Effects of Collagen Crosslink Augmentation on Mechanism of Compressive Load Sharing in Intervertebral Discs.

Exogenous crosslinking has been shown to have potential for treating disc degeneration and back pain due to its ability to increase the strength and toughness of the annulus fibrosus, increase intervertebral joint stability, decrease intradiscal pressure, and increase fluid flow through the disc. Some results imply that crosslink augmentation may also lead to changes in the compressive load sharing properties of the disc. The objective of the present study was to evaluate directional stress distribution changes of the disc following genipin crosslinking treatment. Bovine lumbar motion segments were randomly divided into control and crosslinked groups. Annular strains were determined from simultaneous deformation measurements at various time points during compressive creep testing. Four stress components of the annulus were then calculated according to the previously measured modulus data. Immediately after the application of a 750-N compressive load, mean axial and radial compressive stresses in the crosslinked group were twofold higher than control means. Conversely, mean lamellae-aligned and circumferential tensile stresses of the crosslinked discs were 8- and threefold lower, respectively, compared to control means. After 1-h creep loading, the two compressive mean stresses in both the control and genipin-crosslinked specimens increased approximately threefold from their initial 750-N-loaded values. The two tensile mean stresses in the crosslinked group remained lower than the respective levels of the control means after creep loading. A greater proportion of annular compressive load support under compressive creep loading, with a commensurate decrease in both tensile stresses and strains, was seen in the discs following exogenous crosslink augmentation.

Interspinous-Interbody Fusion via a Strictly Lateral Surgical Approach: A Biomechanical Stabilization Comparison to Constructs Requiring Both Lateral and Posterior Approaches.

Objective Lumbar fusion performed through lateral approaches is becoming more common. The interbody devices are generally supported by supplemental posterior fixation implanted through a posterior approach, potentially requiring a second incision and intraoperative repositioning of the patient. A minimally invasive lateral interspinous fixation device may eliminate the need for intraoperative repositioning and avoid disruption of the supraspinous ligament. The objective of this in vitrobiomechanical study was to investigate segmental multidirectional stability and maintenance of foraminal distraction of a lateral interspinous fixation device compared to commonly used pedicle screw and facet screw posterior fixation constructs when combined with lumbar interbody cages. Methods Six human cadaver lumbar spine specimens were subjected to nondestructive quasistatic loading in the following states: (1) intact; (2) interspinous fixation device alone and (3) with lateral interbody cage; (4) lateral lumbar interbody cage with bilateral pedicle screws; (5) lateral lumbar interbody cage with unilateral pedicle screws; and (6) lateral lumbar interbody cage with facet screws. Multidirectional pure bending in 1.5 Nm increments to 7.5 Nm, and 7.5 Nm flexion-extension bending with a 700 N compressive follower load were performed separately with optoelectronic segmental motion measurement. Relative angular motions of L2-L3, L3-L4, and L4-L5 functional spinal units were evaluated, and the mean instantaneous axis of rotation in the sagittal plane was calculated for the index level. Foraminal height was assessed during combined flexion-extension and compression loading for each test construct. Results All implant configurations significantly restricted flexion-extension motion compared with intact (p < 0.05). No significant differences were found in flexion-extension when comparing the different posterior implants combined with lateral lumbar interbody cages. All posterior fixation devices provided comparable neuroforaminal distraction and maintained distraction during flexion and extension. Conclusions When combinedwith lateral lumbar interbody cages, the minimally invasive lateral interspinous fixation device effectively stabilized the spine and maintained neuroforaminal distraction comparable to pedicle screw constructs or facet screws. These results suggest the lateral interspinous fixation device may provide a favorable alternative to other posterior systems that require patient repositioning during surgery and involve a greater disruption of native tissues.

Effects of exogenous crosslinking on in vitro tensile and compressive moduli of lumbar intervertebral discs.

