00b5m; (D) 25 \u00b5m; (E) top line 10 \u00b5m and bottom line 5 \u00b5m.\">\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\/2023\/07\/ijms-24-11581-g004-200x200.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\/2023\/07\/ijms-24-11581-g005.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-855]\" title=\"&lt;strong&gt;Figure 5&lt;\/strong&gt; Transmission electron microscopy (TEM) images showing (A) platelet aggregates scattered throughout the three-dimensional PRGF matrix after formation and (B) after three weeks of culture. (C) Typical striation of the fibrin fibres after three weeks of in vitro culture. (D\u2013F) Live chondroid cells within the cartilage fragments were observed during the whole incubation period. Scale bars: 500 nm (C); 1 \u00b5m (B); 2 \u00b5m (A,D\u2013F).\">\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\/2023\/07\/ijms-24-11581-g005-200x200.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\/2023\/07\/ijms-24-11581-g006.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-855]\" title=\"&lt;strong&gt;Figure 6&lt;\/strong&gt; Release kinetics of growth factors from the constructs consisting of cartilage fragments and PRGF and PRGF matrix alone. (A) IGF-1, (B) HDGF, COMP release in (C) the PRGF matrix, and in (D) the constructs of cartilage and PRGF. The COMP release kinetics are shown in two separate graphs due to the different order of magnitudes.\">\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\/2023\/07\/ijms-24-11581-g006-200x200.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\/2023\/07\/ijms-24-11581-g007.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-855]\" title=\"&lt;strong&gt;Figure 7&lt;\/strong&gt; Detail of sample collection and preparation. (A) Cartilage samples were examined, and fragments of normal macroscopic appearance (*) were selected. (B) The cartilage was cut into slices, avoiding the subchondral bone. (C) These slices were then mechanically minced into smaller fragments using a scalpel, similar to those used in the clinical setting. (D) Cartilage fragments were kept submerged and homogeneously distributed in Petri dishes. (E) Each dish was photographed independently to obtain the size distribution of the cartilage fragments. (F) Finally, three-dimensional matrices of PRGF and PRGF-embedded cartilage fragments were obtained.\">\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\/2023\/07\/ijms-24-11581-g007-200x200.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\/ijms-24-11581.pdf.pdf\" target=\"_blank\"  class=\"qodef-btn qodef-btn-medium qodef-btn-solid btnWebRosa2\"  >\n    <span class=\"qodef-btn-text\">Download article<\/span>\n    <\/a>[\/vc_column][\/vc_row][vc_row][vc_column][vc_column_text]<\/p><\/div>\n<p>[\/vc_column_text][\/vc_column][\/vc_row]<\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"<p><strong>Int J Mol Sci. 2023 Jul 18;24(14):11581.<\/strong><br \/>\nThe treatment of chondral and osteochondral defects is challenging. These types of lesions are painful and progress to osteoarthritis over time. Tissue engineering offers tools to address this unmet medical need. The use of an autologous cartilage construct consisting of hyaline cartilage chips embedded in plasma rich in growth factors (PRGF) has been proposed as a therapeutic alternative. The purpose of this study was to dig into the potential mechanisms behind the in vitro remodelling process that might explain the clinical success of this technique and facilitate its optimisation. <\/p>\n","protected":false},"author":2002,"featured_media":40313,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[150],"tags":[],"class_list":["post-40322","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-regenerative-medicine"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/posts\/40322","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/users\/2002"}],"replies":[{"embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/comments?post=40322"}],"version-history":[{"count":2,"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/posts\/40322\/revisions"}],"predecessor-version":[{"id":40324,"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/posts\/40322\/revisions\/40324"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/media\/40313"}],"wp:attachment":[{"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/media?parent=40322"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/categories?post=40322"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/en\/wp-json\/wp\/v2\/tags?post=40322"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}