LED red light up-regulates MAPK signal to promote osteogenic/odontogenic differentiation of human dental pulp stem cells in an inflammatory environment
Objective To study the effect of light-emitting diode (LED) red light on the osteogenic/odontogenic differentiation of human dental pulp stem cells (hDPSCs) and its mechanism were discussed. Methods This study has been reviewed and approved by the Ethics Committee. hDPSCs were cultured by tissue blo...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | zho |
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Editorial Department of Journal of Prevention and Treatment for Stomatological Diseases
2023-10-01
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| Series: | 口腔疾病防治 |
| Subjects: | |
| Online Access: | https://www.kqjbfz.com/CN/10.12016/j.issn.2096-1456.2023.10.003 |
| _version_ | 1797755738698809344 |
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| author | LIU Yuan HUI Yining JIANG Bing ZHENG Genzi WANG Yao |
| author_facet | LIU Yuan HUI Yining JIANG Bing ZHENG Genzi WANG Yao |
| author_sort | LIU Yuan |
| collection | DOAJ |
| description | Objective To study the effect of light-emitting diode (LED) red light on the osteogenic/odontogenic differentiation of human dental pulp stem cells (hDPSCs) and its mechanism were discussed. Methods This study has been reviewed and approved by the Ethics Committee. hDPSCs were cultured by tissue block enzyme digestion. The proliferative capacity of hDPSCs was detected by the CCK-8 at days 1, 3, 5 and 7 under stimulation with 0, 1, 5 and 10 μg/mL lipopolysaccharide (LPS), and the LPS stimulatory concentration was screened. The CG group (mineralization induction), LPS+CG group, and LPS+CG+ (2, 4, 6, 8, and 10 J/cm2) LED red light groups were set. On day 7, alkaline phosphatase (ALP) staining and ALP activity were determined. Relative expression levels of the ALP, osterix (OSX), dentin matrix protein-1 (DMP-1) and dentin sialophosphoprotein (DSPP) genes were measured by qRT-PCR. On day 21, alizarin red staining and calcium nodule quantitative determination were performed to screen the best light energy. The LPS+CG group and LPS+CG+LED group (optimal energy) were set up, and the secretion and expression levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by ELISAs on days 1, 3, 5 and 7. The relative expression levels of the extracellular regulated protein kinases 1/2 (ERK1/2), p38, c-Jun N-terminal kinase (JNK), and extracellular regulated protein kinases 5 (ERK5) proteins and their phosphorylated proteins in the MAPK signaling pathway were detected by Western blots. After the pathway was blocked, the relative expression levels of the ALP, OSX, DMP-1, and DSPP proteins after LED red light irradiation on day 7 were detected by Western blots. Results CCK-8 assays showed that the proliferation of hDPSCs induced by 10 μg/mL LPS was lower than that of the 0, 1, and 5 μg/mL groups on the 5th and 7th days (P<0.05), and 10 μg/mL was selected as the LPS stimulatory concentration in the follow-up experiment. ALP staining, ALP activity, gene expression levels of ALP, OSX, DMP-1 and DSPP and calcium nodule quantification in the LPS+CG+4 J/cm2 group were higher than those in the other treatment groups (P<0.05). 