BACKGROUND: Collagen crosslinks may play a vital role in preventing ongoing disc degeneration. Age-accumulating crosslinks have been thought to increase brittleness and reduce fatigue resistance. However recent studies have demonstrated increases in fatigue resistance, joint stability and nutritional flow properties resulting from crosslink augmentation. In this study, multi-directional moduli of bovine lumbar intervertebral discs were measured in vitro, including circumferential tension, radial compression, axial tension, and axial compression in control and crosslinked specimens. METHODS: Four types of annulus fibrosus specimens were dissected from control and crosslinked discs. Cross-sectional areas were measured using a non-contact laser measurement system and then four separate mechanical tests were conducted using a materials testing machine with custom-made loading fixtures. FINDINGS: The circumferential specimens demonstrated the highest moduli in both low stiffness and linear elastic regions. After a crosslink treatment, the modulus increased more in circumferential tension compared to axial tension and more in axial compression compared to radial compression. Other tensile properties had higher increases in circumferential tension compared to axial tension after crosslinking. INTERPRETATION: Assuming form follows function, circumferential tension is the predominant type of stress experienced by non-degenerated annulus fibrosus. The anisotropic mechanical properties of the annulus fibrosus is non-uniformly affected by crosslink augmentation. Dominant effects were in the directions with greater inherent stiffnesses. These results suggest some beneficial effects of crosslink augmentation on the mechanical properties of the annulus fibrosus: increase in ultimate strength, yield strength, toughness, and modulus in the principal stress directions.

Exogenous Crosslinking Restores Intradiscal Pressure of Injured Porcine Intervertebral Discs: An In Vivo Examination Using Quantitative Discomanometry.

STUDY DESIGN: In vivo examination of intradiscal pressure by quantitative discomanometry (QD). OBJECTIVE: To determine whether an injectable, exogenous crosslinking could acutely restore intradiscal pressure of stab-injured discs in vivo by short-term treatment. SUMMARY OF BACKGROUND DATA: Disc biomechanical performance depends on its integrity associated with the intradiscal pressure and mechanical properties. Genipin crosslink augmentation has demonstrated the in vitro biomechanical capability to improve intervertebral joint stability and increase mechanical properties of the annulus fibrosus. METHODS: 4 lumbar discs on each of 8 swine were randomly assigned to 4 groups: intact, injured, untreated, and crosslinked. A 16G needle was stabbed into the annulus fibrosus to create the disc injury model. An injection of 0.33% genipin solution was delivered into the annulus to treat the injury. QD technique was performed to examine the intradiscal pressure for the intact and injured discs at the time of surgery, while untreated and crosslinked discs were measured 1-week postsurgery. 4 QD parameters were analyzed and compared across the 4 groups: leakage pressure and volume, and saturation pressure and volume. RESULTS: The leakage and saturation pressures of the injured group were significantly lower than those of the intact group (P = 0.004 and P = 0.01, respectively). The leakage and saturation pressures of untreated discs were statistically equivalent to the injured levels, but with a 2-times higher saturation volume. Relative to the untreated group, the leakage pressure and saturation pressure of genipin-crosslinked discs had a 617% (P = 0.008) and a 473% increase (P = 0.007), respectively. CONCLUSION: A large disc injury produced by annular puncture immediately lowered intradiscal pressure when left untreated. Genipin crosslinking can restore intradiscal pressure acutely in vivo without any obvious morbidity associated with the injection.

Radiofrequency thermal effects on the human meniscus. An in vitro study of systems with monopolar and bipolar electrodes.

BACKGROUND: No data exist on the cutting efficiency of monopolar versus bipolar radiofrequency energy application systems on human meniscal tissue. PURPOSE: To compare the effects of monopolar and bipolar thermal energy systems on human meniscal tissue. STUDY DESIGN: Controlled laboratory study. METHODS: Fresh-frozen menisci were cut in cross-section into 180 pie-shaped specimens. A specially designed jig was used to consistently apply radiofrequency energy to the tissue under a constant 30-g force. Three different systems were tested at the low, middle, and high ranges, with application times of 1 and 3 seconds. Thermal effects were measured by image analysis microscopy. RESULTS: No significant differences in thermal effects were found with respect to energy output for each system. Both the individual system tested and the application time had statistically significant effects on thermal damage, with the individual system tested having a greater effect. The mean depths of thermal change produced by the Mitek (bipolar) device were 564 and 648 microm at 1 and 3 seconds applications, respectively. The Arthrocare device (bipolar) produced depths of 1444 and 1697 microm at 1 and 3 seconds. The Oratec device (monopolar) produced depths of 895 and 1057 microm, respectively. CONCLUSIONS: A differential thermal effect was created in the meniscal tissue by three commercially available radiofrequency systems. Within the parameters of the experiment, all three systems limited thermal damage to a depth of less than 2 mm. The results appeared to depend more on the particular system used, not whether it had monopolar or bipolar electrodes. CLINICAL RELEVANCE: These data imply reasonably safe (less than 2 mm) thermal changes in the meniscus after radiofrequency energy application from these three systems.

Thermal profile of radiofrequency energy in the inferior glenohumeral ligament.

