Multilineage Differentiation Potential of Human Dental Pulp Stem Cells-Impact of 3D and Hypoxic Environment on Osteogenesis In Vitro.

Anna Labedz-Maslowska,Andrzej Kubiak,Zbigniew Madeja,Sylwia Noga,Ewa Zuba-Surma,Dariusz Boruczkowski,Tomasz Kaczmarzyk,Natalia Bryniarska,Malgorzata Sekula-Stryjewska

doi:10.3390/ijms21176172

Abstract

Human dental pulp harbours unique stem cell population exhibiting mesenchymal stem/stromal cell (MSC) characteristics. This study aimed to analyse the differentiation potential and other essential functional and morphological features of dental pulp stem cells (DPSCs) in comparison with Wharton’s jelly-derived MSCs from the umbilical cord (UC-MSCs), and to evaluate the osteogenic differentiation of DPSCs in 3D culture with a hypoxic microenvironment resembling the stem cell niche. Human DPSCs as well as UC-MSCs were isolated from primary human tissues and were subjected to a series of experiments. We established a multiantigenic profile of DPSCs with CD45−/CD14−/CD34−/CD29+/CD44+/CD73+/CD90+/CD105+/Stro-1+/HLA-DR− (using flow cytometry) and confirmed their tri-lineage osteogenic, chondrogenic, and adipogenic differentiation potential (using qRT-PCR and histochemical staining) in comparison with the UC-MSCs. The results also demonstrated the potency of DPSCs to differentiate into osteoblasts in vitro. Moreover, we showed that the DPSCs exhibit limited cardiomyogenic and endothelial differentiation potential. Decreased proliferation and metabolic activity as well as increased osteogenic differentiation of DPSCs in vitro, attributed to 3D cell encapsulation and low oxygen concentration, were also observed. DPSCs exhibiting elevated osteogenic potential may serve as potential candidates for a cell-based product for advanced therapy, particularly for bone repair. Novel tissue engineering approaches combining DPSCs, 3D biomaterial scaffolds, and other stimulating chemical factors may represent innovative strategies for pro-regenerative therapies.

Highlights

The mesenchymal stem/stromal cells (MSCs) represent an adult stem cell (SCs) population that exhibits tri-lineage differentiation potential [1]
Accumulating evidence indicates that the dental pulp contains a unique population of adherent cells with mesenchymal characteristics, including adhesion to plastic surfaces, fibroblast-like morphology, lack of expression of CD14, CD34 or CD45 and potential to differentiate into osteoblasts in vitro [18,43]
The results indicate that 3D cell encapsulation as well as the low concentration of O2 resembling conditions in the stem cell niches may favour osteogenic differentiation of dental pulp stem cells (DPSCs) in an in vitro environment

Summary

Introduction

The mesenchymal stem/stromal cells (MSCs) represent an adult stem cell (SCs) population that exhibits tri-lineage differentiation potential [1]. Recent evidence indicates that murine BM-MSCs may differentiate into hepatocytes, lung epithelial cells, myofibroblasts, and renal tubular cells in vivo [8]. Based on their wide differentiation potential and their pro-regenerative potential in injured tissues, MSCs are promising “candidates” as medicinal products employed in in vivo applications for advanced therapy in humans. Recent evidence demonstrates the biological variability (including differentiation capacity) within the MSCs of various origins [11]. The selection of the appropriate source of MSCs might be critical for the effective regeneration of injured tissues following their application in vivo

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