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Functional probe for annulus fibrosus-targeted intervertebral disc degeneration imaging.
Kim HY, Mcclincy M, Vo NV, Sowa GA, Kang JD, Bai M
(2013) J Biomed Opt 18: 101308
MeSH Terms: Animals, Cell Survival, Cells, Cultured, Fluorescent Dyes, Intervertebral Disc, Intervertebral Disc Degeneration, Microscopy, Fluorescence, Molecular Imaging, Optical Imaging, Rabbits
Show Abstract · Added April 2, 2019
Intervertebral disc degeneration (IDD) is closely associated with low back pain. Typically nonsurgical treatment of IDD is the most effective when detected early. As such, establishing reliable imaging methods for the early diagnosis of disc degeneration is critical. The cellular and tissue localization of a facile functional fluorescent probe, HYK52, that labels disc annulus fibrosus is reported. HYK52 was synthesized with high yield and purity via a two-step chemical reaction. Rabbit disc cell studies and ex vivo tissue staining images indicated intracellular localization and intervertebral disc (IVD) tissue binding of HYK52 with negligible cytotoxicity. Moreover, HYK52 is purposefully designed with a functional terminal carboxyl group to allow for coupling with various signaling molecules for multimodal imaging applications. These results suggest that this IVD-targeted probe may have great potential in early diagnosis of IDD.
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MeSH Terms
Cumulative multiple freeze-thaw cycles and testing does not affect subsequent within-day variation in intervertebral flexibility of human cadaveric lumbosacral spine.
Tan JS, Uppuganti S
(2012) Spine (Phila Pa 1976) 37: E1238-42
MeSH Terms: Aged, Biomechanical Phenomena, Cadaver, Cryopreservation, Female, Freezing, Humans, Intervertebral Disc, Lumbar Vertebrae, Pliability, Range of Motion, Articular, Sacrum, Time Factors
Show Abstract · Added October 31, 2013
STUDY DESIGN - An in vitro biomechanical study on 3-dimensional flexibility of human lumbosacral motion segments after multiple freeze-thaw cycles and cumulative testing.
OBJECTIVE - To determine the significance of multiple freeze-thaw cycles and extended testing duration on between-day and within-day variations in motion segment flexibility.
SUMMARY OF BACKGROUND DATA - Previous studies have found no significant effect of single freeze-thaw cycle on creep behavior of human spinal motion segments. Up to 3 freeze-thaw cycles were found to not affect flexibility of porcine spines and viscoelastic properties of human tendons, but more than 5 freeze-thaw cycles resulted in declined structural properties of human tendons.
METHODS - Three lumbosacral motion segments were subjected to repeated flexibility tests to determine both the effects of within-day ambient exposure and between-day multiple freeze-thaw cycles on range of motion (ROM) and neutral zone (NZ). Repeated measures analysis of variance was carried out to evaluate within-day and between-day effects at α = .05.
RESULTS - Significant between-day effects were found for intervertebral ROM and NZ in flexion-extension (FE), lateral bending (LB), and axial rotation (AR) (all P < 0.001). Post hoc analysis indicated that significant differences from test day 1 become apparent after 4 freeze-thaw cycles with cumulative testing. There were no within-day variations from repeated testing on intervertebral FE ROM (P = 0.10), LB ROM (P = 0.36), AR ROM (P = 0.46), FE NZ (P = 0.83), LB NZ (P = 0.42), and AR NZ (P = 0.72).
CONCLUSION - The flexibility of the human cadaveric lumbosacral motion segments between test days was significantly affected after repeated freeze-thaw and cumulative testing cycles. Multiple freeze-thaw cycles and cumulative testing, however, does not affect flexibility data for tests carried out within a single test day.
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13 MeSH Terms
Mice lacking Nf1 in osteochondroprogenitor cells display skeletal dysplasia similar to patients with neurofibromatosis type I.
