Angiogenic Effects of Human Dental Pulp and Bone Marrow-Derived Mesenchymal Stromal Cells and their Extracellular Vesicles

Abstract

Blood vessel formation or angiogenesis is a key process for successful tooth regeneration. Bone marrow-derived mesenchymal stromal cells (BM-MSCs) possess paracrine proangiogenic properties, which are, at least partially, induced by their extracellular vesicles (EVs). However, the isolation of BM-MSCs is associated with several drawbacks, which could be overcome by MSC-like cells of the teeth, called dental pulp stromal cells (DPSCs). This study aims to compare the angiogenic content and functions of DPSC and BM-MSC EVs and conditioned medium (CM). The angiogenic protein profile of DPSC- and BM-MSC-derived EVs, CM and EV-depleted CM was screened by an antibody array and confirmed by ELISA. Functional angiogenic effects were tested in transwell migration and chicken chorioallantoic membrane assays. All secretion fractions contained several pro- and anti-angiogenic proteins and induced in vitro endothelial cell motility. This chemotactic potential was higher for (EV-depleted) CM, compared to EVs with a stronger effect for BM-MSCs. Finally, BM-MSC CM, but not DPSC CM, nor EVs, increased in ovo angiogenesis. In conclusion, we showed that DPSCs are less potent in relation to endothelial cell chemotaxis and in ovo neovascularization, compared to BM-MSCs, which emphasizes the importance of choice of cell type and secretion fraction for stem cell-based regenerative therapies in inducing angiogenesis.

Evolving applications of the egg: chorioallantoic membrane assay andex vivoorganotypic culture of materials for bone tissue engineering

“…The main use in research of the CAM is in the assessment of angiogenic, 21 , 41 anti-angiogenic or toxic 42 responses to drugs, 43 cells, extracellular vesicles 44 or materials, 3 biocompatibility of materials, 45 the growth or treatment of cancer 46 – 48 and xenograft application 49 – 52 for tissue growth and testing treatment modalities. Oncology research remains, to date, the most common field using the CAM assay.…”

Pancreas-derived mesenchymal stromal cells share immune response-modulating and angiogenic potential with bone marrow mesenchymal stromal cells and can be grown to therapeutic scale under Good Manufacturing Practice conditions

“…Supernatants were frozen at À80°C for Luminex analysis of protein expression (see following). Expression of chemokine, cytokine, chemokine receptor and cytokine receptor genes was assessed using quantitative polymerase chain reaction and human chemokine and receptor RT 2 Profiler polymerase chain reaction array (Qiagen) as previously described [12].…”

Therapeutic Functions of Stem Cells from Oral Cavity: An Update

“…The ectopic expression of angiogenic and neurogenic factors within the dental MSC-CM would contribute to dental tissue regeneration [165][166][167]. Interestingly, exosomes isolated from DPSC-CM could also exert similar beneficial actions as CM itself in terms of odontogenic induction and dental pulp regeneration [168,169]. Considering their convenience of manipulation, storage, handling, and management compared to whole CM or cells, exosomes could serve as a novel potent option in the dental MSC therapeutic field.…”