Intradiscal injections of osteogenic protein-1 restore the viscoelastic properties of degenerated intervertebral discs

BACKGROUND CONTEXT: Using biochemical, histological, and radiological parameters in a rabbit model of intervertebral disc (IVD) degeneration, the intradiscal injection of a growth factor, such as osteogenic protein-1 (OP-1), has been shown to regenerate the IVD. However, very little is known about how such a biological therapeutic approach affects the biomechanical properties of the degenerated IVD. PURPOSE: To investigate the effects of an intradiscal injection of OP-1 on the biomechanical properties of IVDs in the rabbit annular-puncture disc degeneration model and to determine their relationship to biochemical properties. STUDY DESIGN/SETTING: In vivo study on the effects of intradiscally administered OP-1 on the biomechanical and biochemical properties of IVDs in the rabbit annular-puncture disc degeneration model. METHODS: New Zealand White rabbits (n=16) underwent annulus fibrosus (AF) puncture, using an 18-gauge needle, at L2-L3 and L4-L5 (L3-L4: nonpunctured control). Four weeks later, the punctured discs received an injection of either 5% lactose (10 microL) or OP-1 (100 microg/10 microL of 5% lactose) into the nucleus pulposus (NP). The disc height was radiographically monitored biweekly. After sacrifice and removal of bone-disc-bone complexes 8 weeks postinjection, the dynamic viscoelastic properties of the IVDs were tested by applying a cycle of sinusoidal strain in uniaxial compression at six loading frequencies (0.05 to 2 Hz). The biochemical properties of the dissected IVDs were then analyzed and correlated with the biomechanical properties. RESULTS: A single injection of OP-1 significantly restored disc height when compared with the lactose-injected discs (OP-1 vs. lactose, p<.001). The elastic modulus of the IVDs in the OP-1-injected discs was significantly higher than that in the lactose-injected discs at all frequencies (mean: +43%, p<.001). The viscous modulus in the OP-1-injected discs was significantly higher at 0.05, 0.2, 0.5, and 1 Hz (mean: +55%, p<.001) and showed higher tendencies at other frequencies (p=.08-.09). For both moduli, no significant differences were observed between the OP-1-injected and the nonpunctured control discs. The OP-1 injection significantly increased the proteoglycan (PG) content in the NP and AF, and the collagen content in the NP (p<.001-.05). Both elastic and viscous moduli showed significant positive correlations with PG content in the NP and collagen content in the NP and AF (Rho=.357-.466, p=.010-.047). CONCLUSIONS: We have shown for the first time that an injection of the growth factor, OP-1, restored the biomechanical properties of IVDs in a rabbit model of IVD degeneration. Comparing biomechanical with biochemical data suggests that the OP-1-induced biomechanical restoration was a consequence of increased activities of anabolic pathways that resulted in biochemical changes in the IVD.     

Effects of Intradiscal Injection of Local Anesthetics on Intervertebral Disc Degeneration in Rabbit Degenerated Intervertebral Disc

Analgesic discoblock is widely used for the diagnosis or treatment of discogenic low back pain by injecting local anesthetics. The purpose of this study was to investigate the deleterious effects of local anesthetics on degenerated rabbit intervertebral disks (IVDs) using an organotypic culture model and in vivo long‐term follow‐up model. To induce IVD degeneration, a rabbit annular puncture model was used. For the organotypic culture model, degenerated IVDs were harvested 1 month after the initial annular puncture and cultured for 3 or 7 days after intradiscal injection of local anesthetics (1% lidocaine and 0.5% bupivacaine). To perform in vivo analysis, local anesthetics were injected into degenerated IVDs, and IVDs were prepared for histological analysis after 6 or 12 months. In the organotypic model, terminal deoxynucleotidyl transferase dUTP nick end labeling‐positive nucleus pulposus (NP) cells were significantly increased in the bupivacaine group compared with the other groups. In the in vivo study, the number of NP cells was significantly decreased in the saline and local anesthetics groups compared with the untreated control and puncture‐only groups. However, there was no significant difference among the saline, lidocaine, and bupivacaine groups. In addition, histological analysis showed no significant difference of IVD degeneration among the puncture‐only, saline, lidocaine, and bupivacaine groups. Although bupivacaine induced apoptotic NP cell death in the organotypic culture model, in vivo observations did not show any definitive proof to suggest that local anesthetics were capable of promoting degeneration in the degenerated IVD, except for pressurized injection‐induced damage. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1963–1971, 2019     