4 J/cm2 LED red light had the strongest ability to promote osteogenic/odontogenic differentiation and was used as the LED light energy density in subsequent experiments. ELISA showed that the secretion and expression levels of TNF-α and IL-1β in the LPS+CG+LED group were lower than those in the LPS+CG group on the 5th and 7th days (P<0.05). Western blot analysis showed that 4 J/cm2 LED red light promoted the expression levels of the p-ERK1/2, p-p38, p-JNK and p-ERK5 proteins. After the MAPK pathway was blocked, the expression levels of the ALP, OSX, DMP-1, and DSPP proteins in the LPS+CG+LED+U0126 (ERK1/2 inhibitor), SP600125 (JNK inhibitor), and BIX02189 (ERK5 inhibitor) groups were lower than those in the LPS+CG+LED group (P<0.001). The protein expression levels of ALP, OSX and DMP-1 in the LPS+CG+LED+SB203580 (p38 inhibitor) group were not significantly different from those in the LPS+CG+LED group (P>0.05). Conclusion In inflammatory conditions, LED red light promotes osteogenic/odontogenic differentiation of hDPSCs. This effect may be attributed to enhancement of the ERK1/2, JNK, and ERK5 signaling pathways, which reduces the production of the inflammatory cytokines TNF-α and IL-1β. |
| first_indexed | 2024-03-12T17:51:32Z |
| format | Article |
| id | doaj.art-6c38976f5af74c67b20b26bbd62a385c |
| institution | Directory Open Access Journal |
| issn | 2096-1456 |
| language | zho |
| last_indexed | 2024-03-12T17:51:32Z |
| publishDate | 2023-10-01 |
| publisher | Editorial Department of Journal of Prevention and Treatment for Stomatological Diseases |
| record_format | Article |
| series | 口腔疾病防治 |
| spelling | doaj.art-6c38976f5af74c67b20b26bbd62a385c2023-08-03T06:46:20ZzhoEditorial Department of Journal of Prevention and Treatment for Stomatological Diseases口腔疾病防治2096-14562023-10-01311070171110.12016/j.issn.2096⁃1456.2023.10.003LED red light up-regulates MAPK signal to promote osteogenic/odontogenic differentiation of human dental pulp stem cells in an inflammatory environmentLIU Yuan0 HUI Yining1 JIANG Bing2 ZHENG Genzi 3 WANG Yao41.Department of Preventive Health Care, the Affiliated Stomatological Hospital of Southwest Medical University 2. InstituteofStomatology, SouthwestMedicalUniversity 3.Oral and Maxillofacial Reconstruction and Regeneration of Luzhou Key LaboratorySouthwest Medical University1. InstituteofStomatology, SouthwestMedicalUniversity 2.Oral and Maxillofacial Reconstruction and Regeneration of Luzhou Key Laboratory1. InstituteofStomatology, SouthwestMedicalUniversity 2.Oral and Maxillofacial Reconstruction and Regeneration of Luzhou Key LaboratoryDepartment of Preventive Health Care, the Affiliated Stomatological Hospital of Southwest Medical UniversityObjective To study the effect of light-emitting diode (LED) red light on the osteogenic/odontogenic differentiation of human dental pulp stem cells (hDPSCs) and its mechanism were discussed. Methods This study has been reviewed and approved by the Ethics Committee. hDPSCs were cultured by tissue block enzyme digestion. The proliferative capacity of hDPSCs was detected by the CCK-8 at days 1, 3, 5 and 7 under stimulation with 0, 1, 5 and 10 μg/mL lipopolysaccharide (LPS), and the LPS stimulatory concentration was screened. The CG group (mineralization induction), LPS+CG group, and LPS+CG+ (2, 4, 6, 8, and 10 J/cm2) LED red light groups were set. On day 7, alkaline phosphatase (ALP) staining and ALP activity were determined. Relative expression levels of the ALP, osterix (OSX), dentin matrix protein-1 (DMP-1) and dentin sialophosphoprotein (DSPP) genes were measured by qRT-PCR. On day 21, alizarin red staining and calcium nodule quantitative determination were performed to screen the best light energy. The LPS+CG group and LPS+CG+LED group (optimal energy) were set up, and the secretion and expression levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were detected by ELISAs on days 1, 3, 5 and 7. The relative expression levels of the extracellular regulated protein kinases 1/2 (ERK1/2), p38, c-Jun N-terminal kinase (JNK), and extracellular regulated protein kinases 5 (ERK5) proteins and their phosphorylated proteins in the MAPK signaling pathway were detected by Western blots. After the pathway was blocked, the relative expression levels of the ALP, OSX, DMP-1, and DSPP proteins after LED red light irradiation on day 7 were detected by Western blots. Results CCK-8 assays showed that the proliferation of hDPSCs induced by 10 μg/mL LPS was lower than that of the 0, 1, and 5 μg/mL groups on the 5th and 7th days (P<0.05), and 10 μg/mL was selected as the LPS stimulatory concentration in the follow-up experiment. ALP staining, ALP activity, gene expression levels of ALP, OSX, DMP-1 and DSPP and calcium nodule quantification in the LPS+CG+4 J/cm2 group were higher than those in the other treatment groups (P<0.05). 