Abstract
Altered mechanical loading, secondary to biochemical changes in the nucleus pulposus, is a potential mechanism in disc degeneration. An understanding of the role of this altered mechanical loading is only possible by separating the mechanical and biological effects of early nucleus pulposus changes. The objective of this study was to quantify the mechanical effect of decreased glycosaminoglycans (GAG) and increased crosslinking in the nucleus pulposus using in vitro rat lumbar discs. Following initial mechanical testing the discs were injected according to the four treatment groups: PBS control, chondroitinase-ABC (ChABC) for GAG degradation, genipin (Gen) for crosslinking, or a combination of chondroitinase and genipin (ChABC+Gen). After treatment the discs were again mechanically tested, followed by histology or biochemistry. Neutral zone mechanical properties were changed by approximately 20% for PBS, ChABC, and ChABC+Gen treatments (significant only for PBS in a paired comparison). These trends were reversed with genipin crosslinking alone. With ChABC treatment the effective compressive modulus increased and the GAG content decreased; with the combination of ChABC+Gen the mechanics and GAG content were unchanged. Degradation of nucleus pulposus GAG alters disc axial mechanics, potentially contributing to the degenerative cascade. Crosslinking is unlikely to contribute to degeneration, but may be a potential avenue of treatment.
Similar content being viewed by others
References
Adams MA, McNally DS, Dolan P (1996) ‘Stress’ distributions inside intervertebral discs - The effects of age and degeneration. J Bone Joint Surg-Br 78B(6):965–972
Andersson GBJ, Schultz AB (1979) Effects of fluid injection on mechanical-properties of intervertebral disks. J Biomech 12(6):453–458
Ando T, Kato F, Mimatsu K, Iwata H (1995) Effects of chondroitinase Abc on degenerative intervertebral discs. Clin Orthop Relat Res (318):214–221
Boxberger JI, Sen S, Auerbach JA, Yerramalli CS, Elliott DM (2005) Glycosaminoglycan content affects intervertebral disc neutral zone mechanics in axial loading. Annual Meeting of Biomedical Engineering Society, Baltimore, Maryland, USA
Boxberger JI, Sen S, Yerramalli CS, Elliott DM (2006) Nucleus pulposus glycosaminoglycan content is correlated with axial mechanics in ratlumbar motin segments. J Orthop Res (in press)
Buckwalter JA (1995) Spine update - aging and degeneration of the human intervertebral disc. Spine 20(11):1307–1314
Buckwalter JA, Thomas A, Einhorn MD, Sheldon R Ed. (2000) Orthopaedic basic science, American Academy of Orthopaedic Surgeons
Difabio JL, Pearce RH, Caterson B, Hughes H (1987) The heterogeneity of the non-aggregating proteoglycans of the human intervertebral-disk. Biochem J 244(1):27–33
Duance VC, Crean JKG, Sims TJ, Avery N, Smith S, Menage J, Eisenstein SM, Roberts S (1998) Changes in collagen cross-linking in degenerative disc disease and scoliosis. Spine 23(23):2545–2551
Elliott DM, Sarver JJ (2004) Young investigator award winner: validation of the mouse and rat disc as mechanical models of the human lumbar disc. Spine 29(7):713–722
Eurell JAC, Brown MD, Ramos M (1990) The effects of chondroitinase abc on the rabbit intervertebral-disk - a roentgenographic and histologic-study. Clin Orthop Relat Res (256):238–243
Farndale RW, Sayers CA, Barrett AJ (1982) A direct spectrophotometric micro-assay for sulfated glycosaminoglycans in cartilage cultures. Connect Tissue Res 9(4):247–248
Fry TR, Eurell JC, Johnson AL, Brown MD, Losonsky JM, Schaeffer DJ (1991) Radiographic and histologic effects of chondroitinase abc on normal canine lumbar intervertebral-disk. Spine 16(7): 816–819
Hedman TP, Chuang S-Y, Syed B, Gray D (2003) Biomechanical benefits of crosslink augmentation in spinal discs. 2003 ASME International Mechanical Engineering Congress, Nov 15–21 2003, American Society of Mechanical Engineers, Washington, DC
Henderson N, Stanescu V, Cauchoix J (1991) Nucleolysis of the rabbit intervertebral-disk using chondroitinase abc. Spine 16(2):203–208
Herbage D, Bouillet J, Bernengo JC (1977) Biochemical and physicochemical characterization of pepsin-solubilized type-Ii collagen from bovine articular-cartilage. Biochem J 161(2):303–312
Hiyama K, Okada S (1975) Crystallization and some properties of chondroitinase from arthrobacter-aurescens. J Biol Chem 250(5): 1824–1828
Iatridis JC, Weidenbaum M, Setton LA, Mow VC (1996) Is the nucleus pulposus a solid or a fluid? mechanical behaviors of the nucleus pulposus of the human intervertebral disc. Spine 21(10):1174–1184
Johannessen W, Vresilovic EJ, Wright AC, Elliott DM (2004) Intervertebral disc mechanics are restored following cyclic loading and unloaded recovery. Ann Biomed Eng 32(1):70–76
Johannessen W, Vresilovic EJ, Mills JR, Cloyd JM, Guerin HL, Elliott DM (2005) Effect of nucleotomy on tension-compression behavior of the intervertebral disc. Transactions of Orthopaedic Research Society Meeting, Orthopaedic Research Society, Washington D.C.
