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Hydrostatic Pressure Differentially Regulates Outer and Inner Annulus Fibrosus Cell Matrix Production in 3D Scaffolds

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Abstract

Mechanical stimulation may be used to enhance the development of engineered constructs for the replacement of load bearing tissues, such as the intervertebral disc. This study examined the effects of dynamic hydrostatic pressure (HP) on outer and inner annulus (OA, IA) fibrosus cells seeded on fibrous poly(glycolic acid)-poly(l-lactic acid) scaffolds. Constructs were pressurized (5 MPa, 0.5 Hz) for 4 h/day from day 3 to day 14 of culture and analyzed using ELISAs and immunohistochemistry (IHC) to assess extracellular matrix (ECM) production. Both cell types were viable, with OA cells exhibiting more infiltration into the scaffold, which was enhanced by HP. ELISA analyses revealed that HP had no effect on type I collagen production while a significant increase in type II collagen (COL II) was measured in pressurized OA constructs compared to day 14 unloaded controls. Both OA and IA dynamically loaded scaffolds exhibited more uniform COL II elaboration as shown by IHC analyses, which was most pronounced in OA-seeded scaffolds. Overall, HP resulted in enhanced ECM elaboration and organization by OA-seeded constructs, while IA-seeded scaffolds were less responsive. As such, hydrostatic pressurization may be beneficial in annulus fibrosus tissue engineering when applied in concert with an appropriate cell source and scaffold material.

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Acknowledgments

This work was supported by an NSF Graduate Fellowship and NIH Grants DE14228 (S. Nicoll) and EB002425 (PI: D. Elliott). The authors would also like to thank Christopher Hee and Jay Sy for their custom-written MATLAB and LabVIEW programs.

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  1. Department of Bioengineering, University of Pennsylvania, Room 240 Skirkanich Hall, 210 S. 33rd Street, Philadelphia, PA, 19104, USA

    Anna T. Reza & Steven B. Nicoll

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  1. Anna T. Reza
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  2. Steven B. Nicoll
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Correspondence to Steven B. Nicoll.

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Reza, A.T., Nicoll, S.B. Hydrostatic Pressure Differentially Regulates Outer and Inner Annulus Fibrosus Cell Matrix Production in 3D Scaffolds. Ann Biomed Eng 36, 204–213 (2008). https://doi.org/10.1007/s10439-007-9407-6

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  • DOI: https://doi.org/10.1007/s10439-007-9407-6

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