ToF-SIMS study of differentiation of human bone-derived stromal cells: new insights into osteoporosis

Abstract

Lipids have numerous important functions in the human body, as they form the cells' plasma membranes and play a key role in many disease states, presumably also in osteoporosis. Here, the fatty acid composition of the outer plasma membranes of cells differentiated into the osteogenic and adipogenic direction is studied with surface-sensitive time-of-flight secondary ion mass spectrometry (ToF-SIMS). For data evaluation, principal component analysis (PCA) is applied. Human (bone-derived) mesenchymal stromal cells (hMSCs) from an osteoporotic donor and a control donor are compared to reveal differences in the fatty acid composition of the membranes. The chemical information is correlated to staining and real-time quantitative polymerase chain reaction (rt-qPCR) results to provide insight into the gene expression of several differentiation markers on the RNA level. Adipogenic differentiation of hMSCs from a non-osteoporotic donor correlates with increased relative intensities of all fatty acids under investigation. After osteogenic differentiation of non-osteoporotic cells, the relative mass signal intensities of unsaturated fatty acids such as oleic and linoleic acids are increased. However, the osteoporotic cells show increased levels of palmitic acid in the plasma membrane after exposure to osteogenic differentiation conditions, which correlates to an immature differentiation state relative to non-osteoporotic osteogenic cells. This immature differentiation state is confirmed by increased early osteogenic differentiation factor Runx2 on RNA level and by less calcium mineralization spots seen in von Kossa staining and ToF-SIMS images. Graphical abstract Time-of-flight secondary ion mass spectrometry is applied to analyze the fatty acid composition of the outer plasma membranes of cells differentiated into the adipogenic and osteogenic direction. Cells from an osteoporotic and a control donor are compared to reveal differences due to differentiation and disease stage of the cells.

Study of the diacylglycerol composition in the liver and serum of mice with prediabetes and diabetes using MeV TOF-SIMS

“…Our results are in agreement with the TOF-SIMS results obtained on the tissue samples of people suffering from non-alcoholic fatty liver disease [15,32,33]. Additionally, the method is successfully applied to image the location and to quantify a single compound or even a set of compounds in diseases such as colon dysplasia, osteoporosis, breast cancer or chronic obstructive pulmonary disease [34][35][36][37]. To our knowledge, the only work in which TOF-SIMS was used to identify the changes in the lipid composition within mouse tissue (skeletal muscle, liver and adipose tissue) in response to the dietary intake of omega-3 fatty acids was published by Sjovall et al [2].…”

LncRNA KCNQ1OT1 promotes osteogenic differentiation to relieve osteolysis via Wnt/β-catenin activation

“…It has been well documented that bone remodeling was supported by dynamic equilibrium between bone resorption and bone formation which were regulated and maintained by osteoblasts throughout lifelong [ 1 ]. Imbalance especially induced by inhibition of osteogenic differentiation and osteolysis aggravation would result in pathological bone defects including aseptic loosening of the implant during postoperation of total joint arthroplasty (TJA) [ 2 ], osteoporosis [ 3 ] as well as rheumatoid arthritis [ 4 ] and other bone diseases. Recently, a great deal of studies provided evidence that particulate wear particles were the leading causes of periprosthetic osteolysis, which mainly consisted of PMMA [ 5 ], ultra-high molecular weight polyethylene (UHMWPE) [ 6 ] and titanium [ 7 ].…”

Visfatin alters the cytokine and matrix-degrading enzyme profile during osteogenic and adipogenic MSC differentiation

“…Primary hMSCs (n ¼ 4) or commercially obtained donor MSCs (n ¼ 3) were cultured at 4 Â 10 4 cells per 9.6 cm 2 in alphaMEM Glutamax expansion medium supplemented with 10% FCS (Sig-maeAldrich), 100 U/ml penicillin (PAA), 10 mg/ml streptomycin (PAA) and osteogenic supplements, 0.1 mM dexamethasone (SigmaeAldrich), 10 mM b-glycerophosphate (SigmaeAldrich), and 0.06 mM ascorbic acid (SigmaeAldrich) for 21 days 23,25 . Cells were permanently stimulated with/without visfatin (100 ng/ml, Biovendor, Brno, Czech Republic), resistin (20 ng/ml, PeproTech, Rocky Hill, USA), and leptin (10 ng/ml, Biovendor) during differentiation.…”