IADR Abstract Archives
Effects of Hypergravity on Proliferation, Differentiation and Circadian Rhythm of SCAP
Objectives: Aim of this study was to investigate the effect of hypergravity, applied for bone mass recovery after space flights or for treatment of chronic musculoskeletal diseases (e.g. osteoporosis), on viability/proliferation, osteo/odontogenic differentiation potential and circadian rhythm of oral stem cells.
Methods: Stem cells from apical papilla (SCAP) cultures were established from third molars of young healthy donors and characterized for stem cell-SC markers with flow cytometry. SCAP were exposed to hypergravity of 5g, 10g and 20g by utilizing centrifugation (Centrifuge 5804 R, Eppendorf) for 30 and 45 minutes. Viability/proliferation was evaluated by MTT assay compared with the control (1g). Real-time PCR was used to evaluate osteo/odontogenic markers' expression (DSPP, RUNX2, BMP-2, ALP), as well as circadian rhythm-related genes (CLOCK, BMAL1, CRY1, PER1, PER2).
Results: Results showed that application of 5g for 30 min had a significantly higher impact on increasing cell viability/proliferation compared with 1g. Application of 5g and 10g for 45 min also exhibited a significant increase in cell viability/proliferation at 72h. Gene expression indicated pronounced osteo/odontogenic shift of SCAP, evidenced by upregulation of DSPP, BMP-2, ALP, and RUNX2 particularly at 5g conditions. The expression of circadian-related genes showed differential, gravity-related alterations during differentiation, while the expression of CLOCK, BMAL1 and PER2 was significantly changed compared to 1g. Specifically, 5g and 20g induced downregulation of CLOCK and upregulation of PER2 compared with 1g, while BMAL1 increased significant at 5g. The expression of PER1 and CRY1 significantly decreased in all groups compared with baseline expression.
Conclusions: This is the first study validating the potential of hypergravity to influence proliferation, osteo/odontogenic differentiation, and circadian rhythmicity of SCAP. These results reveal that hypergravity engaging oral stem cells can be considered a promising new strategy towards dynamic tissue engineering of mineralized tissues, in addition to therapeutic application in management of bone-related diseases.
Methods: Stem cells from apical papilla (SCAP) cultures were established from third molars of young healthy donors and characterized for stem cell-SC markers with flow cytometry. SCAP were exposed to hypergravity of 5g, 10g and 20g by utilizing centrifugation (Centrifuge 5804 R, Eppendorf) for 30 and 45 minutes. Viability/proliferation was evaluated by MTT assay compared with the control (1g). Real-time PCR was used to evaluate osteo/odontogenic markers' expression (DSPP, RUNX2, BMP-2, ALP), as well as circadian rhythm-related genes (CLOCK, BMAL1, CRY1, PER1, PER2).
Results: Results showed that application of 5g for 30 min had a significantly higher impact on increasing cell viability/proliferation compared with 1g. Application of 5g and 10g for 45 min also exhibited a significant increase in cell viability/proliferation at 72h. Gene expression indicated pronounced osteo/odontogenic shift of SCAP, evidenced by upregulation of DSPP, BMP-2, ALP, and RUNX2 particularly at 5g conditions. The expression of circadian-related genes showed differential, gravity-related alterations during differentiation, while the expression of CLOCK, BMAL1 and PER2 was significantly changed compared to 1g. Specifically, 5g and 20g induced downregulation of CLOCK and upregulation of PER2 compared with 1g, while BMAL1 increased significant at 5g. The expression of PER1 and CRY1 significantly decreased in all groups compared with baseline expression.
Conclusions: This is the first study validating the potential of hypergravity to influence proliferation, osteo/odontogenic differentiation, and circadian rhythmicity of SCAP. These results reveal that hypergravity engaging oral stem cells can be considered a promising new strategy towards dynamic tissue engineering of mineralized tissues, in addition to therapeutic application in management of bone-related diseases.