PURPOSE: Currently, two different methods of applying radiofrequency (RF) energy (monopolar and bipolar) are available to the surgeon for thermal shortening of the shoulder capsule. The objective of this study was to investigate the temperature changes and the thermal conduction across the human inferior glenohumeral ligament (IGHL) during radiofrequency energy application. METHODS: Thermistors were secured onto both the intra-articular and extra-articular surfaces of human IGHL. Monopolar RF energy and bipolar RF energy were delivered to the intra-articular surface at the manufacturer's recommended settings. Pre-treatment and post-treatment ligament lengths, widths, heating times, and temperatures were measured and compared. RESULTS: For the monopolar devices, temperature spikes to 89 degrees C were recorded for the set temperature of 67 degrees C, averaging 77 degrees C +/- 10 degrees C. Temperatures across the ligament averaged 48 degrees C +/- 3 degrees C. For both devices, the IGHL became thicker with higher RF settings. Recorded temperatures decreased as distance increased from the point of application. Maximum temperatures occurred at least 6 to 7 seconds after cessation of energy application. CONCLUSIONS: The bipolar and monopolar devices had similar conduction times across the ligament, suggesting that this occurs by simple diffusion of heat. Bipolar and monopolar devices were equally efficacious for capsular shrinkage if the extent of the shortening is tightly defined. CLINICAL RELEVANCE: The thermal probe should not rest in one position for an extended period of time during RF energy application because, as our study showed, the monitoring of temperature or the visualization of tissue change is not efficacious for determining the end point of thermal shrinkage of the shoulder capsule.

Radiofrequency thermal effects on the human meniscus: an in vitro analysis.

PURPOSE: The purpose of this study was to evaluate the thermal effects produced in meniscal tissue with different radiofrequency (RF) energy levels and exposure times using a bipolar device. TYPE OF STUDY: An anatomic in vitro analysis of the thermal effects of an RF device. METHODS: A specially designed jig was used to apply RF energy under a constant force to cadaveric menisci. Three different energy levels were applied for 4 different contact times. RESULTS: The overall mean depth of thermal change was 547 microm (range, 468 to 650 microm). There were no significant differences or trends when contact time and energy were varied. CONCLUSIONS: These data suggest that increased contact times and energy outputs are not associated with increased thermal change in the meniscus. The submillimeter thermal denaturation was consistent with published reports using other thermal devices, such as laser.

The dose response relationship between intervertebral disc flexion-extension neutral zone metrics and injected genipin concentration.

PURPOSE: Quantify changes in the flexion--extension neutral zone of the intervertebral disc with injections of increasing genipin concentration. METHODS: Bovine motion segments were treated with varying concentrations of genipin using bilateral injections of constant volume. After overnight static compression loading of the treated segments, anterior-posterior offset loading was used to simulate flexion-extension motion. Range of motion, neutral zone length, neutral zone stiffness, and an instability score were measured. RESULTS: Injection of the disc annulus with increasing concentrations of genipin resulted in corresponding changes in flexion-extension neutral zone. A minimum concentration of 40 mM was needed to observe a significant change. The largest changes were observed with the 400 mM injection. Netural zone stability was the most sensitive of the metrics with a percent change of 48% at 40 mM and over 200% at 400 mM. CONCLUSION: This study establishes the efficacy of using injection delivery to affect disc joint mechanics and quantifies the dose response between injected genipin and the flexion-extension stability of the disc.

Lumbar facet joint and intervertebral disc loading during simulated pelvic obliquity.

BACKGROUND CONTEXT: Intervertebral disc and facet joints are the two primary load-bearing structures of the lumbar spine, and altered loading to these structures may be associated with frontal plane spinal deviations. PURPOSE: To determine the load on the lumbar facet joint and intervertebral disc under simulated frontal plane pelvic obliquity combined loading, an in vitro biomechanical study was conducted. STUDY DESIGN/SETTING: An in vitro biomechanical study using a repeated-measures design was used to compare L4-L5 facet joint and intervertebral disc loading across pure moment and combined loading conditions. METHODS: Eight fresh-frozen lumbosacral specimens were tested under five loading conditions: flexion/extension, lateral bending, axial rotation using pure moment bending (±10 Nm), and two additional tests investigating frontal plane pelvic obliquity and axial rotation (sacrum tilted left 5° and at 10° followed by a ±10-Nm rotation moment). Three-dimensional kinematics, facet load, and intradiscal pressures were recorded from the L4-L5 functional spinal unit. RESULTS: Sagittal and frontal plane loading resulted in significantly smaller facet joint forces compared with conditions implementing a rotation moment (p<.05). The facet joint had the highest peak load during the 10° combined loading condition (124.0±30.2 N) and the lowest peak load in flexion (26.8±16.1 N). Intradiscal pressure was high in lateral flexion (495.6±280.9 kPa) and flexion (429.0±212.9 kPa), whereas intradiscal pressures measured in rotation (253.2±135.0 kPa) and 5° and 10° combined loading conditions were low (255.5±132.7 and 267.1±127.1 kPa, respectively). CONCLUSIONS: Facet loading increased during simulated pelvic obliquity in frontal and transverse planes, whereas intradiscal pressures were decreased compared with sagittal and frontal plane motions alone. Altered spinopelvic alignment may increase the loads experienced by spinal tissue, especially the facet joints.

