Tensile strain as a regulator of mesenchymal stem cell osteogenesis

Annals of Biomedical Engineering

Volumn 38, Issue 5, 2010, Pages 1767-1779

  Kearney, E M ^a   Farrell, E ^a   Prendergast, P J ^a   Campbell, V A ^a,b  

^a TRINITY COLLEGE DUBLIN   (Ireland)

^b TRINITY COLLEGE DUBLIN   (Ireland)

Author keywords

Mechanical strain; Mesenchymal stem cells; Mitogen activated protein kinase; Osteogenic differentiation; PI3 kinase; Proliferation; Stretch activated cation channels

Indexed keywords

CATION CHANNELS; MECHANICAL STRAIN; MESENCHYMAL STEM CELL; MITOGEN ACTIVATED PROTEIN KINASE; OSTEOGENIC DIFFERENTIATION; PI3-KINASE;

CELL CULTURE; CHLORINE COMPOUNDS; COLLAGEN; ENZYME ACTIVITY; FLOWCHARTING; GADOLINIUM; MECHANISMS; POSITIVE IONS; SILICONES; STEM CELLS; STRAIN RATE; SYNTHESIS (CHEMICAL); TISSUE ENGINEERING;

TENSILE STRAIN;

2 MORPHOLINO 8 PHENYLCHROMONE; 2-(4-MORPHOLINYL)-8-PHENYL-4H-1-BENZOPYRAN-4-ONE; CHROMONE DERIVATIVE; COLLAGEN; COLLAGEN TYPE 1; MORPHOLINE DERIVATIVE; OSTEOCALCIN; PHOSPHATIDYLINOSITOL 3 KINASE;

ANIMAL; ARTICLE; BONE DEVELOPMENT; CELL DIFFERENTIATION; CYTOLOGY; DRUG ANTAGONISM; EXTRACELLULAR MATRIX; MALE; MESENCHYMAL STEM CELL; METABOLISM; METHODOLOGY; PHYSIOLOGY; RAT; TISSUE ENGINEERING; WISTAR RAT;

1-PHOSPHATIDYLINOSITOL 3-KINASE; ANIMALS; CELL DIFFERENTIATION; CHROMONES; COLLAGEN; COLLAGEN TYPE I; EXTRACELLULAR MATRIX; MALE; MESENCHYMAL STEM CELLS; MORPHOLINES; OSTEOCALCIN; OSTEOGENESIS; RATS; RATS, WISTAR; TISSUE ENGINEERING;

EID: 77952008742     PISSN: 00906964     EISSN: 15739686     Source Type: Journal    
DOI: 10.1007/s10439-010-9979-4     Document Type: Article

References (65)

