Enamel Matrix Derivative Decreases Pyroptosis-Related Genes in Macrophages

<b>Background</b>: Pyroptosis is a caspase-dependent catabolic process relevant to periodontal disorders for which inflammation is central to the pathophysiology of the disease. Although enamel matrix derivative (EMD) has been applied to support periodontal regeneration, its capacity to...

Full description

Bibliographic Details
Main Authors: Mariane Beatriz Sordi, Ariadne Cristiane Cabral da Cruz, Layla Panahipour, Reinhard Gruber
Format: Article
Language:English
Published: MDPI AG 2022-05-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:https://www.mdpi.com/1422-0067/23/9/5078
_version_ 1797504448817266688
author Mariane Beatriz Sordi
Ariadne Cristiane Cabral da Cruz
Layla Panahipour
Reinhard Gruber
author_facet Mariane Beatriz Sordi
Ariadne Cristiane Cabral da Cruz
Layla Panahipour
Reinhard Gruber
author_sort Mariane Beatriz Sordi
collection DOAJ
description <b>Background</b>: Pyroptosis is a caspase-dependent catabolic process relevant to periodontal disorders for which inflammation is central to the pathophysiology of the disease. Although enamel matrix derivative (EMD) has been applied to support periodontal regeneration, its capacity to modulate the expression of pyroptosis-related genes remains unknown. Considering EMD has anti-inflammatory properties and pyroptosis is linked to the activation of the inflammasome in chronic periodontitis, the question arises whether EMD could reduce pyroptosis signalling. <b>Methods</b>: To answer this question, primary macrophages obtained from murine bone marrow and RAW 264.7 macrophages were primed with EMD before being challenged by lipopolysaccharide (LPS). Cells were then analysed for pyroptosis-signalling components by gene expression analyses, interleukin-1β (IL-1β) immunoassay, and the detection of caspase-1 (CAS1). The release of mitochondrial reactive oxygen species (ROS) was also detected. <b>Results</b>: We report here that EMD, like the inflammasome (NLRP3) and CAS1 specific inhibitors—MCC950 and Ac-YVAD-cmk, respectively—lowered the LPS-induced expression of NLRP3 in primary macrophages (EMD: <i>p</i> = 0.0232; MCC950: <i>p</i> = 0.0426; Ac-YVAD-cmk: <i>p</i> = 0.0317). EMD further reduced the LPS-induced expression of NLRP3 in RAW 264.7 cells (<i>p</i> = 0.0043). There was also a reduction in CAS1 and IL-1β in RAW 264.7 macrophages on the transcriptional level (<i>p</i> = 0.0598; <i>p</i> = 0.0283; respectively), in IL-1β protein release (<i>p</i> = 0.0313), and CAS1 activity. Consistently, EMD, like MCC950 and Ac-YVAD-cmk, diminished the ROS release in activated RAW 264.7 cells. In ST2 murine mesenchymal cells, EMD could not be tested because LPS, saliva, and IL-1β + TNF-α failed to provoke pyroptosis signalling. <b>Conclusion</b>: These findings suggest that EMD is capable of dampening the expression of pyroptosis-related genes in macrophages.
first_indexed 2024-03-10T04:04:43Z
format Article
id doaj.art-d85fe96f6fc442b7ad0eb9abb8a494cd
institution Directory Open Access Journal
issn 1661-6596
1422-0067
language English
last_indexed 2024-03-10T04:04:43Z
publishDate 2022-05-01
publisher MDPI AG
record_format Article
series International Journal of Molecular Sciences
