Conditioned Medium from Bone Marrow Mesenchymal Stem Cells Restored Oxidative Stress-Related Impaired Osteogenic Differentiation

Abstract

Oxidative stress from high levels of intracellular reactive oxygen species (ROS) has been linked to various bone diseases. Previous studies indicate that mesenchymal stem cells (MSC) secrete bioactive factors (conditioned medium (MSC-CM)) that have antioxidant effects. However, the antioxidant role of MSC-CM on osteogenesis has not been fully studied. We aimed to identify antioxidant proteins in MSC-CM using mass spectrometry-based proteomics and to explore their effects on osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSC) exposed to oxidative stress induced by hydrogen peroxide (H2O2). Our analysis revealed that MSC-CM is comprised of antioxidant proteins that are involved in several biological processes, including negative regulation of apoptosis and positive regulation of cell proliferation. Then, hBMSC exposed to H2O2 were treated with MSC-CM, and the effects on their osteogenic differentiation were evaluated. MSC-CM restored H2O2-induced damage to hBMSC by increasing the antioxidant enzyme-SOD production and the mRNA expression level of the anti-apoptotic BCL-2. A decrease in ROS production and cellular apoptosis was also shown. MSC-CM also modulated mRNA expression levels of osteogenesis-related genes, runt-related transcription factor 2, collagen type I, bone morphogenic protein 2, and osteopontin. Furthermore, collagen type I protein secretion, alkaline phosphatase activity, and in vitro mineralization were increased. These results indicate that MSC-CM contains several proteins with antioxidant and anti-apoptotic properties that restored the impaired hBMSC osteogenic differentiation associated with oxidative stress.

Brief communication: Effects of conditioned media from human platelet lysate cultured MSC on osteogenic cell differentiation in vitro

“…Cell metabolism and survival are controlled by the action of growth factors and cytokines through inhibition of apoptosis or promotion of cell survival. In previous studies, we have shown that CM from FBS-cultured BMSC contains several growth factors and cytokines with antiapoptotic and antioxidant properties, including platelet-derived growth factor (PDGF) and vascular endothelial growth factor A (VEGFA), and enhances in vitro osteogenic differentiation of MSC ( Al-Sharabi et al, 2014 ; Al-Sharabi et al, 2016 ; Saleem et al, 2021 ). In context, HPL also contains several growth factors and cytokines, including PDGF, epidermal growth factor (EGF), insulin-like growth factor (IGF), transforming growth factor (TGF), FGF2, stem cell-growth factor-beta (SCGF), interleukin (IL)-1β, IL-2, -6, -10, -12p70, and IL-17A, tumor necrosis factor (TNF)-α and interferon (IFN) ( Cañas-Arboleda et al, 2020 ; Shanbhag et al, 2020a ).…”

Oxidative Stress Response in Adipose Tissue-Derived Mesenchymal Stem/Stromal Cells

“…Atashi et al (2015) concluded that ROS positively influence adipogenesis in MSC and other adipogenic progenitor cells, whereas osteogenesis is inhibited by increased amounts of ROS [ 47 ]. Osteogenic differentiation of bone marrow MSC is shown to be suppressed by oxidative stress upon exposure to H 2 O 2 [ 26 , 54 ]. Oxidative stress is associated with many bone diseases, including osteoporosis, inflammatory joint disease, and impaired healing of fractures [ 55 ], and may, therefore, partly explain the development of these diseases.…”

“…The effects of oxidative stress are often illustrated in vitro by exposure to the oxidative stressor H 2 O 2 [ 24 , 25 , 26 ]. H 2 O 2 is one of the most important compounds for the study of ROS signaling due to its physicochemical properties, which are relatively low reactivity and the ability to diffuse through membranes [ 27 ].…”

Effect of Passage Number of Conditioned Medium Collected from Equine Amniotic Fluid Mesenchymal Stem Cells: Porcine Oocyte Maturation and Embryo Development

“…Such media have anti-inflammatory, antioxidant, and anti-apoptotic properties [ 17 ]. Conditioned media also increase cell viability and antioxidant enzyme activity, while reducing reactive oxygen species (ROS) levels and apoptosis [ 18 ].…”