Effect of advanced oxidation protein products on the proliferation and osteogenic differentiation of rat mesenchymal stem cells

Abstract

Abstract. Advanced oxidation protein products (AOPPs) as a novel marker of oxidative stress, are involved in a variety of diseases, including osteoporosis. Although a number of studies have shown the possible functions of AOPPs in biological processes, little is known about the role of AOPPs in the pathogenesis of osteoporosis. In this study, we aimed to investigate the effect of AOPPs on the proliferation and osteogenic differentiation of rat mesenchymal stem cells (MSCs). MSCs, isolated from bone marrow, were cultured in the absence or presence of AOPPs (50, 100, 200 and 400 mg/ml). MTT assay was used to determine the proliferative ability of the cells. Alkaline phosphatase (ALP) activity, the mRNA expression of ALP and collagen I and bone nodule formation were detected to assess osteogenic differentiation. Reactive oxygen species (ROS) generation was analyzed with the probe 2' ,7'-dichlorodihydrofluorescein diacetate (DCFH-DA). The expression of receptor of advanced glycation end-products (RAGE) at the mRNA and protein level was detected by real-time PCR and western blot analysis, respectively. Compared with the control group, AOPPs inhibited MSC proliferation in a dose-and timedependent manner. Moreover, AOPPs induced a significant reduction in ALP activity, as well as a decrease in ALP and collagen I mRNA levels in the MSCs; bone nodule formation was also inhibited. Furthermore, AOPPs increased ROS generation in the MSCs, and upregulated the expression of RAGE at the mRNA and protein level. These results suggest that AOPPs inhibit the proliferation and osteogenic differentiation of MSCs, possibly through the AOPPs-RAGE-ROS pathway; this may be an important mechanism in the development of osteoporosis.

Regulation of the mitochondrial reactive oxygen species: Strategies to control mesenchymal stem cell fates ex vivo and in vivo

“…In general, undifferentiated MSCs have fewer mitochondria than differentiated MSCs, though the mitochondrial copy number, OXPHOS supercomplex, SOD expression, mitochondrial biogenesis and ROS levels are all significantly increased after specific differentiation . ROS imbalance will lead to a significantly reduced MSC differentiation capacity, thus limiting MSC applications in vitro and in vivo. When adult MSCs are incubated under stress‐inducing conditions, they are considered to be in a state of cyclic stretch and produce more ROS and have weaker differentiation capacities .…”

“…In general, undifferentiated MSCs have fewer mitochondria than differentiated MSCs, though the mitochondrial copy number, OXPHOS supercomplex, SOD expression, mitochondrial biogenesis and ROS levels are all significantly increased after specific differentiation. 51,52 ROS imbalance will lead to a significantly reduced MSC differentiation capacity, 53 thus limiting MSC applications in vitro and in vivo.…”

Osteogenic Differentiation of Periosteal Cells During Fracture Healing

“…However unregulated levels of ROS are harmful, which lead to the induction of apoptosis (Martindale and Holbrook, 2002). Some reports suggested that ROS may be involved in the proliferation of osteoprogenitors and their subsequent differentiation (Imhoff and Hansen, 2011;Sun et al, 2013). The ROS level is increased after fracture, and oxidative stress may be proportional to the number of bones fractured (Turgut et al, 1999;Prasad et al, 2003).…”

Advanced oxidation protein products induce apoptosis, and upregulate sclerostin and RANKL expression, in osteocytic MLO-Y4 cells via JNK/p38 MAPK activation

“…Previous studies have demonstrated that AOPPs may inhibit the proliferation and differentiation of rat osteoblastic cells and rat mesenchymal stem cells (8,9). As the most abundant cell type in bone (90–95%), osteocytes function as more than just mechanosensors in bone homeostasis.…”