Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro
Abstract Different structures and cell types of the periodontium respond to orthodontic tooth movement (OTM) individually. Cementoblasts (OC/CM) located in the immediate vicinity of the fibroblasts on the cement have found way to the centre of actual research. Here, we identify and validate possible...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2020-07-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-020-67449-w |
| _version_ | 1818683373347930112 |
|---|---|
| author | Christian Niederau Rogerio B. Craveiro Irma Azraq Julia Brockhaus Asisa Bastian Christian Kirschneck Michael Wolf |
| author_facet | Christian Niederau Rogerio B. Craveiro Irma Azraq Julia Brockhaus Asisa Bastian Christian Kirschneck Michael Wolf |
| author_sort | Christian Niederau |
| collection | DOAJ |
| description | Abstract Different structures and cell types of the periodontium respond to orthodontic tooth movement (OTM) individually. Cementoblasts (OC/CM) located in the immediate vicinity of the fibroblasts on the cement have found way to the centre of actual research. Here, we identify and validate possible reference genes for OC/CM cells by RT-qPCR with and without static compressive loading. We investigated the suitability of 3 reference genes in an in vitro model of cementoblast cells using four different algorithms (Normfinder, geNorm, comparative delta-Ct method and BestKeeper) under different confluences and time. Comparable to our previous publications about reference genes in OTM in rats and human periodontal ligament fibroblasts (hPDLF), Rpl22 in murine OC/CM cells appears as the least regulated gene so that it represents the most appropriate reference gene. Furthermore, unlike to the expression of our recommended reference genes, the expression of additionally investigated target genes changes with confluence and under loading compression. Based on our findings for future RT-qPCR analyses in OC/CM cells, Rpl22 or the combination Rpl22/Tbp should be favored as reference gene. According to our results, although many publications propose the use of Gapdh, it does not seem to be the most suitable approach. |
| first_indexed | 2024-12-17T10:33:42Z |
| format | Article |
| id | doaj.art-758b4d980e3b457091f5d5dfdc522e55 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-17T10:33:42Z |
| publishDate | 2020-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-758b4d980e3b457091f5d5dfdc522e552022-12-21T21:52:27ZengNature PortfolioScientific Reports2045-23222020-07-0110111010.1038/s41598-020-67449-wSelection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitroChristian Niederau0Rogerio B. Craveiro1Irma Azraq2Julia Brockhaus3Asisa Bastian4Christian Kirschneck5Michael Wolf6Department of Orthodontics, Dental Clinic, University of AachenDepartment of Orthodontics, Dental Clinic, University of AachenDepartment of Orthodontics, Dental Clinic, University of AachenDepartment of Orthodontics, Dental Clinic, University of AachenDepartment of Orthodontics, Dental Clinic, University of AachenDepartment of Orthodontics, University Medical Centre of RegensburgDepartment of Orthodontics, Dental Clinic, University of AachenAbstract Different structures and cell types of the periodontium respond to orthodontic tooth movement (OTM) individually. Cementoblasts (OC/CM) located in the immediate vicinity of the fibroblasts on the cement have found way to the centre of actual research. Here, we identify and validate possible reference genes for OC/CM cells by RT-qPCR with and without static compressive loading. We investigated the suitability of 3 reference genes in an in vitro model of cementoblast cells using four different algorithms (Normfinder, geNorm, comparative delta-Ct method and BestKeeper) under different confluences and time. Comparable to our previous publications about reference genes in OTM in rats and human periodontal ligament fibroblasts (hPDLF), Rpl22 in murine OC/CM cells appears as the least regulated gene so that it represents the most appropriate reference gene. Furthermore, unlike to the expression of our recommended reference genes, the expression of additionally investigated target genes changes with confluence and under loading compression. Based on our findings for future RT-qPCR analyses in OC/CM cells, Rpl22 or the combination Rpl22/Tbp should be favored as reference gene. According to our results, although many publications propose the use of Gapdh, it does not seem to be the most suitable approach.https://doi.org/10.1038/s41598-020-67449-w |
| spellingShingle | Christian Niederau Rogerio B. Craveiro Irma Azraq Julia Brockhaus Asisa Bastian Christian Kirschneck Michael Wolf Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro Scientific Reports |
| title | Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro |
| title_full | Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro |
| title_fullStr | Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro |
| title_full_unstemmed | Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro |
| title_short | Selection and validation of reference genes by RT-qPCR for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro |
| title_sort | selection and validation of reference genes by rt qpcr for murine cementoblasts in mechanical loading experiments simulating orthodontic forces in vitro |
| url | https://doi.org/10.1038/s41598-020-67449-w |
| work_keys_str_mv | AT christianniederau selectionandvalidationofreferencegenesbyrtqpcrformurinecementoblastsinmechanicalloadingexperimentssimulatingorthodonticforcesinvitro AT rogeriobcraveiro selectionandvalidationofreferencegenesbyrtqpcrformurinecementoblastsinmechanicalloadingexperimentssimulatingorthodonticforcesinvitro AT irmaazraq selectionandvalidationofreferencegenesbyrtqpcrformurinecementoblastsinmechanicalloadingexperimentssimulatingorthodonticforcesinvitro AT juliabrockhaus selectionandvalidationofreferencegenesbyrtqpcrformurinecementoblastsinmechanicalloadingexperimentssimulatingorthodonticforcesinvitro AT asisabastian selectionandvalidationofreferencegenesbyrtqpcrformurinecementoblastsinmechanicalloadingexperimentssimulatingorthodonticforcesinvitro AT christiankirschneck selectionandvalidationofreferencegenesbyrtqpcrformurinecementoblastsinmechanicalloadingexperimentssimulatingorthodonticforcesinvitro AT michaelwolf selectionandvalidationofreferencegenesbyrtqpcrformurinecementoblastsinmechanicalloadingexperimentssimulatingorthodonticforcesinvitro |