TY - JOUR
T1 - Micropatterned hydrogels and cell alignment enhance the odontogenic potential of stem cells from apical papilla in-vitro
AU - Ha, Michael
AU - Athirasala, Avathamsa
AU - Tahayeri, Anthony
AU - Menezes, Paula P.
AU - Bertassoni, Luiz E.
N1 - Funding Information:
The authors thank Dr. Anibal Diogenes from University of Texas Health Center, for the donation of SCAP used in this study. This project was supported by funding from the National Institute of Dental and Craniofacial Research (R01DE026170 and 3R01DE026170-03S1 to LEB), the Oregon Clinical & Translational Research Institute (OCTRI) - Biomedical Innovation Program (BIP). MH acknowledge funding from the American Association for Dental Research (AADR) through their Student Fellowship program. The authors declare no conflict of interest.
Funding Information:
The authors thank Dr. Anibal Diogenes from University of Texas Health Center, for the donation of SCAP used in this study. This project was supported by funding from the National Institute of Dental and Craniofacial Research ( R01DE026170 and 3R01DE026170-03S1 to LEB), the Oregon Clinical & Translational Research Institute (OCTRI) - Biomedical Innovation Program (BIP). MH acknowledge funding from the American Association for Dental Research (AADR) through their Student Fellowship program. The authors declare no conflict of interest.
Publisher Copyright:
© 2019
PY - 2020/1
Y1 - 2020/1
N2 - Introduction: An understanding of the extracellular matrix characteristics which stimulate and guide stem cell differentiation in the dental pulp is fundamental for the development of enhanced dental regenerative therapies. Our objectives, in this study, were to determine whether stem cells from the apical papilla (SCAP) responded to substrate stiffness, whether hydrogels providing micropatterned topographical cues stimulate SCAP self-alignment, and whether the resulting alignment could influence their differentiation towards an odontogenic lineage in-vitro. Methods: Experiments utilized gelatin methacryloyl (GelMA) hydrogels of increasing concentrations (5, 10 and 15%). We determined their compressive modulus via unconfined compression and analyzed cell spreading via F-actin/DAPI immunostaining. GelMA hydrogels were micropatterned using photolithography, in order to generate microgrooves and ridges of 60 and 120 μm, onto which SCAP were seeded and analyzed for self-alignment via fluorescence microscopy. Lastly, we analyzed the odontogenic differentiation of SCAP using alkaline phosphatase protein expression (ANOVA/Tukey α = 0.05). Results: SCAP appeared to proliferate better on stiffer hydrogels. Both 60 and 120 μm micropatterned hydrogels guided the self-alignment of SCAP with no significant difference between them. Similarly, both 60 and 120 μm micropattern aligned cells promoted higher odontogenic differentiation than non-patterned controls. Significance: In summary, both substrate mechanics and geometry have a statistically significant influence on SCAP response, and may assist in the odontogenic differentiation of dental stem cells. These results may point toward the fabrication of cell-guiding scaffolds for regenerative endodontics, and may provide cues regarding the development of the pulp-dentin interface during tooth formation.
AB - Introduction: An understanding of the extracellular matrix characteristics which stimulate and guide stem cell differentiation in the dental pulp is fundamental for the development of enhanced dental regenerative therapies. Our objectives, in this study, were to determine whether stem cells from the apical papilla (SCAP) responded to substrate stiffness, whether hydrogels providing micropatterned topographical cues stimulate SCAP self-alignment, and whether the resulting alignment could influence their differentiation towards an odontogenic lineage in-vitro. Methods: Experiments utilized gelatin methacryloyl (GelMA) hydrogels of increasing concentrations (5, 10 and 15%). We determined their compressive modulus via unconfined compression and analyzed cell spreading via F-actin/DAPI immunostaining. GelMA hydrogels were micropatterned using photolithography, in order to generate microgrooves and ridges of 60 and 120 μm, onto which SCAP were seeded and analyzed for self-alignment via fluorescence microscopy. Lastly, we analyzed the odontogenic differentiation of SCAP using alkaline phosphatase protein expression (ANOVA/Tukey α = 0.05). Results: SCAP appeared to proliferate better on stiffer hydrogels. Both 60 and 120 μm micropatterned hydrogels guided the self-alignment of SCAP with no significant difference between them. Similarly, both 60 and 120 μm micropattern aligned cells promoted higher odontogenic differentiation than non-patterned controls. Significance: In summary, both substrate mechanics and geometry have a statistically significant influence on SCAP response, and may assist in the odontogenic differentiation of dental stem cells. These results may point toward the fabrication of cell-guiding scaffolds for regenerative endodontics, and may provide cues regarding the development of the pulp-dentin interface during tooth formation.
KW - GelMA
KW - Mechanotransduction
KW - Micropatterning
KW - Odontogenic differentiation
KW - Regenerative dentistry
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UR - http://www.scopus.com/inward/citedby.url?scp=85076797767&partnerID=8YFLogxK
U2 - 10.1016/j.dental.2019.10.013
DO - 10.1016/j.dental.2019.10.013
M3 - Article
C2 - 31780101
AN - SCOPUS:85076797767
VL - 36
SP - 88
EP - 96
JO - Dental Materials
JF - Dental Materials
SN - 0109-5641
IS - 1
ER -