title: Selective enrichment of microRNAs in extracellular matrix vesicles produced by growth plate chondrocytes.
abstract: 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±0.78ng 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 RNAs 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.
authors.affiliation: Department of Periodontics, Virginia Commonwealth University, Richmond, VA, United States; Department of Biomedical Engineering, School of Engineering, Virginia Commonwealth University, Richmond, VA, United States.
authors.forename: Zhao
authors.initials: Z
authors.lastname: Lin
authors.affiliation: School of Biology, Virginia Commonwealth University, Richmond, VA, United States.
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Rodriguez
authors.affiliation: Department of Periodontics, Virginia Commonwealth University, Richmond, VA, United States; Department of Biomedical Engineering, School of Engineering, Virginia Commonwealth University, Richmond, VA, United States; General Dentistry, 9th People's Hospital, College of Stomatology, Shanghai JiaoTong University School of Medicine, Shanghai, China.
authors.forename: Junjun
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Biomedical Engineering, School of Engineering, Virginia Commonwealth University, Richmond, VA, United States.
authors.forename: Allison N
authors.initials: AN
authors.lastname: Ramey
authors.affiliation: Department of Biomedical Engineering, School of Engineering, Virginia Commonwealth University, Richmond, VA, United States.
authors.forename: Sharon L
authors.initials: SL
authors.lastname: Hyzy
authors.affiliation: Department of Biomedical Engineering, School of Engineering, Virginia Commonwealth University, Richmond, VA, United States; Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, United States. Electronic address: bboyan@vcu.edu.
authors.forename: Barbara D
authors.initials: BD
authors.lastname: Boyan
authors.affiliation: Department of Biomedical Engineering, School of Engineering, Virginia Commonwealth University, Richmond, VA, United States; Department of Periodontics, The University of Texas Health Science Center at San Antonio, San Antonio, TX, United States.
authors.forename: Zvi
authors.initials: Z
authors.lastname: Schwartz
mesh_terms: D000818:Animals; D019902:Chondrocytes / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D000067128:Extracellular Vesicles / Q000378:metabolism*; D006132:Growth Plate / Q000378:metabolism*; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D058977:Molecular Sequence Annotation; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012260:Ribonucleases / Q000378:metabolism
chemical_list: D035683:MicroRNAs; D012260:Ribonucleases
keywords: Bone formation; Cell membrane; Chondrocyte; Endochondral calcification; Exosomes; Matrix vesicles; MiRNA