Recently, several studies have indicated that circular RNAs (circRNAs) play significant roles in various disease; however, little is known about the chronology of estrogen receptor beta (ER beta) deficiency and altered circRNA expression, or their relationship with osteogenesis. Herein, we show through western blot and quantitative real-time PCR assays, that when ER beta is silenced, the expression of osteogenesis-related proteins and mRNAs were down-regulated. We then performed RNA-Seq to analyze differential circRNA expression between the control and ER beta knockdown group. This analysis revealed that, 146 circRNAs were differentially expressed by fold-change >= 2.0, p <= 0.05, and, among this group, 68 circRNAs were down-regulated, while 78 were up-regulated. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and PANTHER pathway analyses were performed to predict the function of these differentially expressed circRNAs. Finally, co-expressed targets gene, and circRNA-microRNA network were constructed for predicted miRNA sponges. This research suggested that ER beta may through 2:27713879 vertical bar 27755789/2:240822115 vertical bar 240867796-miR-328-5p-mRNA axis to regulate osteogenic differentiation. (C) 2017 Published by Elsevier Inc.