Comparison of 3.0-T MR vs 3.0-T MR arthrography of the hip for detection of acetabular labral tears and chondral defects in the same patient population.

OBJECTIVE: We report our experience in diagnostic sensitivity of 3.0-T conventional MR vs 3.0-T MR arthrography of the hip for detection of acetabular labral tears and chondral defects in the same patient population. METHODS: 43 consecutive patients had both conventional hip MR and MR arthrography examinations performed. These examinations were reviewed retrospectively by independent reading of two musculoskeletal radiologists who read the MR and MR arthrogram examinations in a randomized fashion (i.e. MR and MR arthrogram examinations were read at separate sittings and in a randomized fashion so as not to bias reviewers). Scans were assessed for acetabular labral tears and chondral defects. All patients went on to arthroscopy. RESULTS: Of these 43 patients, 40 had acetabular labral tears read by Reader 1 and 39 had acetabular labral tears read by Reader 2 on MR arthrogram, 39 had acetabular labral tears read by Reader 1 and 38 had acetabular labral tears read by Reader 2 on conventional MR examination. There were 42 labral tears in 43 patients at arthroscopy. There were four false-negative labral tears compared with arthroscopy on MR and three false negatives on MR arthrography for Reader 1 and five false negatives on MR and four false negatives on MR arthrography for Reader 2. Each reader had one false-positive labral tear compared with arthroscopy on both MR and MR arthrography. There were 32 acetabular chondral defects at arthroscopy. Reader 1 saw 21 acetabular chondral defects on conventional MR and 27 chondral defects at MR arthrography. Reader 2 saw 19 acetabular chondral defects at conventional MR and 25 acetabular chondral defects on MR arthrography. There were no false-positive readings of chondral defects compared with arthroscopy on MR and one false positive for Reader 1 and two false positives for Reader 2 on MR arthrography as compared with arthroscopy. On conventional MR examination, sensitivities and specificities as compared with arthroscopy were as follows: Reader 1 acetabular labral tear (90% sensitivity, 0% specificity) and Reader 2 acetabular labral tear (88% sensitivity, 0% sensitivity). On MR arthrogram, sensitivities and specificities as compared with arthroscopy for Reader 1 were 93%, 0% and for Reader 2 were 90%, 0%, respectively. Sensitivities and specificities for detection of acetabular chondral defects as compared with arthroscopy were Reader 1 conventional MR (65% sensitivity, 100% specificity), Reader 1 MR arthrography (81% sensitivity, 91% specificity), Reader 2 conventional MR (59% sensitivity, 100% specificity) and Reader 2 MR arthrography (71% sensitivity, 82% specificity). CONCLUSION: In this series, 3.0-T MR demonstrated sensitivity for detection of acetabular labral tears that rivals the sensitivity of 3.0-T MR arthrography of the hip. In this series, 3.0-T MR arthrography was more sensitive than conventional 3.0-T MR for detection of acetabular chondral defects. ADVANCES IN KNOWLEDGE: 3.0-T MR and MR arthrography are near equivalent in the diagnosis of acetabular labral tears. This information is useful for pre-operative planning.

Culture temperature affects redifferentiation and cartilaginous extracellular matrix formation in dedifferentiated human chondrocytes.

To date, there have been few studies on how temperature affects the phenotype and metabolism of human chondrocytes. Thus, the purpose of this study was to elucidate the effects of culture temperature on chondrocyte redifferentiation and extracellular matrix (ECM) formation using dedifferentiated mature human chondrocytes in vitro. Dedifferentiated chondrocytes were cultured in a pellet culture system for up to 21 days. The pellets were randomly divided into three groups with different culture temperature (32, 37, and 41°C). Chondrocyte redifferentiation and ECM formation were evaluated by wet weight, messenger ribonucleic acid (mRNA), histological, and biochemical analyses. The results showed that the wet weight and the mRNA expressions of collagen type II A1 and cartilage oligomeric matrix protein at 37°C were higher than the corresponding values at 32°C. The histological and biochemical analyses revealed that the syntheses of type II collagen and proteoglycan were promoted at 37°C compared to those at 32°C, whereas they were considerably inhibited at 41°C. In conclusion, the results obtained herein indicated that temperature affects chondrocyte redifferentiation and ECM formation, and modulation of temperature might thus represent an advantageous means to regulate the phenotype and biosynthetic activity of chondrocytes.

