Bone marrow mesenchymal stem cells that are committed to adipogenesis were exposed daily to high-frequency low-intensity mechanical vibrations to understand molecular, morphological and ultrastructural adaptations to mechanical signals during adipogenesis. D1-ORL-UVA mouse bone marrow mesenchymal stem cells were cultured with either growth or adipogenic medium for 1 week. Low-intensity vibration signals (15 min/day, 90 Hz, 0.1 g) were applied to one group of adipogenic cells, while the other adipogenic group served as a sham control. Cellular viability, lipid accumulation, ultrastructure and morphology were determined with MTT, Oil-Red-O staining, phalloidin staining and atomic force microscopy. Semiquantitative reverse transcription polymerase chain reaction showed expression profile of the genes responsible for adipogenesis and ultrastructure of cells. Low-intensity vibration signals increased viability of the cells in adipogenic culture that was reduced significantly compared to quiescent controls. Low-intensity vibration signals also normalized the effects of adipogenic condition on cell morphology, including area, perimeter, circularization and actin cytoskeleton. Furthermore, low-intensity vibration signals reduced the expression of some adipogenic markers significantly. Mesenchymal stem cells are sensitive and responsive to mechanical loads, but debilitating conditions such as aging or obesity may steer mesenchymal stem cells toward adipogenesis. Here, daily application of low-intensity vibration signals partially neutralized the effects of adipogenic induction on mesenchymal stem cells, suggesting that these signals may provide an alternative and/or complementary option to reduce fat deposition.
DeitelM. Overweight and obesity worldwide now estimated to involve 1.7 billion people. Obes Surg2003; 13: 329–330.
2.
TanabeYKogaMSaitoM. Inhibition of adipocyte differentiation by mechanical stretching through ERK-mediated downregulation of PPARγ2. J Cell Sci2004; 117: 3605–3614.
3.
FarooqiISO’RahillyS. Genetic factors in human obesity. Obes Rev2007; 8(Suppl. 1): 37–40.
4.
BouretSLevinBEOzanneSE. Gene-environment interactions controlling energy and glucose homeostasis and the developmental origins of obesity. Physiol Rev2015; 95: 47–82.
5.
WrightSMAronneLJ. Causes of obesity. Abdom Imaging2012; 37: 730–732.
6.
VitaliAMuranoIZingarettiMC. The adipose organ of obesity-prone C57BL/6J mice is composed of mixed white and brown adipocytes. J Lipid Res2012; 53: 619–629.
7.
StynerMThompsonWRGaliorK. Bone marrow fat accumulation accelerated by high fat diet is suppressed by exercise. Bone2014; 64: 39–46.
TanSHSenarath-YapaKChungMT. Wnts produced by Osterix-expressing osteolineage cells regulate their proliferation and differentiation. Proc Natl Acad Sci USA2014; 111: E5262–E5271.
10.
GautamJChoudharyDKhedgikarV. Micro-architectural changes in cancellous bone differ in female and male C57BL/6 mice with high-fat diet-induced low bone mineral density. Br J Nutr2014; 111: 1811–1821.
11.
XiaoYCuiJLiYX. Dyslipidemic high-fat diet affects adversely bone metabolism in mice associated with impaired antioxidant capacity. Nutrition2011; 27: 214–220.
12.
HamrickMWPenningtonCNewtonD. Leptin deficiency produces contrasting phenotypes in bones of the limb and spine. Bone2004; 34: 376–383.
13.
Di IorgiNMoAOGrimmK. Bone acquisition in healthy young females is reciprocally related to marrow adiposity. J Clin Endocrinol Metab2010; 95: 2977–2982.
14.
QiuWAndersenTEBollerslevJ. Patients with high bone mass phenotype exhibit enhanced osteoblast differentiation and inhibition of adipogenesis of human mesenchymal stem cells. J Bone Miner Res2007; 22: 1720–1731.
15.
WrenTAChungSADoreyFJ. Bone marrow fat is inversely related to cortical bone in young and old subjects. J Clin Endocrinol Metab2011; 96: 782–786.
16.
PinoAMRiosSAstudilloP. Concentration of adipogenic and proinflammatory cytokines in the bone marrow supernatant fluid of osteoporotic women. J Bone Miner Res2010; 25: 492–498.
17.
KöllmerMBuhrmanJSZhangY. Markers are shared between adipogenic and osteogenic differentiated mesenchymal stem cells. J Dev Biol Tissue Eng5: 18–25.
18.
OzciviciELuuYKAdlerB. Mechanical signals as anabolic agents in bone. Nat Rev Rheumatol2010; 6: 50–59.
19.
OzciviciE. Effects of spaceflight on cells of bone marrow origin. Turk J Haematol2013; 30: 1–7.
20.
GaoJFuSZengZ. Cyclic stretch promotes osteogenesis-related gene expression in osteoblast-like cells through a cofilin-associated mechanism. Mol Med Rep2016; 14: 218–224.
21.
ZhangLLiuWZhaoJ. Mechanical stress regulates osteogenic differentiation and RANKL/OPG ratio in periodontal ligament stem cells by the Wnt/β-catenin pathway. Biochim Biophys Acta2016; 1860: 2211–2219.
22.