spelling doaj.art-d85fe96f6fc442b7ad0eb9abb8a494cd2023-11-23T08:26:48ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-01239507810.3390/ijms23095078Enamel Matrix Derivative Decreases Pyroptosis-Related Genes in MacrophagesMariane Beatriz Sordi0Ariadne Cristiane Cabral da Cruz1Layla Panahipour2Reinhard Gruber3Department of Oral Biology, University Clinic of Dentistry, Medical University of Vienna, 1090 Vienna, AustriaDepartment of Dentistry, Federal University of Santa Catarina, Florianopolis 88040-900, BrazilDepartment of Oral Biology, University Clinic of Dentistry, Medical University of Vienna, 1090 Vienna, AustriaDepartment of Oral Biology, University Clinic of Dentistry, Medical University of Vienna, 1090 Vienna, Austria<b>Background</b>: Pyroptosis is a caspase-dependent catabolic process relevant to periodontal disorders for which inflammation is central to the pathophysiology of the disease. Although enamel matrix derivative (EMD) has been applied to support periodontal regeneration, its capacity to modulate the expression of pyroptosis-related genes remains unknown. Considering EMD has anti-inflammatory properties and pyroptosis is linked to the activation of the inflammasome in chronic periodontitis, the question arises whether EMD could reduce pyroptosis signalling. <b>Methods</b>: To answer this question, primary macrophages obtained from murine bone marrow and RAW 264.7 macrophages were primed with EMD before being challenged by lipopolysaccharide (LPS). Cells were then analysed for pyroptosis-signalling components by gene expression analyses, interleukin-1β (IL-1β) immunoassay, and the detection of caspase-1 (CAS1). The release of mitochondrial reactive oxygen species (ROS) was also detected. <b>Results</b>: We report here that EMD, like the inflammasome (NLRP3) and CAS1 specific inhibitors—MCC950 and Ac-YVAD-cmk, respectively—lowered the LPS-induced expression of NLRP3 in primary macrophages (EMD: <i>p</i> = 0.0232; MCC950: <i>p</i> = 0.0426; Ac-YVAD-cmk: <i>p</i> = 0.0317). EMD further reduced the LPS-induced expression of NLRP3 in RAW 264.7 cells (<i>p</i> = 0.0043). There was also a reduction in CAS1 and IL-1β in RAW 264.7 macrophages on the transcriptional level (<i>p</i> = 0.0598; <i>p</i> = 0.0283; respectively), in IL-1β protein release (<i>p</i> = 0.0313), and CAS1 activity. Consistently, EMD, like MCC950 and Ac-YVAD-cmk, diminished the ROS release in activated RAW 264.7 cells. In ST2 murine mesenchymal cells, EMD could not be tested because LPS, saliva, and IL-1β + TNF-α failed to provoke pyroptosis signalling. <b>Conclusion</b>: These findings suggest that EMD is capable of dampening the expression of pyroptosis-related genes in macrophages.https://www.mdpi.com/1422-0067/23/9/5078enamel matrix derivativepyroptosisinflammasomesperiodontal diseasesmacrophagesmesenchymal cells
spellingShingle Mariane Beatriz Sordi
Ariadne Cristiane Cabral da Cruz
Layla Panahipour
Reinhard Gruber
Enamel Matrix Derivative Decreases Pyroptosis-Related Genes in Macrophages
International Journal of Molecular Sciences
enamel matrix derivative
pyroptosis
inflammasomes
periodontal diseases
macrophages
mesenchymal cells
title Enamel Matrix Derivative Decreases Pyroptosis-Related Genes in Macrophages
title_full Enamel Matrix Derivative Decreases Pyroptosis-Related Genes in Macrophages
title_fullStr Enamel Matrix Derivative Decreases Pyroptosis-Related Genes in Macrophages
title_full_unstemmed Enamel Matrix Derivative Decreases Pyroptosis-Related Genes in Macrophages
title_short Enamel Matrix Derivative Decreases Pyroptosis-Related Genes in Macrophages
title_sort enamel matrix derivative decreases pyroptosis related genes in macrophages
topic enamel matrix derivative
pyroptosis
inflammasomes
periodontal diseases
macrophages
mesenchymal cells
url https://www.mdpi.com/1422-0067/23/9/5078
work_keys_str_mv AT marianebeatrizsordi enamelmatrixderivativedecreasespyroptosisrelatedgenesinmacrophages
AT ariadnecristianecabraldacruz enamelmatrixderivativedecreasespyroptosisrelatedgenesinmacrophages
AT laylapanahipour enamelmatrixderivativedecreasespyroptosisrelatedgenesinmacrophages
AT reinhardgruber enamelmatrixderivativedecreasespyroptosisrelatedgenesinmacrophages