Collagen and chondrocyte concentrations control ultrasound scattering in agarose scaffolds.

Ultrasound imaging has been proposed for diagnostics of osteoarthritis and cartilage injuries in vivo. However, the specific contribution of chondrocytes and collagen to ultrasound scattering in articular cartilage has not been systematically studied. We investigated the role of these tissue structures by measuring ultrasound scattering in agarose scaffolds with varying collagen and chondrocyte concentrations. Ultrasound catheters with center frequencies of 9 MHz (7.1-11.0 MHz, -6 dB) and 40 MHz (30.1-45.3 MHz, -6 dB) were applied using an intravascular ultrasound device. Ultrasound backscattering quantified in a region of interest starting right below sample surface differed significantly (p < 0.05) with the concentrations of collagen and chondrocytes. An ultrasound frequency of 40 MHz, as compared with 9 MHz, was more sensitive to variations in collagen and chondrocyte concentrations. The present findings may improve diagnostic interpretation of arthroscopic ultrasound imaging and provide information necessary for development of models describing ultrasound propagation within cartilage.

Destabilization of the medial meniscus leads to subchondral bone defects and site-specific cartilage degeneration in an experimental rat model.

OBJECTIVE: This study aimed to investigate subchondral bone changes using micro-computed tomography (micro-CT) and regional differences in articular cartilage degeneration, focusing on changes of cartilage covered by menisci, in the early phase using a destabilization of the medial meniscus (DMM) model. METHOD: The DMM model was created as an experimental rat osteoarthritis (OA) model (12 weeks old; n = 24). At 1, 2, and 4 weeks after surgery, the rats were sacrificed, and knee joints were scanned using a Micro-CT system. Histological sections of the medial tibial plateau, which was divided into inner, middle, and outer regions, were prepared and scored using the modified OARSI scoring system. The cartilage thickness was also calculated, and matrix metalloproteinase 13 (MMP13), Col2-3/4c, and vascular endothelial growth factor (VEGF) expression was assessed immunohistochemically. RESULTS: Subchondral bone defects were observed in the middle region, in which the cartilage thickness decreased over time after surgery, and these defects were filled with MMP13- and VEGF-expressing fibrous tissue. The OARSI score increased over time in the middle region, and the score was significantly higher in the middle region than in the inner and outer regions at 1, 2, and 4 weeks after surgery. Col2-3/4c and MMP13 expression was observed primarily in the meniscus-covered outer region, in which the cartilage thickness increased over time. CONCLUSION: Loss of meniscal function caused cartilage degeneration and subchondral bone defects in the early phase site-specifically in the middle region. Furthermore, our results might indicate cartilage covered by menisci is easily degraded resulting in osmotic swelling of the cartilage in early OA.

FK506 protects against articular cartilage collagenous extra-cellular matrix degradation.

OBJECTIVE: Osteoarthritis (OA) is a non-rheumatologic joint disease characterized by progressive degeneration of the cartilage extra-cellular matrix (ECM), enhanced subchondral bone remodeling, activation of synovial macrophages and osteophyte growth. Inhibition of calcineurin (Cn) activity through tacrolimus (FK506) in in vitro monolayer chondrocytes exerts positive effects on ECM marker expression. This study therefore investigated the effects of FK506 on anabolic and catabolic markers of osteoarthritic chondrocytes in 2D and 3D in vitro cultures, and its therapeutic effects in an in vivo rat model of OA. METHODS: Effects of high and low doses of FK506 on anabolic (QPCR/histochemistry) and catabolic (QPCR) markers were evaluated in vitro on isolated (2D) and ECM-embedded chondrocytes (explants, 3D pellets). Severe cartilage damage was induced unilaterally in rat knees using papain injections in combination with a moderate running protocol. Twenty rats were treated with FK506 orally and compared to twenty untreated controls. Subchondral cortical and trabecular bone changes (longitudinal microCT) and macrophage activation (SPECT/CT) were measured. Articular cartilage was analyzed ex vivo using contrast enhanced microCT and histology. RESULTS: FK506 treatment of osteoarthritic chondrocytes in vitro induced anabolic (mainly collagens) and reduced catabolic ECM marker expression. In line with this, FK506 treatment clearly protected ECM integrity in vivo by markedly decreasing subchondral sclerosis, less development of subchondral pores, depletion of synovial macrophage activation and lower osteophyte growth. CONCLUSION: FK506 protected cartilage matrix integrity in vitro and in vivo. Additionally, FK506 treatment in vivo reduced OA-like responses in different articular joint tissues and thereby makes Cn an interesting target for therapeutic intervention of OA.