KimKMChoiYJHwangJH. Shear stress induced by an interstitial level of slow flow increases the osteogenic differentiation of mesenchymal stem cells through TAZ activation. PLoS ONE2014; 9: e92427.
23.
ZhengLChenLChenY. The effects of fluid shear stress on proliferation and osteogenesis of human periodontal ligament cells. J Biomech2016; 49: 572–579.
24.
NikukarHReidSTsimbouriPM. Osteogenesis of mesenchymal stem cells by nanoscale mechanotransduction. ACS Nano2013; 7: 2758–2767.
25.
PembertonGDChildsPReidS. Nanoscale stimulation of osteoblastogenesis from mesenchymal stem cells: nanotopography and nanokicking. Nanomedicine2015; 10: 547–560.
26.
LuuYKCapillaERosenCJ. Mechanical stimulation of mesenchymal stem cell proliferation and differentiation promotes osteogenesis while preventing dietary-induced obesity. J Bone Miner Res2009; 24: 50–61.
27.
RubinCTCapillaELuuYK. Adipogenesis is inhibited by brief, daily exposure to high-frequency, extremely low-magnitude mechanical signals. Proc Natl Acad Sci USA2007; 104: 17879–17884.
28.
OlcumMBaskanOKaradasO. Application of low intensity mechanical vibrations for bone tissue maintenance and regeneration. Turk J Biol2016; 40: 300–307.
29.
OzciviciELuuYKRubinCT. Low-level vibrations retain bone marrow’s osteogenic potential and augment recovery of trabecular bone during reambulation. PLoS ONE2010; 5: e11178.
30.
ChanMEAdlerBJGreenDE. Bone structure and B-cell populations, crippled by obesity, are partially rescued by brief daily exposure to low-magnitude mechanical signals. FASEB J2012; 26: 4855–4863.
31.
DemirayLOzciviciE. Bone marrow stem cells adapt to low-magnitude vibrations by altering their cytoskeleton during quiescence and osteogenesis. Turk J Biol2015; 39: 88–97.
32.
UzerGPongkitwitoonSEte ChanM. Vibration induced osteogenic commitment of mesenchymal stem cells is enhanced by cytoskeletal remodeling but not fluid shear. J Biomech2013; 46: 2296–2302.
33.
UzerGPongkitwitoonSIanC. Gap junctional communication in osteocytes is amplified by low intensity vibrations in vitro. PLoS ONE2014; 9: e90840.
34.
OlcumMOzciviciE. Daily application of low magnitude mechanical stimulus inhibits the growth of MDA-MB-231 breast cancer cells in vitro. Cancer Cell Int2014; 14: 102.
35.
SenBXieZCaseN. Mechanical signal influence on mesenchymal stem cell fate is enhanced by incorporation of refractory periods into the loading regimen. J Biomech2011; 44: 593–599.
36.
BatraNNLiYJYellowleyCE. Effects of short-term recovery periods on fluid-induced signaling in osteoblastic cells. J Biomech2005; 38: 1909–1917.
37.
PreDCeccarelliGGastaldiG. The differentiation of human adipose-derived stem cells (hASCs) into osteoblasts is promoted by low amplitude, high frequency vibration treatment. Bone2011; 49: 295–303.
38.
SenBXieZCaseN. Mechanical strain inhibits adipogenesis in mesenchymal stem cells by stimulating a durable beta-catenin signal. Endocrinology2008; 149: 6065–6075.
39.
LuuYOzciviciECapillaE. Development of diet-induced fatty liver disease in the aging mouse is suppressed by brief daily exposure to low-magnitude mechanical signals. Int J Obes2009; 34: 401–405.
40.
CaseNThomasJXieZ. Mechanical input restrains PPARγ2 expression and action to preserve mesenchymal stem cell multipotentiality. Bone2013; 52: 454–464.
41.
RosenBSCookKSYaglomJ. Adipsin and complement factor D activity: an immune-related defect in obesity. Science1989; 244: 1483–1487.
42.
RosenEDMacDougaldOA. Adipocyte differentiation from the inside out. Nat Rev Mol Cell Biol2006; 7: 885–896.
43.
LoweCEO’RahillySRochfordJJ. Adipogenesis at a glance. J Cell Sci2011; 124: 2681–2686.
44.
MuruganandanSRomanASinalC. Adipocyte differentiation of bone marrow-derived mesenchymal stem cells: cross talk with the osteoblastogenic program. Cell Mol Life Sci2009; 66: 236–253.
ZhaoLGregoireFSulHS. Transient induction of ENC-1, a Kelch-related actin-binding protein, is required for adipocyte differentiation. J Biol Chem2000; 275: 16845–16850.
48.
SureshS. Biomechanics and biophysics of cancer cells. Acta Biomater2007; 3: 413–438.
49.
DarlingEMTopelMZauscherS. Viscoelastic properties of human mesenchymally-derived stem cells and primary osteoblasts, chondrocytes, and adipocytes. J Biomech2008; 41: 454–464.
50.
McAndrewsKMMcGrailDJQuachND. Spatially coordinated changes in intracellular rheology and extracellular force exertion during mesenchymal stem cell differentiation. Phys Biol2014; 11: 056004.
