Mechano-Biological Regulation of Human Mesenchymal Stem Cell Osteogenic Differentiation

Micromotion around bone implant and healing fractures results in the formation of a variety of skeletal tissues instead of bone in locations that are biochemically similar. Mechanical deformation including tensile strain has been implicated in altering the osteogenic potential and subsequent activity of bone marrow derived pluripotent human mesenchymal stem cells (hMSCs), however, the pathway remains unclear.

In this investigation, mechano-biological regulation of hMSC differentiation into the osteogenic lineage was evaluated. Once gradient and biaxial strain loading models were established and characterized, hMSCs, seeded on type I collagen coated silicone elastic membranes, were exposed to cyclic gradient and biaxial strain at 0.1 Hz in the presence and absence of dexamethasone for 2 hours daily up to 21 days.

It was found that the regions of high compression (-3<-2%) and low compression (-2%<0) enhanced extracellular matrix (ECM) mineralization in hMSC cultures without dexamethasone, while regions of low compression (-2<0%), low tension (0<3%), and high tension (3<10%) inhibited ECM mineralization in hMSC cultures with dexamethasone. Biaxial strain (3%) downregulated ALP mRNA expression levels in hMSC cultured with dexamethasone, while production of extracellular matrix decreased, and the accumulation of calcium-containing mineral increased for hMSC cultured in the presence and absence of dexamethasone.

The strain-enhanced progression of hMSCs into the osteogenic lineage was demonstrated by significant decreases in procollagen in conjunction with significant increases in calcium for hMSCs exposed to strain. Maximal expression of osteopontin (OPN) and alkaline phosphatase (ALP) occurred at day 4 and day 5, respectively, for hMSCs exposed to strain. In contrast, the maximal expression of OPN and ALP in hMSCs exposed to control conditions progressed more slowly at day 8 and day 7, respectively. Cyclic strain also demonstrated reorganization of key extracellular components including type I collagen.

Mechanical deformation of hMSCs, in the absence of dexamethasone, increased bone formation. The appearance of osteogenic markers at earlier time points suggests that cyclic strain (0<3%) was capable of accelerating stem cell progression into the osteogenic lineage. The results support and are consistent with predictions made by computational models associating low tensile loading with bone formation around implants and during fracture healing.