Bone marrow stimulation induces greater chondrogenesis in trochlear vs condylar cartilage defects in skeletally mature rabbits.

OBJECTIVE: The aim of this study was to compare the early repair response of cartilage defects in trochlea (TR) and medial femoral condyle (MFC) at 2-3 weeks after bone marrow stimulation. DESIGN: Bilateral full-thickness cartilage defects were generated in central trochlear groove and MFC of skeletally mature rabbits. Four subchondral perforations were made on each defect, either by microfracture to 2 mm deep, or by drilling to 2 mm or 6 mm deep. Rabbits were sacrificed either on Day 14 post-operatively or on Day 21. Defects were analyzed by histology, stereology, histomorphometry and micro-computed tomography (CT). Intact femurs (N = 4) served as controls. RESULTS: Stromal cell density recruitment was similar in all defects, irrespective of defect location and surgical techniques used. There was a robust appearance of chondrocytes at Day 21 in TR defects with significantly higher volume fraction of chondrocytes in TR compared to MFC (P = 0.013). Chondrogenic foci were observed in marrow penetrating holes, with a significantly higher frequency and larger foci in TR vs MFC defects at Day 21 (P = 0.043 and P = 0.0014, respectively). Micro-CT analysis showed that deep drilling elicited significantly more mineralized bone fill compared to shallower perforations at 2 and 3 weeks repair (all at P ≤ 0.0008). CONCLUSIONS: Bone marrow stimulation induced greater chondrogenesis in TR vs MFC defects in adult rabbits, with more chondrocytes and larger chondrogenic foci appearing in TR vs MFC on Day 21 post-operation.

Quantitative imaging of murine osteoarthritic cartilage by phase-contrast micro-computed tomography.

OBJECTIVE: The mouse is an optimal model organism in which gene-environment interactions can be used to study the pathogenesis of osteoarthritis (OA). The gold standard for arthritis research in mice is based on histopathology and immunohistochemistry, which are labor-intensive, prone to sampling bias and technical variability, and limited in throughput. The aim of this study was to develop a new technique that assesses mouse cartilage by integrating quantitative volumetric imaging techniques. METHODS: A novel mouse model of OA was generated by cruciate ligament transection (CLT) and evaluated by histopathology and immunohistochemistry. Knee joint specimens were then imaged using a new technique that combines high-resolution micro-computed tomography (micro-CT) and phase-contrast optics followed by quantitative analyses. A comparative analysis was also performed in a previously established mouse model of OA generated by destabilization of the medial meniscus (DMM). RESULTS: Phase-contrast micro-CT achieved cellular resolution of chondrocytes and quantitative assessment of parameters such as articular cartilage volume and surface area. In mouse models of OA generated by either CLT or DMM, we showed that phase-contrast micro-CT distinguished control and OA cartilage by providing quantitative measures with high reproducibility and minimal variability. Features of OA at the cellular or tissue level could also be observed in images generated by phase-contrast micro-CT. CONCLUSION: We established an imaging technology that comprehensively assessed and quantified the 2-dimensional and 3-dimensional changes of articular cartilage. Application of this technology will facilitate the rapid and high-throughput assessment of genetic and therapeutic models of OA in mice.

The effect of ultrasound stimulation on the cytoskeletal organization of chondrocytes seeded in three-dimensional matrices.

