Glycosaminoglycan Content Of Cartilage Research Articles

Clodronate disodium does not produce measurable effects on bone metabolism in an exercising, juvenile, large animal model.

Bisphosphonates are commonly used to treat and prevent bone loss, but their effects in active, juvenile populations are unknown. This study examined the effects of intramuscular clodronate disodium (CLO) on bone turnover, serum bone biomarkers (SBB), bone mineral density (BMD), bone microstructure, biomechanical testing (BT), and cartilage glycosaminoglycan content (GAG) over 165 days. Forty juvenile sheep (253 ± 6 days of age) were divided into four groups: Control (saline), T0 (0.6 mg/kg CLO on day 0), T84 (0.6 mg/kg CLO on day 84), and T0+84 (0.6 mg/kg CLO on days 0 and 84). Sheep were exercised 4 days/week and underwent physical and lameness examinations every 14 days. Blood samples were collected for SBB every 28 days. Microstructure and BMD were calculated from tuber coxae (TC) biopsies (days 84 and 165) and bone healing was assessed by examining the prior biopsy site. BT and GAG were evaluated postmortem. Data, except lameness data, were analyzed using a mixed-effects model; lameness data were analyzed as ordinal data using a cumulative logistic model. CLO did not have any measurable effects on the skeleton of sheep. SBB showed changes over time (p ≤ 0.03), with increases in bone formation and decreases in some bone resorption markers. TC biopsies showed increasing bone volume fraction, trabecular spacing and thickness, and reduced trabecular number on day 165 versus day 84 (p ≤ 0.04). These changes may be attributed to exercise or growth. The absence of a treatment effect may be explained by the lower CLO dose used in large animals compared to humans. Further research is needed to examine whether low doses of bisphosphonates may be used in active juvenile populations for analgesia without evidence of bone changes.

Material properties in regenerating axolotl limbs using inverse finite element analysis

BackgroundThe extracellular mechanical environment plays an important role in the skeletal development process. Characterization of the material properties of regenerating tissues that recapitulate development, provides insights into the mechanical environment experienced by the cells and the maturation of the matrix. In this study, we estimated the viscoelastic material properties of regenerating forelimbs in the axolotl (Ambystoma mexicanum) at three different regeneration stages: 27 days post-amputation (mid-late bud) and 41 days post-amputation (palette stage), and fully-grown time points. A stress-relaxation indentation test followed by two-term Prony series viscoelastic inverse finite element analysis was used to obtain material parameters. Glycosaminoglycan (GAG) content was estimated using a 1,9- dimethyl methylene blue assay. ResultsThe instantaneous and equilibrium shear moduli significantly increased with regeneration while the short-term stress relaxation time significantly decreased with limb regeneration. The long-term stress relaxation time in the fully-grown time point was significantly lower than 27 and 41 DPA groups. The GAG content was not significantly different between 27 and 41 DPA but the GAG content of cartilage in the fully-grown group was significantly greater than in 27 and 41 DPA. ConclusionsThe mechanical environment of the proliferating cells changes drastically during limb regeneration. Understanding how the tissue's mechanical properties change during limb regeneration is critical for linking molecular-level matrix production of the cells to tissue-level behavior and mechanical signals.

Age-Related Histologic and Biochemical Changes in Auricular and Nasal Cartilages.

Analyze age-related changes in histologic features and biochemical properties of human auricular cartilage and two subsites of nasal cartilages (quadrangular cartilage and dorsal septal articulation with upper lateral cartilages). Prospective cross-sectional study of nasal and auricular cartilages from seventy-three (73) live donors. Auricular cartilage (AC), quadrangular cartilage (QC), and dorsal septal cartilage articulation (DSA) with the upper lateral cartilage (ULCs) were collected intraoperatively. Histochemical staining was used: Safranin O for glycosaminoglycans (GAGs), Verhoeff's for elastin, and Masson's trichrome for collagen. ImageJ2 software was used to calculate cell count and percent stained for each cartilage type. R studio "ggplot" package was used to visualize age versus cell count or percent stained. Participant ages ranged from 20 to 77 years, average 46.5 years. There was a significant decline in GAGs with age for the DSA subsite, (n = 64, p < 0.001). Significant increase in collagen content with age was observed for DSA subsite (n = 66, p < 0.001) and the QC subsite (n = 64, p < 0.05). There was a statistically insignificant decline in elastin with age (n = 41, p = 0.309) for AC. Cell count declined with age at all cartilage subsites. Our findings confirm that there were age-related decreases in cartilage glycosaminoglycan content, and chondrocyte cell count in both auricular and nasal cartilages. We have also confirmed that collagen content increases with age for both auricular and nasal cartilage. The histologic findings while not statistically significant in all comparisons, provides additional evidence that there is some loss of structural integrity and flexibility in nasal and auricular cartilage with aging. NA Laryngoscope, 134:1220-1226, 2024.

