Translating proteome and transcriptome dynamics of periodontal ligament stem cell-derived secretome/conditioned medium in an in vitro model of periodontitis
Abstract Background Periodontal ligament stem cells (PDLSCs) have been proposed as therapeutic candidates in periodontal diseases and periodontium defects. Paracrine factors of PDLSCs, namely, secretome, can contribute to tissue regeneration comparable to direct stem cell application. This study exp...
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | BMC Oral Health |
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| Online Access: | https://doi.org/10.1186/s12903-024-04167-z |
| _version_ | 1797233218225700864 |
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| author | Han Na Suh Ju Young Ji Jung Sun Heo |
| author_facet | Han Na Suh Ju Young Ji Jung Sun Heo |
| author_sort | Han Na Suh |
| collection | DOAJ |
| description | Abstract Background Periodontal ligament stem cells (PDLSCs) have been proposed as therapeutic candidates in periodontal diseases and periodontium defects. Paracrine factors of PDLSCs, namely, secretome, can contribute to tissue regeneration comparable to direct stem cell application. This study explored restoration effects of PDLSC-derived secretome/conditioned medium (PDLSC-CM) on PDLSCs themselves in an inflammatory microenvironment and identified its action mechanisms using proteomics and transcriptomic profiling. Methods PDLSC-CM was prepared from cells under healthy culture conditions. Mass spectrometry and liquid chromatography–tandem mass spectrometry (LC–MS/MS) were then performed to analyze the PDLSC-CM proteome. Osteogenic differentiation of PDLSCs under inflammatory conditions or in the presence of PDLSC-CM was then characterized in assays of alkaline phosphatase activity, intracellular calcium levels, protein expression of osteogenic markers, and matrix mineralization. Furthermore, the transcriptomic profile was assessed to identify significantly enriched signaling pathways and associated molecular networks by RNA sequencing. Results LC–MS/MS proteomics identified a total of 203 proteins and distinguished 187 significant protein changes in PDLSC-CM compared to control-CM. LPS-treated PDLSCs significantly attenuated osteogenic differentiation. When PDLSCs were treated with PDLSC-CM alone, their osteogenic activity was significantly upregulated compared to the control group. Moreover, the LPS-impaired osteogenesis of PDLSCs was reconstituted by PDLSC-CM treatment. RNA sequencing revealed 252, 1,326, and 776 differentially expressed genes in the control vs. LPS, control vs. PDLSC-CM, and LPS vs. LPS + PDLSC-CM groups, respectively. Conclusion This study suggest that PDLSC-CM restores the osteogenic potential of PDLSCs in an inflammatory environment through secretory functions representing potential repair and regenerative mechanisms. |
| first_indexed | 2024-04-24T16:12:40Z |
| format | Article |
| id | doaj.art-e225177bab3c47f498a89b74fe90d77c |
| institution | Directory Open Access Journal |
| issn | 1472-6831 |
| language | English |
| last_indexed | 2024-04-24T16:12:40Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Oral Health |
| spelling | doaj.art-e225177bab3c47f498a89b74fe90d77c2024-03-31T11:37:20ZengBMCBMC Oral Health1472-68312024-03-0124111310.1186/s12903-024-04167-zTranslating proteome and transcriptome dynamics of periodontal ligament stem cell-derived secretome/conditioned medium in an in vitro model of periodontitisHan Na Suh0Ju Young Ji1Jung Sun Heo2Korea Institute of ToxicologyDepartment of Maxillofacial Biomedical Engineering, College of Dentistry, Kyung Hee UniversityDepartment of Maxillofacial Biomedical Engineering, College of Dentistry, Kyung Hee UniversityAbstract Background Periodontal ligament stem cells (PDLSCs) have been proposed as therapeutic candidates in periodontal diseases and periodontium defects. Paracrine factors of PDLSCs, namely, secretome, can contribute to tissue regeneration comparable to direct stem cell application. This study explored restoration effects of PDLSC-derived secretome/conditioned medium (PDLSC-CM) on PDLSCs themselves in an inflammatory microenvironment and identified its action mechanisms using proteomics and transcriptomic profiling. Methods PDLSC-CM was prepared from cells under healthy culture conditions. Mass spectrometry and liquid chromatography–tandem mass spectrometry (LC–MS/MS) were then performed to analyze the PDLSC-CM proteome. Osteogenic differentiation of PDLSCs under inflammatory conditions or in the presence of PDLSC-CM was then characterized in assays of alkaline phosphatase activity, intracellular calcium levels, protein expression of osteogenic markers, and matrix mineralization. Furthermore, the transcriptomic profile was assessed to identify significantly enriched signaling pathways and associated molecular networks by RNA sequencing. Results LC–MS/MS proteomics identified a total of 203 proteins and distinguished 187 significant protein changes in PDLSC-CM compared to control-CM. LPS-treated PDLSCs significantly attenuated osteogenic differentiation. When PDLSCs were treated with PDLSC-CM alone, their osteogenic activity was significantly upregulated compared to the control group. Moreover, the LPS-impaired osteogenesis of PDLSCs was reconstituted by PDLSC-CM treatment. RNA sequencing revealed 252, 1,326, and 776 differentially expressed genes in the control vs. LPS, control vs. PDLSC-CM, and LPS vs. LPS + PDLSC-CM groups, respectively. Conclusion This study suggest that PDLSC-CM restores the osteogenic potential of PDLSCs in an inflammatory environment through secretory functions representing potential repair and regenerative mechanisms.https://doi.org/10.1186/s12903-024-04167-zPeriodontal ligament stem cellsSecretome/conditioned mediumOsteogenic differentiationProteomic and transcriptomic profiling |
| spellingShingle | Han Na Suh Ju Young Ji Jung Sun Heo Translating proteome and transcriptome dynamics of periodontal ligament stem cell-derived secretome/conditioned medium in an in vitro model of periodontitis BMC Oral Health Periodontal ligament stem cells Secretome/conditioned medium Osteogenic differentiation Proteomic and transcriptomic profiling |
| title | Translating proteome and transcriptome dynamics of periodontal ligament stem cell-derived secretome/conditioned medium in an in vitro model of periodontitis |
| title_full | Translating proteome and transcriptome dynamics of periodontal ligament stem cell-derived secretome/conditioned medium in an in vitro model of periodontitis |
| title_fullStr | Translating proteome and transcriptome dynamics of periodontal ligament stem cell-derived secretome/conditioned medium in an in vitro model of periodontitis |
| title_full_unstemmed | Translating proteome and transcriptome dynamics of periodontal ligament stem cell-derived secretome/conditioned medium in an in vitro model of periodontitis |
| title_short | Translating proteome and transcriptome dynamics of periodontal ligament stem cell-derived secretome/conditioned medium in an in vitro model of periodontitis |
| title_sort | translating proteome and transcriptome dynamics of periodontal ligament stem cell derived secretome conditioned medium in an in vitro model of periodontitis |
| topic | Periodontal ligament stem cells Secretome/conditioned medium Osteogenic differentiation Proteomic and transcriptomic profiling |
| url | https://doi.org/10.1186/s12903-024-04167-z |
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