1. GH Altman RL Horan I Martin J Farhadi PR Stark V Volloch JC Richmond G Vunjak-Novakovic DL Kaplan 2002 Cell differentiation by mechanical stress FASEB J. 16 2 270 272 11772952 1:CAS:528:DC%2BD38XhsVCjsb8%3D
2. JS Bae S Gutierrez R Narla J Pratap R Devados AJ van Wijnen JL Stein GS Stein JB Lian A Javed 2007 Reconstitution of Runx2/Cbfa1-null cells identifies a requirement for BMP2 signaling through a Runx2 functional domain during osteoblast differentiation J. Cell. Biochem. 100 2 434 449 10.1002/jcb.21039 16927309 1:CAS:528:DC%2BD2sXpsVyktg%3D%3D (Pubitemid 46146092)
3. CT Brighton B Strafford SB Gross DF Leatherwood JL Williams SR Pollack 1991 The proliferative and synthetic response of isolated calvarial bone cells of rats to cyclic biaxial mechanical strain J. Bone Joint Surg. Am. 73 3 320 331 1848246 1:STN:280:DyaK3M7mtFKgug%3D%3D
4. SP Bruder DJ Fink AI Caplan 1994 Mesenchymal stem cells in bone development, bone repair, and skeletal regeneration therapy J. Cell. Biochem. 56 3 283 294 10.1002/jcb.240560303 7876320 1:CAS:528:DyaK2MXitFyhu70%3D (Pubitemid 24360045)
5. J Caetano-Lopes H Canhao JE Fonseca 2007 Osteoblasts and bone formation Acta Reumatol. Port. 32 2 103 110 17572649
6. AI Caplan 1987 Bone development and repair Bioessays 6 4 171 175 10.1002/bies.950060406 3593327 1:STN:280:DyaL2s3ktFCkug%3D%3D
7. EY Chao N Inoue 2003 Biophysical stimulation of bone fracture repair, regeneration and remodelling Eur. Cell. Mater. 6 72 84 14722904 discussion 84-5
8. GT Charras BA Williams SM Sims MA Horton 2004 Estimating the sensitivity of mechanosensitive ion channels to membrane strain and tension Biophys. J. 87 4 2870 2884 10.1529/biophysj.104.040436 15454477 1:CAS:528:DC%2BD2cXot12mtrg%3D
9. TE Danciu RM Adam K Naruse MR Freeman PV Hauschka 2003 Calcium regulates the PI3K-Akt pathway in stretched osteoblasts FEBS Lett. 536 1-3 193 197 10.1016/S0014-5793(03)00055-3 12586362 1:CAS:528:DC%2BD3sXhtVGgsLk%3D (Pubitemid 36206430)
10. P Ducy C Desbois B Boyce G Pinero B Story C Dunstan E Smith J Bonadio S Goldstein C Gundberg A Bradley G Karsenty 1996 Increased bone formation in osteocalcin-deficient mice Nature 382 6590 448 452 10.1038/382448a0 8684484 1:CAS:528:DyaK28Xks1Olur0%3D (Pubitemid 26264860)
11. P Ducy R Zhang V Geoffroy AL Ridall G Karsenty 1997 Osf2/Cbfa1: A transcriptional activator of osteoblast differentiation Cell 89 5 747 754 10.1016/S0092-8674(00)80257-3 9182762 1:CAS:528:DyaK2sXjs1ejsbg%3D
12. E Farrell FJ O'Brien P Doyle J Fischer I Yannas BA Harley B O'Connell PJ Prendergast VA Campbell 2006 A collagen-glycosaminoglycan scaffold supports adult rat mesenchymal stem cell differentiation along osteogenic and chondrogenic routes Tissue Eng. 12 3 459 468 10.1089/ten.2006.12.459 16579679 1:CAS:528:DC%2BD28XjtVaksLk%3D
13. RT Franceschi G Xiao 2003 Regulation of the osteoblast-specific transcription factor, Runx2: Responsiveness to multiple signal transduction pathways J. Cell. Biochem. 88 3 446 454 10.1002/jcb.10369 12532321 1:CAS:528:DC%2BD3sXhtFKjtr4%3D (Pubitemid 36158266)
14. G Friedl H Schmidt I Rehak G Kostner K Schauenstein R Windhager 2007 Undifferentiated human mesenchymal stem cells (hMSCs) are highly sensitive to mechanical strain: Transcriptionally controlled early osteo-chondrogenic response in vitro Osteoarthritis Cartilage 15 11 1293 1300 10.1016/j.joca.2007. 04.002 17977755 1:STN:280:DC%2BD2snmsF2qtg%3D%3D (Pubitemid 350029038)