Acute loading has minor influence on human articular cartilage gene expression and glycosaminoglycan composition in late-stage knee osteoarthritis: a randomised controlled trial

Osteoarthritis (OA) remains clinically challenging. Regular physical exercise improves symptoms though it is unclear whether exercise influences cartilage at the molecular level. Thus, we aimed to determine the effect of acute loading on gene expression and glycosaminoglycan (GAG) content in human OA cartilage. Patients with primary knee OA participated in this single-blind randomised controlled trial initiated 3.5h prior to scheduled joint replacement surgery with or without loading by performing one bout of resistance exercise (one-legged leg press). Cartilage from the medial tibia condyle was sampled centrally, under the meniscus, and from peripheral osteophytes. Samples were analysed for gene expression by real-time reverse transcriptase polymerase chain reaction, and hyaluronidase-extracted matrix was analysed for GAG composition by immuno- and dimethyl-methylene blue assays. Of 32 patients randomised, 31 completed the intervention: mean age 69±7.5 years (SD), 58% female, BMI 29.4±4.4kg/m2. Exercise increased chondroitin sulphate extractability [95% CI: 1.01 to 2.46; P=0.0486] but cartilage relevant gene expression was unchanged. Regionally, the submeniscal area showed higher MMP-3, MMP-13, IGF-1Ea, and CTGF, together with lower lubricin and COMP expression compared to the central condylar region. Further, osteophyte expression of MMP-1, MMP-13, IGF-1Ea, and TGF-β3 was higher than articular cartilage and lower for aggrecan, COMP, and FGF-2. Hyaluronidase-extracted matrix from central condylar cartilage contained more GAGs but less chondroitin sulphate compared to submeniscal cartilage. Acute exercise had minor influence on cartilage GAG dynamics, indicating that osteoarthritic cartilage is not significantly affected by acute exercise. However, the regional differences suggest a chronic mechanical influence on human cartilage. NCT03410745.

Human Adipose- and Amnion-Derived Mesenchymal Stromal Cells Similarly Mitigate Osteoarthritis Progression in the Dunkin Hartley Guinea Pig

Background: Clinical trials are currently underway to investigate the efficacy of intra-articular administration of mesenchymal stromal cells (MSCs) to mitigate osteoarthritis (OA) progression in the knee. Although multiple MSC sources exist, studies have yet to determine whether differences in therapeutic efficacy exist between them. Purpose: To compare the ability of intra-articularly injected adipose-derived MSCs (AD-MSCs) and amnion-derived MSCs (AM-MSCs) to mitigate the progression of knee OA in a small animal model of spontaneous OA, as well as to compare the therapeutic potential of MSCs in hyaluronic acid (HA) and in HA only with saline (OA) controls. Study Design: Controlled laboratory study. Methods: Injections of AD-MSCs or AM-MSCs suspended in HA or HA only were performed in the rear stifle joints of 3-month-old Dunkin Hartley guinea pigs (DHGPs). Repeat injections occurred at 2 and 4 months after the initial injection in each animal. Contralateral limbs received saline injections and served as untreated controls. Subsequently, joints were analyzed for osteoarthritic changes of the cartilage and subchondral bone via histologic and biochemical analyses. To evaluate MSC retention time in the joint space, DHGPs received a single intra-articular injection of fluorescently labeled AD-MSCs or AM-MSCs, and the fluorescence intensity was longitudinally tracked via an in vivo imaging system. Results: No statistically significant differences in outcomes were found when comparing the ability of AD-MSCs and AM-MSCs to mitigate OA. However, the injection of AD-MSCs, AM-MSCs, and HA-only treatments more effectively mitigated cartilage damage compared with that of saline controls by demonstrating higher amounts of cartilage glycosaminoglycan content and improved histological proteoglycan scoring while reducing the percentage of osteophytes present. Conclusion: Intra-articular injection of AD-MSCs, AM-MSCs, or HA only was able to similarly mitigate the progression of cartilage damage and reduce the percentage of osteophytes compared with that of saline controls in the DHGP. However, this study was unable to establish the superiority of AD-MSCs versus AM-MSCs as a treatment to mitigate spontaneous OA. Clinical Relevance: MSCs demonstrate the ability to mitigate the progression of knee OA and thus may be used in a prophylactic approach to delay the need for end-stage treatment strategies.

The Impact of Circular Exercise Diameter on Bone and Joint Health of Juvenile Animals.

