Engineered Extracellular Vesicles From Human Periodontal-Ligament Stem Cells Increase VEGF/VEGFR2 Expression During Bone Regeneration

Abstract

Bone regeneration represents still a challenge, in particular for calvarium defects. Recently, the development of biomaterials with the addiction of stem cells is giving promising results for the treatment of bone defects. In particular, it was demonstrated that scaffolds enriched with mesenchymal stem cells (MSCs) and/or their derivatives, such as conditioned medium (CM) and extracellular vesicles (EVs), may improve bone regeneration. Moreover, given the deep link between osteogenesis and angiogenesis, a successful approach must also take into consideration the development of vascularization. In this work we evaluated the bone regeneration capacity of a collagen membrane (3D-COL) enriched with human periodontal-ligament stem cells (hPDLSCs) and CM or EVs or EVs engineered with polyethylenimine (PEI-EVs) in rats subjected to a calvarial defect. We evaluated also their capacity to induce angiogenic factors. At first, in vitro results showed an increased expression of osteogenic markers in hPDLSCs cultured with the 3D-COL and PEI-EVs, associated also with the increased protein levels of Vascular endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2). The increased expression of these proteins was confirmed also in vivo in rats implanted with the 3D-COL enriched with hPDLSCs and PEI-EVs. Moreover, histological examination evidenced in this group of rats the activation of bone regeneration and of the vascularization process. Also MicroCT imaging with morphometric analysis confirmed in rats transplanted with 3D-COL enriched with hPDLSCs and PEI-EVs an important regenerative process and a better integration level. All together, these results evidenced that the 3D-COL enriched with hPDLSCs and PEI-EVs may promote bone regeneration of calvaria defects, associated also with an increased vascularization.

Extracellular Vesicles Derived from Human Gingival Mesenchymal Stem Cells: A Transcriptomic Analysis

“…In rats subjected to calvarial defects, the scaffolds enriched with EVs/PEI-EVs and hPDLSCs improved also the vascularization process. Indeed, the pro-angiogenic factor VEGF resulted more expressed in the calvaria of the rats grafted with the scaffold [55]. Therefore, the expression of the pro-angiogenic factors in our EVs, as demonstrated in this work, supports the angiogenetic properties of EVs and their possible therapeutic role in the field of tissue repair.…”

A Novel Role of Ascorbic Acid in Anti-Inflammatory Pathway and ROS Generation in HEMA Treated Dental Pulp Stem Cells

“…Mohammed and colleagues showed that AS could promote wound healing process through a variety of mechanisms for example it protects vascular endothelium function through vascular endothelial growth factor (VEGF) upregulation, a fundamental protein in damaged tissue regeneration [68,69]. Furthermore, in our experimental conditions, AS inhibited the HEMA-induced effect on wound healing, accelerating the processes by reducing the scar area in hDPSCs.…”

Dental Stem Cell-Derived Secretome/Conditioned Medium: The Future for Regenerative Therapeutic Applications

“…The increased expression of these proteins was confirmed in clavarial defects implanted with the 3D-COL loaded with human periodontal ligament MSCs and PEI-EVs. Moreover, histological examination and micro-CT imaging confirmed this regenerative ability [193]. Likewise, Evolution (Evo) (a commercially available collagen membrane) loaded with human periodontal ligament MSCs enriched with EVs and PEI-EVs demonstrated high biocompatibility and osteogenic properties in vitro and in rats' calvarial defects.…”

“…Dental MSC-CM promotes osteogenesis through enhancing the migration and mineralization potential of MSCs by TGF-β1 [153] as well as the upregulation of their osteoblastic and chondrogenic marker expression (Osterix, SOX-5, factor 8) [154]. In this context, TGF-β-BMP signaling pathway plays a pivot role in osseous regeneration induced by dental MSCs and their secretome through upregulating the expression of TGF-β1, TGF-β2, BMP2, BMP4, MMP8, TUFT1, TFIP11, RUNX2, and SOX-9 was detected [194], as well as VEGF, VEGFR2, and COL1A1 [193]. The osteoblastic differentiation potential is primarily mediated by TGF-βR1, SMAD1, BMP2, MAPK1, MAPK14, and RUNX2 through the TGF-β signaling pathway [177].…”