The impact of low-intensity diffuse ultrasound (LIDUS) stimulation on the cytoskeletal organization of chondrocytes seeded in three-dimensional (3D) scaffolds was evaluated. Chondrocytes seeded on 3D chitosan matrices were exposed to LIDUS at 5.0 MHz (approx. 15 kPa, 51 s, 4 applications/day) in order to study the organization of actin, tubulin and vimentin. The results showed that actin presented a punctate cytosolic distribution and tubulin presented a quasiparallel organization of microtubules, whereas vimentin distribution was unaffected. Chondrocytes seeded on 3D scaffolds responded to US stimulation by the disruption of actin stress fibers and were sensitive to the presence of Rho-activated kinase (ROCK) inhibitor (Y27632). The gene expression of ROCK-I, a key element in the formation of stress fibers and mDia1, was significantly upregulated under the application of US. We conclude that the results of both the cytoskeletal analyses and gene expression support the argument that the presence of punctate actin upon US stimulation was accompanied by the upregulation of the RhoA/ROCK pathway.

Assessment of bone dysplasia by micro-CT and glycosaminoglycan levels in mouse models for mucopolysaccharidosis type I, IIIA, IVA, and VII.

Mucopolysaccharidoses (MPS) are a group of lysosomal storage diseases caused by mutations in lysosomal enzymes involved in degradation of glycosaminoglycans (GAGs). Patients with MPS grow poorly and become physically disabled due to systemic bone disease. While many of the major skeletal effects in mouse models for MPS have been described, no detailed analysis that compares GAGs levels and characteristics of bone by micro-CT has been done. The aims of this study were to assess severity of bone dysplasia among four MPS mouse models (MPS I, IIIA, IVA and VII), to determine the relationship between severity of bone dysplasia and serum keratan sulfate (KS) and heparan sulfate (HS) levels in those models, and to explore the mechanism of KS elevation in MPS I, IIIA, and VII mouse models. Clinically, MPS VII mice had the most severe bone pathology; however, MPS I and IVA mice also showed skeletal pathology. MPS I and VII mice showed severe bone dysplasia, higher bone mineral density, narrowed spinal canal, and shorter sclerotic bones by micro-CT and radiographs. Serum KS and HS levels were elevated in MPS I, IIIA, and VII mice. Severity of skeletal disease displayed by micro-CT, radiographs and histopathology correlated with the level of KS elevation. We showed that elevated HS levels in MPS mouse models could inhibit N-acetylgalactosamine-6-sulfate sulfatase enzyme. These studies suggest that KS could be released from chondrocytes affected by accumulation of other GAGs and that KS could be useful as a biomarker for severity of bone dysplasia in MPS disorders.

Effect of low-intensity pulsed ultrasound on MMP-13 and MAPKs signaling pathway in rabbit knee osteoarthritis.

We evaluated the effect of low-intensity pulsed ultrasound (LIPUS) on MMP-13 and MAPKs expression in rabbit knee osteoarthritis (OA). For this purpose, 18 New Zealand white rabbits were randomly and equally divided into O + L, O - L, and SO groups. In O + L group, animals underwent right back leg ACLT operation and LIPUS radiation. In O - L group, animals underwent ACLT but no LIPUS treatment. In SO (control) group, animals underwent sham operation without LIPUS. After 6 weeks, we assessed the pathologic changes in the articular surface of femoral condyle and compared using Mankin scores. Also, expression of type-II collagen, MMP-13, ERK1/2, p38, and JNK was measured by Western blot. Compared with controls, Mankin scores were higher in O + L (P < 0.05)/O - L (P < 0.01) groups. Compared with O + L group, score was higher in O - L group (P < 0.05). Compared with controls, type-II collagen expression was less in O + L/O - L groups, with more significant decrease in O - L group (P < 0.05). Contrarily, expression of MMP-13, p-ERK1/2, and p-p38 was enhanced in O + L/O - L groups as compared with controls, with more significant increase in O - L group (P < 0.01). Compared with O + L group, expression was higher in O - L group (P < 0.05). We, therefore, concluded that LIPUS application promoted cartilage repair in OA through the downregulation of MMP-13, ERK1/2, and p38.

Magnetization transfer imaging provides a quantitative measure of chondrogenic differentiation and tissue development.