Simple SummaryIn many equestrian disciplines, circular exercise is utilized to train, exercise, and compete with horses and can vary in gait, as well as diameter. This study aimed to determine if circular exercise diameter impacts animal health. Calves have previously been used as a terminal skeletal model of juvenile horses, allowing for the collection of musculoskeletal samples that are not acceptable from horses. Calves exercised on a small circle (12-m clockwise), large circle (18-m clockwise), treadmill, or served as non-exercised controls. Exercise was performed at a walking speed, starting at 5 min per day and increasing 5 min weekly until reaching 30 min per day during the 7-week study. The response to exercise was monitored in the forelimb bones and joints. The small circular exercise group was found to have bone diameters that differed between the right and left fused third and fourth metacarpi, and between lateral and medial proximal phalanx bones. Cartilage glycosaminoglycan content was greater in the outside leg of the small circle exercise calves than the inside leg, with no differences noted within other treatments. These differences suggest that altering circular exercise diameter can impact bone and joint health, and that larger diameter circles may prevent asymmetric loading between inside and outside legs.Circular exercise is used in many equestrian disciplines and this study aimed to determine if circle diameter impacts juvenile animal forelimb bone and joint health. On day 0, 24 calves at 9 weeks of age were assigned the following exercise treatments: small circle (12 m clockwise), large circle (18-m clockwise), treadmill, or non-exercised control. Exercise was initiated at 1.1–1.5 m/s for 5 min/d and increased 5 min weekly until reaching 30 min/d. On day 49, synovial fluid was collected from multiple joints, cartilage was collected from the proximal surface of fused third and fourth metacarpi (MC III and IV), and forelimbs underwent computed tomography scans. A statistical analysis (PROC mixed) was performed in SAS 9.4. The inside leg of the small circle treatment had a larger MC III and IV dorsopalmar external diameter than the outside (p = 0.05). The medial proximal phalanx had a greater mediolateral diameter than the lateral proximal phalanx of the small circle treatment (p = 0.01). Fetlock nitric oxide was greater in the large circle and treadmill treatments (p < 0.0001). Cartilage glycosaminoglycan concentration was greater in the outside leg of the small circle exercise treatment than the inside leg (p = 0.03). Even at slow speeds, circular exercise diameter can impact joint and bone health, but faster speeds may have greater alterations.

Spaceflight and hind limb unloading induces an arthritic phenotype in knee articular cartilage and menisci of rodents

Reduced knee weight-bearing from prescription or sedentary lifestyles are associated with cartilage degradation; effects on the meniscus are unclear. Rodents exposed to spaceflight or hind limb unloading (HLU) represent unique opportunities to evaluate this question. This study evaluated arthritic changes in the medial knee compartment that bears the highest loads across the knee after actual and simulated spaceflight, and recovery with subsequent full weight-bearing. Cartilage and meniscal degradation in mice were measured via microCT, histology, and proteomics and/or biochemically after: (1) ~ 35 days on the International Space Station (ISS); (2) 13-days aboard the Space Shuttle Atlantis; or (3) 30 days of HLU, followed by a 49-day weight-bearing readaptation with/without exercise. Cartilage degradation post-ISS and HLU occurred at similar spatial locations, the tibial-femoral cartilage-cartilage contact point, with meniscal volume decline. Cartilage and meniscal glycosaminoglycan content were decreased in unloaded mice, with elevated catabolic enzymes (e.g., matrix metalloproteinases), and elevated oxidative stress and catabolic molecular pathway responses in menisci. After the 13-day Shuttle flight, meniscal degradation was observed. During readaptation, recovery of cartilage volume and thickness occurred with exercise. Reduced weight-bearing from either spaceflight or HLU induced an arthritic phenotype in cartilage and menisci, and exercise promoted recovery.

Biochemical and Morphological Abnormalities of Subchondral Bone and Their Association with Cartilage Degeneration in Spontaneous Osteoarthritis.

