Src blockage by siRNA inhibits VEGF-induced vascular hyperpemeability and osteoclast activity – an in vitro mechanism study for preventing destructive repair of osteonecrosis

Cao, Huijuan; Zheng, Lizhen; Wang, Nan; Wang, Lin-Ying; Li, Ye; Li, Dan; Wang, Xinluan; Qin, Ling

doi:10.1016/j.bone.2014.12.060

Cited by 24 publications

(16 citation statements)

References 36 publications

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“…We also observed that ONFH stimulated maturation, migration, adhesion, and pit formation (bone resorption) of osteoclasts. In many skeletal diseases including osteoporosis, osteoarthritis and osteonecrosis, osteoclast activities increase [43–45], and our results accord with them. Furthermore, geometric inspection revealed that the samples in the osteonecrosis group exhibited a shorter height and width of the femoral head than the sham operated control group.…”

Section: Discussionsupporting

confidence: 87%

Knee loading protects against osteonecrosis of the femoral head by enhancing vessel remodeling and bone healing

Liu

et al. 2015

Bone

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Osteonecrosis of the femoral head is a serious orthopedic problem. Moderate loads with knee loading promote bone formation, but its effects on osteonecrosis have not been investigated. Using a rat model, we examined a hypothesis that knee loading enhances vessel remodeling and bone healing through the modulation of the fate of bone marrow-derived cells. In this study, osteonecrosis was induced by transecting the ligamentum teres followed by a tight ligature around the femoral neck. For knee loading, 5 N loads were laterally applied to the knee at 15 Hz for 5 min/day for 5 weeks. Changes in bone mineral density (BMD) and bone mineral content (BMC) of the femur were measured by pDEXA, and ink infusion was performed to evaluate vessel remodeling. Femoral heads were harvested for histomorphometry, and bone marrow-derived cells were isolated to examine osteoclast development and osteoblast differentiation. The results showed that osteonecrosis significant induced bone loss, and knee loading stimulated both vessel remodeling and bone healing. The osteonecrosis group exhibited the lowest trabecular BV/TV (p < 0.001) in the femoral head, and lowest femoral BMD and BMC (both p < 0.01). However, knee loading increased trabecular BV/TV (p < 0.05) as well as BMD and BMC (both p < 0.05). Osteonecrosis decreased the vessel volume, vessel number and VEGF expression (all p < 0.01), and knee loading increased them (all p < 0.01). Osteonecrosis activated osteoclast development, and knee loading reduced its formation, migration, adhesion and the level of “pit” formation (all p < 0.001). Furthermore, knee loading significantly increased osteoblast differentiation and CFU-F (both p < 0.001). A significant positive correlation was observed between vessel remodeling and bone healing (both p < 0.01). These results indicate that knee loading could be effective in repair osteonecrosis of the femoral head in a rat model. This effect might be attributed to promoting vessel remodeling, suppressing osteoclast development, and increasing osteoblast and fibroblast differentiation. In summary, the current study suggests that knee loading might potentially be employed as a non-invasive therapy for osteonecrosis of the femoral head.

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Section: Discussionsupporting

confidence: 87%

Knee loading protects against osteonecrosis of the femoral head by enhancing vessel remodeling and bone healing

Liu

et al. 2015

Bone

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“…(30,31) We hypothesize that blockade of aberrant VEGF-Src signaling can inhibit both the vascular component (as evidenced by persistent marrow edema) and the skeletal component (ie, persistent bone resorption) of the destructive repair, but preserve VEGF-induced neovascularization in SAON. (32) We aim to determine the role of VEGF in destructive repair by supplementing and neutralizing VEGF, respectively, and define the role of Src through blocking Src by means of in vivo gene knockdown technology-RNAi using siRNA (33) with and without supplement of VEGF in a rabbit SAON model. (30,34) …”

Section: Introductionmentioning

confidence: 99%

Blockage of Src by Specific siRNA as a Novel Therapeutic Strategy to Prevent Destructive Repair in Steroid-Associated Osteonecrosis in Rabbits

