Journal
BIOENGINEERING-BASEL
Volume 10, Issue 3, Pages -Publisher
MDPI
DOI: 10.3390/bioengineering10030323
Keywords
tissue engineering; dental pulp; stem cells; elasticity; scaffold; cellular mechanotransduction; regeneration
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This study investigated the differentiation of dental pulp stem cells (DPSCs) towards soft tissue using extracellular elasticity. DPSCs cultured on substrates with a low elasticity of 1.5 kPa showed a soft tissue-like phenotype and gene expression, while cells cultured on substrates with a higher elasticity of 28 kPa exhibited a differentiation signature resembling hard tissues. Cells cultured on substrates with an intermediate elasticity of 15 kPa showed the highest cytokine expression, suggesting their potential role as trophic mediators.
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.
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