This study aims to investigate how biochemical composition in subchondral bone (SB) relates to the sulfated glycosaminoglycan (sGAG) content of articular cartilage (AC) in the knee joint of guinea pigs from the early to moderate osteoarthritis (OA). Male Dunkin Hartley strain guinea pigs were grouped according to age (1, 3, 6, and 9months, with 10 guinea pigs in each group). The biochemical properties of the AC and SB in the tibial plateau of the guinea pigs were determined through histology and Raman spectroscopy, respectively. Furthermore, the microstructures of the SB were investigated using micro-computed tomography (micro-CT) and histology. Increased thickness and bone mineral density (BMD) and decreased porosity were observed in the subchondral plate (SP) with the progression of spontaneous OA, accompanied by a decreasing trend in sGAG integrated optical density (IOD) of AC. Compared with the changes in the microstructure of subchondral bone, the content of sGAG was more correlated to the changes in the mineral/matrix ratio of subchondral bone. The mineralization of the matrix was significantly correlated to the content of sGAG compared with crystallinity/maturity and Type B carbonate substitution. PO43- ν1/Amide III was more correlated to the content of sGAG than PO43- ν1/Amide I, PO43- ν1/CH2 wag during the progression of spontaneous osteoarthritis. This study demonstrated that the mineralization of subchondral bone plays a crucial role in the pathogenesis of OA. Future studies may access to the mineralization of subchondral bone in addition to its microstructure in the study for pathogenesis and early diagnosis of osteoarthritis.

Sprint Exercise of Juvenile Animals Does Not Impact Cartilage Glycosaminoglycan or Synovial Fluid Neopeptide Collagenase Cleavage of Type I and II Collagen Content

Short sprints performed during growth can increase bone strength, mineral density, and cortical widths, but their impact on joint health is not fully understood. Some studies have found joints of young animals are damaged by forced sprints, while others found confinement hindered joint development. This study aimed to determine the impact of short sprints on synovial fluid neopeptide collagenase cleavage of type I and II collagen (C1,2C) and cartilage glycosaminoglycan (GAG) content. Calves were used as a model for young horses in this terminal study. Twenty-four Holstein bull calves were assigned to treatment groups of 1, 3, 5, or 0 days/week of sprinting (n = 6). A sprint consisted of an individual calf being walked from its stall and down a 71-m concrete aisle, at the end of which the calf was released, and encouraged to sprint towards the barn. Calves remained on treatment for 6 weeks and were euthanized on day 42. Immediately posteuthanasia synovial fluid was sampled from the middle carpal joint and cartilage discs sampled from 9 locations on the carpal bones. Analysis was performed in SAS 9.4 with fixed effects of treatment and bone. There were no differences in C1,2C or GAG concentrations between treatments (P = .73 and P = .96, respectively), but there was a difference between individual carpal bone GAG concentration (P = .03). Short sprints for 6 weeks during growth did not impact middle carpal joint C1,2C or carpal bone GAG content, and should be permitted given the benefits to bone health.

Effects of solute size and tissue composition on molecular and macromolecular diffusivity in human knee cartilage

Articular cartilage is an avascular tissue. Accordingly, diffusivity represents a fundamental transport mechanism for nutrients and other molecular signals regulating its cell metabolism and maintenance of the extracellular matrix. Understanding how solutes spread into articular cartilage is crucial to elucidating its pathologies, and to designing treatments for repair and restoration of its extracellular matrix. As in other connective tissues, diffusivity in articular cartilage may vary depending both its composition and the specific diffusing solute. Hence, this study investigated the roles of solute size and tissue composition on molecular diffusion in knee articular cartilage. FRAP tests were conducted to measure diffusivity of five molecular probes, with size ranging from ~332Da to 70,000Da, in human knee articular cartilage. The measured diffusion coefficients were related to molecular size, as well as water and glycosaminoglycan (GAG) content of femoral and tibial condyle cartilage. Diffusivity was affected by molecular size, with the magnitude of the diffusion coefficients decreasing as the Stokes radius of the probe increased. The values of diffusion coefficients in tibial and femoral samples were not significantly different from one another, despite the fact that tibial samples exhibited significantly higher water content and lower GAG content of the femoral specimens. Water content did not affect diffusivity. In contrast, diffusivities of large molecules were sensitive to GAG content. This study provides new knowledge on the mechanisms of diffusion in articular cartilage. Our findings can be leveraged to further investigate osteoarthritis and to design treatments for cartilage restoration or replacement.

Contrast-enhanced micro-computed tomography of articular cartilage morphology with ioversol and iomeprol.

Non-ionic, low-osmolar contrast agents (CAs) used for computed tomography, such as Optiray (ioversol) and Iomeron (iomeprol), are associated with the reduced risk of adverse reactions and toxicity in comparison with ionic CAs, such as Hexabrix. Hexabrix has previously been used for imaging articular cartilage but has been commercially discontinued. This study aimed to evaluate the efficacy of Optiray and Iomeron as alternatives for visualisation of articular cartilage in small animal joints using contrast-enhanced micro-computed tomography (CECT). For this purpose, mouse femora were immersed in different concentrations (20%-50%) of Optiray 350 or Iomeron 350 for periods of time starting at five minutes. The femoral condyles were scanned ex vivo using CECT, and regions of articular cartilage manually contoured to calculate mean attenuation at each time point and concentration. For both CAs, a 30% CA concentration produced a mean cartilage attenuation optimally distinct from both bone and background signal, whilst 5-min immersion times were sufficient for equilibration of CA absorption. Additionally, plugs of bovine articular cartilage were digested by chondroitinase ABC to produce a spectrum of glycosaminoglycan (GAG) content. These samples were immersed in CA and assessed for any correlation between mean attenuation and GAG content. No significant correlation was found between attenuation and cartilage GAG content for either CAs. In conclusion, Optiray and Iomeron enable high-resolution morphological assessment of articular cartilage in small animals using CECT; however, they are not indicative of GAG content.