Cao

Chen

et al. 2015

Journal of Bone and Mineral Research

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Vascular hyperpermeability and highly upregulated bone resorption in the destructive repair progress of steroid-associated osteonecrosis (SAON) are associated with a high expression of VEGF and high Src activity (Src is encoded by the cellular sarcoma [c-src] gene). This study was designed to prove our hypothesis that blocking the VEGF-Src signaling pathway by specific Src siRNA is able to prevent destructive repair in a SAON rabbit model. Destructive repair in SAON was induced in rabbits. At 2, 4, and 6 weeks after SAON induction, VEGF, anti-VEGF, Src siRNA, Src siRNAþVEGF, control siRNA, and saline were introduced via intramedullary injection into proximal femora for each group, respectively. Vascularization and permeability were quantified by dynamic contrast-enhanced (DCE) MRI. At week 6 after SAON induction, proximal femurs were dissected for micro-computed tomography (mCT)-based trabecular architecture with finite element analysis (FEA), mCT-based angiography, and histological analysis. Histological evaluation revealed that VEGF enhanced destructive repair, whereas anti-VEGF prevented destructive repair and Src siRNA and Src siRNAþVEGF prevented destructive repair and enhanced reparative osteogenesis. Findings of angiography and histomorphometry were consistent with those determined by DCE MRI. Src siRNA inhibited VEGF-mediated vascular hyperpermeability but preserved VEGF-induced neovascularization. Bone resorption was enhanced in the VEGF group and inhibited in the anti-VEGF, Src siRNA, Src siRNAþVEGF groups as determined by both 3D mCT and 2D histomorphometry. FEA showed higher estimated failure load in the Src siRNA and Src siRNAþVEGF groups when compared to the vehicle control group. Blockage of VEGF-Src signaling pathway by specific Src siRNA was able to prevent steroid-associated destructive repair while improving reconstructive repair in SAON, which might become a novel therapeutic strategy.

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“…While the mechanisms orchestrating endothelial repair, particularly considering those of the BBB, are not entirely defined, some molecular effectors and interconnecting pathways have been identified in the literature for their demonstrated involvement in repair processes in various model systems. In particular, cellular Src kinase (c-Src), a ubiquitously expressed member of the Src family of non-receptor tyrosine kinases, has a defined role in endothelial cell regulation and repair, both in vitro and in vivo (Takenaga et al, 2009; Liu et al, 2010; Franco et al, 2013; Bai et al, 2014; Cao et al, 2015), the specifics of which are described below. Importantly, c-Src is involved in pathways linked to the activation of vascular endothelial growth factor receptor 2 (VEGFR2) and N-methyl D-aspartate receptor (NMDAR), both of which are expressed on human BMECs and with demonstrated roles in BBB regulation and integrity (Sharp et al, 2003, 2005; Andras et al, 2007; Holmes et al, 2007; Davis et al, 2010; Reijerkerk et al, 2010; Hudson et al, 2014; Chen et al, 2016; Fearnley et al, 2016).…”

mentioning

confidence: 99%

“…To further support the role of c-Src in BBB health and repair, it has been reported that inhibition of c-Src by siRNA limited permeability of human umbilical vein endothelial cells (HUVEC) exposed to vascular endothelial growth factor (VEGF, a known inducer of permeability of the BBB; Holmes et al, 2007; Davis et al, 2010; Hudson et al, 2014; Cao et al, 2015; Fearnley et al, 2016). In addition, chemical inhibition of c-Src with the inhibitor 1-(1,1-dimethylethyl)-3-(4-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (PP1) accelerated healing of wounded HUVEC (Franco et al, 2013), in vitro .…”

mentioning

confidence: 99%

“…Furthermore, in vivo treatment with the broad Src family inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl)-pyrazolo[3,4-d]pyrimidine (PP2) in rats subjected to ischemic insult resulted in the rescue of ischemic BBB leakage (Takenaga et al, 2009) and improved neurological deficit scores (Bai et al, 2014) in the presence of the inhibitor. Moreover, c-Src has been identified as an upstream regulator of a number of tight junction complex components, including occludin (Takenaga et al, 2009), claudin-5 (Bai et al, 2014), and zona occludens-1 (Morin-Brureau et al, 2011), as well as a modulator of NMDAR activity in neurons (Lu et al, 1999; Yu and Salter, 1999; Rong et al, 2001; Heidinger et al, 2002; Hossain et al, 2012; Tang et al, 2012; Krogh et al, 2014), and a downstream effector of the VEGFR2 signaling pathway (He et al, 1999; Morin-Brureau et al, 2011; Sun et al, 2012; Cao et al, 2015), in addition to its role in cell cycle regulation and proliferation (Boggon and Eck, 2004; Parsons and Parsons, 2004; Hu et al, 2008; Sen and Johnson, 2011; Reinecke and Caplan, 2014). …”

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confidence: 99%

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Opinion: Inhibition of Blood-Brain Barrier Repair as a Mechanism in HIV-1 Disease

2017

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Src blockage by siRNA inhibits VEGF-induced vascular hyperpemeability and osteoclast activity – an in vitro mechanism study for preventing destructive repair of osteonecrosis

Cited by 24 publications

References 36 publications

Knee loading protects against osteonecrosis of the femoral head by enhancing vessel remodeling and bone healing

Knee loading protects against osteonecrosis of the femoral head by enhancing vessel remodeling and bone healing

Blockage of Src by Specific siRNA as a Novel Therapeutic Strategy to Prevent Destructive Repair in Steroid-Associated Osteonecrosis in Rabbits

Opinion: Inhibition of Blood-Brain Barrier Repair as a Mechanism in HIV-1 Disease

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