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Conference Paper: Injectable Highly Tunable Collagen Hydrogel Scaffold for Pulp-Dentin Tissue Regeneration

TitleInjectable Highly Tunable Collagen Hydrogel Scaffold for Pulp-Dentin Tissue Regeneration
Authors
Han, YuanyuanXu, JinpingZhang, ZhaochengChopra, HiteshDissanayaka, WarunaNör, JacquesBottino, Marco
Issue Date15-May-2023
Abstract

Objectives: To investigate the regenerative capacity of highly tunable (stiffness) dental pulp stem cells/DPSCs-laden collagen hydrogel for pulp-dentin regeneration. We explored the role of collagen matrix stiffness to modulate DPSCs’ differentiation to enable simultaneous pulp-dentin regeneration.
Methods: A commercial collagen hydrogel was prepared at two concentrations: 3mg/ml(Col3) and 10mg/ml(Col10). Lithium-phenyl(2,4,6-trimethylbenzoyl)phosphinate was added as a photocrosslinker. Stiffness of light-cured Col3 and Col10 was assessed by rheology (n=3) and microstructure by electron microscopy/SEM. Isolated dog DPSCs were evaluated for CD34, CD44, and CD90 by flow-cytometry. DPSCs viability in Col3 and Col10 was determined by live/dead assay (n=3). RT-PCR was performed to quantify the mRNA expression of endothelial and odontogenic markers within DPSCs-laden Col3 and Col10 in-vitro. For in-vivo experiments, the synergistic effect of growth factor (GF) incorporation was evaluated by encapsulating vascular endothelial growth factor (VEGF[50ng/mL]) into Col3 and bone morphogenetic protein (BMP2[50ng/mL]) into Col10. 5 groups were established (n=6): blank, Col10, Col10+Col3, Col10+Col3+DPSCs, and Col10+Col3+DPSCs+GFs. DPSCs-encapsulated collagen matrices were injected into tooth slices pulp chambers and implanted subcutaneously in SCID mice for 6 weeks. Histological and immunofluorescent/IF staining were performed to evaluate pulp-dentin regeneration. Statistical analysis was performed using Two-way ANOVA (α=0.05).
Results: Stiffness and SEM data revealed that Col10 has 6305Pa stiffness with a more condensed, less porous structure, whereas Col3 has 565Pa with a looser microstructure. Both Col3 and Col10 supported DPSCs viability and proliferation. RT-PCR results showed Col3 promoted significantly higher vWF and CD31 expression after 7 and 14 days of endothelial differentiation (P<0.05), whereas Col10 enhanced the expression of DSPP, ALP, Runx2, and Col1 after 7, 14, and 21 days of odontogenic differentiation (P<0.05). H&E and IF (CD31 and vWF) staining revealed Col10+Col3+DPSCs+GFs supported more pronounced pulp-dentin tissue regeneration.
Conclusions: The collagen-based concentric construct modified by growth factors guides the specific lineage differentiation of DPSCs and promotes pulp-dentin tissue regeneration.


Persistent Identifierhttp://hdl.handle.net/10722/337812

 

DC FieldValueLanguage
dc.contributor.authorHan, Yuanyuan-
dc.contributor.authorXu, Jinping-
dc.contributor.authorZhang, Zhaocheng-
dc.contributor.authorChopra, Hitesh-
dc.contributor.authorDissanayaka, Waruna-
dc.contributor.authorNör, Jacques-
dc.contributor.authorBottino, Marco-
dc.date.accessioned2024-03-11T10:24:04Z-
dc.date.available2024-03-11T10:24:04Z-
dc.date.issued2023-05-15-
dc.identifier.urihttp://hdl.handle.net/10722/337812-
dc.description.abstract<p>Objectives: To investigate the regenerative capacity of highly tunable (stiffness) dental pulp stem cells/DPSCs-laden collagen hydrogel for pulp-dentin regeneration. We explored the role of collagen matrix stiffness to modulate DPSCs’ differentiation to enable simultaneous pulp-dentin regeneration.<br>Methods: A commercial collagen hydrogel was prepared at two concentrations: 3mg/ml(Col3) and 10mg/ml(Col10). Lithium-phenyl(2,4,6-trimethylbenzoyl)phosphinate was added as a photocrosslinker. Stiffness of light-cured Col3 and Col10 was assessed by rheology (n=3) and microstructure by electron microscopy/SEM. Isolated dog DPSCs were evaluated for CD34, CD44, and CD90 by flow-cytometry. DPSCs viability in Col3 and Col10 was determined by live/dead assay (n=3). RT-PCR was performed to quantify the mRNA expression of endothelial and odontogenic markers within DPSCs-laden Col3 and Col10 <em>in-vitro</em>. For <em>in-vivo</em> experiments, the synergistic effect of growth factor (GF) incorporation was evaluated by encapsulating vascular endothelial growth factor (VEGF[50ng/mL]) into Col3 and bone morphogenetic protein (BMP2[50ng/mL]) into Col10. 5 groups were established (n=6): blank, Col10, Col10+Col3, Col10+Col3+DPSCs, and Col10+Col3+DPSCs+GFs. DPSCs-encapsulated collagen matrices were injected into tooth slices pulp chambers and implanted subcutaneously in SCID mice for 6 weeks. Histological and immunofluorescent/IF staining were performed to evaluate pulp-dentin regeneration. Statistical analysis was performed using Two-way ANOVA (α=0.05).<br>Results: Stiffness and SEM data revealed that Col10 has 6305Pa stiffness with a more condensed, less porous structure, whereas Col3 has 565Pa with a looser microstructure. Both Col3 and Col10 supported DPSCs viability and proliferation. RT-PCR results showed Col3 promoted significantly higher vWF and CD31 expression after 7 and 14 days of endothelial differentiation (P<0.05), whereas Col10 enhanced the expression of DSPP, ALP, Runx2, and Col1 after 7, 14, and 21 days of odontogenic differentiation (P<0.05). H&E and IF (CD31 and vWF) staining revealed Col10+Col3+DPSCs+GFs supported more pronounced pulp-dentin tissue regeneration.<br>Conclusions: The collagen-based concentric construct modified by growth factors guides the specific lineage differentiation of DPSCs and promotes pulp-dentin tissue regeneration.<br></p>-
dc.languageeng-
dc.relation.ispartof2023 AADOCR/CADR Annual Meeting (15/05/2023-18/05/2023, Portland, Oregon)-
dc.titleInjectable Highly Tunable Collagen Hydrogel Scaffold for Pulp-Dentin Tissue Regeneration-
dc.typeConference_Paper-

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