
{"id":48133,"date":"2021-02-26T16:03:19","date_gmt":"2021-02-26T15:03:19","guid":{"rendered":"https:\/\/www.btitrainingcenter.com\/the-effect-of-plasma-rich-in-growth-factors-on-microglial-migration-macroglial-gliosis-and-proliferation-and-neuronal-survival\/"},"modified":"2026-04-13T17:44:53","modified_gmt":"2026-04-13T16:44:53","slug":"the-effect-of-plasma-rich-in-growth-factors-on-microglial-migration-macroglial-gliosis-and-proliferation-and-neuronal-survival","status":"publish","type":"post","link":"https:\/\/www.btitrainingcenter.com\/fr\/the-effect-of-plasma-rich-in-growth-factors-on-microglial-migration-macroglial-gliosis-and-proliferation-and-neuronal-survival\/","title":{"rendered":"The Effect of Plasma Rich in Growth Factors on Microglial Migration, Macroglial Gliosis and Proliferation, and Neuronal Survival"},"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>Plasma rich in growth factors (PRGF) is a subtype of platelet-rich plasma that has being employed in the clinic due to its capacity to accelerate tissue regeneration. Autologous PRGF has been used in ophthalmology to repair a range of retinal pathologies with some efficiency. In the present study, we have explored the role of PRGF and its effect on microglial motility, as well as its possible pro-inflammatory effects. Organotypic cultures from adult pig retinas were used to test the effect of the PRGF obtained from human as well as pig blood. Microglial migration, as well as gliosis, proliferation and the survival of retinal ganglion cells (RGCs) were analyzed by immunohistochemistry. The cytokines present in these PRGFs were analyzed by multiplex ELISA. In addition, we set out to determine if blocking some of the inflammatory components of PRGF alter its effect on microglial migration. In organotypic cultures, PRGF induces microglial migration to the outer nuclear layers as a sign of inflammation. This phenomenon could be due to the presence of several cytokines in PRGF that were quantified here, such as the major pro-inflammatory cytokines IL-1\u03b2, IL-6 and TNF\u03b1. Heterologous PRGF (human) and longer periods of cultured (3 days) induced more microglia migration than autologous porcine PRGF. Moreover, the migratory effect of microglia was partially mitigated by: 1) heat inactivation of the PRGF; 2) the presence of dexamethasone; or 3) anti-cytokine factors. Furthermore, PRGF seems not to affect gliosis, proliferation or RGC survival in organotypic cultures of adult porcine retinas. PRGF can trigger an inflammatory response as witnessed by the activation of microglial migration in the retina. This can be prevented by using autologous PRGF or if this is not possible due to autoimmune diseases, by mitigating its inflammatory effect. In addition, PRGF does not increase either the proliferation rate of microglial cells or the survival of neurons. We cannot discard the possible positive effect of microglial cells on retinal function. Further studies should be performed to warrant the use of PRGF on the nervous system.<\/p>\n<\/div>\n<p><strong class=\"sub-title\">Keywords:\u00a0<\/strong>cytokines; glia; inflammation; microglia; neuron; ophthalmology; plasma rich in growth factors; retina.<\/p>\n<\/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\/fphar-12-606232-g001.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-490]\" title=\"&lt;strong&gt;Figure 1.&lt;\/strong&gt; Scheme to analyze the properties of the PRGF in organotypic cultures of adult porcine retinas. First, PRGF was obtained from human and porcine blood by separation and plasma extraction. Subsequently, the effect of the PRGF was evaluated on retinal explants by assessing microglial activation, gliosis and proliferation, and RGC survival. In addition, the cytokines in the human and porcine PRGF was quantified using a Q-plex assay. Finally, given the implication of PRGF in promoting inflammation, the effects of exposing the explants exposed to dexamethasone (Dex), cytokines (Cyt: IL-1\u03b2, IL-6 and TNF\u03b1) or cytokines plus antibodies raised against these cytokines (anti-cyt: anti-IL-1\u03b2, IL-6 and TNF\u03b1) were also evaluated. Finally, we examined the effect of heat-inactivated PRGF, as well as PRGF combined with Dex or anti-cyt, analyze their effects on microglial migration.\">\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\/fphar-12-606232-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\/2021\/08\/fphar-12-606232-g002.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-490]\" title=\"&lt;strong&gt;Figure 2.&lt;\/strong&gt; Microglia in a control retina and in organotypic retinal cultures maintained in the presence or absence of human PRGF. Representative images of an adult pig retina from a control eye (A), or of explants of organotypic cultures maintained in the absence (B,D) or in the presence (C,E) of 10% human PRGF. The explants were cultured for 1 (B,C) or 3 (D,E) days, labeling the microglial cells with antibodies against Iba1 (red, anti-Iba1 rabbit antibody, 1:2,000, WAKO, Osaka, Japan) and the nuclei with DAPI (blue). The white arrows point to microglial cells in the ONL: ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner nuclear layer; GCL, ganglion cell layer; NFL, nerve fiber layer. Scale bar = 100 \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\/fphar-12-606232-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\/2021\/08\/fphar-12-606232-g003.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-490]\" title=\"&lt;strong&gt;Figure 3.&lt;\/strong&gt; Quantification of microglia migration in the presence of autologous and heterologous porcine PRGF or human PRGF. Microglial migration on day 1 and 3 in retinal explants treated with 10% PRGF from the same pig as the retinal explant (autologous) (n = 5), from another pig (heterologous) (n = 5) or from human blood (n = 5). Microglial migration is represented as the percentage of microglial cells located in the outer nuclear layer (ONL) relative to the total number of microglia: *p &lt; 0.05, **p &lt; 0.01 relative to the control.