Biomechanical Modulation of Dental Pulp Stem Cell (DPSC) Properties for Soft Tissue Engineering

Abstract

Dental pulp regeneration strategies frequently result in hard tissue formation and pulp obliteration. The aim of this study was to investigate whether dental pulp stem cells (DPSCs) can be directed toward soft tissue differentiation by extracellular elasticity. STRO-1-positive human dental pulp cells were magnetically enriched and cultured on substrates with elasticities of 1.5, 15, and 28 kPa. The morphology of DPSCs was assessed visually. Proteins relevant in mechanobiology ACTB, ITGB1, FAK, p-FAK, TALIN, VINCULIN, PAXILLIN, ERK 1/2, and p-ERK 1/2 were detected by immunofluorescence imaging. Transcription of the pulp marker genes BMP2, BMP4, MMP2, MMP3, MMP13, FN1, and IGF2 as well as the cytokines ANGPT1, VEGF, CCL2, TGFB1, IL2, ANG, and CSF1 was determined using qPCR. A low stiffness, i.e., 1.5 kPa, resulted in a soft tissue-like phenotype and gene expression, whereas DPSCs on 28 kPa substrates exhibited a differentiation signature resembling hard tissues with a low cytokine expression. Conversely, the highest cytokine expression was observed in cells cultured on intermediate elasticity, i.e., 15 kPa, substrates possibly allowing the cells to act as “trophic mediators”. Our observations highlight the impact of biophysical cues for DPSC fate and enable the design of scaffold materials for clinical pulp regeneration that prevent hard tissue formation.

In Vivo Evaluation of Collagen and Chitosan Scaffold, Associated or Not with Stem Cells, in Bone Repair

“…These cells can be obtained from postnatal teeth, extracted wisdom teeth, or exfoliated deciduous teeth [39,40]. Depending on their multipotency and sensitivity to local paracrine activity, DPSCs could be used for therapeutic applications in tissue regeneration [41,42], in addition to other advantages such as immunomodulatory properties, good proliferation capacity and osteogenic differentiation [37,43,44]. Furthermore, there is an abundant source of these cells as they are obtained from discarded or removed teeth [45].…”

“…Collagen and chitosan are easy to obtain, but few studies correlate the effectiveness of these natural biopolymers combined with human DPSCs in the treatment of craniomaxillofacial defects, which require rapid recovery in order to morpho-physiologically restore the bone tissue. Considering these uncertainties about the use of DPSCs [31,[41][42][43], and using the hypothesis that these matrices can contribute to the repair process and support for stem cells, the aim of this study was to analyze the repair of mandibular defects, filled with a new bovine collagen/chitosan scaffold, associated or not with DPSCs.…”

Polyelectrolyte‐Cation Complexes Using PAsp‐Sr Complexes Induce Biomimetic Mineralization with Antibacterial Ability

“…The culture of hDPSCs is described in detail in Supporting Information. [ 30 ] Cells from passages 3 to 5 were seeded in 24‐well culture plates (2.5 × 10 4 cells well −1 ) and cultured for 1 day before use.…”

Biology, Preclinical and Clinical Uses of Mesenchymal Dental Pulp Stem Cells

“…At the moment that cells reach the 70% of confluence several passages are performed until we obtain the desired number of cells. Then, cells are cyoprotected using agents that maintain the integrity of the cells while the temperature is gradually reduced to be transferred to the liquid nitrogen tank, where they are stored until use (Figure 2) [13][14][15][16].…”