Cartilage tissues regulate systemic aging via ectonucleotide pyrophosphatase/phosphodiesterase 1 in mice.

Aging presents fundamental health concerns worldwide; however, mechanisms underlying how aging is regulated are not fully understood. Here, we show that cartilage regulates aging by controlling phosphate metabolism via ectonucleotide pyrophosphatase/phosphodiesterase 1 (Enpp1). We newly established an Enpp1 reporter mouse, in which an EGFP-luciferase sequence was knocked-in at the Enpp1 gene start codon (Enpp1/EGFP-luciferase), enabling detection of Enpp1 expression in cartilage tissues of resultant mice. We then established a cartilage-specific Enpp1 conditional knockout mouse (Enpp1 cKO) by generating Enpp1 flox mice and crossing them with cartilage-specific type 2 collagen Cre mice. Relative to WT controls, Enpp1 cKO mice exhibited phenotypes resembling human aging, such as short life span, ectopic calcifications, and osteoporosis, as well as significantly lower serum pyrophosphate levels. We also observed significant weight loss and worsening of osteoporosis in Enpp1 cKO mice under phosphate overload conditions, similar to global Enpp1-deficient mice. Aging phenotypes seen in Enpp1 cKO mice under phosphate overload conditions were rescued by a low vitamin D diet, even under high phosphate conditions. These findings suggest overall that cartilage tissue plays an important role in regulating systemic aging via Enpp1.

Transthyretin amyloid deposition in ligamentum flavum (LF) is significantly correlated with LF and epidural fat hypertrophy in patients with lumbar spinal stenosis.

Lumbar spinal stenosis (LSS) is a degenerative disease characterized by intermittent claudication and numbness in the lower extremities. These symptoms are caused by the compression of nerve tissue in the lumbar spinal canal. Ligamentum flavum (LF) hypertrophy and spinal epidural lipomatosis in the spinal canal are known to contribute to stenosis of the spinal canal: however, detailed mechanisms underlying LSS are still not fully understood. Here, we show that surgically harvested LFs from LSS patients exhibited significantly increased thickness when transthyretin (TTR), the protein responsible for amyloidosis, was deposited in LFs, compared to those without TTR deposition. Multiple regression analysis, which considered age and BMI, revealed a significant association between LF hypertrophy and TTR deposition in LFs. Moreover, TTR deposition in LF was also significantly correlated with epidural fat (EF) thickness based on multiple regression analyses. Mesenchymal cell differentiation into adipocytes was significantly stimulated by TTR in vitro. These results suggest that TTR deposition in LFs is significantly associated with increased LF hypertrophy and EF thickness, and that TTR promotes adipogenesis of mesenchymal cells. Therapeutic agents to prevent TTR deposition in tissues are currently available or under development, and targeting TTR could be a potential therapeutic approach to inhibit LSS development and progression.

Angiopoietin-Like Protein 2 Induces Synovial Inflammation in the Facet Joint Leading to Degenerative Changes via Interleukin-6 Secretion.