Wang W, Nyman JS, Ono K, Stevenson DA, Yang X, Elefteriou F
(2011) Hum Mol Genet 20: 3910-24
MeSH Terms: Animals, Bone Diseases, Developmental, Bone Remodeling, Bone and Bones, Chondrocytes, Collagen Type II, Enzyme Activation, Extracellular Signal-Regulated MAP Kinases, Female, Intervertebral Disc, Lovastatin, Mice, Mice, Knockout, Neurofibromatosis 1, Neurofibromin 1, Osteoblasts, Osteocytes, Osteogenesis, Phenotype, Porosity, Promoter Regions, Genetic, Recombination, Genetic, Stem Cells, ras Proteins
Show Abstract · Added November 14, 2013
Mutations in NF1 cause neurofibromatosis type I (NF1), a disorder characterized, among other clinical manifestations, by generalized and focal bony lesions. Dystrophic scoliosis and tibial pseudoarthrosis are the most severe skeletal manifestations for which treatment is not satisfactory, emphasizing the dearth of knowledge related to the biology of NF1 in bone cells. Using reporter mice, we report here that the mouse Col2α1-Cre promoter (collagen, type II, alpha 1) is active not only in chondrocytes but also in adult bone marrow osteoprogenitors giving rise to osteoblasts. Based on this finding, we crossed the Col2α1-Cre transgenic and Nf1(flox/flox) mice to determine whether loss of Nf1 in axial and appendicular osteochondroprogenitors recapitulates the skeletal abnormalities of NF1 patients. By microtomographic and X-rays studies, we show that Nf1(Col2)(-/-) mice display progressive scoliosis and kyphosis, tibial bowing and abnormalities in skull and anterior chest wall formation. These defects were accompanied by a low bone mass phenotype, high bone cortical porosity, osteoidosis, increased osteoclastogenesis and decreased osteoblast number, as quantified by histomorphometry and 3D-microtomography. Loss of Nf1 in osteochondroprogenitors also caused severe short stature and intervertebral disc defects. Blockade of the RAS/ERK activation characteristic of Nf1(-/-) osteoprogenitors by lovastatin during embryonic development could attenuate the increased cortical porosity observed in mutant pups. These data and the skeletal similarities between this mouse model and NF1 patients thus suggest that activation of the RAS/ERK pathway by Nf1 loss-of-function in osteochondroprogenitors is responsible for the vertebral and tibia lesions in NF1 patients, and that this molecular signature may represent a good therapeutic target.
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24 MeSH Terms
Frequency response of pig intervertebral disc cells subjected to dynamic hydrostatic pressure.
Kasra M, Merryman WD, Loveless KN, Goel VK, Martin JD, Buckwalter JA
(2006) J Orthop Res 24: 1967-73
MeSH Terms: Animals, Cell Culture Techniques, Cells, Cultured, Extracellular Matrix Proteins, Hydrostatic Pressure, Intervertebral Disc, Spinal Diseases, Swine, Vibration
Show Abstract · Added February 12, 2015
The pathogenesis of vibration-induced disorders of intervertebral disc at the cellular level is largely unknown. Dynamic loads with frequencies close to that of the in vivo human spine resonant frequency (4-6 Hz) have a destructive effect, which may induce extracellular disc matrix (ECM) degradation. To investigate this issue, three-dimensional (3D) alginate cultures of normal pig intervertebral disc nucleus and inner annulus cells were tested under dynamic hydrostatic loading. Alginate cultures of each region were divided into six groups; five groups were exposed to cyclic hydrostatic pressures of frequencies 1, 3, 5, 8, and 10 Hz with the same amplitude (1 MPa), and group 6 was the control group (no loading). Cultures of different groups were loaded for 3 days (30 min daily) in a hydraulic chamber. Effects of loading frequency on disc collagen and protein metabolism were investigated by measuring 3H-proline-labeled proteins associated with the cells in the extracellular matrix and release of 3H-proline-labeled molecules into culture medium. The results indicated a poor synthesis rate and more degradation near the 5 Hz frequency. The repeatability of experiments was verified by performing two experiments with the same protocol. Both experiments indicated that a threshold frequency of around 5 Hz disrupted protein metabolism.
Copyright (c) 2006 Orthopaedic Research Society.