15. S Gallea F Lallemand A Atfi G Rawadi V Ramez S Spinella-Jaegle S Kawai C Faucheu L Huet R Baron S Roman-Roman 2001 Activation of mitogen-activated protein kinase cascades is involved in regulation of bone morphogenetic protein-2-induced osteoblast differentiation in pluripotent C2C12 cells Bone 28 5 491 498 10.1016/S8756-3282(01)00415-X 11344048 1:CAS:528:DC%2BD3MXjtlChsL0%3D (Pubitemid 32410000)
16. C Ge G Xiao D Jiang RT Franceschi 2007 Critical role of the extracellular signal-regulated kinase-MAPK pathway in osteoblast differentiation and skeletal development J. Cell Biol. 176 5 709 718 10.1083/jcb.200610046 17325210 1:CAS:528:DC%2BD2sXisVymtbw%3D (Pubitemid 46333984)
17. S Ghazanfari M Tafazzoli-Shadpour MA Shokrgozar 2009 Effects of cyclic stretch on proliferation of mesenchymal stem cells and their differentiation to smooth muscle cells Biochem. Biophys. Res. Commun. 388 3 601 605 10.1016/j.bbrc.2009.08.072 19695226 1:CAS:528:DC%2BD1MXhtFSnsr3J
18. N Ghosh-Choudhury SL Abboud R Nishimura A Celeste L Mahimainathan GG Choudhury 2002 Requirement of BMP-2-induced phosphatidylinositol 3-kinase and Akt serine/threonine kinase in osteoblast differentiation and Smad-dependent BMP-2 gene transcription J. Biol. Chem. 277 36 33361 33368 10.1074/jbc. M205053200 12084724 1:CAS:528:DC%2BD38Xnt1SksrY%3D (Pubitemid 34984859)
19. TS Gross SL Poliachik BJ Ausk DA Sanford BA Becker S Srinivasan 2004 Why rest stimulates bone formation: A hypothesis based on complex adaptive phenomenon Exerc. Sport Sci. Rev. 32 1 9 13 10.1097/00003677-200401000-00003 14748543 (Pubitemid 38082439)
20. BK Hall T Miyake 2000 All for one and one for all: Condensations and the initiation of skeletal development Bioessays 22 2 138 147 10.1002/(SICI)1521- 1878(200002)22:2<138::AID-BIES5>3.0.CO;2-4 10655033 1:CAS:528: DC%2BD3MXmslSht7w%3D (Pubitemid 30092080)
21. DW Hamilton TM Maul DA Vorp 2004 Characterization of the response of bone marrow-derived progenitor cells to cyclic strain: Implications for vascular tissue-engineering applications Tissue Eng. 10 3-4 361 369 10.1089/ 107632704323061726 15165453 1:CAS:528:DC%2BD2cXjvVKgtbw%3D (Pubitemid 38580257)
22. RA Hipskind G Bilbe 1998 MAP kinase signaling cascades and gene expression in osteoblasts Front. Biosci. 3 d804 d816 9682034 1:CAS:528:DyaK1cXlvVWlsLs%3D
23. M Jagodzinski M Drescher J Zeichen S Hankemeier C Krettek U Bosch M van Griensven 2004 Effects of cyclic longitudinal mechanical strain and dexamethasone on osteogenic differentiation of human bone marrow stromal cells Eur. Cell. Mater. 7 35 41 15095254 1:STN:280:DC%2BD2c7ps1equw%3D%3D discussion 41
24. RK Jaiswal N Jaiswal SP Bruder G Mbalaviele DR Marshak MF Pittenger 2000 Adult human mesenchymal stem cell differentiation to the osteogenic or adipogenic lineage is regulated by mitogen-activated protein kinase J. Biol. Chem. 275 13 9645 9652 10.1074/jbc.275.13.9645 10734116 1:CAS:528: DC%2BD3cXitlCqs70%3D (Pubitemid 30185197)
25. JH Jansen FA Weyts I Westbroek H Jahr H Chiba HA Pols JA Verhaar JP van Leeuwen H Weinans 2004 Stretch-induced phosphorylation of ERK1/2 depends on differentiation stage of osteoblasts J. Cell. Biochem. 93 3 542 551 10.1002/jcb.20162 15378606 1:CAS:528:DC%2BD2cXosVyqs7o%3D (Pubitemid 44264189)
26. G Karsenty 2003 The complexities of skeletal biology Nature 423 6937 316 318 10.1038/nature01654 12748648 1:CAS:528:DC%2BD3sXjs1ynu7w%3D (Pubitemid 40852704)
