Study Design: In vitro experiment using degenerated human ligamentum flavum (LF) and various inflammatory cytokines. Objectives: To examine the effect of inflammatory cytokines on LF cells and to identify their roles in the pathogenesis of LF hypertrophy and ossification. Summary of Background Data: Spinal stenosis is caused, in part, by hypertrophy and ossification of the LF, which are induced by the degenerative processes (ie, increased collagen synthesis and chondroid metaplasia) of ligament fibroblasts. Degenerated intervertebral disk spontaneously produces inflammatory cytokines, which might affect the adjacent LF through local milieu of the spinal canal. Methods: The interlaminar portion of the LF was collected during surgical spinal procedures in 15 patients (age range, 49–78 y) with lumbar spinal stenosis. LF fibroblasts were isolated by enzymatic digestion of LF tissue. LF cell cultures were treated with various inflammatory cytokines: interleukin (IL)-1α, IL-6, tumor necrosis factor-α (TNF-α), prostaglandin E₂ (PGE₂), and nitric oxide (NO). Cytotoxicity was analyzed by MTT assays. DNA synthesis was measured with ³H-thymidine incorporation, and mRNA expression of types I, III, V, and XI collagen and osteocalcin were performed by reverse transcription-polymerase chain reaction. Histochemical stains such as Von Kossa were also performed to detect bone nodule formation. Results: There was no cytotoxicity in the LF cells treated with each cytokine. There were significant increases in DNA synthesis and upregulated mRNA expression of types I, V, XI collagen and osteocalcin in LF cultures treated with various cytokines. LF cultures treated with IL-6, TNF-α, PGE₂, and NO showed positive Von Kossa staining, indicating bone nodule formation from LF cells. Conclusions: Inflammatory cytokines (IL-6, TNF-α, PGE₂, and NO) seem to play a crucial role in hypertrophy and ossification of LF. Degenerated, herniated intervertebral disks, and facet arthrosis may influence LF through inflammatory cytokines and cause hypertrophy and ossification of LF.

Significance: Meniscus damage is a common musculoskeletal injury and a major risk factor for knee osteoarthritis (OA). An important knowledge gap exists about meniscus cell homeostasis mechanisms and how their impairment contributes to meniscus pathology following trauma and in aging. Forkhead Box O (FoxO) transcription factors are essential regulators of cellular homeostasis. We performed analysis of expression patterns and function of FoxO in human and mouse menisci. Our results show that FoxOs are suppressed in degenerated meniscus and that FoxO1 regulates meniscus growth and maturation, while both FoxO1 and 3 have protective functions in meniscus during aging and OA. These discoveries provide insight into mechanisms of meniscus degeneration and targets for therapeutic intervention.

A prospective case control study. To investigate the risk of a fall by using functional mobility tests in patients with lumbar spinal stenosis (LSS) via a comparison with patients with knee osteoarthritis (KOA). LSS is a degenerative arthritic disease in the spine that results in decreasing function, impaired balance, and gait deficit, with increased levels of leg and back pain. This physical impairment may result in an increased risk of fall later in the disease process, as shown in KOA. However, there has been no study regarding the association between the risk of a fall and LSS. The study was an age- and weight-matched case control study consisting of two groups: one group consisting of 40 patients with LSS who were scheduled to undergo spine surgery (LSS group) and the other group consisting of 40 patients with advanced osteoarthritis in both knees, scheduled to undergo TKA on both knees (KOA group). For both groups, four functional mobility tests, such as a Six-Meter-Walk Test (SMT), Sit-to-Stand test (STS), Alternative-Step Test (AST), and Timed Up and Go Test (TUGT), were performed. There was no difference in demographic data between both groups except for body mass index. For the SMT and STS, the patients in the LSS group spent significantly more time performing these tests than the patients in the KOA. For the AST, however, patients in the KOA group presented a statistically worse performance in functional mobility, compared with the LSS group. The mean TUGT time was not statistically different between the two groups. The current study highlights that patients with symptomatic LSS have a risk of a fall comparable with the patients who had degenerative KOA based on the results of functional mobility tests (SMT, STS, AST, and TUGT).

In vitro experiment using rabbit nucleus pulposus (NP) cells seeded in atelocollagen scaffolds under the stimulation of growth factors. To demonstrate the effect of anabolic growth factors in rabbit NP cells cultured in atelocollagen type I and type II. Atelocollagen provides intervertebral disc (IVD) cells for a biocompatible environment to produce extracellular matrix. IVD cells with exogenous transforming growth factor-beta 1 (TGF-β1) and bone morphogenetic protein-2 (BMP-2) also render an increase in matrix synthesis. However, the effect of anabolic growth factors in NP cells cultured in atelocollagens was not elucidated before. Rabbit NP cell was harvested, enzymatically digested, and cultured. The NP cells were seeded to atelocollagen type I and type II scaffolds, and then cultures were exposed to TGF-β1 (10 ng/mL) and/or BMP-2 (100 ng/mL). DNA synthesis was measured using [4H]-thymidine incorporation. Newly synthesized proteoglycan was measured using [35S]-sulfate incorporation. Reverse transcription-polymerase chain reactions (RT-PCRs) for mRNA expression of aggrecan, collagen type I, collagen type II, and osteocalcin were performed. Rabbit NP cells cultured in atelocollagen type I scaffold showed an increase (1.7 to 2.4-fold) in DNA synthesis in response to TGF-β1 and/or BMP-2 (P < 0.05), whereas NP cultures in atelocollagen type II demonstrated a 30% increase in DNA synthesis only with combination of both growth factors compared with control (P < 0.05). Rabbit NP cells in atelocollagen type II scaffold with TGF-β1 and combination of both growth factors exhibited robust 5.3- and 5.4-fold increases in proteoglycan synthesis (P < 0.05), whereas any cultures in atelocollagen type I failed to show any significant increase compared with control. Rabbit NP cells in atelocollagen type I and type II scaffolds with TGF-β1 and/or BMP-2 demonstrated the upregulation of aggrecan, collagen type I, and collagen type II mRNA expression compared with saline control (P < 0.05). The response in transcriptional level was more robust in atelocollagen type II than in type I. In any event, there is no recognizable expression of osteocalcin (P < 0.05). NP cells in atelocollagens under the stimulation of TGF-β1 and BMP-2 exhibited anabolic responses in transcriptional and translational levels. Hence, such an approach can provide a suitable engineered tissue for IVD regeneration with potential for robust refurbishment of matrix.