Research Papers

Changes in Intervertebral Disk Mechanical Behavior During Early Degeneration

[+] Author and Article Information
Cornelis P. L. Paul, Kaj S. Emanuel

Department of Orthopedic Surgery,
Academic Medical Center,
University of Amsterdam,
Amsterdam Movement Sciences,
Amsterdam 1105 AZ, The Netherlands

Idsart Kingma

Department of Human Movement Sciences,
Vrije Universiteit,
Amsterdam Movement Sciences,
Amsterdam 1081 BT, The Netherlands

Albert J. van der Veen

Department of Physics and
Medical Technology,
VU University Medical Center,
Amsterdam 1081 HV, The Netherlands

Roderick M. Holewijn, Pieter-Paul A. Vergroesen

Department of Orthopedic Surgery,
VU University Medical Center,
Amsterdam 1081 HV, The Netherlands

Peter M. van de Ven

Department of Epidemiology and Biostatistics,
VU University Medical Center,
Amsterdam 1081 BT, The Netherlands

Margriet G. Mullender

Department of Plastic, Reconstructive and
Hand Surgery,
VU University Medical Center,
Amsterdam 1081 HV, The Netherlands

Marco N. Helder

Department of Oral and Maxillofacial Surgery,
VU University Medical Center,
Amsterdam 1081 HV, The Netherlands

Theodoor H. Smit

Department of Orthopedic Surgery,
Academic Medical Center,
University of Amsterdam,
Amsterdam Movement Sciences,
Amsterdam 1105 AZ, The Netherlands;
Department of Medical Biology,
Academic Medical Center,
University of Amsterdam,
Amsterdam 1105 AZ, The Netherlands
e-mail: t.h.smit@amc.uva.nl

1Corresponding author.

Manuscript received September 22, 2017; final manuscript received March 22, 2018; published online May 24, 2018. Assoc. Editor: James C. Iatridis.

J Biomech Eng 140(9), 091008 (May 24, 2018) (9 pages) Paper No: BIO-17-1423; doi: 10.1115/1.4039890 History: Received September 22, 2017; Revised March 22, 2018

Intervertebral disk (IVD) degeneration is commonly described by loss of height and hydration. However, in the first stage of IVD degeneration, this loss has not yet occurred. In the current study, we use an ex vivo degeneration model to analyze the changes in IVDs mechanical behavior in the first phase of degeneration. We characterize these changes by stretched-exponential fitting, and suggest the fitted parameters as markers for early degeneration. Enzymatic degeneration of healthy lumbar caprine IVDs was induced by injecting 100 μL of Chondroïtinase ABC (Cabc) into the nucleus. A no-intervention and phosphate buffered saline (PBS) injected group were used as controls. IVDs were cultured in a bioreactor for 20 days under diurnal, simulated-physiological loading (SPL) conditions. Disk deformation was continuously monitored. Changes in disk height recovery behavior were quantified using stretched-exponential fitting. Disk height, histological sections, and water- and glycosaminoglycan (GAG)-content measurements were used as gold standards for the degenerative state. Cabc injection caused significant GAG loss from the nucleus and had detrimental effects on poro-elastic mechanical properties of the IVDs. These were progressive over time, with a propensity toward more linear recovery behavior. On histological sections, both PBS and Cabc injected IVDs showed moderate degeneration. A small GAG loss yields changes in IVD recovery behavior, which can be quantified with stretched-exponential fitting. Parameters changed significantly compared to control. Studies on disk degeneration and biomaterial engineering for degenerative disk disease (DDD) could benefit from focusing on IVD biomechanical behavior rather than height and water-content, as a marker for early disk degeneration.

Copyright © 2018 by ASME
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Grahic Jump Location
Fig. 1

Scheme of the daily SPL regimes. On the Y-axis the axial load (MPa) as applied on the IVDs. All IVDs start with 8 h of LDL around 0.1 MPa, after which a 16 h SPL loading regime is applied as indicated in the caption.

Grahic Jump Location
Fig. 2

Line graph with a typical example of an IVD's displacement curve during culture and loading. The most left square depicts the ∼0.1 MPa initial LDL load curve (day 0; subsidence), and the other squares the follow-up recovery curves during low ∼0.1 MPa LDL load (day 1, 4, and 20) after SPL loading.

Grahic Jump Location
Fig. 3

(a) Height loss (mm; mean±SEM) of all three experimental groups from hour 1 to 8 (relative to t = 0) under LDL load in the LDCS, directly after dissection and either no intervention (culture control) or 100 μL PBS injection or 100 μL Cabc at 0.5 μ/ml conc. In the culture control group, an equilibrium was reach at hour 3, after which no further significant height loss was measured between time points, for PBS this was after 4 h and the Cabc group after 5 h. Asterisk (*) indicates p-values below 0.05. (b) Line graph with height loss (mm; mean±SEM) of all three experimental groups from day 1 to day 20 in culture (relative to t = 0). Notice how after day four difference between groups do not change during the remaining culture period; there is no additional height loss in subsequent days due to Cabc injection. Asterisk (*) indicates p-values below 0.05. See color figure online.

Grahic Jump Location
Fig. 4

(a) Typical examples of recovery curves during the LDL recovery phase in culture at day 1, day 4, and day 20 of a healthy noninjected culture control IVD (left), and a PBS (middle) and Cabc injected IVD (right). We can see the recovery arches of the degenerated IVD are progressively flatter (more linear) and shows only a minute loss of recovery of height from day 1 to 20 (0.44–0.40 mm). (b) Stretch fit characteristics quantifying changes in poro-elastic recovery behavior of the IVDs during culture at day 1, 4, and 20. The brackets above the boxplots depict the significant differences between groups. The brackets below the boxplot depict the differences within groups between time points. All parameters are influenced by a settling effect (differences between day 1 and 4), as IVDs need to adjust to culture and loading condition. Especially, β and δ differ between day 1 and 20, exceeding the settling effect and the relative difference between groups at each time point. The time constant increases significantly more in the Cabc group when compared to culture control and PBS. β and δ decrease significantly more and progressively due to Cabc injection when compared to controls. see color figure online.

Grahic Jump Location
Fig. 5

Typical examples of histological sections (scale indicates millimeters) used for scoring of IVD degeneration after the culture and loading experiment as used for the Rutges scale. From top to bottom, section are: midsagittal H&E stained sections to score endplate damage (left posterior, right anterior); transverse H&E stained sections to score annulus and nucleus matrix morphology (all left half IVDs: above anterior; left lateral; bottom posterior); transverse sections stained with Safranin-O and Alcian-blue to observe changes in proteoglycan and GAG distribution. All sections and stainings combined depict the characteristics of the transition zone. Changes due to Cabc injection ranged from mild to moderate degeneration. The overall effect of Cabc injection can be most clearly observed in the Alcian-blue stainings with a lighter, less homogeneous staining of the nucleus region and additional (blue) staining in especially the anterior outer annulus showing GAG leakage from the NP outward through the annulus. See color figure online.

Grahic Jump Location
Fig. 6

Water and GAG content in the nucleus region of all three experimental groups after 21 days of culture. The brackets above the boxplots depict the significant differences between groups with the asterisk (*) indicating p-values below 0.05. See color figure online.



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