Biomechanical assessment of the effects of decompressive surgery in non-chondrodystrophic and chondrodystrophic canine multisegmented lumbar spines

Purpose

Dogs are often used as an animal model in spinal research, but consideration should be given to the breed used as chondrodystrophic (CD) dog breeds always develop IVD degeneration at an early age, whereas non-chondrodystrophic (NCD) dog breeds may develop IVD degeneration, but only later in life. The aim of this study was to provide a mechanical characterization of the NCD [non-degenerated intervertebral discs (IVDs), rich in notochordal cells] and CD (degenerated IVDs, rich in chondrocyte-like cells) canine spine before and after decompressive surgery (nucleotomy).

Methods

The biomechanical properties of multisegmented lumbar spine specimens (T13–L5 and L5–Cd1) from 2-year-old NCD dogs (healthy) and CD dogs (early degeneration) were investigated in flexion/extension (FE), lateral bending (LB), and axial rotation (AR), in the native state and after nucleotomy of L2–L3 or dorsal laminectomy and nucleotomy of L7–S1. The range of motion (ROM), neutral zone (NZ), and NZ stiffness (NZS) of L1–L2, L2–L3, L6–L7, and L7–S1 were calculated.

Results

In native spines in both dog groups, the greatest mobility in FE was found at L7–S1, and the greatest mobility in LB at L2–L3. Surgery significantly increased the ROM and NZ, and significantly decreased the NZS in FE, LB, and AR in both breed groups. However, surgery at L2–L3 resulted in a significantly larger increase in NZ and decrease in NZS in the CD spines compared with the NCD spines, whereas surgery at L7–S1 induced a significantly larger increase in ROM and decrease in NZS in the NCD spines compared with the CD spines.

Conclusions

Spinal biomechanics significantly differ between NCD and CD dogs and researchers should consider this aspect when using the dog as a model for spinal research.     

Influence of degenerative changes of intervertebral disc on its material properties and pathology

Objective: To investigate the material properties of normal and degenerated intervertebral discs (IVDs) and examine the effect of degenerative changes on IVD pathology.Methods: A computer-based online ...     

Transplantation of mesenchymal stem cells in a canine disc degeneration model

Transplantation of mesenchymal stem cells (MSCs) is effective in decelerating disc degeneration in small animals; much remains unknown about this new therapy in larger animals or humans. Fas‐ligand (FasL), which is only found in tissues with isolated immune privilege, is expressed in IVDs, particularly in the nucleus pulposus (NP). Maintaining the FasL level is important for IVD function. This study evaluated whether MSC transplantation has an effect on the suppression of disc degeneration and preservation of immune privilege in a canine model of disc degeneration. Mature beagles were separated into a normal control group (NC), a MSC group, and the disc degeneration (nucleotomy‐only) group. In the MSC group, 4 weeks after nucleotomy, MSCs were transplanted into the degeneration‐induced discs. The animals were followed for 12 weeks after the initial operation. Subsequently, radiological, histological, biochemical, immunohistochemical, and RT‐PCR analyses were performed. MSC transplantation effectively led to the regeneration of degenerated discs. FACS and RT‐PCR analyses of MSCs before transplantation demonstrated that the MSCs expressed FasL at the genetic level, not at the protein level. GFP‐positive MSCs detected in the NP region 8 weeks after transplantation expressed FasL protein. The results of this study suggest that MSC transplantation may contribute to the maintenance of IVD immune privilege by the differentiation of transplanted MSCs into cells expressing FasL. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:589–600, 2008     

Alterations in the mechanical behavior of the human lumbar nucleus pulposus with degeneration and aging

