Article

Annals of Biomedical Engineering

, Volume 36, Issue 5, pp 813-820

First online:

Hydrostatic Pressure Enhances Chondrogenic Differentiation of Human Bone Marrow Stromal Cells in Osteochondrogenic Medium

  • Diane R. WagnerAffiliated withDepartment of Mechanical Engineering, Stanford UniversityDepartment of Surgery, Stanford UniversityAerospace and Mechanical Engineering Email author 
  • , Derek P. LindseyAffiliated withBone and Joint Center of Excellence, VA Palo Alto Health Care System
  • , Kelvin W. LiAffiliated withBone and Joint Center of Excellence, VA Palo Alto Health Care System
  • , Padmaja TummalaAffiliated withBone and Joint Center of Excellence, VA Palo Alto Health Care System
  • , Sheena E. ChandranAffiliated withDepartment of Mechanical Engineering, Stanford University
  • , R. Lane SmithAffiliated withBone and Joint Center of Excellence, VA Palo Alto Health Care System
  • , Michael T. LongakerAffiliated withDepartment of Surgery, Stanford University
  • , Dennis R. CarterAffiliated withDepartment of Mechanical Engineering, Stanford UniversityBone and Joint Center of Excellence, VA Palo Alto Health Care System
  • , Gary S. BeaupreAffiliated withDepartment of Mechanical Engineering, Stanford UniversityBone and Joint Center of Excellence, VA Palo Alto Health Care System

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Abstract

This study demonstrated the chondrogenic effect of hydrostatic pressure on human bone marrow stromal cells (MSCs) cultured in a mixed medium containing osteogenic and chondrogenic factors. MSCs seeded in type I collagen sponges were exposed to 1 MPa of intermittent hydrostatic pressure at a frequency of 1 Hz for 4 h per day for 10 days, or remained in identical culture conditions but without exposure to pressure. Afterwards, we compared the proteoglycan content of loaded and control cell/scaffold constructs with Alcian blue staining. We also used real-time PCR to evaluate the change in mRNA expression of selected genes associated with chondrogenic and osteogenic differentiation (aggrecan, type I collagen, type II collagen, Runx2 (Cbfa-1), Sox9, and TGF-β1). With the hydrostatic pressure loading regime, proteoglycan staining increased markedly. Correspondingly, the mRNA expression of chondrogenic genes such as aggrecan, type II collagen, and Sox9 increased significantly. We also saw a significant increase in the mRNA expression of type I collagen, but no change in the expression of Runx2 or TGF-β1 mRNA. This study demonstrated that hydrostatic pressure enhanced differentiation of MSCs in the presence of multipotent differentiation factors in vitro, and suggests the critical role that this loading regime may play during cartilage development and regeneration in vivo.

Keywords

Mechanotransduction Cartilage tissue engineering Collagen scaffold Chondrogenesis