Selective enrichment of microRNAs in extracellular matrix vesicles produced by growth plate chondrocytes

Matrix vesicles (MVs) are membrane organelles found in the extracellular matrix of calcifying cells, which contain matrix processing enzymes and regulate the extracellular environment via action of these enzymes. It is unknown whether MVs are also exosomic mediators of cell-cell communication via transfer of RNA material, and specifically, microRNA (miRNA). We investigated the presence of RNA in MVs isolated from cultures of costochondral growth zone chondrocytes. Our results showed that the average yield of MV RNA was 1.93 +/- 0.78 ng RNA/10(4) cells, which was approximately 0.1% of the parent cell's total RNA. MV RNA was well protected from RNase by the lipid membrane and was highly enriched in small RNA molecules compared to cells. Moreover, coding and non-coding small RNA5 in MVs were in proportions that differed from parent cells. Enrichment of specific miRNAs was consistently observed in all three miRNA detection platforms that we used, suggesting that miRNAs are selectively packaged into MVs. MV-enriched miRNAs were related to different signaling pathways associated with bone formation. This study suggests a significant role for MVs as matrisomes in cell-cell communication in cartilage and bone development via transfer of specific miRNAs. (C) 2016 Elsevier Inc. Allrights reserved.