This study tested the hypothesis that changes in the morphology and composition of the nucleus pulposus with age and degeneration have associated changes in its mechanical properties. A torsional shear experiment was used to determine viscoelastic shear properties of cylindrical samples of human nucleus pulposus with large ranges of grades of morphological degeneration (normal to severely degenerated) and ages (range: 16–88 years; average: 57 ± 21.5 years). Viscoclastic shear properties were determined from stress-relaxation and dynamic sinusoidal tests. A linear viscoelastic law with a variable-amplitude relaxation spectrum was used to model experimental behaviors of nucleus pulposus specimens. A statistically significant increase in the instantaneous and dynamic shear moduli was found with increasing age and grade of degeneration; the values for moduli ranged from 5.0 to 60 kPa. A significant decrease in tanδ was also detected; the values ranged from 0.43 to 0.33, indicating a decreased capacity for the nucleus pulposus to dissipate energy. The dynamic modulus and tanδ were also significantly affected by frequency. It was generally concluded that the nucleus pulposus undergoes a transition from “fluid-like” behavior to more “solid-like” behavior with aging and degeneration.     

An injectable nucleus pulposus implant restores compressive range of motion in the ovine disc

STUDY DESIGN.: Investigation of injectable nucleus pulposus (NP) implant. OBJECTIVE.: To assess the ability of a recently developed injectable hydrogel implant to restore nondegenerative disc mechanics through support of NP functional mechanics. SUMMARY OF BACKGROUND DATA.: Although surgical intervention for low back pain is effective for some patients, treated discs undergo altered biomechanics and adjacent levels are at increased risk for accelerated degeneration. One potential treatment as an alternative to surgery for degenerated disc includes the percutaneous delivery of agents to support NP functional mechanics. The implants are delivered in a minimally invasive fashion, potentially on an outpatient basis, and do not preclude later surgical options. One of the challenges in designing such implants includes the need to match key NP mechanical behavior and mimic the role of native nondegenerate NP in spinal motion. METHODS.: The oxidized hyaluronic acid gelatin implant material was prepared. In vitro mechanical testing was performed in mature ovine bone-disc-bone units in 3 stages: intact, discectomy, and implantation versus sham. Tested samples were cut axially for qualitative structural observations. RESULTS.: Discectomy increased axial range of motion (ROM) significantly compared with intact. Hydrogel implantation reduced ROM 17% (P < 0.05) compared with discectomy and returned ROM to intact levels (ROM intact 0.71 mm, discectomy 0.87 mm, postimplantation 0.72 mm). Although ROM for the hydrogel implant group was statistically unchanged compared with the intact disc, ROM for sham discs, which received a discectomy and no implant, was significantly increased compared with intact. The compression and tension stiffness were decreased with discectomy and remained unchanged for both implant and sham groups as expected because the annulus fibrosus was not repaired. Gross morphology images confirmed no ejection of NP implant. CONCLUSION.: An injectable implant that mimics nondegenerate NP has the potential to return motion segment ROM to normal subsequent to injury.     

Needle puncture injury causes acute and long‐term mechanical deficiency in a mouse model of intervertebral disc degeneration

Low back pain is a significant socioeconomic burden and intervertebral disc degeneration has been implicated as a cause. A reliable animal model of disc degeneration is necessary to evaluate therapeutics, and functional metrics are essential to quantify their benefit. To this end, needle puncture injuries were created in the caudal intervertebral discs of mice to induce disc degeneration. Compression, torsion, and creep mechanics were assessed both immediately and after eight weeks to distinguish between the effects of injury and the subsequent reparative or degenerative response. Two needle sizes (29 and 26 gauge) were used to determine injury size‐dependence. Compressive stiffness (62%), torsional stiffness (60%), and early damping stiffness (84%) decreased immediately after injury with the large needle (26G). These mechanical properties did not change over time despite structural and compositional changes. At 8 weeks following large needle injury, disc height decreased (37%), nucleus pulposus (NP) glycosaminoglycan content decreased (41%), and NP collagen content increased (45%). The small needle size had no significant effect on mechanics and did not initiate degenerative changes in structure and composition. Thus, the injection of therapeutics into the NP with a minimal needle size may limit damage due to the needle insertion. These findings, along with the wide commercial availability of mouse‐specific biological probes, indicate that the mouse caudal disc model can be a powerful tool for investigating disc degeneration and therapy. © 2013 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 31:1276–1282, 2013     

Disc cell clusters in pathological human intervertebral discs are associated with increased stress protein immunostaining

