Odontogenic Differentiation of Human Dental Pulp Stem Cells on Hydrogel Scaffolds Derived from Decellularized Bone Extracellular Matrix and Collagen Type I

Abstract

ObjectivesThe aim of this study was to evaluate the level of odontogenic differentiation of dental pulp stem cells (DPSCs) on hydrogel scaffolds derived from bone extracellular matrix (bECM) in comparison to those seeded on collagen I (Col-I), one of the main components of dental pulp ECM.MethodsDPSCs isolated from human third molars were characterized for surface marker expression and odontogenic potential prior to seeding into bECM or Col-I hydrogel scaffolds. The cells were then seeded onto bECM and Col-I hydrogel scaffolds and cultured under basal conditions or with odontogenic and growth factor (GF) supplements. DPSCs cultivated on tissue culture polystyrene (TCPS) with and without supplements were used as controls. Gene expression of dentin sialophosphoprotein (DSPP), dentin matrix protein 1 (DMP-1) and matrix extracellular phosphoglycoprotein (MEPE) was evaluated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and mineral deposition was observed by Von Kossa staining.ResultsWhen DPSCs were cultured on bECM hydrogels, the mRNA expression levels of DSPP, DMP-1 and MEPE genes were significantly upregulated with respect to those cultured on Col-I scaffolds or TCPS in the absence of extra odontogenic inducers. In addition, more mineral deposition was observed on bECM hydrogel scaffolds as demonstrated by Von Kossa staining. Moreover, DSPP, DMP-1 and MEPE mRNA expressions of DPSCs cultured on bECM hydrogels were further upregulated by the addition of GFs or osteo/odontogenic medium compared to Col-I treated cells in the same culture conditions.SignificanceThese results demonstrate the potential of the bECM hydrogel scaffolds to stimulate odontogenic differentiation of DPSCs.

Applications of decellularized extracellular matrix in bone and cartilage tissue engineering

“…Cells encapsulated in dECM hydrogels showed a higher proliferation rate than those in demineralized bone or collagen type I hydrogels. Paduano et al demonstrated that dental pulp stem cells (DPSCs), when seeded on dECM derived from bovine tibia, underwent odontogenic differentiation in vitro . Adding exogenous growth factors such as basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) to dECM hydrogels had a synergistic effect on the degree of differentiation .…”

Extracellular matrix hydrogels from decellularized tissues: Structure and function

“…The viability of cells cultured on the surface of ECM hydrogels in vitro has been consistently shown for cell lines [23, 54, 63, 64, 70, 71, 83], primary cells [57, 63, 69, 71, 75, 83, 85], and stem cells [44, 49, 50, 73, 82, 86]. In addition, the innate bioactivity of soluble factors within the ECM has been demonstrated using in vitro culture with media supplemented with solubilized ECM to remove the influence of hydrogel structure on the function of cells.…”

Vitamin D Effects on Osteoblastic Differentiation of Mesenchymal Stem Cells from Dental Tissues

“…The differentiation into osteogenic lineage has already been demonstrated in DPSCs [27–29, 34] and a considerable mineral matrix deposition has been observed using innovative scaffolds for their culturing [35, 36]; moreover these cells, if cultured on β -tricalcium phosphate/poly (l-lactic acid/caprolactone) three-dimensional scaffolds, showed an increased osteogenesis in the presence of Vit D compared to dexamethasone [37]. …”