Tyrosinase-crosslinked, tissue adhesive and biomimetic alginate sulfate hydrogels for cartilage repair

The native cartilage extracellular matrix (ECM) is enriched in sulfated glycosaminoglycans with important roles in the signaling and phenotype of resident chondrocytes. Recapitulating the key ECM components within engineered tissues through biomimicking strategies has potential to improve the regenerative capacity of encapsulated cells and lead to better clinical outcome. Here, we developed a double-modified, biomimetic and tissue adhesive hydrogel for cartilage engineering. We demonstrated sequential modification of alginate with first sulfate moieties to mimic the high glycosaminoglycan content of native cartilage and then tyramine moieties to allow in situ enzymatic crosslinking with tyrosinase under physiological conditions. Tyrosinase-crosslinked alginate sulfate tyramine (ASTA) hydrogels showed strong adhesion to native cartilage tissue with higher bond strength compared to alginate tyramine (AlgTA). Both ASTA and AlgTA hydrogels supported the viability of encapsulated bovine chondrocytes and induced a strong increase in the expression of chondrogenic genes such as collagen 2, aggrecan and Sox9. Aggrecan and Sox9 gene expression of chondrocytes in ASTA hydrogels were significantly higher than those in AlgTA. Chondrocytes in both ASTA and AlgTA hydrogels showed potent deposition of cartilage matrix components collagen 2 and aggrecan after 3 weeks of culture whereas a decreased collagen 1 deposition was observed in the sulfated hydrogels. ASTA and AlgTA hydrogels with encapsulated human chondrocytes showed in vivo stability as well as cartilage matrix deposition upon subcutaneous implantation into mice for 4 weeks. Our data is the first demonstration of a double-modified alginate with sulfation and tyramination that allows in situ enzymatic crosslinking, strong adhesion to native cartilage and chondrogenic re-differentiation.

Pre-contrast T1 and cartilage thickness as confounding factors in dGEMRIC when evaluating human cartilage adaptation to physical activity

BackgroundThe dGEMRIC (delayed Gadolinium-Enhanced MRI of Cartilage) technique has been used in numerous studies for quantitative in vivo evaluation of the relative glycosaminoglycan (GAG) content in cartilage. The purpose of this study was to determine the influence of pre-contrast T1 and cartilage thickness when assessing knee joint cartilage quality with dGEMRIC.MethodsCartilage thickness and T1 relaxation time were measured in the central part of the femoral condyles before and two hours after intravenous Gd-DTPA2− administration in 17 healthy volunteers from a previous study divided into two groups: 9 sedentary volunteers and 8 exercising elite runners. Results were analyzed in superficial and a deep weight-bearing, as well as in non-weight-bearing regions of interest.ResultsIn the medial compartment, the cartilage was thicker in the exercising group, in weight-bearing and non-weight-bearing segments. In most of the segments, the T1 pre-contrast value was longer in the exercising group compared to the sedentary group. Both groups had a longer pre-contrast T1 in the superficial cartilage than in the deep cartilage. In the superficial cartilage, the gadolinium concentration was independent of cartilage thickness. In contrast, there was a linear correlation between the gadolinium concentration and cartilage thickness in the deep cartilage region.ConclusionCartilage pre-contrast T1 and thickness are sources of error in dGEMRIC that should be considered when analysing bulk values. Our results indicate that differences in cartilage structure due to exercise and weight-bearing may be less pronounced than previously demonstrated.

