Gene expression profile of mouse masseter muscle after repetitive electrical stimulation.

PURPOSE: To examine gene expression profile changes in the mouse masseter muscle tissue after repetitive electrical stimulation by using a DNA microarray technique. METHODS: Nine male ICR mice aged 10 weeks were used. Each anesthetized mouse was secured on a platform in a supine position and the masseter muscle tissues on both sides were exposed. Bipolar electrodes were set on the right masseteric fascia to electrically stimulate the masseter muscle (8 V, 10 Hz, 20 ms) for 30 min. After cessation of stimulation bilateral masseter muscle tissues were sampled at 0 h (n=3), 1h (n=3), 2h (n=3). Total RNA was isolated from the homogenized muscle tissues and purified mRNA samples (50 microg) were processed and hybridized with microarray slides. Probe arrays were then scanned and analyzed to calculate the signal density. Gene expression profiles were compared at each time point between the right (stimulation side) and left (control side) masseter. When the gene expression levels were different more than 2-fold, the difference was regarded as positive. RESULTS: Of the 6400 genes assessed, 1733 genes were up-regulated and 515 genes were down-regulated in the stimulation side at least once during the experimental time course. These up- or down-regulated genes were associated with autoimmune/inflammatory disease (28/114), cardiovascular disease (17/61), neuroscience (12/50), apoptosis (27/93), diabetes/obesity (9/28), signal transduction (66/250) and others. 28 genes were up-regulated and 25 genes were down-regulated at all time points. CONCLUSIONS: Dramatic gene expression changes were induced by the repetitive electrical muscle stimulation in mouse masseter.

Regeneration of defects in articular cartilage in rat knee joints by CCN2 (connective tissue growth factor).

UNLABELLED: CTGF/CCN2, a hypertrophic chondrocyte-specific gene product, possessed the ability to repair damaged articular cartilage in two animal models, which were experimental osteoarthritis and full-thickness defects of articular cartilage. These findings suggest that CTGF/CCN2 may be useful in regeneration of articular cartilage. INTRODUCTION: Connective tissue growth factor (CTGF)/CCN2 is a unique growth factor that stimulates the proliferation and differentiation, but not hypertrophy, of articular chondrocytes in vitro. The objective of this study was to investigate the therapeutic use of CTGF/CCN2. MATERIALS AND METHODS: The effects of recombinant CTGF/CCN2 (rCTGF/CCN2) on repair of damaged cartilage were evaluated by using both the monoiodoacetic acid (MIA)-induced experimental rat osteoarthritis (OA) model and full-thickness defects of rat articular cartilage in vivo. RESULTS: In the MIA-induced OA model, quantitative real-time RT-PCR assays showed a significant increase in the level of CTGF/CCN2 mRNA, and immunohistochemical analysis and in situ hybridization revealed that the clustered chondrocytes, in which clustering indicates an attempt to repair the damaged cartilage, produced CTGF/CCN2. Therefore, CTGF/CCN2 was suspected to play critical roles in cartilage repair. In fact, a single injection of rCTGF/CCN2 incorporated in gelatin hydrogel (rCTGF/CCN2-hydrogel) into the joint cavity of MIA-induced OA model rats repaired their articular cartilage to the extent that it became histologically similar to normal articular cartilage. Next, to examine the effect of rCTGF/CCN2 on the repair of articular cartilage, we created defects (2 mm in diameter) on the surface of articular cartilage in situ and implanted rCTGF/CCN2-hydrogel or PBS-hydrogel therein with collagen sponge. In the group implanted with rCTGF/CCN2-hydrogel collagen, new cartilage filled the defect 4 weeks postoperatively. In contrast, only soft tissue repair occurred when the PBS-hydrogel collagen was implanted. Consistent with these in vivo effects, rCTGF/CCN2 enhanced type II collagen and aggrecan mRNA expression in mouse bone marrow-derived stromal cells and induced chondrogenesis in vitro. CONCLUSION: These findings suggest the utility of CTGF/CCN2 in the regeneration of articular cartilage.

Soluble tumour necrosis factor receptors in synovial fluids from temporomandibular joints with painful anterior disc displacement without reduction and osteoarthritis.

The objective of this study was to detect soluble-form tumour necrosis factor receptors (sTNFRs) in temporomandibular joint (TMJ) synovial fluid aspirates, and to compare the sTNFR concentrations between painful anterior disc displacement without reduction and osteoarthritis (ADDwoR/OA) and asymptomatic TMJs. Synovial fluid was sampled from the superior TMJ cavity of 11 painful ADDwoR/OA cases (mean age: 36.9 years) and 10 asymptomatic females (mean age: 24.7 years) by diluted aspiration. The concentrations of sTNFR-I and -II in the synovial fluid were measured using human sTNFR-I and -II enzyme-linked immunosorbent assays. The total protein concentrations in synovial fluids were measured using a bicinchoninic acid protein assay kit. All data were normalised to the total protein concentration of each sample.Two-way factorial analysis of variance and post hoc multiple comparison revealed that: (1). mean normalised sTNFR-I and -II concentrations were higher in TMJ synovial aspirates from ADDwoR/OA patients than from healthy controls; (2). in the ADDwoR/OA patients and the healthy controls, the sTNFR-I concentration in TMJ synovial aspirates was higher than the sTNFR-II concentration; and (3). high TMJ synovial aspirate sTNFR-II seemed to be associated with less TMJ pain and a less restricted range of mouth opening in the ADDwoR/OA patients. The concentrations of sTNFRs in TMJ synovial fluid are higher in the presence of painful ADDwoR/OA, which could modulate intracapsular inflammation.