Limitation of finite element analysis of poroelastic behavior of biological tissues undergoing rapid loading

The finite element method is used in biomechanics to provide numerical solutions to simulations of structures having complex geometry and spatially differing material properties. Time-varying load deformation behaviors can result from solid viscoelasticity as well as viscous fluid flow through porous materials. Finite element poroelastic analysis of rapidly loaded slow-draining materials may be ill-conditioned, but this problem is not widely known in the biomechanics field. It appears as instabilities in the calculation of interstitial fluid pressures, especially near boundaries and between different materials. Accurate solutions can require impractical compromises between mesh size and time steps. This article investigates the constraints imposed by this problem on tissues representative of the intervertebral disc, subjected to moderate physiological rates of deformation. Two test cylindrical structures were found to require over 10(4) linear displacement-constant pressure elements to avoid serious oscillations in calculated fluid pressure. Fewer Taylor–Hood (quadratic displacement–linear pressure elements) were required, but with complementary increases in computational costs. The Vermeer–Verruijt criterion for 1D mesh size provided guidelines for 3D mesh sizes for given time steps. Pressure instabilities may impose limitations on the use of the finite element method for simulating fluid transport behaviors of biological soft tissues at moderately rapid physiological loading rates

Challenges and strategies in the repair of ruptured annulus fibrosus

Lumbar discectomy is the surgical procedure most frequently performed for patients suffering from low back pain and sciatica. Disc herniation as a consequence of degenerative or traumatic processes is commonly encountered as the underlying cause for the painful condition. While discectomy provides favourable outcome in a majority of cases, there are conditions where unmet requirements exist in terms of treatment, such as large disc protrusions with minimal disc degeneration; in these cases, the high rate of recurrent disc herniation after discectomy is a prevalent problem. An effective biological annular repair could improve the surgical outcome in patients with contained disc herniations but otherwise minor degenerative changes. An attractive approach is a tissue-engineered implant that will enable/stimulate the repair of the ruptured annulus. The strategy is to develop three-dimensional scaffolds and activate them by seeding cells or by incorporating molecular signals that enable new matrix synthesis at the defect site, while the biomaterial provides immediate closure of the defect and maintains the mechanical properties of the disc. This review is structured into (1) introduction, (2) clinical problems, current treatment options and needs, (3) biomechanical demands, (4) cellular and extracellular components, (5) biomaterials for delivery, scaffolding and support, (6) pre-clinical models for evaluation of newly developed cell- and material-based therapies, and (7) conclusions. This article highlights that an interdisciplinary approach is necessary for successful development of new clinical methods for annulus fibrosus repair. This will benefit from a close collaboration between research groups with expertise in all areas addressed in this review

Harmonization and standardization of nucleus pulposus cell extraction and culture methods

Background In vitro studies using nucleus pulposus (NP) cells are commonly used to investigate disc cell biology and pathogenesis, or to aid in the development of new therapies. However, lab-to-lab variability jeopardizes the much-needed progress in the field. Here, an international group of spine scientists collaborated to standardize extraction and expansion techniques for NP cells to reduce variability, improve comparability between labs and improve utilization of funding and resources. Methods The most commonly applied methods for NP cell extraction, expansion, and re-differentiation were identified using a questionnaire to research groups worldwide. NP cell extraction methods from rat, rabbit, pig, dog, cow, and human NP tissue were experimentally assessed. Expansion and re-differentiation media and techniques were also investigated. Results Recommended protocols are provided for extraction, expansion, and re-differentiation of NP cells from common species utilized for NP cell culture. Conclusions This international, multilab and multispecies study identified cell extraction methods for greater cell yield and fewer gene expression changes by applying species-specific pronase usage, 60–100 U/ml collagenase for shorter durations. Recommendations for NP cell expansion, passage number, and many factors driving successful cell culture in different species are also addressed to support harmonization, rigor, and cross-lab comparisons on NP cells worldwide

