Cytotoxicity and bioactivity of various pulpotomy materials on stem cells from human exfoliated primary teeth.
OBJECTIVE
To investigate the cytotoxicity and bioactivity of several pulpotomy materials: Biodentine (Septodont, Saint-Maur-des-Fosses, France) MTA (Angelus, Londrina, PR, Brazil), Theracal LC (Bisco Inc., Schamburg, IL, USA) and IRM (Dentsply DeTrey GmbH, Konstanz, Germany), after contact with stem cells isolated from human exfoliated primary teeth (SHEDs).


METHODS
SHEDs were cultured in the presence of the eluates of various pulpotomy materials for 24, 48 and 72 h. Cell viability was determined by mitochondrial dehydrogenase enzymatic (MTT) assay. Apoptosis and changes in cell phenotype were evaluated by flow cytometry. Also, an in vitro scratch wound-healing assay was used to determine their effects on cell migration. To assess cell morphology and attachment to the different pulpotomy materials, SHEDs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). Finally, the deposition of a calcified matrix in presence of these materials was verified by Alizarin Red staining. Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post-test (α = 0.05).


RESULTS
Cell viability in the presence of Biodentine eluates was significantly higher to that obtained using complete medium alone (control; P < 0.01) and was also significantly higher than using MTA Angelus from 48 h of incubation (P < 0.01). However, Theracal LC and IRM were associated with low rates of cell viability (P < 0.001). Similar results were obtained in an apoptosis assay. In addition, SHEDs maintained their mesenchymal phenotype in all conditions although their capacity to migrate was higher in the presence of Biodentine. SEM studies revealed a suitable proliferation rate, cell spreading and attachment, especially when using Biodentine and MTA Angelus discs. Finally, Biodentine eluates significantly induced calcified matrix deposition from 7 days of culture (P < 0.01).


CONCLUSIONS
Biodentine exhibited better cytocompatibility and bioactivity than MTA Angelus, Theracal LC and IRM.
Cellular Therapy and Hematopoietic Transplant Unit, Hematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain.
M
M
Collado-González
Cellular Therapy and Hematopoietic Transplant Unit, Hematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain.
D
D
García-Bernal
School of Dentistry, Faculty of Medicine, University of Murcia, Murcia, Spain.
R E
RE
Oñate-Sánchez
Department of Stomatology, Universitat de Valencia, Valencia, Spain.
P S
PS
Ortolani-Seltenerich
Dental Clinic Esdens, Alicante, Spain.
T
T
Álvarez-Muro
Department of Stomatology, Universitat de Valencia, Valencia, Spain.
A
A
Lozano
Department of Stomatology, Universitat de Valencia, Valencia, Spain.
L
L
Forner
Department of Stomatology, Universitat de Valencia, Valencia, Spain.
C
C
Llena
Cellular Therapy and Hematopoietic Transplant Unit, Hematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain.
J M
JM
Moraleda
Cellular Therapy and Hematopoietic Transplant Unit, Hematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain.|School of Dentistry, Faculty of Medicine, University of Murcia, Murcia, Spain.
F J
http://orcid.org/0000-0002-0623-740X
FJ
Rodríguez-Lozano
D017607:Aluminum Compounds / Q000494:pharmacology / Q000633:toxicity; D017209:Apoptosis / Q000187:drug effects; D017610:Calcium Compounds / Q000494:pharmacology / Q000633:toxicity; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004338:Drug Combinations; D005434:Flow Cytometry; D006801:Humans; D066298:In Vitro Techniques; D008422:Materials Testing; D008768:Methylmethacrylates / Q000494:pharmacology / Q000633:toxicity; D008855:Microscopy, Electron, Scanning; D010087:Oxides / Q000494:pharmacology / Q000633:toxicity; D010641:Phenotype; D057872:Pulp Capping and Pulpectomy Agents / Q000494:pharmacology* / Q000633:toxicity; D011672:Pulpotomy; D017640:Silicates / Q000494:pharmacology / Q000633:toxicity; D013234:Stem Cells / Q000187:drug effects*; D013997:Time Factors; D014094:Tooth, Deciduous; D015035:Zinc Oxide-Eugenol Cement / Q000494:pharmacology / Q000633:toxicity
D017607:Aluminum Compounds; D017610:Calcium Compounds; D004338:Drug Combinations; D008768:Methylmethacrylates; D010087:Oxides; D057872:Pulp Capping and Pulpectomy Agents; D017640:Silicates; D015035:Zinc Oxide-Eugenol Cement; C086631:mineral trioxide aggregate; C506393:tricalcium silicate; C031667:IRM cement
cytotoxicity; mineral trioxide aggregate; pulpotomy materials