STUDY DESIGN: Experimental human study. PURPOSE: To determine whether angiopoietin-like protein 2 (ANGPTL2) is highly expressed in the hyperplastic facet joint (FJ) synovium and whether it activates interleukin-6 (IL-6) secretion in FJ synoviocytes. OVERVIEW OF LITERATURE: Mechanical stress-induced synovitis is partially, but significantly, responsible for degenerative and subsequently osteoarthritic changes in the FJ tissues in patients with lumbar spinal stenosis (LSS). However, the underlying molecular mechanism remains unclear. IL-6 is highly expressed in degenerative FJ synovial tissue and is responsible for local chronic inflammation. ANGPTL2, an inflammatory and mechanically induced mediator, promotes the expression of IL-6 in many cells. METHODS: FJ tissues were harvested from five patients who had undergone lumbar surgery. Immunohistochemistry for ANGPTL2, IL-6, and cell markers was performed in the FJ tissue samples. After cultured synoviocytes from the FJ tissues were subjected to mechanical stress, ANGPTL2 expression and secretion were measured quantitatively using real-time quantitative reverse-transcription-polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA), respectively. Following ANGPTL2 administration in the FJ synoviocytes, anti-nuclear factor-κB (NF-κB) activation was investigated using immunocytochemistry, and IL-6 expression and secretion were assayed quantitatively with or without NF-κB inhibitor. Moreover, we assessed whether ANGPTL2-induced IL-6 modulates leucocyte recruitment in the degenerative process by focusing on the monocyte chemoattractant protein-1 (MCP-1) expression. RESULTS: ANGPTL2 and IL-6 were highly expressed in the hyperplastic FJ synovium samples. ANGPTL2 was co-expressed in both, fibroblast-like and macrophage-like synoviocytes. Further, the expression and secretion of ANGPTL2 in the FJ synoviocytes increased in response to stimulation by mechanical stretching. ANGPTL2 protein promoted the nuclear translocation of NF-κB and induced IL-6 expression and secretion in the FJ synoviocytes. This effect was reversed following treatment with NF-κB inhibitor. Furthermore, ANGPTL2-induced IL-6 upregulated the MCP-1 expression in the FJ synoviocytes. CONCLUSIONS: Mechanical stress-induced ANGPTL2 promotes chronic inflammation in the FJ synovium by activating IL-6 secretion, leading to FJ degeneration and subsequent LSS.

Matrix metalloproteinase promotes elastic fiber degradation in ligamentum flavum degeneration.

Ligamentum flavum (LF) hypertrophy in lumbar spinal canal stenosis (LSCS) is characterized by a loss of elastic fibers and fibrosis. Chronic inflammation is thought to be responsible for the histological change but the mechanism underlying elastic fiber degradation remains unclear. Given that matrix metalloproteinase (MMP)-2 and -9 have elastolytic activity and are partly regulated by inflammatory cytokines such as interleukin (IL)-6, in this study, we investigated whether MMPs mediate LF degeneration using 52 LF samples obtained during lumbar surgery, including 31 LSCS and 21 control specimens. We confirmed by histological analysis that the LSCS samples exhibited severe degenerative changes compared with the controls. We found that MMP-2 was upregulated in LF tissue from patients with LSCS at the mRNA and protein levels, whereas MMP-9 expression did not differ between the two groups. The MMP-2 level was positively correlated with LF thickness and negatively correlated with the area occupied by elastic fibers. IL-6 mRNA expression was also increased in LF tissue from patients with LSCS and positively correlated with that of MMP-2. Signal transducer and activator of transcription (STAT)3, a component of the IL-6 signaling pathway, was activated in hypertrophied LF tissues. Our in vitro experiments using fibroblasts from LF tissue revealed that IL-6 increased MMP-2 expression, secretion, and activation via induction of STAT3 signaling, and this effect was reversed by STAT3 inhibitor treatment. Moreover, elastin degradation was promoted by IL-6 stimulation in LF fibroblast culture medium. These results indicate that MMP-2 induction by IL-6/STAT3 signaling in LF fibroblasts can degrade elastic fibers, leading to LF degeneration in LSCS.

TGF-ß1 Improves Biomechanical Strength by Extracellular Matrix Accumulation Without Increasing the Number of Tenogenic Lineage Cells in a Rat Rotator Cuff Repair Model.