27. D Kaspar W Seidl C Neidlinger-Wilke A Beck L Claes A Ignatius 2002 Proliferation of human-derived osteoblast-like cells depends on the cycle number and frequency of uniaxial strain J. Biomech. 35 7 873 880 10.1016/S0021- 9290(02)00058-1 12052389 (Pubitemid 34615986)
28. A Kawarizadeh C Bourauel W Gotz A Jager 2005 Early responses of periodontal ligament cells to mechanical stimulus in vivo J. Dent. Res. 84 10 902 906 10.1177/154405910508401006 16183788 1:CAS:528:DC%2BD2MXhtFGiu7zM (Pubitemid 43827797)
29. EM Kearney PJ Prendergast VA Campbell 2008 Mechanisms of strain-mediated mesenchymal stem cell apoptosis J. Biomech. Eng. 130 6 061004 10.1115/1.2979870 19045533 1:STN:280:DC%2BD1cjnvVSmsQ%3D%3D
30. RL Kent JK Hoober GT Cooper 1989 Load responsiveness of protein synthesis in adult mammalian myocardium: Role of cardiac deformation linked to sodium influx Circ. Res. 64 1 74 85 2909303 1:CAS:528:DyaL1MXntlamtQ%3D%3D
31. YJ Kim RL Sah JY Doong AJ Grodzinsky 1988 Fluorometric assay of DNA in cartilage explants using Hoechst 33258 Anal. Biochem. 174 1 168 176 10.1016/0003-2697(88)90532-5 2464289 1:CAS:528:DyaL1cXmt1Kju7o%3D (Pubitemid 18261664)
32. PJ Kostenuik BP Halloran ER Morey-Holton DD Bikle 1997 Skeletal unloading inhibits the in vitro proliferation and differentiation of rat osteoprogenitor cells Am. J. Physiol. 273 6 Pt 1 E1133 E1139 9435529 1:CAS:528: DyaK1cXjvFSkug%3D%3D
33. CF Lai SL Cheng 2002 Signal transductions induced by bone morphogenetic protein-2 and transforming growth factor-beta in normal human osteoblastic cells J. Biol. Chem. 277 18 15514 15522 10.1074/jbc.M200794200 11854297 1:CAS:528:DC%2BD38Xjs1Squ7s%3D (Pubitemid 34967818)
34. KS Lee HJ Kim QL Li XZ Chi C Ueta T Komori JM Wozney EG Kim JY Choi HM Ryoo SC Bae 2000 Runx2 is a common target of transforming growth factor beta1 and bone morphogenetic protein 2, and cooperation between Runx2 and Smad5 induces osteoblast-specific gene expression in the pluripotent mesenchymal precursor cell line C2C12 Mol. Cell. Biol. 20 23 8783 8792 10.1128/MCB.20.23. 8783-8792.2000 11073979 1:CAS:528:DC%2BD3MXitFGrsr4%3D (Pubitemid 32245911)
35. WC Lee TM Maul DA Vorp JP Rubin KG Marra 2007 Effects of uniaxial cyclic strain on adipose-derived stem cell morphology, proliferation, and differentiation Biomech. Model. Mechanobiol. 6 4 265 273 10.1007/s10237-006- 0053-y 16906436 (Pubitemid 47000763)
36. EJ Mackie 2003 Osteoblasts: Novel roles in orchestration of skeletal architecture Int. J. Biochem. Cell Biol. 35 9 1301 1305 10.1016/S1357-2725(03) 00107-9 12798343 1:CAS:528:DC%2BD3sXksValt74%3D (Pubitemid 36703168)
37. M Moretti A Prina-Mello AJ Reid V Barron PJ Prendergast 2004 Endothelial cell alignment on cyclically-stretched silicone surfaces J. Mater. Sci. Mater. Med. 15 10 1159 1164 10.1023/B:JMSM.0000046400.18607.72 15516879 1:CAS:528:DC%2BD2cXptV2hurs%3D (Pubitemid 39491723)
38. ER Morey DJ Baylink 1978 Inhibition of bone formation during space flight Science 201 4361 1138 1141 10.1126/science.150643 150643 1:CAS:528: DyaE1cXlvFCks7w%3D
39. GB Muller 2003 Embryonic motility: Environmental influences and evolutionary innovation Evol. Dev. 5 1 56 60 10.1046/j.1525-142X.2003.03009.x 12492410 (Pubitemid 36174978)
40. A Nieponice TM Maul JM Cumer L Soletti DA Vorp 2007 Mechanical stimulation induces morphological and phenotypic changes in bone marrow-derived progenitor cells within a three-dimensional fibrin matrix J. Biomed. Mater. Res. A 81 3 523 530 17133453