Kato F, Iwata H, Mimatsu K, Miura T (1990) Experimental chemonucleolysis with chondroitinase-abc. Clin Orthop Relat Res (253): 301–308
Lu DS, Shono Y, Oda I, Abumi K, Kaneda K (1997) Effects of chondroitinase ABC and chymopapain on spinal motion segment biomechanics - An in vivo biomechanical, radiologic, and histologic canine study. Spine 22(16):1828–1834
Luoma K, Riihimaki H, Luukkonen R, RAininko R, Viikari-Juntura E, Lamminen A (2000) Low back pain in relation to lumbar disc degeneration. Spine 25(4):487–492
Martinez JB, Oloyede VOA, Broom ND (1997) Biomechanics of load-bearing of the intervertebral disc: an experimental and finite element model. Med Eng Phys 19(2):145–156
Mimura M, Panjabi MM, Oxlan TR, Crisco JJ, Yamamoto I, Vasavada A (1994) Disc degeneration affects the multidirectional flexibility of the lumbar spine. Spine 19(12):1371–1380
Monnier VM, Kohn RR, Cerami A (1984) Accelerated age-related browning of human collagen in diabetes-mellitus. In: Proceedings of the national academy of sciences of the United States of America-biological sciences 81(2):583–587
Nachemson A, Morris JM (1964) Invivo measurements of intradiscal pressure - discometry, a method for the determination of pressure in the lower lumbar discs. J Bone Joint Surg-Am 46(5):1077–1092
Natarajan RN, Andersson GBJ (1999) The influence of lumbar disc height and cross-sectional area on the mechanical response of the disc to physiologic loading. Spine 24(18):1873–1881
Norcross JP, Lester GE, Weinhold P, Dahners LE (2003) An in vivo model of degenerative disc disease. J Orthop Res 21(1):183–188
Pal S, Tang LH, Choi H, Habermann E, Rosenberg L, Roughley P, Poole AR (1981) Structural-changes during development in bovine fetal epiphyseal cartilage. Coll Relat Res 1(2):151–176
Panjabi MM (2003) Clinical spinal instability and low back pain. J Electromyogr Kinesiol 13(4):371–379
Pokharna HK, Phillips FM (1998) Collagen crosslinks in human lumbar intervertebral disc aging. Spine 23(15):1645–1648
Riches PE, Dhillon N, Lotz J, Woods AW, McNally DS (2002) The internal mechanics of the intervertebral disc under cyclic loading. J Biomech 35(9):1263–1271
Roughley PJ, Alini M, Antoniou J (2002) The role of proteoglycans in aging, degeneration and repair of the intervertebral disc. Biochem Soc Trans 30:869–874
Sarver JJ, Elliott DM (2004) Altered disc mechanics in mice genetically engineered for reduced type I collagen. Spine 29(10):1094–1098
Sarver JJ, Elliott DM (2005) Mechanical differences between lumbar and tail discs in the mouse. J Orthop Res 23(1):150–155
Sasaki M, Takahashi T, Miyahara K, Hirose T (2001) Effects of chondroitinase ABC on intradiscal pressure in sheep - An in vivo study. Spine 26(5):463–468
Saunders EC (1964) Treatment of canine intervertebral disc syndrome with chymopapain. J Am Vet Med Assoc 145(9):893–896
Schnider SL, Kohn RR (1981) Effects of age and diabetes-mellitus on the solubility and non-enzymatic glucosylation of human-skin collagen. J Clin Invest 67(6):1630–1635
Smith L (1964) Enzyme dissolution of nucleus pulposus in humans. JAMA 187(2):137–140
Smith L, Brown JE (1967) Treatment of lumbar intervertebral disc lesions by direct injection of chymopapain. J Bone Joint Surg Am 49B:502
Spencer DL, Miller JAA, Schultz AB (1985) The effects of chemonucleolysis on the mechanical-properties of the canine lumbar-disk. Spine 10(6):555–561
Stegeman H, Stalder K (1967) Determination of hydroxyproline. Clinica Chimica Acta 18(2):267–273
Sugimura T, Kato F, Mimatsu K, Takenaka O, Iwata H (1996) Experimental chemonucleolysis with chondroitinase ABC in monkeys. Spine 21(2):161–165
Sung HW, Chang Y, Chiu CT, Chen CN, Liang HC (1999a) Crosslinking characteristics and mechanical properties of a bovine pericardium fixed with a naturally occurring crosslinking agent. J Biomed Mat Res 47(2):116–126
Sung HW, Huang DM, Chang WH, Huang RN, Hsu JC (1999b) Evaluation of gelatin hydrogel crosslinked with various crosslinking agents as bioadhesives: in vitro study. J Biomed Mat Res 46(4):520–530
Sung HW, Chang Y, Liang IL, Chang WH, Chen YC (2000) Fixation of biological tissues with a naturally occurring crosslinking agent: fixation rate and effects of pH, temperature, and initial fixative concentration. J Biomed Mat Res 52(1):77–87
Takahashi K, Inoue SI, Takada SI, Nishiyama H, Mimura M, Wada Y (1986) Experimental-study on chemonucleolysis with special reference to the change of intradiscal pressure. Spine 11(6):617–620
Thompson JP, Pearce RH, Schechter MT, Adams ME, Tsang IKY, Bishop PB (1990) Preliminary evaluation of a scheme for grading the gross morphology of the human intervertebral-disk. Spine 15(5): 411–415
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yerramalli, C.S., Chou, A.I., Miller, G.J. et al. The Effect of Nucleus Pulposus Crosslinking and Glycosaminoglycan Degradation on Disc Mechanical Function. Biomech Model Mechanobiol 6, 13–20 (2007). https://doi.org/10.1007/s10237-006-0043-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10237-006-0043-0