The goal of the present investigation was to test whether quantitative magnetization transfer imaging can be used as a noninvasive evaluation method for engineered cartilage. In this work, we used magnetic resonance imaging (MRI) to monitor the chondrogenesis of stem-cell-based engineered tissue over a 3-week period by measuring on a pixel-by-pixel basis the relaxation times (T1 and T2), the apparent diffusion coefficient, and the magnetization transfer parameters: bound proton fraction and cross-relaxation rate (k). Tissue-engineered constructs for generating cartilage were created by seeding mesenchymal stem cells in a gelatin sponge. Every 7 days, tissue samples were analyzed using MRI, histological, and biochemical methods. The MRI measurements were verified by histological analysis, and the imaging data were correlated with biochemical analysis of the developing cartilage matrix for glycosaminoglycan content. The MRI analysis for bound proton fraction and k showed a statistically significant increase that was correlated with the increase of glycosaminoglycan (R = 0.96 and 0.87, respectively, p < 0.05), whereas T1, T2, and apparent diffusion coefficient results did not show any significant changes over the 3-week measurement period.

Going beyond histology. Synchrotron micro-computed tomography as a methodology for biological tissue characterization: from tissue morphology to individual cells.

Current light microscopic methods such as serial sectioning, confocal microscopy or multiphoton microscopy are severely limited in their ability to analyse rather opaque biological structures in three dimensions, while electron optical methods offer either a good three-dimensional topographic visualization (scanning electron microscopy) or high-resolution imaging of very thin samples (transmission electron microscopy). However, sample preparation commonly results in a significant alteration and the destruction of the three-dimensional integrity of the specimen. Depending on the selected photon energy, the interaction between X-rays and biological matter provides semi-transparency of the specimen, allowing penetration of even large specimens. Based on the projection-slice theorem, angular projections can be used for tomographic imaging. This method is well developed in medical and materials science for structure sizes down to several micrometres and is considered as being non-destructive. Achieving a spatial and structural resolution that is sufficient for the imaging of cells inside biological tissues is difficult due to several experimental conditions. A major problem that cannot be resolved with conventional X-ray sources are the low differences in density and absorption contrast of cells and the surrounding tissue. Therefore, X-ray monochromatization coupled with a sufficiently high photon flux and coherent beam properties are key requirements and currently only possible with synchrotron-produced X-rays. In this study, we report on the three-dimensional morphological characterization of articular cartilage using synchrotron-generated X-rays demonstrating the spatial distribution of single cells inside the tissue and their quantification, while comparing our findings to conventional histological techniques.

Calcium pyrophosphate dehydrate crystal deposition in the ligamentum flavum of the cervical spine: histopathological and immunohistochemical findings.

OBJECTIVE: To investigate the histopathological and immunohistochemical properties of degenerative changes in the ligamentum flavum of the cervical spine with calcium crystal deposition. METHODS: Sections of the calcified ligamentum flavum harvested from 26 patients who required cervical decompression were examined by scanning electron microscopy (SEM), energy dispersive X-ray microanalysis, immunohistochemical staining [for transforming growth factor (TGF)-Beta, vascular endothelial growth factor (VEGF), Sox9, and Msx2] and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labelling (TUNEL) method (for cell apoptosis). RESULTS: Energy dispersive x-ray microanalysis and SEM confirmed the deposited calcium to be calcium pyrophosphate dihydrate (CPPD) crystals. The calcified ligamentum flavum showed disorganisation of the elastic fibre bundles together with increased collagen fibrils in the matrix. Abundant hypertrophic chondrocytes were noted around the calcified lesions, which were strongly immunoreactive to TGF-Beta and VEGF. Staining for Sox9 was positive in metaplastic chondrocytes but negative in hypertrophic chondrocytes. Both chondrocytes and mesenchymal cells were positive for Msx2. TUNEL-positive hypertrophic chondrocytes were significantly more noticeable in nodular than diffusely scattered type of CPPD deposition. CONCLUSIONS: Calcium crystal deposition in the cervical ligamentum flavum seems to progress with reduction in elastic fibres, increase in collagen fibrils in the matrix, and migration of metaplastic hypertrophic chondrocytes, whose differentiation is controlled by cytokines and transcriptional factors, and potentially regulate crystal formation. The presence of abundant TUNEL-positive hypertrophic chondrocytes around CPPD deposition suggests that materials from apoptotic cells play some role in crystal deposition.

Imprint smear cytology of chondroblastic osteosarcoma: a case report.

