title: Tooth Tissue Engineering: The Importance of Blood Products as a Supplement in Tissue Culture Medium for Human Pulp Dental Stem Cells.
abstract: One of the goals in using cells for tissue engineering (TE) and cell therapy consists of optimizing the medium for cell culture. The present study compares three different blood product supplements for improved cell proliferation and protection against DNA damage in cultured human dental pulp stem cells for tooth TE applications. Human cells from dental pulp were first characterized as adult stem cells (ectomesenchymal mixed origin) by flow cytometry. Next, four different cell culture conditions were tested: I, supplement-free; II, supplemented with fetal bovine serum; III, allogeneic human serum; and IV, autologous human serum. Cultured cells were then characterized for cell proliferation, mineralized nodule formation, and colony-forming units (CFU) capability. After 28 days in culture, the comet assay was performed to assess possible damage in cellular DNA. Our results revealed that Protocol IV achieved higher cell proliferation than Protocol I (p = 0.0112). Protocols II and III resulted in higher cell proliferation than Protocol I, but no statistical differences were found relative to Protocol IV. The comet assay revealed less cell damage in cells cultured using Protocol IV as compared to Protocols II and III. The damage percentage observed on Protocol II was significantly higher than all other protocols. CFUs capability was highest using Protocol IV (p = 0.0018) and III, respectively, and the highest degree of mineralization was observed using Protocol IV as compared to Protocols II and III. Protocol IV resulted in significantly improved cell proliferation, and no cell damage was observed. These results demonstrate that human blood product supplements can be used as feasible supplements for culturing adult human dental stem cells.
authors.affiliation: 1 CTCMol, Center of Cellular and Molecular Therapy, UNIFESP, São Paulo, Brazil .|2 Translational Surgery, Surgery Department, UNIFESP, São Paulo, Brazil .
authors.forename: Ricardo Luiz
authors.initials: RL
authors.lastname: Pisciolaro
authors.affiliation: 1 CTCMol, Center of Cellular and Molecular Therapy, UNIFESP, São Paulo, Brazil .|2 Translational Surgery, Surgery Department, UNIFESP, São Paulo, Brazil .|3 National Institute of Science and Technology, Biofabrication Institute, BIOFABRIS, Campinas, São Paulo, Brazil .
authors.forename: Monica Talarico
authors.initials: MT
authors.lastname: Duailibi
authors.affiliation: 2 Translational Surgery, Surgery Department, UNIFESP, São Paulo, Brazil .|4 Health Science Department, UNISA, São Paulo, Brazil .
authors.forename: Neil Ferreira
authors.initials: NF
authors.lastname: Novo
authors.affiliation: 2 Translational Surgery, Surgery Department, UNIFESP, São Paulo, Brazil .|4 Health Science Department, UNISA, São Paulo, Brazil .|5 Dentistry Department, UNISA, São Paulo, Brazil .
authors.forename: Yara
authors.initials: Y
authors.lastname: Juliano
authors.affiliation: 5 Dentistry Department, UNISA, São Paulo, Brazil .
authors.forename: Debora
authors.initials: D
authors.lastname: Pallos
authors.affiliation: 6 Department of Oral and Maxillofacial Pathology, Tufts University, Boston, Massachusetts.
authors.forename: Pamela Crotty
authors.initials: PC
authors.lastname: Yelick
authors.affiliation: 7 Laboratory for Tissue Engineering and Organ Fabrication, Massachusetts General Hospital and Department of Surgery, and Harvard Medical School, Boston, Massachusetts.
authors.forename: Joseph Phillip
authors.initials: JP
authors.lastname: Vacanti
authors.affiliation: 2 Translational Surgery, Surgery Department, UNIFESP, São Paulo, Brazil .
authors.forename: Lydia Masako
authors.initials: LM
authors.lastname: Ferreira
authors.affiliation: 1 CTCMol, Center of Cellular and Molecular Therapy, UNIFESP, São Paulo, Brazil .|2 Translational Surgery, Surgery Department, UNIFESP, São Paulo, Brazil .|3 National Institute of Science and Technology, Biofabrication Institute, BIOFABRIS, Campinas, São Paulo, Brazil .
authors.forename: Silvio Eduardo
authors.initials: SE
authors.lastname: Duailibi
mesh_terms: D000293:Adolescent; D053687:Adult Stem Cells / Q000166:cytology / Q000502:physiology*; D061249:Batch Cell Culture Techniques; D001769:Blood / Q000378:metabolism*; D049109:Cell Proliferation; D002648:Child; D003470:Culture Media / Q000737:chemistry*; D003782:Dental Pulp / Q000166:cytology* / Q000502:physiology; D005260:Female; D006801:Humans; D008297:Male; D009924:Organ Culture Techniques / Q000379:methods; D046509:Tissue Culture Techniques / Q000379:methods; D023822:Tissue Engineering / Q000379:methods*; D014070:Tooth / Q000166:cytology* / Q000254:growth & development
chemical_list: D003470:Culture Media