The effects of varus degree on the early metabolic changes of the lateral compartment cartilage in knees with medial unicompartmental osteoarthritis

Objective To find the effects of varus degree on the early metabolic changes of the lateral compartment cartilage in knees with medial unicompartmental osteoarthritis by detecting glycosaminoglycan (GAG) in varus knees. Methods From June 2016 to December 2017, twenty middle-aged volunteers without osteoarthritis or coronal deformities were recruited as the control group. Sixty patients diagnosed as medial unicompartmental osteoarthritis were recruited as the osteoarthritis group. The patients were further divided into four groups according to the degrees of varus angle, namely 2°-5° varus group, 5°-10° varus group, 10°-15° varus group and >15° varus group with 15 patients in each group. Delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) was performed in all participants. The dGEMRIC indices (T1Gd) representing GAG content were calculated in the weight-bearing femoral cartilage (wbFC), the posterior non-weight-bearing femoral cartilage (pFC), the lateral femoral cartilage (FC, wbFC+pFC) and the tibial cartilage (TC) in the lateral compartment by using Matlab 7.1 and MRIMapper software. Results T1Gd of wbFC, pFC, FC and TC were 400.3±51.5 ms, 393.6±57.9 ms, 397.5±52.3 ms and 448.6±62.5 ms in the control group, 391.8±41.5 ms, 407.2±43.8 ms, 400.1±37.8 ms and 461.3±41.6 ms in 2°-5° varus group and 386.9±57.1 ms, 401.3±73.5 ms, 397.7±59.6 ms and 438.9±42.8 ms in 5°-10° varus group. There was no significant difference among the above three groups in T1Gd in any of the analyzed cartilage regions (P>0.05). In 10°-15° varus group, T1Gd of wbFC, pFC, FC and TC were 380.1±45.5 ms, 385.5±76.6 ms, 384.0±53.5 ms and 400.2±43.8 ms, respectively. Although T1Gd of wbFC, pFC and FC in 10°-15° varus group were similar with that in the control group, 2°-5° varus group and 5°-10° varus (P>0.05), T1Gd of TC in 10°-15° varus group decreased significantly (P 15° varus group were 327.7±54.3 ms, 340.1±33.0 ms, 334.9±36.0 ms and 363.6±48.6 ms, respectively. T1Gd of all regions of interest in >15° varus group were significantly lower than that informer four groups (P<0.05). Conclusion In medial unicompartmental knee osteoarthritis, there is a relationship between varus degree and GAG content of the lateral compartment cartilage. If varus angle ≤10°, the GAG content of the lateral compartment cartilage was similar with the similar aged subjects without osteoarthritis. If varus angle >10°, GAG content of the lateral compartment decreases significantly. Key words: Osteoarthritis, knee; Genu varum; Glycosaminoglycans; Magnetic resonance imaging

Mitigation of Articular Cartilage Degeneration and Subchondral Bone Sclerosis in Osteoarthritis Progression Using Low-Intensity Ultrasound Stimulation

The purpose of this study was to evaluate the effect of low-intensity ultrasound on articular cartilage and subchondral bone alterations in joints under normal and functional disuse conditions during osteoarthritis (OA) progression. Total of thirty 5-mo-old female Sprague–Dawley rats were randomly assigned to six groups (n = 5/group): age-matched group, OA group, OA + ultrasound (US) group, hindlimb suspension (HLS) group, HLS + OA group and HLS + OA + US group. The surgical anterior cruciate ligament was used to induce OA in the right knee joints. After 2 wk of OA induction, low-intensity ultrasound generated with a 3-MHz transducer with 20% pulse duty cycle and 30 mW/cm2 acoustic intensity was delivered to the right knee joints for 20 min a day, 5 d a week for a total of 6 wk. Then, the right tibias were harvested for micro-computed tomography, histologic and mechanical analysis. Micro-computed tomography results indicated that the thickness and sulfated glycosaminoglycan content of cartilage decreased, but the thickness of the subchondral cortical bone plate and the formation of subchondral trabecular bone increased in the OA group under the normal joint use condition. Furthermore, histologic results revealed that chondrocyte density and arrangement in cartilage corrupted and the underlying subchondral bone increased during OA progression. These changes were accompanied by reductions in mechanical parameters in OA cartilage. However, fewer OA symptoms were observed in the HLS + OA group under the joint disuse condition. The cartilage degeneration and subchondral bone sclerosis were alleviated in the US treatment group, especially under normal joint use condition. In conclusion, low-intensity ultrasound could improve cartilage degeneration and subchondral sclerosis during OA progression. Also, it could provide a promising strategy for future clinical treatment for OA patients.

The Glycosaminoglycan Content of Hip Cartilage in Osteonecrosis of Femoral Head: Evaluation with Delayed Gadolinium-Enhanced Magnetic Resonance Imaging of Cartilage.