Intervertebral disc (IVD) cells within the annulus fibrosus (AF) and nucleus pulposus (NP) maintain distinct functional extracellular matrices and operate within a potentially noxious and stressful environment. How disc cells respond to stress and whether stress is responsible for triggering degeneration is unknown. Disc cell proliferation and cluster formation are most marked in degenerate IVDs, possibly indicating attempts at matrix repair. In other tissues, stress proteins increase rapidly after stress protecting cell function and, although implicated in degeneration of articular cartilage, have received little attention in degenerative IVD pathologies. We have compared the distribution of stress protein immunolocalization in pathological and control IVDs. Disc tissues were obtained at surgery from 43 patients with degenerative disc disease (DDD) and herniation, and 12 controls at postmortem. Tissues were immunostained with a polyclonal antibody for heat shock factor 1 (HSF-1) and monoclonal antibodies for the heat shock proteins, Hsp27 and Hsp72, using an indirect immunoperoxidase method. Positively stained cells were expressed as a percentage of the total. Cell cluster formation was also assessed. The proportion of cells in clusters was similar in the AF (both 2%) and NP (8 and 9%) of control and DDD samples, whereas in herniated tissues this was increased (AF 12%, NP 14%). Stress antigen staining tended to be more frequent in clustered rather than in single/doublet cells, and this was significant (P < 0.005) in both the AF and NP of herniated discs. Clustered cells, which are most common in herniated discs, may be mounting a protective response to abnormal environmental factors associated with disc degeneration. A better understanding of the stress response in IVD cells may allow its utilization in disc cell therapies.     

Mechanical behavior of the human lumbar spine. I. Creep analysis during static compressive loading

The in vitro viscoelastic "creep" behavior was examined in 18 cadaveric human lumbar motion segments subjected to static axial compressive loads. Axial deformation was followed for 30 min under constant applied load. Compressive material constants (moduli and viscosity coefficients) were then determined for each intervertebral disc using a linearization method based on a Taylor series expansion of experimental data for the "three parameter" viscoelastic creep model. The degree of disc degeneration and bone mineral content (BMC) were also assessed. Good correlation between the experimentally determined and model predicted strain values were found, with the average error less than 1%. We found that motion segments from older and more degenerated lumbar discs were less stable and had lower material constants than segments from younger and less degenerated discs. Material constants and BMC correlated closely, suggesting that an interdependency of disc and vertebral body properties exists. No correlation between the creep characteristics and disc height, disc area, segment level, or sex were noted.     

Use of Allogeneic Hypoxic Mesenchymal Stem Cells For Treating Disc Degeneration in Rabbits

Intervertebral discs (IVDs) are important biomechanical components of the spine. Once degenerated, mesenchymal stem cell (MSC)‐based therapies may aid in the repair of these discs. Although hypoxic preconditioning enhances the chondrogenic potential of MSCs, it is unknown whether bone marrow MSCs expanded under hypoxic conditions (1% O₂, here referred to as hypoxic MSCs) are better than bone marrow MSCs expanded under normoxic conditions (air, here referred to as normoxic MSCs) with regards to disc regeneration capacity. The purpose of this study was to compare the therapeutic effects of hypoxic and normoxic MSCs in a rabbit needle puncture degenerated disc model after intra‐disc injection. Six weeks after needle puncture, MSCs were injected into the IVD. A vehicle‐treated group and an un‐punctured sham‐control group were included as controls. The tissues were analyzed by histological and immunohistochemical methods 6 and 12 weeks post‐injection. At 6 and 12 weeks, less disc space narrowing was evident in the hypoxic MSC‐treated group compared to the normoxic MSC‐treated group. Significantly better histological scores were observed in the hypoxic MSC group. Discs treated with hypoxic MSCs also demonstrated significantly better extracellular matrix deposition in type II and XI collagen. Increased CD105 and BMP‐7 expression were also observed upon injection of hypoxic MSCs. In conclusion, hypoxic MSC injection was more effective than normoxic MSC injection for reducing IVD degeneration progression in vivo. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1440–1450, 2019.     

