title: Mineralization of dense collagen hydrogel scaffolds by human pulp cells.
abstract: While advances in biomineralization have been made in recent years, unanswered questions persist on bone- and tooth-cell differentiation, on outside-in signaling from the extracellular matrix, and on the link between protein expression and mineral deposition. In the present study, we validate the use of a bioengineered three-dimensional (3D) dense collagen hydrogel scaffold as a cell-culture model to explore these questions. Dental pulp progenitor/stem cells from human exfoliated deciduous teeth (SHEDs) were seeded into an extracellular matrix-like collagen gel whose fibrillar density was increased through plastic compression. SHED viability, morphology, and metabolic activity, as well as scaffold mineralization, were investigated over 24 days in culture. Additionally, measurements of alkaline phosphatase enzymatic activity, together with immunoblotting for mineralized tissue cell markers ALPL (tissue-non-specific alkaline phosphatase), DMP1 (dentin matrix protein 1), and OPN (osteopontin), demonstrated osteo/odontogenic cell differentiation in the dense collagen scaffolds coincident with mineralization. Analyses of the mineral phase by electron microscopy, including electron diffraction and energy-dispersive x-ray spectroscopy, combined with Fourier-transform infrared spectroscopy and biochemical analyses, were consistent with the formation of apatitic mineral that was frequently aligned along collagen fibrils. In conclusion, use of a 3D dense collagen scaffold promoted SHED osteo/odontogenic cell differentiation and mineralization.
authors: Faculty of Dentistry, McGill University, Montreal, QC, Canada.
authors: B R
authors: BR
authors: Coyac
authors: F
authors: F
authors: Chicatun
authors: B
authors: B
authors: Hoac
authors: V
authors: V
authors: Nelea
authors: C
authors: C
authors: Chaussain
authors: S N
authors: SN
authors: Nazhat
authors: M D
authors: MD
authors: McKee
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D001031:Apatites / Q000032:analysis; D015415:Biomarkers / Q000032:analysis; D002113:Calcification, Physiologic / Q000502:physiology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D048430:Cell Shape / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002648:Child; D002675:Child, Preschool; D003782:Dental Pulp / Q000166:cytology*; D005109:Extracellular Matrix / Q000737:chemistry; D016326:Extracellular Matrix Proteins / Q000032:analysis; D024022:Fibrillar Collagens / Q000737:chemistry; D005782:Gels; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D009805:Odontogenesis / Q000502:physiology; D010012:Osteogenesis / Q000502:physiology; D053495:Osteopontin / Q000032:analysis; D010750:Phosphoproteins / Q000032:analysis; D011312:Pressure; D013234:Stem Cells / Q000502:physiology*; D013997:Time Factors; D023822:Tissue Engineering / Q000295:instrumentation; D054457:Tissue Scaffolds / Q000737:chemistry; D014094:Tooth, Deciduous / Q000166:cytology
chemical_list: D001031:Apatites; D015415:Biomarkers; C513309:DMP1 protein, human; D016326:Extracellular Matrix Proteins; D024022:Fibrillar Collagens; D005782:Gels; D010750:Phosphoproteins; D053495:Osteopontin; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; C548267:ALPL protein, human; D000469:Alkaline Phosphatase
keywords: bioengineering; cell differentiation; deciduous teeth; dense collagen hydrogels; dental pulp stem cells; plastic compression; tissue engineering