41. A Nohe S Hassel M Ehrlich F Neubauer W Sebald YI Henis P Knaus 2002 The mode of bone morphogenetic protein (BMP) receptor oligomerization determines different BMP-2 signaling pathways J. Biol. Chem. 277 7 5330 5338 10.1074/jbc.M102750200 11714695 1:CAS:528:DC%2BD38XhsFCjsLc%3D (Pubitemid 34968578)
42. TA Owen M Aronow V Shalhoub LM Barone L Wilming MS Tassinari MB Kennedy S Pockwinse JB Lian GS Stein 1990 Progressive development of the rat osteoblast phenotype in vitro: Reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix J. Cell. Physiol. 143 3 420 430 10.1002/jcp.1041430304 1694181 1:CAS:528:DyaK3cXksFKmsbc%3D (Pubitemid 20201656)
43. SA Park JW Shin YI Yang YK Kim KD Park JW Lee IH Jo YJ Kim 2004 In vitro study of osteogenic differentiation of bone marrow stromal cells on heat-treated porcine trabecular bone blocks Biomaterials 25 3 527 535 10.1016/S0142-9612(03) 00553-2 14585702 1:CAS:528:DC%2BD3sXotlynt7s%3D (Pubitemid 37329705)
44. MG Petroff SH Kim S Pepe C Dessy E Marban JL Balligand SJ Sollott 2001 Endogenous nitric oxide mechanisms mediate the stretch dependence of Ca2+ release in cardiomyocytes Nat. Cell Biol. 3 10 867 873 10.1038/ncb1001-867 11584267 1:STN:280:DC%2BD3Mrjs1KqtA%3D%3D (Pubitemid 32952250)
45. MF Pittenger AM Mackay SC Beck RK Jaiswal R Douglas JD Mosca MA Moorman DW Simonetti S Craig DR Marshak 1999 Multilineage potential of adult human mesenchymal stem cells Science 284 5411 143 147 10.1126/science.284.5411.143 10102814 1:CAS:528:DyaK1MXitlCnu7o%3D (Pubitemid 29282067)
46. J Pratap M Galindo SK Zaidi D Vradii BM Bhat JA Robinson JY Choi T Komori JL Stein JB Lian GS Stein AJ van Wijnen 2003 Cell growth regulatory role of Runx2 during proliferative expansion of preosteoblasts Cancer Res. 63 17 5357 5362 14500368 1:CAS:528:DC%2BD3sXntlekur8%3D
47. PJ Prendergast R Huiskes K Soballe 1997 ESB Research Award 1996. Biophysical stimuli on cells during tissue differentiation at implant interfaces J. Biomech. 30 6 539 548 10.1016/S0021-9290(96)00140-6 9165386 1:STN:280:DyaK2szgvVWlsg%3D%3D (Pubitemid 27214187)
48. PG Robey JD Termine 1985 Human bone cells in vitro Calcif. Tissue Int. 37 5 453 460 10.1007/BF02557826 2998572 1:STN:280:DyaL28%2Flt1yhsQ%3D%3D (Pubitemid 15250110)
49. AG Robling DB Burr CH Turner 2001 Recovery periods restore mechanosensitivity to dynamically loaded bone J. Exp. Biol. 204 Pt 19 3389 3399 11606612 1:STN:280:DC%2BD3MnhtFCitQ%3D%3D
50. AG Robling FM Hinant DB Burr CH Turner 2002 Improved bone structure and strength after long-term mechanical loading is greatest if loading is separated into short bouts J. Bone Miner. Res. 17 8 1545 1554 10.1359/jbmr.2002.17.8.1545 12162508
51. GY Rochefort P Vaudin N Bonnet JC Pages J Domenech P Charbord V Eder 2005 Influence of hypoxia on the domiciliation of mesenchymal stem cells after infusion into rats: Possibilities of targeting pulmonary artery remodeling via cells therapies? Respir. Res. 6 125 10.1186/1465-9921-6-125 16253136 1:CAS:528:DC%2BD2MXht1KhurzI (Pubitemid 41631829)
52. J Rubin TC Murphy X Fan M Goldschmidt WR Taylor 2002 Activation of extracellular signal-regulated kinase is involved in mechanical strain inhibition of RANKL expression in bone stromal cells J. Bone Miner. Res. 17 8 1452 1460 10.1359/jbmr.2002.17.8.1452 12162499 1:CAS:528:DC%2BD38XmsVWktLg%3D
53. IM Sebastine DJ Williams 2006 The role of mechanical stimulation in engineering of extracellular matrix (ECM) Conf. Proc. IEEE Eng. Med. Biol. Soc. 1 3648 3651 10.1109/IEMBS.2006.4398238 17945786 1:STN:280:DC%2BD2snisFCisw%3D%3D