Complex loading affects intervertebral disc mechanics and biology

SummaryBackgroundComplex loading develops in multiple spinal motions and in the case of hyperflexion is known to cause intervertebral disc (IVD) injury. Few studies have examined the interacting biologic and structural alterations associated with potentially injurious complex loading, which may be an important contributor to chronic progressive degeneration.ObjectiveThis study tested the hypothesis that low magnitudes of axial compression loading applied asymmetrically can induce IVD injury affecting cellular and structural responses in a large animal IVD ex-vivo model.MethodsBovine caudal IVDs were assigned to either a control or wedge group (15°) and placed in organ culture for 7 days under static 0.2MPa load. IVD tissue and cellular responses were assessed through confined compression, qRT-PCR, histology and structural and compositional measurements, including Western blot for aggrecan degradation products.ResultsComplex loading via asymmetric compression induced cell death, an increase in caspase-3 staining (apoptosis), a loss of aggrecan and an increase in aggregate modulus in the concave annulus fibrosis. While an up-regulation of MMP-1, ADAMTS4, IL-1β, and IL-6 mRNA, and a reduced aggregate modulus were induced in the convex annulus.ConclusionAsymmetric compression had direct deleterious effects on both tissue and cells, suggesting an injurious loading regime that could lead to a degenerative cascade, including cell death, the production of inflammatory mediators, and a shift towards catabolism. This explant model is useful to assess how injurious mechanical loading affects the cellular response which may contribute to the progression of degenerative changes in large animal IVDs, and results suggest that interventions should address inflammation, apoptosis, and lamellar integrity

Effects of immobilization and dynamic compression on intervertebral disc cell gene expression in vivo

AB Study Design. In vitro and in vivo studies to assess the effect of direct anular repair on subsequent degeneration of intervertebral discs (IVDs). Objective. To assess whether a new suturing method could provide sealing effect on IVD after discectomy, and influence degenerative process of IVD. Summary of Background Data. Recurrent disc herniation and subsequent disc degeneration are major problems after discectomy. Anular repair can reduce the risk of recurrence, but its effect on disc degeneration needs more investigation. Methods. A new suturing technique, the modified purse-string suture (MPSS), was designed for direct closure of anular incision. Intact motion segments of porcine lumbar spine were used to validate this technique in resisting disc pressure under mechanical loadings. A transverse slit incision was made in the anterior anulus of porcine cervical discs, with or without sealing of the anular defect by this suturing method. Magnetic resonance imaging grading was recorded before and after surgery. Anular healing was assessed histologically and gene expression of aggrecan, collagen type I, II, and matrix metalloproteinase-13 in nucleus pulposus were investigated. Results. The average failure force of axial compression was 1150.3 +/- 121.1 N for a simple suture, and 2917.9 +/- 627.6 N for a MPSS. Cyclic loading test showed that the repaired discs succeeded against repeated compression forces. Magnetic resonance imaging and gross appearances showed lesser degenerative changes in repaired discs than in injured discs at each time period. In repaired discs, mRNA expression of aggrecan and type II collagen downregulated slightly with time, whereas it decreased rapidly and persistently in unrepaired discs. Histologic findings showed primary healing of outer anular tract in repaired discs. Conclusion. In this pilot study, the MPSS can provide effectively sealing for damaged anulus to withstand stresses. Direct repair of anular incision by this suturing method does significantly slow down degenerative process within discs after discectomy

Streaming potential of human lumbar anulus fibrosus is anisotropic and affected by disc degeneration

The streaming potential responses of non-degenerate and degenerate human anulus fibrosus were measured in a one-dimensional permeation configuration under static and dynamic loading conditions. The goal of this study was to investigate the influence of the changes in tissue structure and composition on the electrokinetic behavior of intervertebral disc tissues. It was found that the static streaming potential of the anulus fibrosus depended on the degenerative grade of the discs ( p=0.0001) and on the specimen orientation in which the fluid flows ( p=0.0001). For a statically applied pressure of 0.07 MPa, the ratio of streaming potential to applied pressure ranged from 5.3 to 6.9 mV/MPa and was largest for Grade I tissue with axial orientation and lowest for Grade III tissue with circumferential orientation. The dynamic streaming potential responses of anulus fibrosus were sensitive to the degeneration of the disc: the total harmonic distortion factor increased by 108%, from 3.92±0.66% (mean±SD) for Grade I specimens to 8.15±3.05% for Grades II and III specimens. The alteration of streaming potential reflects the changes in tissue composition and structure with degeneration. To our knowledge, this is the first reported data for the streaming potential of human intervertebral disc tissues. Knowledge of the streaming potential response of the intervertebral disc provides an understanding of potentially important signal transduction mechanisms in the disc and of the etiology of intervertebral disc degeneration