4 J/cm2 LED red light had the strongest ability to promote osteogenic/odontogenic differentiation and was used as the LED light energy density in subsequent experiments. ELISA showed that the secretion and expression levels of TNF-α and IL-1β in the LPS+CG+LED group were lower than those in the LPS+CG group on the 5th and 7th days (P<0.05). Western blot analysis showed that 4 J/cm2 LED red light promoted the expression levels of the p-ERK1/2, p-p38, p-JNK and p-ERK5 proteins. After the MAPK pathway was blocked, the expression levels of the ALP, OSX, DMP-1, and DSPP proteins in the LPS+CG+LED+U0126 (ERK1/2 inhibitor), SP600125 (JNK inhibitor), and BIX02189 (ERK5 inhibitor) groups were lower than those in the LPS+CG+LED group (P<0.001). The protein expression levels of ALP, OSX and DMP-1 in the LPS+CG+LED+SB203580 (p38 inhibitor) group were not significantly different from those in the LPS+CG+LED group (P>0.05). Conclusion In inflammatory conditions, LED red light promotes osteogenic/odontogenic differentiation of hDPSCs. This effect may be attributed to enhancement of the ERK1/2, JNK, and ERK5 signaling pathways, which reduces the production of the inflammatory cytokines TNF-α and IL-1β.https://www.kqjbfz.com/CN/10.12016/j.issn.2096-1456.2023.10.003human dental pulp stem cellslipopolysaccharideinflammatory environmentlight-emitting diodered lightosteogenic/odontogenic differentiationmapk signal pathwayosterixdentin matrix protein-1 |
| spellingShingle | LIU Yuan HUI Yining JIANG Bing ZHENG Genzi WANG Yao LED red light up-regulates MAPK signal to promote osteogenic/odontogenic differentiation of human dental pulp stem cells in an inflammatory environment 口腔疾病防治 human dental pulp stem cells lipopolysaccharide inflammatory environment light-emitting diode red light osteogenic/odontogenic differentiation mapk signal pathway osterix dentin matrix protein-1 |
| title | LED red light up-regulates MAPK signal to promote osteogenic/odontogenic differentiation of human dental pulp stem cells in an inflammatory environment |
| title_full | LED red light up-regulates MAPK signal to promote osteogenic/odontogenic differentiation of human dental pulp stem cells in an inflammatory environment |
| title_fullStr | LED red light up-regulates MAPK signal to promote osteogenic/odontogenic differentiation of human dental pulp stem cells in an inflammatory environment |
| title_full_unstemmed | LED red light up-regulates MAPK signal to promote osteogenic/odontogenic differentiation of human dental pulp stem cells in an inflammatory environment |
| title_short | LED red light up-regulates MAPK signal to promote osteogenic/odontogenic differentiation of human dental pulp stem cells in an inflammatory environment |
| title_sort | led red light up regulates mapk signal to promote osteogenic odontogenic differentiation of human dental pulp stem cells in an inflammatory environment |
| topic | human dental pulp stem cells lipopolysaccharide inflammatory environment light-emitting diode red light osteogenic/odontogenic differentiation mapk signal pathway osterix dentin matrix protein-1 |
| url | https://www.kqjbfz.com/CN/10.12016/j.issn.2096-1456.2023.10.003 |
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