Subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice: An in vivo pilot study
Objectives: This study aimed to explore the potential of tooth germ stem cells for regenerating tooth-like structures by subcutaneously implanting first molar tooth germ stem cells over the masseter muscle in mice. Methods: Five pairs of house mice, Mus musculus, were selected for mating. At gestati...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | Regenerative Therapy |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352320425000215 |
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| author | Nosrat Nourbakhsh Ghazaleh Baniebrahimi Sahar Talebi Ardeshir Talebi Mohammad Hossein Nasr Esfahani Bijan Movahedian Maziar Manshayi Navid Naghdi Fatemeh Ejeian Elahe Masaeli Seyed Ali Mosaddad |
| author_facet | Nosrat Nourbakhsh Ghazaleh Baniebrahimi Sahar Talebi Ardeshir Talebi Mohammad Hossein Nasr Esfahani Bijan Movahedian Maziar Manshayi Navid Naghdi Fatemeh Ejeian Elahe Masaeli Seyed Ali Mosaddad |
| author_sort | Nosrat Nourbakhsh |
| collection | DOAJ |
| description | Objectives: This study aimed to explore the potential of tooth germ stem cells for regenerating tooth-like structures by subcutaneously implanting first molar tooth germ stem cells over the masseter muscle in mice. Methods: Five pairs of house mice, Mus musculus, were selected for mating. At gestational day 14 (E14), the fetuses were extracted, and the first molar tooth germ at the cap stage was isolated. Tooth germ stem cells were prepared into a suspension and seeded onto scaffolds, which were then implanted subcutaneously over the masseter muscle in male mice. The control group (n = 5 male mice) received acellular scaffolds implanted at the same site. After 20 days, the regenerated tissues were resected and analyzed histologically using hematoxylin and eosin (H & E) staining, Masson's trichrome staining, and immunohistochemical (IHC) staining for cytokeratin (CK) and vimentin markers. Results: H & E staining showed the formation of integrated oval structures at the implant site in all samples. Masson's trichrome staining identified dispersed accumulations of cellular mineralized matrix within the connective tissue. IHC staining was positive for vimentin, confirming the mesenchymal origin of the loose tissue at the center, indicating future dental pulp development. Positive CK staining indicated the ectodermal origin of dense peripheral tissues, suggesting the future formation of inner enamel epithelium. The combined immunohistochemical results for vimentin and CK confirmed the ecto-mesenchymal origin of the regenerated tissue, which resembled a late bell-stage tooth germ observed around gestational days 17.5–18 and showed early indications of dentin formation (D0). Conclusion: The study indicates that tooth germ stem cells may have the potential to produce dense, tooth-like structures when implanted subcutaneously in mice. These findings provide preliminary insights into the possible applications of tooth germ stem cells in regenerative dental tissue engineering. |
| format | Article |
| id | doaj-art-0612c446a40445e79dfa962f89fae7d9 |
| institution | Kabale University |
| issn | 2352-3204 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Regenerative Therapy |
| spelling | doaj-art-0612c446a40445e79dfa962f89fae7d92025-02-12T05:31:17ZengElsevierRegenerative Therapy2352-32042025-03-0128536543Subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice: An in vivo pilot studyNosrat Nourbakhsh0Ghazaleh Baniebrahimi1Sahar Talebi2Ardeshir Talebi3Mohammad Hossein Nasr Esfahani4Bijan Movahedian5Maziar Manshayi6Navid Naghdi7Fatemeh Ejeian8Elahe Masaeli9Seyed Ali Mosaddad10Department of Pediatric Dentistry, Dental Research Center, Dental Research Institute, School of Dentistry, Isfahan University of Medical Sciences, Isfahan, IranDepartment of Pediatric Dentistry, School of Dentistry, Tehran University of Medical Sciences, Tehran, IranResearch Committee, School of Dentistry, Isfahan University of Medical Sciences, Isfahan, Iran; Corresponding author. Research Committee, School of Dentistry, Isfahan University of Medical Sciences, Isfahan, Iran.Department of Pathology, Medical School, Dental Research Center, Dental Research Institute, Isfahan University of Medical Sciences, Isfahan, Iran; Corresponding author. Department of Pathology, Medical