Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration
Objectives: To evaluate the effect of SHED-CM on the proliferation, differentiation, migration ability, cell death, gene expression and production of VEGF of HUVEC in vitro and in a rodent orthotopic dental pulp regeneration. Methods: Three culture media [M199, DMEM/Ham's F12 and DMEM/Ham'...
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| Format: | Article |
| Language: | English |
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Elsevier
2019-04-01
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| Series: | Heliyon |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844018378447 |
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| author | Sueli Patricia Harumi Miyagi de Cara Clarice Silvia Taemi Origassa Fernando de Sá Silva Maria Stella N.A. Moreira Danilo Candido de Almeida Ana Clara Fagundes Pedroni Giovanna Lopes Carvalho Diego Pulzatto Cury Niels Olsen Saraiva Câmara Márcia Martins Marques |
| author_facet | Sueli Patricia Harumi Miyagi de Cara Clarice Silvia Taemi Origassa Fernando de Sá Silva Maria Stella N.A. Moreira Danilo Candido de Almeida Ana Clara Fagundes Pedroni Giovanna Lopes Carvalho Diego Pulzatto Cury Niels Olsen Saraiva Câmara Márcia Martins Marques |
| author_sort | Sueli Patricia Harumi Miyagi de Cara |
| collection | DOAJ |
| description | Objectives: To evaluate the effect of SHED-CM on the proliferation, differentiation, migration ability, cell death, gene expression and production of VEGF of HUVEC in vitro and in a rodent orthotopic dental pulp regeneration. Methods: Three culture media [M199, DMEM/Ham's F12 and DMEM/Ham's F12 conditioned by SHEDs] were used as experimental groups. SHED-CM was prepared maintaining confluent cells in culture without serum for 3 days. The proliferation and cell death marker of HUVECs were assessed using flow cytometry. The capacity of formation of vascular-like structures was analyzed in cells grown over Matrigel® in hypoxic condition. HUVECs migration was followed using the scratch test. VEGF-A expression in HUVECs was assessed using real time RT-qPCR; and VEGF synthesis with ELISA test. SHED-CM was also applied in rodent ortotopic model of dental pulp regeneration in rats. The formed tissue was submitted to histological and immunohistochemical analyses. Results: SHED-CM promoted significantly lower expression of 7AAD in HUVECs; whereas the expression of the Ki67 was similar in all groups. The vascular-like structures were observed in all groups. Migration of SHED-CM group was faster than DMEM/Ham's F12. SHED-CM induced similar expression of VEGF-A than M199, and higher than DMEM/Ham's F12. SHED-CM induced significantly higher VEGF synthesis than other media. SHED-CM induced formation of a vascularized connective tissue inside the root canal. Conclusion: The study showed that SHEDs release angiogenic and cytoprotective factors, which are of great importance for tissue engineering. Clinical significance: SHED-CM could be an option to the use of stem cells in tissue engineering. |
| first_indexed | 2024-12-22T10:54:55Z |
| format | Article |
| id | doaj.art-2724f2fcf7e949c583b110f0d4cb8908 |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-12-22T10:54:55Z |
| publishDate | 2019-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-2724f2fcf7e949c583b110f0d4cb89082022-12-21T18:28:38ZengElsevierHeliyon2405-84402019-04-0154e01560Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regenerationSueli Patricia Harumi Miyagi de Cara0Clarice Silvia Taemi Origassa1Fernando de Sá Silva2Maria Stella N.A. Moreira3Danilo Candido de Almeida4Ana Clara Fagundes Pedroni5Giovanna Lopes Carvalho6Diego Pulzatto Cury7Niels Olsen Saraiva Câmara8Márcia Martins Marques9School of Dentistry, Centro Universitário das Faculdades Metropolitanas Unidas (FMU), São Paulo, SP, BrazilDepartamento de Medicina, Divisão de Nefrologia, Universidade Federal de São Paulo, São Paulo, SP, BrazilInstitute of Life Sciences, Universidade Federal de Juiz de Fora (UFJF), Governador Valadares, MG, BrazilSchool of Dentistry, Universidade Ibirapuera (UNIB), Sao Paulo, SP, BrazilDepartamento de Medicina, Divisão de Nefrologia, Universidade Federal de São Paulo, São Paulo, SP, BrazilDepartment of Restorative Dentistry and Endodontics, School of Dentistry, University of Sao Paulo (USP), Sao Paulo, SP, BrazilDepartment of Restorative Dentistry and Endodontics, School of Dentistry, University of Sao Paulo (USP), Sao Paulo, SP, BrazilSchool of Medicine, UNINOVE University, São Paulo, São Paulo, BrazilDepartamento de Medicina, Divisão de Nefrologia, Universidade Federal de São Paulo, São Paulo, SP, BrazilDepartment of Restorative Dentistry and Endodontics, School of Dentistry, University of Sao Paulo (USP), Sao Paulo, SP, Brazil; Corresponding author.Objectives: To evaluate the effect of SHED-CM on the proliferation, differentiation, migration ability, cell death, gene expression and production of VEGF of HUVEC in vitro and in a rodent orthotopic dental pulp regeneration. Methods: Three culture media [M199, DMEM/Ham's F12 and DMEM/Ham's F12 conditioned by SHEDs] were used as experimental groups. SHED-CM was prepared maintaining confluent cells in culture without serum for 3 days. The proliferation and cell death marker of HUVECs were assessed using flow cytometry. The capacity of formation of vascular-like structures was analyzed in cells grown over Matrigel® in hypoxic condition. HUVECs migration was followed using the scratch test. VEGF-A expression in HUVECs was assessed using real time RT-qPCR; and VEGF synthesis with ELISA test. SHED-CM was also applied in rodent ortotopic model of dental pulp regeneration in rats. The formed tissue was submitted to histological and immunohistochemical analyses. Results: SHED-CM promoted significantly lower expression of 7AAD in HUVECs; whereas the expression of the Ki67 was similar in all groups. The vascular-like structures were observed in all groups. Migration of SHED-CM group was faster than DMEM/Ham's F12. SHED-CM induced similar expression of VEGF-A than M199, and higher than DMEM/Ham's F12. SHED-CM induced significantly higher VEGF synthesis than other media. SHED-CM induced formation of a vascularized connective tissue inside the root canal. Conclusion: The study showed that SHEDs release angiogenic and cytoprotective factors, which are of great importance for tissue engineering. Clinical significance: SHED-CM could be an option to the use of stem cells in tissue engineering.http://www.sciencedirect.com/science/article/pii/S2405844018378447BioengineeringCell biology |
| spellingShingle | Sueli Patricia Harumi Miyagi de Cara Clarice Silvia Taemi Origassa Fernando de Sá Silva Maria Stella N.A. Moreira Danilo Candido de Almeida Ana Clara Fagundes Pedroni Giovanna Lopes Carvalho Diego Pulzatto Cury Niels Olsen Saraiva Câmara Márcia Martins Marques Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration Heliyon Bioengineering Cell biology |
| title | Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration |
| title_full | Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration |
| title_fullStr | Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration |
| title_full_unstemmed | Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration |
| title_short | Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration |
| title_sort | angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration |
| topic | Bioengineering Cell biology |
| url | http://www.sciencedirect.com/science/article/pii/S2405844018378447 |
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