Year	Entry
2003	Simmons CA, Matlis S, Thornton AJ, Chen S, Wang C-Y, Mooney DJ. Cyclic strain enhances matrix mineralization by adult human mesenchymal stem cells via the extracellular signal-regulated kinase (ERK1/2) signaling pathway. J Biomech. August 2003;36(8):1087-1096.
1991	Ingber D. Integrins as mechanochemical transducers. Curr Opin Cell Biol. October 1991;3(5):841-848.
1995	Bay BK. Texture correlation: a method for the measurement of detailed strain distributions within trabecular bone. J Orthop Res. March 1995;13(2):258-267.
1994	Rubin CT, McLeod KJ. Promotion of bony ingrowth by frequency-specific, low-amplitude mechanical strain. Clin Orthop Relat Res. January 1994;298:165-174.
1998	Chow JWM, Wilson AJ, Chambers TJ, Fox SW. Mechanical loading stimulates bone formation by reactivation of bone lining cells in 13‐week‐old rats. J Bone Miner Res. November 1998;13(11):1760-1767.
1997	Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP. Osteogenic differentiation of purified, culture‐expanded human mesenchymal stem cells in vitro. J Cell Biochem. February 1997;64(2):295-312.
1988	Carter DR, Blenman PR, Beaupré GS. Correlations between mechanical stress history and tissue differentiation in initial fracture healing. J Orthop Res. September 1988;6(5):736-748.
2001	Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. April 2001;7(2):211-228.
1999	Claes LE, Heigele CA. Magnitudes of local stress and strain along bony surfaces predict the course and type of fracture healing. J Biomech. March 1999;32(3):255-266.
1979	Perren SM. Physical and biological aspects of fracture healing with special reference to internal fixation. Clin Orthop Relat Res. January–February 1979;138:175-196.
2000	Martin RB. Toward a unifying theory of bone remodeling. Bone. January 2000;26(1):1-6.
1998	Carter DR, Beaupré GS, Giori NJ, Helms JA. Mechanobiology of skeletal regeneration. Clin Orthop Relat Res. October 1998;355(suppl):S41-S55.
1993	Fung YC. Biomechanics: Mechanical Properties of Living Tissues. 2nd ed. New York, NY: Springer-Verlag; 1993.
1997	Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao Y-H, Inada M, Sato M, Okamoto R, Kitamura Y, Yoshiki S, Kishimoto T. Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell. May 30, 1997;89(5):755-764.
1997	Ducy P, Zhang R, Geoffroy V, Ridall AL, Karsent G. Osf2/Cbfa1: a transcriptional activator of osteoblast differentiation. Cell. May 30, 1997;89(5):747-754.
1997	Prendergast PJ, Huiskes R, Søballe K. Biophysical stimuli on cells during tissue differentiation at implant interfaces. J Biomech. June 1997;30(6):539-548.
1998	Jilka RL, Weinstein RS, Bellido T, Parfitt AM, Manolagas SC. Osteoblast programmed cell death (apoptosis): modulation by growth factors and cytokines. J Bone Miner Res. May 1998;13(5):793-802.
2004	McBeath R, Pirone DM, Nelson CM, Bhadriraju K, Chen CS. Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell. April 2004;6(4):483-495.
1998	Pavalko FM, Chen NX, Turner CH, Burr DB, Atkinson S, Hsieh Y-F, Qiu J, Duncan RL. Fluid shear-induced mechanical signaling in MC3T3-E1 osteoblasts requires cytoskeleton-integrin interactions. Am J Physiol. December 1998;275(6):C1591-C1601.
1999	Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR. Multilineage potential of adult human mesenchymal stem cells. Science. April 2, 1999;284(5411):143-147.
1984	Binderman I, Shimshoni Z, Somjen D. Biochemical pathways involved in the translation of physical stimulus into biological message. Calcif Tiss Int. March 1984;36(1):S82-S85.
1998	Mackay AM, Beck SC, Murphy JM, Barry FP, Chichester CO, Pittenger MF. Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. Tissue Eng. Winter 1998;4(4):415-428.
2003	Harada S-i, Rodan GA. Control of osteoblast function and regulation of bone mass. Nature. May 15, 2003;423(6937):349-355.
1998	Johnstone B, Hering TM, Caplan AI, Goldberg VM, Yoo JU. In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res. January 10, 1998;238(1):265-272.
1996	Burr DB, Milgrom C, Fyhrie D, Forwood M, Nyska M, Finestone A, Hoshaw S, Saiag E, Simkin A. In vivo measurement of human tibial strains during vigorous activity. Bone. May 1996;18(5):405-410.
2004	Huang CC, Hagar KL, Frost LE, Sun Y, Cheung HS. Effects of cyclic compressive loading on chondrogenesis of rabbit bone-marrow derived mesenchymal stem cells. Stem Cells. May 2004;22(3):313-323.

Year

Entry

2003

Simmons CA, Matlis S, Thornton AJ, Chen S, Wang C-Y, Mooney DJ. Cyclic strain enhances matrix mineralization by adult human mesenchymal stem cells via the extracellular signal-regulated kinase (ERK1/2) signaling pathway. J Biomech. August 2003;36(8):1087-1096.

1991

Ingber D. Integrins as mechanochemical transducers. Curr Opin Cell Biol. October 1991;3(5):841-848.

1995

Bay BK. Texture correlation: a method for the measurement of detailed strain distributions within trabecular bone. J Orthop Res. March 1995;13(2):258-267.

1994

Rubin CT, McLeod KJ. Promotion of bony ingrowth by frequency-specific, low-amplitude mechanical strain. Clin Orthop Relat Res. January 1994;298:165-174.

1998

Chow JWM, Wilson AJ, Chambers TJ, Fox SW. Mechanical loading stimulates bone formation by reactivation of bone lining cells in 13‐week‐old rats. J Bone Miner Res. November 1998;13(11):1760-1767.