54. JE Shea SC Miller DC Poole JP Mattson 2003 Cortical bone dynamics, strength, and densitometry after induction of emphysema in hamsters J. Appl. Physiol. 95 2 631 634 12851418
55. CA Simmons S Matlis AJ Thornton S Chen CY Wang DJ Mooney 2003 Cyclic strain enhances matrix mineralization by adult human mesenchymal stem cells via the extracellular signal-regulated kinase (ERK1/2) signaling pathway J. Biomech. 36 8 1087 1096 10.1016/S0021-9290(03)00110-6 12831733 (Pubitemid 36794104)
56. G Song Y Ju X Shen Q Luo Y Shi J Qin 2007 Mechanical stretch promotes proliferation of rat bone marrow mesenchymal stem cells Colloids Surf. B Biointerfaces 58 2 271 277 10.1016/j.colsurfb.2007.04.001 17499488 1:CAS:528:DC%2BD2sXmvV2ht78%3D (Pubitemid 46921466)
57. S Srinivasan DA Weimer SC Agans SD Bain TS Gross 2002 Low-magnitude mechanical loading becomes osteogenic when rest is inserted between each load cycle J. Bone Miner. Res. 17 9 1613 1620 10.1359/jbmr.2002.17.9.1613 12211431
58. M Suzawa I Takada J Yanagisawa F Ohtake S Ogawa T Yamauchi T Kadowaki Y Takeuchi H Shibuya Y Gotoh K Matsumoto S Kato 2003 Cytokines suppress adipogenesis and PPAR-gamma function through the TAK1/TAB 1/NIK cascade Nat. Cell Biol. 5 3 224 230 10.1038/ncb942 12598905 1:CAS:528:DC%2BD3sXitFKmsbk%3D (Pubitemid 36322249)
59. K Suzuma K Naruse I Suzuma N Takahara K Ueki LP Aiello GL King 2000 Vascular endothelial growth factor induces expression of connective tissue growth factor via KDR, Flt1, and phosphatidylinositol 3-kinase-akt-dependent pathways in retinal vascular cells J. Biol. Chem. 275 52 40725 40731 10.1074/jbc.M006509200 11018037 1:CAS:528:DC%2BD3MXjsVGnsA%3D%3D (Pubitemid 32054890)
60. CH Turner 1998 Three rules for bone adaptation to mechanical stimuli Bone 23 5 399 407 10.1016/S8756-3282(98)00118-5 9823445 1:STN:280: DyaK1M%2FktVOkuw%3D%3D (Pubitemid 28470608)
61. van Griensven, M., S. Diederichs, and C. Kasper. Mechanical strain of bone marrow stromal cells induces proliferation and differentiation into osteoblast-like cells. In: Topics in Tissue Engineering, edited by N. R. Ashammakhi and R.L. Reis, 2005 (E-book).
62. FS Wang CJ Wang SM Sheen-Chen YR Kuo RF Chen KD Yang 2002 Superoxide mediates shock wave induction of ERK-dependent osteogenic transcription factor (CBFA1) and mesenchymal cell differentiation toward osteoprogenitors J. Biol. Chem. 277 13 10931 10937 10.1074/jbc.M104587200 11784711 1:CAS:528: DC%2BD38Xis1Khtbw%3D (Pubitemid 34952848)
63. DF Ward Jr WA Williams NE Schapiro GL Weber SR Christy M Salt RF Klees A Boskey GE Plopper 2007 Focal adhesion kinase signaling controls cyclic tensile strain enhanced collagen I-induced osteogenic differentiation of human mesenchymal stem cells Mol. Cell. Biomech. 4 4 177 188 18437915 (Pubitemid 351493006)
64. A Yamaguchi T Komori T Suda 2000 Regulation of osteoblast differentiation mediated by bone morphogenetic proteins, hedgehogs, and Cbfa1 Endocr. Rev. 21 4 393 411 10.1210/er.21.4.393 10950158 1:CAS:528:DC%2BD3cXmtlSktLY%3D (Pubitemid 32275586)
65. PG Ziros AP Gil T Georgakopoulos I Habeos D Kletsas EK Basdra AG Papavassiliou 2002 The bone-specific transcriptional regulator Cbfa1 is a target of mechanical signals in osteoblastic cells J. Biol. Chem. 277 26 23934 23941 10.1074/jbc.M109881200 11960980 1:CAS:528:DC%2BD38XltF2msrs%3D (Pubitemid 34952238)