Hip cartilage quality is essential for the success of joint-preserving surgery for osteonecrosis. This study aimed to characterize cartilage changes in osteonecrosis of femoral head (ONFH) using delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC). Fifteen asymptomatic (control) and 60 ONFH subjects were included in this study. The ONFH subjects were stratified in accordance with the Association Research Circulation Osseous (ARCO) classification (n = 15 hips per ARCO stage). All participant hips were investigated using dGEMRIC and theT1Gd data were collected and analyzed. T1Gd value was significantly lower in the ONFH group (365.1 ± 90.5 ms; range 200-498 ms) compared with the control group (546.1 ± 26.0 ms; range 504-580 ms) (P < 0.001). The T1Gd values of ARCO stage I-IV ONFH were 460.2 ± 17.3 ms (439-498 ms), 408.9 ± 43.4 ms (337-472 ms), 359.9 ± 34.5 ms (303-412 ms), 231.5 ± 15.1 ms (200-253 ms), respectively. Decreased T1Gd value was found to correlate significantly with increased ONFH severity (P < 0.001). T1Gd value in collapse stage was significantly lower than that of noncollapse stage (295.7 ± 70.3 ms [range 200-412 ms] vs. 434.6 ± 41.7 ms [range 337-498 ms]; P < 0.001). dGEMRIC identified hip cartilage as abnormal in ONFH, even at early-stage, as represented by decreased T1Gd, and this was further aggravated by ONFH collapse.

Use of a 3-Telsa magnet to perform delayed gadolinium-enhanced magnetic resonance imaging of the distal interphalangeal joint of horses with and without naturally occurring osteoarthritis.

OBJECTIVE To characterize delayed gadolinium-enhanced MRI of cartilage (dGEMRIC) features of healthy hyaline cartilage of the distal interphalangeal joint (DIPJ) of horses, to determine whether dGEMRIC can be used to differentiate various stages of naturally occurring osteoarthritis of the DIPJ, and to correlate relaxation times determined by dGEMRIC with the glycosaminoglycan concentration, water content, and macroscopic and histologic findings of hyaline cartilage of DIPJs with and without osteoarthritis. SAMPLE 1 cadaveric forelimb DIPJ from each of 12 adult warmblood horses. PROCEDURES T1-weighted cartilage relaxation times were obtained for predetermined sites of the DIPJ before (T1preGd) and after (T1postGd) intra-articular gadolinium administration. Corresponding cartilage sites underwent macroscopic, histologic, and immunohistochemical evaluation, and cartilage glycosaminoglycan concentration and water content were determined. Median T1preGd and T1postGd were correlated with macroscopic, histologic, and biochemical data. Mixed generalized linear models were created to evaluate the effects of cartilage site, articular surface, and macroscopic and histologic scores on relaxation times. RESULTS 122 cartilage specimens were analyzed. Median T1postGd was lower than the median T1preGd for normal and diseased cartilage. Both T1preGd and T1postGd were correlated with macroscopic and histologic scores, whereby T1preGd increased and T1postGd decreased as osteoarthritis progressed. There was topographic variation of T1preGd and T1postGd within the DIPJ. Cartilage glycosaminoglycan concentration and water content were significantly correlated with T1preGd and macroscopic and histologic scores but were not correlated with T1postGd. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that dGEMRIC relaxation times varied for DIPJs with various degrees of osteoarthritis. These findings may help facilitate early detection of osteoarthritis.

A novel organ culture model of a joint for the evaluation of static and dynamic load on articular cartilage.

ObjectivesThe purpose of this study was to create a novel ex vivo organ culture model for evaluating the effects of static and dynamic load on cartilage.MethodsThe metatarsophalangeal joints of 12 fresh cadaveric bovine feet were skinned and dissected aseptically, and cultured for up to four weeks. Dynamic movement was applied using a custom-made machine on six joints, with the others cultured under static conditions. Chondrocyte viability and matrix glycosaminoglycan (GAG) content were evaluated by the cell viability probes, 5-chloromethylfluorescein diacetate (CMFDA) and propidium iodide (PI), and dimethylmethylene blue (DMMB) assay, respectively.ResultsChondrocyte viability in the static model decreased significantly from 89.9% (sd 2.5%) (Day 0) to 66.5% (sd 13.1%) (Day 28), 94.7% (sd 1.1%) to 80. 9% (sd 5.8%) and 80.1% (sd 3.0%) to 46.9% (sd 8.5%) in the superficial quarter, central half and deep quarter of cartilage, respectively (p < 0.001 in each zone; one-way analysis of variance). The GAG content decreased significantly from 6.01 μg/mg (sd 0.06) (Day 0) to 4.71 μg/mg (sd 0.06) (Day 28) (p < 0.001; one-way analysis of variance). However, with dynamic movement, chondrocyte viability and GAG content were maintained at the Day 0 level over the four-week period without a significant change (chondrocyte viability: 92.0% (sd 4.0%) (Day 0) to 89.9% (sd 0.2%) (Day 28), 93.1% (sd 1.5%) to 93.8% (sd 0.9%) and 85.6% (sd 0.8%) to 84.0% (sd 2.9%) in the three corresponding zones; GAG content: 6.18 μg/mg (sd 0.15) (Day 0) to 6.06 μg/mg (sd 0.09) (Day 28)).ConclusionDynamic joint movement maintained chondrocyte viability and cartilage GAG content. This long-term whole joint culture model could be of value in providing a more natural and controlled platform for investigating the influence of joint movement on articular cartilage, and for evaluating novel therapies for cartilage repair.Cite this article: Y-C. Lin, A. C. Hall, A. H. R. W. Simpson. A novel organ culture model of a joint for the evaluation of static and dynamic load on articular cartilage. Bone Joint Res 2018;7:205–212. DOI: 10.1302/2046-3758.73.BJR-2017-0320.

