title: A novel combinatorial therapy with pulp stem cells and granulocyte colony-stimulating factor for total pulp regeneration.
abstract: Treatment of deep caries with pulpitis is a major challenge in dentistry. Stem cell therapy represents a potential strategy to regenerate the dentin-pulp complex, enabling conservation and restoration of teeth. The objective of this study was to assess the efficacy and safety of pulp stem cell transplantation as a prelude for the impending clinical trials. Clinical-grade pulp stem cells were isolated and expanded according to good manufacturing practice conditions. The absence of contamination, abnormalities/aberrations in karyotype, and tumor formation after transplantation in an immunodeficient mouse ensured excellent quality control. After autologous transplantation of pulp stem cells with granulocyte-colony stimulating factor (G-CSF) in a dog pulpectomized tooth, regenerated pulp tissue including vasculature and innervation completely filled in the root canal, and regenerated dentin was formed in the coronal part and prevented microleakage up to day 180. Transplantation of pulp stem cells with G-CSF yielded a significantly larger amount of regenerated dentin-pulp complex compared with transplantation of G-CSF or stem cells alone. Also noteworthy was the reduction in the number of inflammatory cells and apoptotic cells and the significant increase in neurite outgrowth compared with results without G-CSF. The transplanted stem cells expressed angiogenic/neurotrophic factors. It is significant that G-CSF together with conditioned medium of pulp stem cells stimulated cell migration and neurite outgrowth, prevented cell death, and promoted immunosuppression in vitro. Furthermore, there was no evidence of toxicity or adverse events. In conclusion, the combinatorial trophic effects of pulp stem cells and G-CSF are of immediate utility for pulp/dentin regeneration, demonstrating the prerequisites of safety and efficacy critical for clinical applications.
authors.affiliation: Department of Dental Regenerative Medicine, National Center for Geriatrics and Gerontology, Research Institute, Obu, Japan.
authors.forename: Koichiro
authors.initials: K
authors.lastname: Iohara
authors.forename: Masashi
authors.initials: M
authors.lastname: Murakami
authors.forename: Norio
authors.initials: N
authors.lastname: Takeuchi
authors.forename: Yohei
authors.initials: Y
authors.lastname: Osako
authors.forename: Masataka
authors.initials: M
authors.lastname: Ito
authors.forename: Ryo
authors.initials: R
authors.lastname: Ishizaka
authors.forename: Shinji
authors.initials: S
authors.lastname: Utunomiya
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nakamura
authors.forename: Kenji
authors.initials: K
authors.lastname: Matsushita
authors.forename: Misako
authors.initials: M
authors.lastname: Nakashima
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000502:physiology; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003131:Combined Modality Therapy / Q000379:methods; D003782:Dental Pulp / Q000166:cytology*; D003786:Dental Pulp Cavity / Q000166:cytology; D004285:Dogs; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D051379:Mice; D008817:Mice, Mutant Strains; D023421:Models, Animal; D012038:Regeneration / Q000187:drug effects / Q000502:physiology*; D033581:Stem Cell Transplantation / Q000379:methods*; D013234:Stem Cells / Q000166:cytology*; D059467:Transcriptome; D014182:Transplantation, Autologous
chemical_list: D016179:Granulocyte Colony-Stimulating Factor
keywords: Autologous stem cell transplantation; Dog model; G-CSF; Mesenchymal stem cells; Preclinical trials; Stem cell culture; Tissue regeneration