\">\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\/fphar-12-606232-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\/2021\/08\/fphar-12-606232-g004.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-490]\" title=\"&lt;strong&gt;Figure 4.&lt;\/strong&gt; Quantification of microglial migration in the presence of human PRGF, inhibiting its inflammatory effect. Microglial migration is represented as the proportion of microglial cells located in the outer nuclear layer (ONL) relative to the total number of microglia, reflecting the migration in control conditions (n = 5), and in the presence of dexamethasone (Dex: n =), cytokines (cyt, IL-1\u03b2, IL-6 and TNF\u03b1: n = 4), cytokines plus anti-cytokine antibodies (anti-cyt: n = 3), with 10% human PRGF (Human: n = 5). The inflammatory effect of PRGF (Human) on microglial migration was inhibited by PRGF heat-inactivation (Human inactive: n = 5), or by adding Dex (n = 5) or anti-cytokine antibodies (n = 5). Significant differences relative to the control conditions and the presence of human PRGF are shown: *p &lt; 0.05, **p &lt; 0.01.\">\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\/fphar-12-606232-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\/2021\/08\/fphar-12-606232-g005.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-490]\" title=\"&lt;strong&gt;Figure 5.&lt;\/strong&gt; Gliosis in control retinas and in organotypic retinal cultures maintained in the presence or absence of human PRGF. Representative images of adult pig retinas from a control eye (n = 5: A), or from explants of organotypic cultures maintained in the absence (n = 4: B) or presence (n = 4: C) of 10% human PRGF. The GFAP in astrocytes and M\u00fcller cells was immunolabeled (green, anti-GFAP mouse antibody, 1:1,000, Sigma, Steinheim, Germany), and the nuclei were labeled with DAPI (blue): ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner nuclear layer; GCL, ganglion cell layer; NFL, nerve fiber layer. Scale bar = 100 \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\/fphar-12-606232-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\/2021\/08\/fphar-12-606232-g006.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-490]\" title=\"&lt;strong&gt;Figure 6.&lt;\/strong&gt; Proliferative cells in control retinas and in organotypic retinal cultures maintained in the presence or absence of human PRGF. Representative images of adult pig retinas from a control eye (n = 5: A), and organotypic cultures maintained in the absence (n = 4: B) or presence (n = 4: C) of 10% human PRGF. The edge of the explant is also shown (C). The proliferative cells were labeled with antibodies against Ki67 (green, anti-Ki67 rabbit antibody, 1:100, Abcam, Cambridge, England) and the nuclei with DAPI (blue). The white arrows indicate cells in an active phase of the cell cycle. The edge of the explant is shown to demonstrate the absence of proliferation (left in picture C): ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner nuclear layer; GCL, ganglion cell layer. Scale bar = 100 \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\/fphar-12-606232-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\/2021\/08\/fphar-12-606232-g007.jpg\" data-rel=\"prettyPhoto[image_gallery_pretty_photo-490]\" title=\"&lt;strong&gt;Figure 7.&lt;\/strong&gt; Quantification of RGCs per millimeter of retina in controls retinas and in explants maintained in the presence or absence of human PRGF. The number of RGCs per linear mm of retina are shown in control retinas (n = 5) and in organotypic cultures at day 3, maintained in the presence (n = 4) or absence of 10% human PRGF (n = 4). Significant differences between the controls and explants are indicated: *p &lt; 0.05.\">\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\/fphar-12-606232-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\/19.-fphar-12-606232.pdf.pdf\" target=\"_blank\"  class=\"qodef-btn qodef-btn-medium qodef-btn-solid btnWebRosa2\"  >\n    <span class=\"qodef-btn-text\">T\u00e9l\u00e9charger l&#039;article<\/span>\n    <\/a>[\/vc_column][\/vc_row]<\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"<p><strong>Front Pharmacol. 2021 Feb 26;12:606232.<\/strong><br \/>\nPlasma rich in growth factors (PRGF) is a subtype of platelet-rich plasma that has being employed in the clinic due to its capacity to accelerate tissue regeneration. Autologous PRGF has been used in ophthalmology to repair a range of retinal pathologies with some efficiency. In the present study, we have explored the role of PRGF and its effect on microglial motility, as well as its possible pro-inflammatory effects. Organotypic cultures from adult pig retinas were used to test the effect of the PRGF obtained from human as well as pig blood. Microglial migration, as well as gliosis, proliferation and the survival of retinal ganglion cells (RGCs) were analyzed by immunohistochemistry.<\/p>\n","protected":false},"author":2002,"featured_media":48065,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[155],"tags":[],"class_list":["post-48133","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-medecine-regenerative"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/posts\/48133","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/users\/2002"}],"replies":[{"embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/comments?post=48133"}],"version-history":[{"count":1,"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/posts\/48133\/revisions"}],"predecessor-version":[{"id":48134,"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/posts\/48133\/revisions\/48134"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/media\/48065"}],"wp:attachment":[{"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/media?parent=48133"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/categories?post=48133"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.btitrainingcenter.com\/fr\/wp-json\/wp\/v2\/tags?post=48133"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}