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9 MeSH Terms
Matrix metalloproteinase-7-dependent release of tumor necrosis factor-alpha in a model of herniated disc resorption.
Haro H, Crawford HC, Fingleton B, Shinomiya K, Spengler DM, Matrisian LM
(2000) J Clin Invest 105: 143-50
MeSH Terms: Animals, Bone Resorption, Cell Migration Inhibition, Cells, Cultured, Coculture Techniques, Diffusion Chambers, Culture, Disease Models, Animal, Enzyme Induction, Intervertebral Disc, Intervertebral Disc Displacement, Macrophages, Peritoneal, Matrix Metalloproteinase 3, Matrix Metalloproteinase 7, Mice, Mice, Inbred Strains, Mice, Knockout, Tumor Necrosis Factor-alpha
Show Abstract · Added March 5, 2014
Herniated disc (HD), one of the major causes of low back pain, is often resolved spontaneously without surgical intervention. Resorption is associated with a marked increase in infiltrating macrophages, and the matrix metalloproteinases (MMP) MMP-3 and MMP-7 have been implicated in this phenomenon. We developed a murine organ culture model in which intact intervertebral discs were cocultured with peritoneal macrophages to investigate the role of MMPs in HD resorption. Using macrophages isolated from MMP-null mice, we report that macrophage-produced MMP-7 was required for proteoglycan degradation, loss of wet weight, and macrophage infiltration of cocultured discs. The inability of MMP-7-deficient macrophages to infiltrate discs could not be attributed to a defect in macrophage migration. MMP-7 was required for the release of the cytokine TNF-alpha from peritoneal macrophages. The generation of soluble TNF-alpha was essential for the induction of MMP-3 in disc cocultures, which in turn is required for the generation of a macrophage chemoattractant and subsequent macrophage infiltration. TNF-alpha release from macrophages was necessary but insufficient for disc resorption, which required macrophage infiltration. We conclude that there is extensive communication between macrophages and chondrocytes in HD resorption and that an essential component of this communication is the requirement for MMPs to release soluble bioactive factors.
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17 MeSH Terms
Matrix metalloproteinase-3-dependent generation of a macrophage chemoattractant in a model of herniated disc resorption.
Haro H, Crawford HC, Fingleton B, MacDougall JR, Shinomiya K, Spengler DM, Matrisian LM
(2000) J Clin Invest 105: 133-41
MeSH Terms: Animals, Blotting, Western, Cell Migration Inhibition, Chondrocytes, Coculture Techniques, Culture Media, Conditioned, Diffusion Chambers, Culture, Disease Models, Animal, Dose-Response Relationship, Drug, Intervertebral Disc, Intervertebral Disc Displacement, Macrophages, Peritoneal, Matrix Metalloproteinase 3, Matrix Metalloproteinase 7, Mice, Mice, Inbred Strains, Mice, Knockout, Organ Culture Techniques, RNA, Messenger, Rats, Reverse Transcriptase Polymerase Chain Reaction, Tumor Necrosis Factor-alpha, Up-Regulation
Show Abstract · Added March 5, 2014
Herniated disc (HD) is a common health problem that is resolved by surgery unless spontaneous resorption occurs. HD tissue contains abundant macrophage infiltration and high levels of matrix metalloproteinases (MMPs) MMP-3 and MMP-7. We developed a model system in which disc tissue or isolated chondrocytes from wild-type or MMP-null mice were cocultured with peritoneal macrophages and used this system to investigate the role of MMPs and chondrocyte/macrophage interactions in disc resorption. We observed a marked enhancement of MMP-3 protein and mRNA in chondrocytes after exposure to macrophages. Chondrocytic MMP-3, but not MMP-7, was required for disc resorption, as determined by assaying for a reduction in wet weight and proteoglycan content after 3 days of coculture. Surprisingly, chondrocyte MMP-3 was required for the generation of a macrophage chemoattractant and the subsequent infiltration of the disc tissue by proteolytically active macrophages. We conclude that macrophage induction of chondrocyte MMP-3 plays a major role in disc resorption by mechanisms that include the generation of a bioactive macrophage chemoattractant.
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23 MeSH Terms