Long-term flexion-extension range of motion of the prodisc total disc replacement

The rationale for total disc replacement is avoidance of the junctional degeneration seen after arthrodesis by preservation of segmental motion. To justify the use of disc prostheses, it is essential to document maintained range of motion (ROM) and sagittal alignment at long-term follow-up. This is a retrospective radiographic study of 42 patients who had placement of 58 first-generation Prodisc prostheses at a mean follow-up of 8.7 years. Flexion-extension ROM was measured by Cobb's method. Junctional levels were evaluated for junctional degeneration. Pre- and postoperative global and segmental lordosis were measured. Prognostic patient factors predicting ROM of <2 degrees were evaluated. We observed ROM of at least 2 degrees in 66% of Prodisc prostheses at 8.7-year follow-up, although ROM was less than that reported in asymptomatic normal individuals. Mean ROM for disc prostheses with motion was 7.5 degrees at L3-L4, 6.2 degrees at L4-L5, and 4.1 degrees at L5-S1. Mean ROM for all prostheses was 3.8 degrees. The incidence of radiographic junctional degeneration was 24%, although no patients required surgery for symptomatic junctional degeneration. Mean ROM of prostheses below a degenerated junctional disc was 1.6 degrees compared with 4.7 degrees below a normal junctional disc (P < 0.035). Females were 3.5 times more likely to have ROM of <2 degrees. This is the longest published follow-up study of a lumbar disc replacement. The data show that ROM is preserved at long-term follow-up in the majority of patients. Global and segmental sagittal alignment improve after surgery. Furthermore, there is an association between ROM of disc prostheses and the development of junctional degeneration.     

Biomechanical characterization of an annulus-sparing spinal disc prosthesis

Background contextCurrent spine arthroplasty devices require disruption of the annulus fibrosus for implantation. Preliminary studies of a unique annulus-sparing intervertebral prosthetic disc (IPD) found that preservation of the annulus resulted in load sharing of the annulus with the prosthesis.PurposeDetermine flexibility of the IPD versus fusion constructs in normal and degenerated human spines.Study design/settingBiomechanical comparison of motion segments in the intact, fusion and mechanical nucleus replacement states for normal and degenerated states.Patient settingThirty lumbar motion segments.Outcomes measuresIntervertebral height; motion segment range of motion, neutral zone, stiffness.MethodsMotion segments had multidirectional flexibility testing to 7.5 Nm for intact discs, discs reconstructed using the IPD (n=12), or after anterior/posterior fusions (n=18). Interbody height and axial compression stiffness changes were determined for the reconstructed discs by applying axial compression to 1,500 N. Analysis included stratifying results to normal mobile versus rigid degenerated intact motion segments.ResultsThe mean interbody height increase was 1.5 mm for IPD reconstructed discs versus 3.0 mm for fused segments. Axial compression stiffness was 3.0±0.9 kN/mm for intact compared with 1.2±0.4 kN/mm for IPD reconstructed segments. Reconstructed disc ROM was 9.0°±3.7° in flexion extension, 10.6°±3.4° in lateral bending, and 2.8°±1.4° in axial torsion that was similar to intact values and significantly greater than respective fusion values (p<.001). Mobile intact segments exhibited significantly greater rotation after fusion versus their more rigid counterparts (p<.05); however, intact motion was not related to motion after IPD reconstruction. The NZ and rotational stiffness followed similar trends. Differences in NZ between mobile and rigid intact specimens tended to decrease in the IPD reconstructed state.ConclusionThe annulus-sparing IPD generally reproduced the intact segment biomechanics in terms of ROM, NZ, and stiffness. Furthermore, the IPD reconstructed discs imparted stability by maintaining a small neutral zone. The IPD reconstructed discs were significantly less rigid than the fusion constructs and may be an attractive alternative for the treatment of degenerative disc disease.     

Rheological characterization of the nucleus pulposus and dense collagen scaffolds intended for functional replacement

Lumbar discectomy is an effective therapy for neurological decompression in patients suffering from sciatica due to a herniated nucleus pulposus (NP). However, high numbers of patients suffering from persisting postoperative low back pain have resulted in many strategies targeting the regeneration of the NP. For successful regeneration, the stiffness of scaffolds is increasingly recognized as a potent mechanical cue for the differentiation and biosynthetic response of (stem) cells. The aim of the current study is to characterize the viscoelastic properties of the NP and to develop dense collagen scaffolds with similar properties. The scaffolds consisted of highly dense (0.5%–12%) type I collagen matrices, prepared by plastic compression. The complex modulus of the NP was 22 kPa (at 10 rad s^?1), which should agree with a scaffold with a collagen concentration of 23%. The loss tangent, indicative of energy dissipation, is higher for the NP (0.28) than for the scaffolds (0.12) and was not dependent of the collagen density. Gamma sterilization of the scaffolds increased the shear moduli but also resulted in more brittle behavior and a reduced swelling capacity. In conclusion, by tuning the collagen density, we can approach the stiffness of the NP. Therefore, dense collagen is a promising candidate for tissue engineering of the NP that deserves further study, such as the addition of other proteins. Published by Wiley Periodicals, Inc. J Orthop Res 27: 620–626, 2009     

