Extracellular Vesicles of Human Periodontal Ligament Stem Cells Contain MicroRNAs Associated to Proto-Oncogenes: Implications in Cytokinesis

Abstract

The human Periodontal Ligament Stem Cells (hPDLSCs) exhibit self-renewal capacity and clonogenicity potential. The Extracellular Vesicles (EVs) secreted by hPDLSCs are particles containing lipids, proteins, mRNAs, and non-coding RNAs, among which microRNAs, that are important in intercellular communication. The purpose of this study was the analysis of the non-coding RNAs contained in the EVs derived from hPDLSCs using Next Generation Sequencing. Moreover, our data were enriched using bioinformatic tools. The analysis highlighted the presence of non-coding RNAs and five microRNAs: MIR24-2, MIR142, MIR335, MIR490, and MIR296. Our results show that these miRNAs target the genes classified in two terms of the Gene Ontology: "Ras protein signal transduction" and "Actin/microtubule cytoskeleton organization." Noteworthy, the in-deep analysis of our EVs highlights that the miRNAs could be implicated in the silencing of proto-oncogenes involved in 12 different types of tumors.

Oral Bone Tissue Regeneration: Mesenchymal Stem Cells, Secretome, and Biomaterials

“…In fact, they can be isolated from MSCs with different origins. OMSCs, such as GMSCs [ 104 ] and PDLSCs [ 105 ], produce EVs that have been shown to be useful in some models for regenerative therapies. Recent studies confirmed that MSC-derived EVs may be promising for bone regeneration [ 106 , 107 ].…”

Periodontal and Dental Pulp Cell-Derived Small Extracellular Vesicles: A Review of the Current Status

“…To date, there are no standardized protocols for sEV isolation and characterization. From the 33 studies that are reported in this review, we have summarized periodontal (dental pulp) cell-derived sEV isolation and characterisation methods [ 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 ]. Various isolation methods have been used for periodontal (dental pulp) cells sEVs, including ultracentrifugation (UC), precipitation-based methods, and ultrafiltration ( Figure 3 a).…”

“…We will define EV size <200 nm as sEV, and unclear EV size as EV. Regarding the EV content, it seems that hPDLCs-EV contain miRNAs [52,53] and circular RNAs [51] that may alter the recipient cells functions. RNA sequencing of hPDLSCs-EV (where EV size was unclear) revealed that hPDLSCs-EV contains 955 non-coding transcripts, with five representative miRNAs, including MIR24-2, MIR142, MIR296, MIR335, and MIR490 [53].…”

“…Current studies mainly focus on small EV biogenesis and function in the periodontal regeneration field; thus, this review summarizes 33 studies [ 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 ] on periodontal cell-, gingival cell- and dental pulp (DPSCs and SHED) cell-derived sEV isolation, characterization, and their therapeutic role in tissue regeneration. Most of this research has focused on the function of sEVs in cell differentiation, and 11 studies investigated the cargos of sEVs (i.e., miRNA [ 52 , 53 , 61 , 63 , 64 , 72 , 75 , 81 ], circRNA [ 51 , 71 ], lncRNA [ 51 ], and EV-mRNA [ 67 , 72 ]) during this process.…”

“…A total of 10 studies investigated sEVs derived from human PDL (stem) cells or fibroblasts [ 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 ], and are summarized in Table 2 . Eight studies isolated sEV from hPDLSCs [ 51 , 52 , 53 , 55 , 56 , 58 , 59 , 60 ], with one study each using sEVs from a human PDL fibroblast (hPDLFs) cell line [ 54 ] and hPDLCs [ 57 ]. Three of these studies investigated hPDLCs sEV function in vivo , using animal models [ 56 , 59 , 60 ].…”