BACKGROUND: Transforming growth factor ß1 (TGF-ß1) positively regulates the tenogenic marker genes scleraxis ( Scx) and tenomodulin ( Tnmd) in mesenchymal progenitors in vitro. However, little is known about the effect of TGF-ß1 on the expression of tenogenic markers during rotator cuff (RC) healing in rats. HYPOTHESIS: TGF-ß1 improves the biomechanical properties and histological maturity of reparative tissue in a rat RC repair model by stimulating the growth of tenogenic cells. STUDY DESIGN: Controlled laboratory study. METHODS: Adult male Sprague-Dawley rats (N = 180) underwent unilateral supraspinatus tendon-to-bone surgical repair and were randomly treated with a gelatin hydrogel presoaked in TGF-ß1 (100 ng) or phosphate-buffered saline. The effects of TGF-ß1 on RC healing were investigated at 2, 4, 6, 8, and 12 weeks postoperatively by immunostaining for proliferating cell nuclear antigen, by real-time reverse transcription polymerase chain reaction and in situ hybridization or immunostaining for enthesis-related markers (SRY-box containing gene 9 [ Sox9], Scx, and Tnmd), and by real-time reverse transcription polymerase chain reaction and immunostaining for type I and III collagen. At 6 and 12 weeks postoperatively, biomechanical testing, micro-computed tomography, and biochemical analysis were also performed. At 2 and 4 weeks postoperatively, mesenchymal stem cell-related markers, phospho-Smad2, and matrix metalloproteinase 9 (MMP-9) and MMP-13 were assessed by immunostaining. RESULTS: The TGF-ß1-treated group had significantly higher ultimate load to failure and tissue volume at 6 and 12 weeks postoperatively and a higher collagen content at 12 weeks compared with the saline group. Tendon-related gene expression, histological maturity, cell proliferation, and mesenchymal stem cell-related marker immunoreactivity were not affected by exogenously administrated TGF-ß1 at all time points. In the TGF-ß1-treated group, the percentage of phospho-Smad2-positive cells within the healing tissue increased, whereas the expression of MMP-9 and MMP-13 significantly decreased at 2 and 4 weeks postoperatively. CONCLUSION: TGF-ß1 enhances formation of tough fibrous tissues at the healing site by inhibiting MMP-9 and MMP-13 expression to increase collagen accumulation but without the growth of tenogenic lineage cells. CLINICAL RELEVANCE: These findings suggest that TGF-ß1 could be used for enhancing biomechanical strength after RC surgical repair.

FGF-2 Stimulates the Growth of Tenogenic Progenitor Cells to Facilitate the Generation of Tenomodulin-Positive Tenocytes in a Rat Rotator Cuff Healing Model.

BACKGROUND: Fibroblast growth factor (FGF)-2 has the potential to enhance tendon-to-bone healing after rotator cuff (RC) injury. HYPOTHESIS: FGF-2 stimulates tenogenic differentiation of progenitors to improve the biomechanical strength and histological appearance of repaired RCs in rats. STUDY DESIGN: Controlled laboratory study. METHODS: Adult male Sprague-Dawley rats (N = 156) underwent unilateral surgery to repair the supraspinatus tendon to insertion sites. The FGF-2-treated group (gelatin hydrogel containing 5 µg of FGF-2) and a control group (gelatin hydrogel only) were compared to investigate the effects of FGF-2 at 2, 4, 6, 8, and 12 weeks postoperatively. Biomechanical testing was performed at 6 and 12 weeks. Semiquantitative histological analysis and immunohistochemical analysis for the proliferating cell nuclear antigen (PCNA) were performed, and the expression of tendon-related markers, including Scleraxis (Scx) and Tenomodulin (Tnmd), was monitored by real-time reverse transcription polymerase chain reaction (RT-PCR) and in situ hybridization. SRY-box containing gene 9 (Sox9) expression was monitored by RT-PCR and immunohistochemical analysis. At 2 and 4 weeks, immunohistochemical analysis for mesenchymal stem cell (MSC) markers was also performed. RESULTS: The FGF-2-treated group demonstrated a significant improvement in mechanical strength at 6 and 12 weeks and significantly higher histological scores than the control group at ≥4 weeks. The average incidence of PCNA-positive cells was significantly higher at 2 and 4 weeks, and more cells expressing MSC markers were detected at the insertion site in the FGF-2-treated group. The expression level of Scx increased significantly in the FGF-2-treated group from 4 to 8 weeks, while the Tnmd level increased significantly from 4 to 12 weeks postoperatively. The localization of Tnmd overlapped with the locations of reparative tissues accompanying collagen fibers with an aligned orientation. Sox9 expression was significantly upregulated at 4 weeks in the FGF-2-treated group. CONCLUSION: FGF-2 promotes growth of the tenogenic progenitor cells, which participate in tendon-to-bone healing, resulting in biomechanical and histological improvement of the repaired RC. CLINICAL RELEVANCE: These findings provide clues regarding the clinical development of regenerative repair strategies for RC injury.