Glycosaminoglycan Content of the Lateral Compartment Cartilage in Knees Conforming to the Indications for Oxford Medial Unicompartmental Knee Arthroplasty.

Background:The quality of the lateral compartment cartilage is important to preoperative evaluation and prognostic prediction of unicompartmental knee arthroplasty (UKA). Delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC) enables noninvasive assessment of glycosaminoglycan (GAG) content in cartilage. This study aimed to determine the GAG content of the lateral compartment cartilage in knees scheduled to undergo Oxford medial UKA.Methods:From December 2016 to May 2017, twenty patients (20 osteoarthritic knees) conforming to the indications for Oxford medial UKA were included as the osteoarthritis (OA) group, and 20 healthy volunteers (20 knees) paired by sex, knee side, age (±3 years), and body mass index (BMI) (±3 kg/m2) were included as the control group. The GAG contents of the weight-bearing femoral cartilage (wbFC), the posterior non-weight-bearing femoral cartilage (pFC), the lateral femoral cartilage (FC), and tibial cartilage (TC) were detected using dGEMRIC. The dGEMRIC indices (T1Gd) were calculated in the middle three consecutive slices of the lateral compartment. Paired t-tests were used to compare the T1Gd in each region of interest between the OA group and control group.Results:The average age and BMI in the two groups were similar. In the OA group, T1Gd of FC and TC was 386.7 ± 50.7 ms and 429.6 ± 59.9 ms, respectively. In the control group, T1Gd of FC and TC was 397.5 ± 52.3 ms and 448.6 ± 62.5 ms, respectively. The respective T1Gd of wbFC and pFC was 380.0 ± 47.8 ms and 391.0 ± 66.3 ms in the OA group and 400.3 ± 51.5 ms and 393.6 ± 57.9 ms in the control group. Although the T1Gd of wbFC and TC tended to be lower in the OA group than the control group, there was no significant difference between groups in the T1Gd in any of the analyzed cartilage regions (P value of wbFC, pFC, FC, and TC was 0.236, 0.857, 0.465, and 0.324, respectively).Conclusions:The GAG content of the lateral compartment cartilage in knees conforming to indications for Oxford medial UKA is similar with those of age- and BMI-matched participants without OA.

Combined effects of ovariectomy and streptozotocin-induced diabetes in the articular cartilage of rats

Aim: To evaluate the combined effects of streptozotocin-induced diabetes (Di) and ovariectomy in the articular cartilage of rats.Methods: Forty adult female Wistar rats were ovariectomized (OVX) or sham-operated. After recovery from surgery, the animals were assigned randomly into four groups: OVX control (OVX-C); OVX treated with 10 µg/kg/day of 17β-estradiol (OVX-E); sham-operated subjected to Di (Sham-Di); and OVX subjected to Di (OVX-Di). After 60 days of treatment, the animals were euthanized and the distal femurs with articular cartilage were processed for paraffin-embedding. Sections were stained with hematoxylin and eosin for histomorphometry, Picro-Sirius Red for collagen, or Alcian Blue for glycosaminoglycan (GAG) content. To detect apoptosis, sections were stained with an antibody to cleaved caspase-3 (casp-3).Results: Articular cartilage thickness and GAG content were significantly lower (p < 0.05) in the OVX-Di group, which also showed a higher number of casp-3-positive chondrocytes than the other groups. Interestingly, the higher percentage (p < 0.05) of mature collagen fibers was seen in the OVX-Di group, may be as a result of a reduced extracellular matrix remodeling of the articular cartilage.Conclusion: Our results indicate that the combination of ovariectomy and streptozotocin-induced diabetes produces more deleterious effects in articular cartilage of rats than either condition alone.