Exogenous collagen crosslinking of the intervertebral disc restores joint stability after lumbar posterior decompression surgery.

STUDY DESIGN: In vitro evaluation of a chemical, injectable intervention for discectomy induced destabilization. OBJECTIVE: To investigate the ability of two collagen crosslinking agents to restore mechanical properties to lumbar joints destabilized by surgical decompression procedures. SUMMARY OF BACKGROUND DATA: Posterior decompression surgery is a common procedure indicated for tissue pathology that interferes with surrounding neural structures. Previous in vitro, analytical, and clinical studies have shown that removal of load-supporting tissue can compromise joint stability mandating some form of postsurgical stabilization. Currently, no nonsurgical treatments are capable of restoring stability and preventing subsequent degeneration. Exogenous crosslinking of intact discs has shown a fourfold increase in joint stability. METHODS: Fifteen bovine lumbar intervertebral joints were randomly separated into methylglyoxal or genipin treatment groups. Flexion-extension flexibility was quantified in three conditions: intact, postdecompression surgery, and after crosslinking reagent injections. Instability was quantified by calculating neutral zone (NZ), percentage of hysteresis, range of motion, and percentage of strain energy. RESULTS: Simulated surgical decompression increased NZ 111% (P = 0.009), 28% (P = 0.004), range of motion 57% (P = 0.003), and decreased strain energy 37% (P = 0.004). For those discs undergoing methylglyoxal treatment NZ was subsequently reduced 68% (P = 0.012), hysteresis 28% (P = 0.018), range of motion 29% (P = 0.012), and strain energy was increased 71% (P = 0.018). For discs subjected to genipin treatment, NZ was reduced 52% (P = 0.018), hysteresis 23% (P = 0.012), range of motion 44% (P = 0.017), and strain energy was increased 66% (P = 0.012). Mean NZ was lower than intact mean after both methylglyoxal and genipin treatments, 10% and 17% less, respectively, but these differences were not significant. Mean values for all other parameters posttreatment were within 6% of the corresponding intact mean values. CONCLUSION: Injections of crosslinking reagents into lumbar intervertebral discs after simulated decompression surgery restored joint stability according to all parameters. Similar results were found for genipin and methylglyoxal reagents. Implementing exogenous collagen crosslinking as an adjunct to current surgical decompression procedures may be beneficial in preventing or delaying subsequent spinal instability and degeneration.

The influence of exogenous cross-linking and compressive creep loading on intradiscal pressure.

This study involves a biomechanical evaluation of a prospective injectable treatment for degenerative discs. The high osmolarity of the non-degenerated nucleus pulposus attracts water contributing to the hydrostatic behavior of the tissue. This intradiscal pressure is known to drop as fluid is exuded from the matrix due to compressive loading. The objective of this study was to compare the changes in intradiscal pressure in control and genipin cross-linked intervertebral discs. Thirty bovine lumbar motion segments were randomly divided into a phosphate-buffered saline control group and a 0.33% genipin group and soaked at room temperature for 2 days. A needle pressure sensor was held in the center of the disc while short-term and static creep compressive loads were applied. The control group demonstrated a 25% higher average intradiscal pressure compared to genipin-treated discs under 750 N compressive load (p=0.029). Depressurization during static compressive creep was 56% higher in the control than in the genipin group (p=0.014). These results suggest cross-linking induced changes in the poroelastic properties of the involved tissues affected the mechanics of compressive load support in the disc with lower levels of nucleus pressure, a corresponding decrease in the elastic expansion of the annulus, and an increased axial compressive loading of the inner and outer annulus tissues. It is possible that concurrent changes in hydraulic permeability and proteoglycan retention known to be associated with genipin cross-linking were also contributors to poroelastic changes. Reduction of peak pressures and moderation of pressure fluctuations could be beneficial relative to discogenic pain.