Functional and structural comparisons of the arthroscopic knotless double-row suture bridge and single-row repair for anterosuperior rotator cuff tears.

BACKGROUND: We compared the outcomes of knotless double-row suture bridge and single-row repairs in patients undergoing arthroscopic repair for anterosuperior rotator cuff tears. METHODS: We included 61 full-thickness anterosuperior rotator cuff tears treated by arthroscopic repair, namely, single-row repair (group 1: 25 shoulders; mean patient age, 64 years) and the knotless double-row suture bridge repair (group 2: 36 shoulders; mean patient age, 62 years). Preoperative and postoperative magnetic resonance imaging was performed for all shoulders. Clinical outcomes were evaluated for mean follow-up periods of 81 months (range, 72-96 months) in group 1 and 34 months (range, 24-42 months) in group 2, using the University of California, Los Angeles and Japanese Orthopaedic Association assessments. RESULTS: At the final follow-up, both groups showed improvement in the average University of California, Los Angeles and Japanese Orthopaedic Association scores and range of motion, although no intergroup differences were observed. Both groups showed improved abduction strength, and the average score was higher in group 2 (P = .0112). The lift-off and belly-press test results were improved in both groups. Postoperatively, the incidence of positive lift-off tests tended to be lower (P = .075) and that of positive belly-press tests was lower in group 2, P = .049). The repair failure rate tended to be lower in group 2 (14% [5 of 36]) than in group 1 (32% [8 of 25]; P = .0839). CONCLUSIONS: Arthroscopic knotless double-row suture bridge repair of anterosuperior rotator cuff tears yielded functional outcomes equivalent to those of single-row repair and may be useful for improving subscapularis function, abduction strength, and tendon healing.

Patient outcomes for a minimally invasive approach to treat lumbar spinal canal stenosis: is microendoscopic or microscopic decompressive laminotomy the less invasive surgery?

OBJECTIVE: We performed a study to compare the severity of surgical stress between microscopic and microendoscopic decompressive laminotomy performed via a unilateral approach in patients with lumbar spinal canal stenosis (LSCS). MATERIALS AND METHODS: A total of 41 patients received decompressive laminotomy for lumbar spinal stenosis. Twenty patients received microscopic decompressive laminotomy (MDL), and 21 patients received microendoscopic decompressive laminotomy (MEDL). The pre- and postoperative Japanese Orthopaedic Association (JOA), and Visual Analogue Scale (VAS) lower leg pain scores were evaluated. The other variables studied were the length of the operation, blood loss, length of hospital stay, the reaction of the CRP and WBC levels, the dosage of non-steroidal anti-inflammatory drugs (NSAIDs) used and surgical complications. RESULTS: The clinical analyses of the surgical outcomes were evaluated after a minimum two-year follow-up. The pre-and postoperative JOA scores and VAS in the MDL and MEDL groups were statistically similar. There were statistically significant differences found between the lengths of the operation time, blood loss, length of hospital stay, the reaction of CRP, and the pain indicated by the dosage of NSAIDs. The length of the operation time was longer in the MEDL group, and the other variables were greater in the MDL group. CONCLUSIONS: The MEDL procedure is less invasive and safer than the MDL procedure. Hence, MEDL is an effective technique for treating symptomatic LSCS patients.

Evaluation of the relationship between T1ρ and T2 values and patella cartilage degeneration in patients of the same age group.

OBJECTIVE: The aim of this study was to investigate the association between the T1ρ and T2 values and the progression of cartilage degeneration in patients of the same age group. MATERIALS AND METHODS: Sagittal T1ρ and T2 mapping and three-dimensional (3D) gradient-echo images were obtained from 78 subjects with medial knee osteoarthritis (OA). The degree of patella cartilage degeneration was classified into four groups using MRI-based grading: apparently normal cartilage, mild OA, moderate OA, and severe OA group. We measured the T1ρ and T2 values (ms) in the regions of interest set on the full-thickness patella cartilage. Then, we analyzed the relationship between the T1ρ and T2 values and the degree of patella cartilage degeneration. RESULTS: There were no significant differences in age among the four groups. Both the T1ρ and T2 values showed a positive correlation with the degree of OA progression (ρ=0.737 and ρ=0.632, respectively). By comparison between the apparently normal cartilage and the mild OA groups, there were significant differences in the T1ρ mapping, but not in the T2 mapping. CONCLUSIONS: Our study confirmed that T1ρ and T2 mapping can quantitatively evaluate the degree of patella cartilage degeneration in patients within the same age group.