Organ culture stability of the intervertebral disc: Rat versus rabbit

There is a need to develop mechanically active culture systems to better understand the role of mechanical stresses in intervertebral disc (IVD) degeneration. Motion segment cultures that preserve the native IVD structure and adjacent vertebral bodies are preferred as model systems, but rapid ex vivo tissue degeneration limits their usefulness. The stability of rat and rabbit IVDs is of particular interest, as their small size makes them otherwise suitable for motion segment culture. The goal of this study was to determine if there are substantial differences in the susceptibility of rat and rabbit IVDs to culture‐induced degeneration. Lumbar IVD motion segments were harvested from young adult male Sprague–Dawley rats and New Zealand White rabbits and cultured under standard conditions for 14 days. Biochemical assays and safranin‐O histology showed that while glycosaminoglycan (GAG) loss was minimal in rabbit IVDs, it was progressive and severe in rat IVDs. In the rat IVD, GAG loss was concomitant with the loss of notochordal cells and the migration of endplate (EP) cells into the nucleus pulposus (NP). None of these changes were evident in the rabbit IVDs. Compared to rabbit IVDs, rat IVDs also showed increased matrix metalloproteinase‐3 (MMP‐3) and sharply decreased collagen type I and II collagen expression. Together these data indicated that the rabbit IVD was dramatically more stable than the rat IVD, which showed culture‐related degenerative changes. Based on these findings we conclude that the rabbit motion segments are a superior model for mechanobiologic studies. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 31: 838–846, 2013     

Morphological characteristics of the kangaroo lumbar intervertebral discs and comparison with other animal models used in spine research

Animal models are frequently used to elucidate pathomechanism and pathophysiology of various disorders of the human intervertebral disc (IVD) and also to develop therapeutic approaches. Here we report morphological characteristics of the kangaroo lumbar IVDs and compare them with other animal models used in spine research. Twenty-five fresh-frozen cadaveric lumbar spines (T12–S1) derived from kangaroo carcases (Macropus giganteus) of undetermined age were first scanned in a C-Arm X-ray machine. A photograph of the axial section of the disc including a calibrated metric scale was also acquired. The digital radiographs and photographs were processed in ImageJ to determine the axial and sagittal plane dimensions for the whole disc (WD) and the nucleus pulposus (NP) and the mid-sagittal disc height for all the lumbar levels. Our results suggest that the L6–S1 IVD in kangaroos is distinctly large compared with the upper lumbar IVDs. Based on previously published data, human lumbar IVDs are the largest of all the animal IVDs used in spine research, with camelid cervical IVDs being the closest relative in absolute dimensions (llamas: 78% in disc height, 40% in WD volume, and 38% in NP volume). Kangaroo L6–S1 IVD was approximately 51% in height, 20% in WD volume, and 20% in NP volume of the human lumbar IVD. We conclude that morphological similarities exist between a kangaroo and human lumbar IVD, especially with the lima bean shape in the axial plane, wedge shape in the sagittal plane, convexity at the cephalad endplates, and percentage volume occupied by the NP in the IVD. These slides can be retrieved under Electronic Supplementary Material.     

Histological and reference system for the analysis of mouse intervertebral disc

A new scoring system based on histo‐morphology of mouse intervertebral disc (IVD) was established to assess changes in different mouse models of IVD degeneration and repair. IVDs from mouse strains of different ages, transgenic mice, or models of artificially induced IVD degeneration were assessed. Morphological features consistently observed in normal, and early/later stages of degeneration were categorized into a scoring system focused on nucleus pulposus (NP) and annulus fibrosus (AF) changes. “Normal NP” exhibited a highly cellularized cell mass that decreased with natural ageing and in disc degeneration. “Normal AF” consisted of distinct concentric lamellar structures, which was disrupted in severe degeneration. NP/AF clefts indicated more severe changes. Consistent scores were obtained between experienced and new users. Altogether, our scoring system effectively differentiated IVD changes in various strains of wild‐type and genetically modified mice and in induced models of IVD degeneration, and is applicable from the post‐natal stage to the aged mouse. This scoring tool and reference resource addresses a pressing need in the field for studying IVD changes and cross‐study comparisons in mice, and facilitates a means to normalize mouse IVD assessment between different laboratories. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:233–243, 2018.     