School, Isfahan University of Medical Sciences, Isfahan, Iran.Department of Animal Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, IranDepartment of Maxillofacial Surgery, Dental Research Center, Dental Research Institute, School of Dentistry, Isfahan University of Medical Sciences, Isfahan, IranDental Science Research Center, Dentistry Faculty, Isfahan University of Medical Sciences, Isfahan, IranDepartment of Maxillofacial Surgery, School of Dentistry, Hamedan University of Medical Sciences, Hamedan, IranDepartment of Animal Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, IranDepartment of Animal Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, IranDepartment of Research Analytics, Saveetha Dental College and Hospitals, Saveetha Institute of Medical and Technical Sciences, Saveetha University, Chennai, India; Department of Conservative Dentistry and Bucofacial Prostheses, Faculty of Odontology, University Complutense of Madrid, Madrid, SpainObjectives: This study aimed to explore the potential of tooth germ stem cells for regenerating tooth-like structures by subcutaneously implanting first molar tooth germ stem cells over the masseter muscle in mice. Methods: Five pairs of house mice, Mus musculus, were selected for mating. At gestational day 14 (E14), the fetuses were extracted, and the first molar tooth germ at the cap stage was isolated. Tooth germ stem cells were prepared into a suspension and seeded onto scaffolds, which were then implanted subcutaneously over the masseter muscle in male mice. The control group (n = 5 male mice) received acellular scaffolds implanted at the same site. After 20 days, the regenerated tissues were resected and analyzed histologically using hematoxylin and eosin (H & E) staining, Masson's trichrome staining, and immunohistochemical (IHC) staining for cytokeratin (CK) and vimentin markers. Results: H & E staining showed the formation of integrated oval structures at the implant site in all samples. Masson's trichrome staining identified dispersed accumulations of cellular mineralized matrix within the connective tissue. IHC staining was positive for vimentin, confirming the mesenchymal origin of the loose tissue at the center, indicating future dental pulp development. Positive CK staining indicated the ectodermal origin of dense peripheral tissues, suggesting the future formation of inner enamel epithelium. The combined immunohistochemical results for vimentin and CK confirmed the ecto-mesenchymal origin of the regenerated tissue, which resembled a late bell-stage tooth germ observed around gestational days 17.5–18 and showed early indications of dentin formation (D0). Conclusion: The study indicates that tooth germ stem cells may have the potential to produce dense, tooth-like structures when implanted subcutaneously in mice. These findings provide preliminary insights into the possible applications of tooth germ stem cells in regenerative dental tissue engineering.http://www.sciencedirect.com/science/article/pii/S2352320425000215Tooth germStem cellsImplantationDental stem cellsTissue engineeringTooth development |
| spellingShingle | Nosrat Nourbakhsh Ghazaleh Baniebrahimi Sahar Talebi Ardeshir Talebi Mohammad Hossein Nasr Esfahani Bijan Movahedian Maziar Manshayi Navid Naghdi Fatemeh Ejeian Elahe Masaeli Seyed Ali Mosaddad Subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice: An in vivo pilot study Regenerative Therapy Tooth germ Stem cells Implantation Dental stem cells Tissue engineering Tooth development |
| title | Subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice: An in vivo pilot study |
| title_full | Subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice: An in vivo pilot study |
| title_fullStr | Subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice: An in vivo pilot study |
| title_full_unstemmed | Subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice: An in vivo pilot study |
| title_short | Subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice: An in vivo pilot study |
| title_sort | subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice an in vivo pilot study |
| topic | Tooth germ Stem cells Implantation Dental stem cells Tissue engineering Tooth development |
| url | http://www.sciencedirect.com/science/article/pii/S2352320425000215 |
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