Validity of E-PASS System for Postoperative Morbidity of Spinal Surgery.

STUDY DESIGN: A single-center retrospective cohort study. OBJECTIVE: To evaluate the ability of the Estimation of Physiologic Ability and Surgical Stress (E-PASS) system to predict postoperative risk in patients scheduled for spinal surgery. SUMMARY OF BACKGROUND DATA: The E-PASS system is a surgical audit to predict postoperative morbidity and mortality in general surgery. It is currently not applied in patients with spinal disorders. METHODS: The E-PASS system is comprised of a preoperative risk score (PRS), a surgical stress score (SSS), and a comprehensive risk score (CRS). The latter reflects both the PRS and SSS. We calculated the E-PASS scores for 275 consecutive patients who underwent spinal surgery and evaluated the relationship between the incidence of postoperative complications and each score of the E-PASS system and their ability to predict postoperative morbidity. RESULTS: Postoperative complications developed in 31 patients (11.3%). All E-PASS scores were significantly higher in patients with postoperative complications and they were linearly correlated with the overall incidence of postoperative complications. In particular, PRS was correlated with complications at nonsurgical sites and SSS with surgical site complications. The area under the receiver operating characteristic curve (AUC) for PRS and SSS was higher in patients with complications at nonsurgical and surgical sites, respectively. The AUC for CRS exhibited good predictive power for both types of complication. CONCLUSIONS: The E-PASS system correctly predicted morbidity. The predictive ability of CRS was good for overall morbidity. The E-PASS system is useful for the accurate prediction of the risk for in-hospital morbidity in individual patients scheduled for spinal surgery.

Effects of the water-holding capability of polyvinyl formal sponges on osteogenic ability in in vivo experiments.

In this study, dextran-coated polyvinyl formal (PVF) sponges with high water-holding capability were developed to increase the osteogenic response in the PVF sponge. The study aimed to estimate the effect of the increased water-holding capability of the sponges on osteogenic capacity at a bone defect site in the rabbit femur epiphysis. Bone formation was evaluated using radiography, microcomputed tomography (CT), and histological analysis at 2, 4, and 6 weeks after implantation. As shown by radiography and micro-CT findings, the dextran-coated PVF sponge without water-holding capability showed little bone formation at all evaluated time points. However, the dextran-coated PVF sponge with high water-holding capability showed increasing bone formation around the implant at 4 and 6 weeks after implantation. Furthermore, as shown by micro-CT quantitative analysis, the grafted PVF sponge with high water-holding capability showed significantly greater values for percentage of bone volume per total volume and mean bone mineral density compared with the grafted PVF sponge without water-holding capability at 4 and 6 weeks after implantation. These results suggest that the dextran-coated PVF sponge with high water-holding capability promoted osteogenesis in vivo. The PVF sponge might be a new biomaterial to be used as a fill material for bone defects.

Angiopoietin-like protein 2 induced by mechanical stress accelerates degeneration and hypertrophy of the ligamentum flavum in lumbar spinal canal stenosis.

