Gensheng Zhang1, Ruiyu Liu2, Xiaoqian Dang2, Jichao Liu1, Haibin Jiao1. 1. Department of Orthopaedics, 3201 Hospital of Xi'an Jiaotong University Health Science Center, Hanzhong Shaanxi, 723000, P.R.China. 2. Department of Orthopaedics, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an Shaanxi, 710004, P.R.China.
Abstract
OBJECTIVE: To investigate whether exosomes derived from miR-27a-overexpressing human umbilical vein endothelial cells (HUVECs)-exo (miR-27a) can promote bone regeneration and improve glucocorticoids (GC) induced osteonecrosis of femoral head (ONFH) (GC-ONFH). METHODS: The exo (miR-27a) were intended to be constructed and identified by transmission electron microscopy, nanoparticle tracking analysis, Western blot, and real-time fluorescent quantitative PCR (qRT-PCR). qRT-PCR was used to evaluate the effect of exo (miR-27a) in delivering miR-27a to osteoblasts (MC3T3-E1 cells). Alkaline phosphatase staining, alizarin red staining, and qRT-PCR were used to evaluate its effect on MC3T3-E1 cells osteogenesis. Dual-luciferase reporter (DLRTM) assay was used to verify whether miR-27a targeting Dickkopf WNT signaling pathway inhibitor 2 (DKK2) was a potential mechanism, and the mechanism was further verified by qRT-PCR, Western blot, and alizarin red staining in MC3T3-E1 cells. Finally, the protective effect of exo (miR-27a) on ONFH was verified by the GC-ONFH model in Sprague Dawley (SD) rats. RESULTS: Transmission electron microscopy, nanoparticle tracking analysis, Western blot, and qRT-PCR detection showed that exo (miR-27a) was successfully constructed. exo (miR-27a) could effectively deliver miR-27a to MC3T3-E1 cells and enhance their osteogenic capacity. The detection of DLRTM showed that miR-27a promoted bone formation by directly targeting DDK2. Micro-CT and HE staining results of animal experiments showed that tail vein injection of exo (miR-27a) improved the osteonecrosis of SD rat GC-ONFH model. CONCLUSION: exo (miR-27a) can promote bone regeneration and protect against GC-ONFH to some extent.
OBJECTIVE: To investigate whether exosomes derived from miR-27a-overexpressing human umbilical vein endothelial cells (HUVECs)-exo (miR-27a) can promote bone regeneration and improve glucocorticoids (GC) induced osteonecrosis of femoral head (ONFH) (GC-ONFH). METHODS: The exo (miR-27a) were intended to be constructed and identified by transmission electron microscopy, nanoparticle tracking analysis, Western blot, and real-time fluorescent quantitative PCR (qRT-PCR). qRT-PCR was used to evaluate the effect of exo (miR-27a) in delivering miR-27a to osteoblasts (MC3T3-E1 cells). Alkaline phosphatase staining, alizarin red staining, and qRT-PCR were used to evaluate its effect on MC3T3-E1 cells osteogenesis. Dual-luciferase reporter (DLRTM) assay was used to verify whether miR-27a targeting Dickkopf WNT signaling pathway inhibitor 2 (DKK2) was a potential mechanism, and the mechanism was further verified by qRT-PCR, Western blot, and alizarin red staining in MC3T3-E1 cells. Finally, the protective effect of exo (miR-27a) on ONFH was verified by the GC-ONFH model in Sprague Dawley (SD) rats. RESULTS: Transmission electron microscopy, nanoparticle tracking analysis, Western blot, and qRT-PCR detection showed that exo (miR-27a) was successfully constructed. exo (miR-27a) could effectively deliver miR-27a to MC3T3-E1 cells and enhance their osteogenic capacity. The detection of DLRTM showed that miR-27a promoted bone formation by directly targeting DDK2. Micro-CT and HE staining results of animal experiments showed that tail vein injection of exo (miR-27a) improved the osteonecrosis of SD rat GC-ONFH model. CONCLUSION: exo (miR-27a) can promote bone regeneration and protect against GC-ONFH to some extent.
Entities:
Keywords:
MiR-27a; exosomes; gene targeted therapy; human umbilical vein endothelial cells; osteogenesis; osteonecrosis of femoral head
Authors: Y Sato; H Bando; M Di Piazza; G Gowing; C Herberts; S Jackman; G Leoni; S Libertini; T MacLachlan; J W McBlane; L Pereira Mouriès; M Sharpe; W Shingleton; B Surmacz-Cordle; K Yamamoto; J W van der Laan Journal: Cytotherapy Date: 2019-11-09 Impact factor: 5.414
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