Imprint cytology is gaining increasing popularity in the diagnosis of musculoskeletal lesions and in many patients; a definitive diagnosis can be rendered from aspiration smears alone. However, evaluation of tissue architecture is a major limitation and therefore cytological diagnosis has to be confirmed on histopathology. The authors take this opportunity to report a case of chondroblastic osteosarcoma occurring in the middle 1/3rd of the left thigh in a 35 year old male, diagnosed on imprint cytology and confirmed on histopatholgy.

Low-intensity pulsed ultrasound activates the phosphatidylinositol 3 kinase/Akt pathway and stimulates the growth of chondrocytes in three-dimensional cultures: a basic science study.

INTRODUCTION: The effect of low-intensity pulsed ultrasound (LIPUS) on cell growth was examined in three-dimensional-cultured chondrocytes with a collagen sponge. To elucidate the mechanisms underlying the mechanical activation of chondrocytes, intracellular signaling pathways through the Ras/mitogen-activated protein kinase (MAPK) and the integrin/phosphatidylinositol 3 kinase (PI3K)/Akt pathways as well as proteins involved in proliferation of chondrocytes were examined in LIPUS-treated chondrocytes. METHODS: Articular cartilage tissue was obtained from the metatarso-phalangeal joints of freshly sacrificed pigs. Isolated chondrocytes mixed with collagen gel and culture medium composites were added to type-I collagen honeycomb sponges. Experimental cells were cultured with daily 20-minute exposures to LIPUS. The chondrocytes proliferated and a collagenous matrix was formed on the surface of the sponge. Cell counting, histological examinations, immunohistochemical analyses and western blotting analysis were performed. RESULTS: The rate of chondrocyte proliferation was slightly but significantly higher in the LIPUS group in comparison with the control group during the 2-week culture period. Western blot analysis showed intense staining of type-IX collagen, cyclin B1 and cyclin D1, phosphorylated focal adhesion kinase, and phosphorylated Akt in the LIPUS group in comparison with the control group. No differences were detected, however, in the MAPK, phosphorylated MAPK and type-II collagen levels. CONCLUSION: LIPUS promoted the proliferation of cultured chondrocytes and the production of type-IX collagen in a three-dimensional culture using a collagen sponge. In addition, the anabolic LIPUS signal transduction to the nucleus via the integrin/phosphatidylinositol 3-OH kinase/Akt pathway rather than the integrin/MAPK pathway was generally associated with cell proliferation.

Intermittent applications of continuous ultrasound on the viability, proliferation, morphology, and matrix production of chondrocytes in 3D matrices.

Chondrocytes, the cellular component of the articular cartilage, have long been recognized as strain-sensitive cells, and have the ability to sense mechanical stimulation through surface receptors and intracellular signaling pathways. This strain-induced biological response of chondrocytes has been exploited to facilitate chondrocyte culture in in vitro systems; examples include the application of hydrostatic pressure, dynamic compression, hydrodynamic shear (i.e., rotating bioreactors), and low-intensity pulsed ultrasound (US). While the ability of US to influence chondrogenesis has been documented, the precise mechanisms of US-induced stimulation continue to be investigated. There remains a critical need to evaluate the impact of US on chondrocytes in 3D culture, which is a necessary microenvironment for maintaining the chondrocyte phenotype. In this study, a continuous US wave for predetermined time intervals was employed, as opposed to pulsed US used in previous studies, to stimulate chondrocytes seeded in 3D scaffolds. The chondrocytes (n = 6) were subjected to US stimulation as follows: 1.5 MHz for 161 seconds, 5.0 MHz for 51 seconds, and 8.5 MHz for 24 seconds, and the US signal was applied twice in a 24-hour period. Scaffolds that are not stimulated by US served as the control. Both the control and the US-stimulated groups were maintained in culture for 10 days, and at the conclusion of the culture period, chondrocytes were assayed for total DNA content, morphology, and cartilage-specific gene expression by reverse transcriptase polymerase chain reaction. Our results show that chondrocytes when stimulated with continuous US for predetermined time intervals possessed higher cellular viability (1.2 to 1.4 times) and higher levels of type II collagen and aggrecan mRNA expression when compared to controls.

Study on the effects of mechanical pressure to the ultrastructure and secretion ability of mandibular condylar chondrocytes.