Chronic inflammation and subsequent fibrosis induced by mechanical stress play an important role in ligamentum flavum (LF) hypertrophy and degeneration in patients with lumbar spinal canal stenosis (LSCS). Angiopoietin-like protein 2 (Angptl2) is a chronic inflammatory mediator induced under various pathological conditions and increases the expression of TGF-ß1, which is a well-characterized mediator in LF hypertrophy. We investigated whether Angptl2 is induced by mechanical stress, and whether it contributes to LF hypertrophy and degeneration by activating the TGF-ß1 signaling cascade. In this study, we investigated human LF tissue and LF fibroblasts isolated from patients who underwent lumbar surgery. We found that Angptl2 was abundantly expressed in fibroblasts of hypertrophied LF tissues at both the mRNA and protein levels. This expression was not only positively correlated with LF thickness and degeneration but also positively correlated with lumbar segmental motion. Our in vitro experiments with fibroblasts from hypertrophied LF tissue revealed that mechanical stretching stress increases the expression and secretion of Angptl2 via activation of calcineurin/NFAT pathways. In hypertrophied LF tissue, expression of TGF-ß1 mRNA was also increased and TGF-ß1/Smad signaling was activated. Angptl2 expression in LF tissue was positively correlated with the expression of TGF-ß1 mRNA, suggesting cooperation between Angptl2 and TGF-ß1 in the pathogenesis of LF hypertrophy. In vitro experiments revealed that Angptl2 increased levels of TGF-ß1 and its receptors, and also activated TGF-ß1/Smad signaling. Mechanical stretching stress increased TGF-ß1 mRNA expression, which was partially attenuated by treatment with a calcineurin/NFAT inhibitor or Angptl2 siRNA, indicating that induction of TGF-ß1 expression by mechanical stretching stress is partially mediated by Angptl2. We conclude that expression of Angptl2 induced by mechanical stress in LF fibroblasts promotes LF tissue degeneration by activation of TGF-ß1/Smad signaling, which results in LF hypertrophy in patients with LSCS.

Predictive value of E-PASS and POSSUM systems for postoperative risk assessment of spinal surgery.

OBJECT: The Estimation of Physiological Ability and Surgical Stress (E-PASS) and Physiological and Operative Severity Score for the enUmeration of Mortality and Morbidity (POSSUM) systems are surgical risk scoring systems that take into account both the patient's preoperative condition and intraoperative variables. While they predict postoperative morbidity and mortality rates for several types of surgery, spinal surgeries are currently not included. The authors assessed the usefulness of E-PASS and POSSUM algorithms and compared the predictive ability of both systems in patients with spinal disorders considered for surgery. METHODS: The E-PASS system includes a preoperative risk score, a surgical stress score, and a comprehensive risk score that is determined by both the preoperative risk score and surgical stress score. The POSSUM system is composed of a physiological score and an operative severity score; its total score is based on both the physiological score and operative severity score. The authors calculated the E-PASS and POSSUM scores for 601 consecutive patients who had undergone spinal surgery and investigated the relationship between the individual scores of both systems and the incidence of postoperative complications. They also assessed the correctness of the predicted morbidity rate of both systems. RESULTS: Postoperative complications developed in 64 patients (10.6%); there were no in-hospital deaths. All EPASS scores (p ≤ 0.001) and the operative severity score and total score of the POSSUM (p < 0.03) were significantly higher in patients with postoperative complications than in those without postoperative complications. The morbidity rates correlated linearly and significantly with all E-PASS scores (p ≤ 0.001); their coefficients (preoperative risk score, ρ = 0.179; surgical stress score, ρ = 0.131; and comprehensive risk score, ρ = 0.198) were higher than those for the POSSUM scores (physiological score, ρ = 0.059; operative severity score, ρ = 0.111; and total score, ρ = 0.091). The area under the receiver operating characteristic curve for the predicted morbidity rate was 0.668 for the E-PASS and 0.588 for the POSSUM system. CONCLUSIONS: As E-PASS predicted morbidity more correctly than POSSUM, it is useful for estimating the postoperative risk of patients considered for spinal surgery.

Effects of water-holding capability of the PVF sponge on the adhesion and differentiation of rat bone marrow stem cell culture.

The aim of the study is to estimate the effects of the water-holding capability of the polyvinyl formal (PVF) sponges on osteogenic response in vitro experiments. The rat bone marrow stem cells (BMCs) were seeded and cultured for up to 4 weeks under static conditions in osteogenic media to evaluate the adhesion, proliferation, differentiation, and mineralization on the Dextran-coated PVF sponges with or without water-holding capability. The BMCs seeded onto the PVF sponges with water-holding capability showed more significant increases in DNA content, alkaline phosphatase (ALP) activity, osteocalcin content, and calcium deposition than those without water-holding capability. These results suggest that the Dextran-coated PVF sponges with high water-holding capability would have potential uses as both a new scaffold to bone tissue engineering and as a new biomaterial.