聚乙烯醇水凝胶人工髓核置入对腰椎运动影响的生物力学研究

目的：研究髓核摘除后聚乙烯醇水凝胶人工髓核置入对腰椎活动度和椎间隙高度的影响.方法：对7具新鲜成人尸体L4/5正常椎间隙、髓核摘除后和置入人工髓核后在8.0Nm扭矩下的屈伸、侧弯和旋转的活动度（ROM）、中性区（NZ）和椎间隙高度变化进行测试、比较.结果：髓核摘除后,L4/5椎间屈伸、侧弯、旋转的ROM和NZ较正常组显著增加（P＜0.05或0.01）,在0和200N的负荷下椎间隙高度较相同情况下正常组分别下降1.2mm和1.7mm;置入人工髓核后,相对于正常椎间隙,L4/5椎间屈伸、侧弯、旋转的ROM和NZ无明显差异,较髓核摘除组明显下降（P＜0.05或0.01）,在0N和200N的负荷下椎间隙高度较相同情况下髓核摘除组分别增加1.6mm和2.0mm.结论：聚乙烯醇水凝胶人工髓核置入椎间盘切除后的椎间隙可有效恢复椎间隙高度,维持腰椎节段正常的三维运动功能稳定性.     

Differential expression of galectin‐1 and its interactions with cells and laminins in the intervertebral disc

Galectin‐1 (Gal‐1), an endogenous β‐galactoside‐binding protein, binds to laminins, which are highly expressed in the nucleus pulposus (NP) of the intervertebral disc (IVD). The objective of this study is to evaluate the expression of Gal‐1 protein in IVD tissues during aging and the effect of Gal‐1 on IVD cell adhesion to laminins. Tissues from rat, porcine, and human (scoliosis or disc degeneration) IVDs were used to evaluate Gal‐1 expression via immunostaining, RT‐PCR, and Western blot analysis. Attachment of isolated IVD cells (porcine and human) on select laminin isoforms (LM‐111 and LM‐511) was compared with/without pre‐incubation with exogenous Gal‐1. A biotinylated Gal‐1(B‐Gal‐1) was used to evaluate for binding to IVD cells and to select for IVD cells by magnetic activated cell sorting (MACS). NP cells expressed high levels of Gal‐1 protein as compared to anulus fibrosus (AF) cells in immature tissues, while exogenous Gal‐1 increased both NP and AF cell attachment to laminins and exhibited a similar binding to both cell types in vitro. With aging, Gal‐1 levels in NP tissue appeared to decrease. In addition, incubation with B‐Gal‐1 was able to promote the retention of more than 50% of IVD cells via MACS. Our results provide new findings for the presence and functional role of Gal‐1 within IVDs. Similar staining patterns for Gal‐1 and LM‐511 in IVD tissue suggest that Gal‐1 may serve as an adhesion molecule to interact with both cells and laminins. This MACS protocol may be useful for selecting pure IVD cells from mixed cells of pathological tissue. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:1923–1931, 2012     

Poroelastic dynamic structural models of rhesus spinal motion segments

Finite element models (FEMs) and analytical and experimental models based on poroelastic constitutive laws were developed for rhesus spinal motion segments (SMSs). Long-time creep, transient creep, and impact were studied for SMSs with normal and simulated degenerated discs. The results suggested that long-time creep observed in excised SMSs may be reduced in the in vivo SMS. The fluid phase included in these FEMs was shown to play a significant role in the mechanical response of SMSs. Relative fluid motion fields predicted in the SMS could be related to nutritional paths to the avascular interior of the disc and were found to be very sensitive to changes in discal stiffness. Reduced disc height, increased discal bulge, altered fluid motion, and stresses were quantified and may be related to mechanical failure, disc degeneration, and low-back pain.