During mandibular movement, condyle is subjected to repetitive compression and the mandibular condylar chondrocytes (MCCs) can detect and respond to this biomechanical environment by altering their metabolism. The present study was undertaken to investigate the effects of pressure to the ultrastructure, aggrecan synthesis, nitric oxide (NO) and prostaglandin F(1)alpha(PGF(1)alpha) secretion in MCCs. In vitro cultured rabbit MCCs were incubated and pressed under continuous pressure of 90kPa for 60min and 360min by hydraulic pressure controlled cellular strain unit. The ultrastructure, aggrecan mRNA expression, activity of nitric oxide synthase (NOS) and PGF(1)alpha secretion were investigated. Besides, nitric oxide inhibitor was used together with pressure to investigate the role of NO in mechanical effects. The appearance of MCC on TEM showed that after been pressed under 90kPa for 60min, the cellular processes became elongated and voluminous, together with aggrecan mRNA increasing. Under 90kPa for 360min, some of the cells showed distinct sign of apotosis and the aggrecan mRNA decreased. Pressure of 90kPa could cause increase of NOS activity and decrease of PGF(1)alpha composition. Inhibitor experiments indicated that pressure-induced upregulation of aggrecan mRNA and inhibition of PGF(1)alpha synthesis was partly mediated by NO. Continuous pressure could cause changes on the ultrastructure and function of MCC, as well as up-regulation of aggrecan synthesis, increase of NO secretion and decrease of PGF(1)alpha composition. NO was the upstream molecule, which mediated the response of aggrecan and PGF(1)alpha to mechanical pressure.

Mechanical and biochemical effect of monopolar radiofrequency energy on human articular cartilage: an in vitro study.

BACKGROUND: There are growing concerns about thermal chondroplasty using radiofrequency energy to treat partial-thickness cartilage defects. However, most studies emphasize effects on chondrocyte viability, and other factors such as mechanical properties are less studied. HYPOTHESIS: Radiofrequency energy may cause significant effects on articular cartilage other than chondrocyte viability. STUDY DESIGN: Controlled laboratory study. METHODS: Human osteoarthritic cartilage samples were obtained from total knee arthroplasty, and monopolar radiofrequency energy was applied using commercially available equipment. Material properties (compressive stiffness, surface roughness, and thickness) just before and after thermal treatment were determined using ultrasound. A series of biochemical analyses were also performed after explant culture of the samples. RESULTS: The cartilage surface became smoother by radiofrequency energy, whereas cartilage stiffness or thickness was not altered significantly. Collagen fibrils, especially in the superficial layers, were converted to denatured form, whereas proteoglycan contents released in the media as well as retained in the tissue remained unchanged. The concentrations of matrix metalloproteinases (MMP-1 and MMP-2) were reduced remarkably. CONCLUSION: Radiofrequency energy is able to create a smooth cartilage surface and reduce catabolic enzymes at the cost of collagen denaturation and chondrocyte death in the superficial layers. The stiffness of the cartilage is not changed at time zero. CLINICAL RELEVANCE: Further animal as well as clinical studies will be necessary to fully evaluate the long-term effects of radiofrequency energy.

The effect of PEGT/PBT scaffold architecture on the composition of tissue engineered cartilage.

A highly interconnecting and accessible pore network has been suggested as one of a number of prerequisites in the design of scaffolds for tissue engineering. In the present study, two processing techniques, compression-molding/particulate-leaching (CM), and 3D fiber deposition (3DF), were used to develop porous scaffolds from biodegradable poly(ethylene glycol)-terephthalate/poly(butylene terephthalate) (PEGT/PBT) co-polymers with varying pore architectures. Three-dimensional micro-computed tomography (microCT) was used to characterize scaffold architectures and scaffolds were seeded with articular chondrocytes to evaluate tissue formation. Scaffold porosity ranged between 75% and 80%. Average pore size of tortuous CM scaffolds (182 microm) was lower than those of organized 3DF scaffolds (525 microm). The weight ratio of glycosaminoglycans (GAG)/DNA, as a measure of cartilage-like tissue formation, did not change after 14 days of culture whereas, following subcutaneous implantation, GAG/DNA increased significantly and was significantly higher in 3DF constructs than in CM constructs, whilst collagen type II was present within both constructs. In conclusion, 3DF PEGT/PBT scaffolds create an environment in vivo that enhances cartilaginous matrix deposition and hold particular promise for treatment of articular cartilage defects.