Detecting ICRS grade 1 cartilage lesions in anterior cruciate ligament injury using T1ρ and T2 mapping.

OBJECTIVE: The purpose of this study was to clarify the detectability of the International Cartilage Repair Society (ICRS) grade 1 cartilage lesions in anterior cruciate ligament (ACL)-injured knees using T1ρ and T2 mapping. MATERIALS AND METHODS: We performed preoperative T1ρ and T2 mapping and 3D gradient-echo with water-selective excitation (WATS) sequences on 37 subjects with ACL injuries. We determined the detectability on 3D WATS based on arthroscopic findings. The T1ρ and T2 values (ms) were measured in the regions of interest that were placed on the weight-bearing cartilage of the femoral condyle. The receiver operating characteristic (ROC) curve based on these values was constructed using the arthroscopic findings as a reference standard. The evaluation of cartilage was carried out only in the weight-bearing cartilage. The cut-off values for determining the presence of a cartilage injury were determined using each ROC curve, and the detectability was calculated for the T1ρ and T2 mapping. RESULTS: The cut-off values for the T1ρ and T2 were 41.6 and 41.2, respectively. The sensitivity and specificity of T1ρ were 91.2% and 89.5%, respectively, while those of T2 were 76.5% and 81.6%, respectively. For the 3D WATS images, the same values were 58.8% and 78.9%, respectively. CONCLUSIONS: Our study demonstrated that the T1ρ and T2 values were significantly higher for ICRS grade 1 cartilage lesions than for normal cartilage and that the two mappings were able to non-invasively detect ICRS grade 1 cartilage lesions in the ACL-injured knee with a higher detectability than were 3D WATS images.

Intracellular accumulation of advanced glycation end products induces apoptosis via endoplasmic reticulum stress in chondrocytes.

Mammalian cells attempt to maintain their homeostasis under endoplasmic reticulum (ER) stress. If the stress cannot be alleviated, cells are led to apoptosis through induction of C/EBP homologous protein (CHOP). ER stress is provoked in osteoarthritis chondrocytes, and intracellular accumulation of advanced glycation end products (AGEs) in chondrocytes is a possible cause. To clarify the role of intracellular AGE accumulation in chondrocytes, the present study investigated the effect of intracellular AGE accumulation on ER stress and apoptosis by in vitro and in vivo analysis. Intracellular AGE accumulation induced by AGE precursors caused apoptosis, induced expression of ER stress markers, and led to co-localization of AGEs with glucose-regulated protein 78, leading to formation of high-molecular-weight complexes in cultured chondrocytes. These reactions were inhibited by an AGE formation inhibitor. CHOP deletion inhibited apoptosis induced by intracellular AGE accumulation. In vivo intracellular AGE accumulation induced by intra-articular injection of AGE precursors caused ER stress and apoptosis in chondrocytes and led to degradation of articular cartilage. Additionally, intracellular AGE accumulation increased the degree of cartilage degradation in an osteoarthritis model. These data indicate that intracellular accumulation of AGEs induces modification of unfolded protein response-related protein by AGEs and apoptosis via ER stress in chondrocytes. Moreover, the in vivo study showed that intracellular AGE accumulation in chondrocytes is involved in the occurrence and progression of osteoarthritis through ER stress. Thus, research on mechanisms of apoptosis via ER stress induced by intracellular AGE accumulation in chondrocytes will lead to a new understanding of osteoarthritis pathology.

Corresponding scapular pain with the nerve root involved in cervical radiculopathy.