The effects of exogenous crosslinking on hydration and fluid flow in the intervertebral disc subjected to compressive creep loading and unloading.

STUDY DESIGN: In vitro study of genipin crosslinking effect on disc water content changes under compressive loading and unloading. OBJECTIVE: To investigate the influence of collagen crosslinking on hydration and fluid flow in different regions of intact discs, and to evaluate the nutritional implications. SUMMARY OF BACKGROUND DATA: Age-related reductions of nutrient supply and waste product removal are critically important factors in disc pathogenesis. Diffusion and fluid flow are blocked by subchondral bone thickening, cartilaginous endplate calcification, loss of hydrophilic proteoglycans, and clogging of anular pores by degraded matrix molecules. Previous studies demonstrated increased hydraulic permeability and macromolecular transport through crosslinked collagenous matrices. Genipin has also demonstrated the capability to increase retention of proteoglycans. METHODS: A total of 57 bovine lumbar motion segments were divided randomly into phosphate buffered saline and 0.33% genipin-soaked treatment groups. Water content changes were measured using a mass-loss technique in 3 intervertebral disc regions following successive stages of compressive loading and unloading (post-treatment, after 1 hour 750 N compression, and after a subsequent 24-hour period of nominal loading). Net flow of fluid into or out of a region was determined from the percentage change in mean water content from successive groups. RESULTS: Fluid flow to and from the nucleus doubled with genipin crosslinking. Relative to the buffer-only controls, overall net fluid flow increased 103% in the nucleus pulposus, 36% in the inner anulus, and was 31% less in the outer anulus of genipin treated discs. CONCLUSION: The effects of genipin crosslinking on matrix permeability and proteoglycan retention can alter hydration levels and fluid flow in the intervertebral disc. Resulting increases in fluid flow, including a doubling of flow to and from the nucleus, could lead to enhanced nutritional inflow and waste product outflow for the disc, and may have implications for emerging cell-based therapies.

The analysis of axisymmetric viscoelasticity, time-dependent recovery, and hydration in rat tail intervertebral discs by radial compression test.

The goal of this study was to develop a nondestructive radial compression technique and to investigate the viscoelastic behavior of the rat tail disc under repeated radial compression. Rat tail intervertebral disc underwent radial compression relaxation testing and creep testing using a custom-made gravitational creep machine. The axisymmetric viscoelasticity and time-dependent recovery were determined. Different levels of hydration (with or without normal saline spray) were supplied to evaluate the effect of changes in viscoelastic properties. Viscoelasticity was found to be axisymmetric in rat-tail intervertebral discs at four equidistant locations. Complete relaxation recovery was found to take 20 min, whereas creep recovery required 25 min. Hydration was required for obtaining viscoelastic axisymmetry and complete viscoelastic recovery.

Exogenous cross-linking increases the stability of spinal motion segments.

STUDY DESIGN: The mechanical stability of cross-linked and control spinal motion segments was evaluated using neutral zone, range of motion (ROM), and instability score metrics. OBJECTIVE: To determine if exogenous cross-linking could increase the stability of spinal motion segments. SUMMARY OF BACKGROUND DATA: The microstructure of the anulus fibrosus extracellular matrix can affect the stability of the intervertebral joint. Parallel testing in our laboratory has shown that exogenous cross-linking can improve the fatigue resistance of anulus fibrosus. METHODS: There were 3 separate experimental protocols conducted. The first study used calf lumbar intervertebral joints randomly divided into a genipin cross-linked group and phosphate buffered saline-soaked controls. After 2 days of soaking, flexion-extension ramp cycles were applied to the specimens. The second study repeated the test protocol using 22 moderately and severely degenerated human lumbar intervertebral joints. The third experiment compared the effect of cross-linking treatment on human discs with known degrees of preexisting mechanical instability. Each data set was used to assess joint instability by 3 calculations: ROM, neutral zone, and an instability score. Joint instability for each data set was evaluated using 3 calculations: ROM, neutral zone, and a novel instability score. RESULTS: These results show that cross-link augmentation can effectively reduce instability of intervertebral discs. The stabilizing effect was observed to be higher in the more mechanically unstable discs. However, cross-linking did not appear to affect the total range of sagittal motion. CONCLUSIONS: By reducing the neutral zone, exogenous cross-linking may help combat the progression of instability in degenerative disc disease.