PURPOSE: To correspond scapular pain with the nerve root involved in cervical radiculopathy. METHODS: In the anatomic study, 11 Japanese adult cadavers were dissected to examine the numbers and courses of the cutaneous nerves from C3 to C8 dorsal rami. In the clinical study, 14 men and 11 women aged 34 to 77 years who presented with scapular pain as well as pain, numbness or motor weakness in the upper limbs secondary to cervical radiculopathy were assessed. The involved nerve roots were identified based on the symptoms and signs in the arm and/ or fingers, the radiological diagnosis, and the pain response to cervical nerve root blocks. The sites and characteristics of radicular pain were assessed. RESULTS: In the anatomic study of 22 cutaneous nerves from medial branches of dorsal rami, 18 involved the C5 nerve root, 0 the C6 root, one the C7 root, and 8 the C8 root. In the clinical study, the radicular pain often occurred in the suprascapular region involving the C5 root, in the suprascapular to posterior deltoid region involving the C6 root, in the interscapular region involving the C7 root, and in the interscapular and scapular regions involving the C8 root. All patients with C5 or C8 radiculopathy had both superficial and deep pain, whereas almost all patients with C6 or C7 radiculopathy had deep pain only. No patient had superficial pain only. CONCLUSION: Cervical radiculopathy can cause scapular pain. Pain sites and characteristics are related to the affected nerve root.

Gas-filled intradural cysts of the lumbar spine and the possible pathogenesis.

BACKGROUND CONTEXT: There have only been four reports of gas-filled intradural cysts, and the pathogenesis is unknown. PURPOSE: To document the radiologic and histopathologic features of gas-filled intradural cysts and to discuss the pathogenesis with a review of the literature. STUDY DESIGN: Case report. METHODS: A 67-year-old woman, admitted to our institute, presented with severe right thigh pain. On admission to the institute, enhanced magnetic resonance imaging, showed a cystic lesion in the spinal canal at the L2-L3 level, with an intensity suggesting the presence of gas. An enhanced region around the cyst was noted. Computed tomography after discography also revealed a water-soluble contrast filled the subarachnoid space and area around the cyst, but not inside. RESULTS: The cyst was surgically resected. One of the nerve roots was firmly adherent to the gas-filled cyst. The cyst wall comprised fibrous tissue, including small granulations and herniated disc material. CONCLUSION: Gas-filled intradural cysts are rare. The pathogenesis appears to involve gas in a degenerated intervertebral disc, and spontaneous absorption of herniated disc material.

L5 radiculopathy caused by a ganglion cyst of the posterior longitudinal ligament in a teenager.

BACKGROUND CONTEXT: There is no previous report on the intraspinal ganglion cyst of the posterior longitudinal ligament in a teenager. PURPOSE: To report a case of radiculopathy caused by a ganglion cyst of the posterior longitudinal ligament in a teenager. STUDY DESIGN: Case report. METHODS: A 17-year-old male with a 4-month history of left L5 radicular pain was found to have an intraspinal cystic lesion causing radicular compression. Magnetic resonance imaging showed a cystic lesion located in the ventral side of the dura. The patient suffered from severe leg pain. As a result, a surgical operation was therefore performed. RESULTS: The cyst containing jelly-like components and a hemorrhage was punctured and then extirpated. It originated from the posterior longitudinal ligament. A histological study revealed the cyst to be without any synovial layers. CONCLUSIONS: This is the first report to describe a ganglion cyst originating from the posterior longitudinal ligament in a teenager. This possible etiology should be kept in mind for any other individuals displaying symptoms of spinal nerve root compression as well as disc herniation.

Characterization of an exchangeable gene trap using pU-17 carrying a stop codon-beta geo cassette.

We have developed a new exchangeable gene trap vector, pU-17, carrying the intron-lox71-splicing acceptor (SA)-beta geo-loxP-pA-lox2272-pSP73-lox511. The SA contains three stop codons in-frame with the ATG of beta galactosidase/neomycin-resistance fusion gene (beta geo) that can function in promoter trapping. We found that the trap vector was highly selective for integrations in the introns adjacent to the exon containing the start codon. Furthermore, by using the Cre-mutant lox system, we successfully replaced the beta geo gene with the enhanced green fluorescent protein (EGFP) gene, established mouse lines with the replaced clones, removed the selection marker gene by mating with Flp-deleter mice, and confirmed that the replaced EGFP gene was expressed in the same pattern as the beta geo gene. Thus, using this pU-17 trap vector, we can initially carry out random mutagenesis, and then convert it to a gain-of-function mutation by replacing the beta geo gene with any gene of interest to be expressed under the control of the trapped promoter through Cre-mediated recombination.