Non-invasive Electroarthrography Measures Load-Induced Cartilage Streaming Potentials via Electrodes Placed on Skin Surrounding an Articular Joint

Objective We aimed to demonstrate that electroarthrography (EAG) measures streaming potentials originating in the cartilage extracellular matrix during load bearing through electrodes adhered to skin surrounding an articular joint. Design Equine metacarpophalangeal joints were subjected to simulated physiological loads while (1) replacing synovial fluid with immersion buffers of different electrolyte concentrations and (2) directly degrading cartilage with trypsin. Results An inverse relationship between ionic strength and EAG coefficient was detected. Compared to native synovial fluid, EAG coefficients increased ( P < 0.05) for 5 of 6 electrodes immersed in 0.1X phosphate-buffered saline (PBS) (0.014 M NaCl), decreased ( P < 0.05) for 4 of 6 electrodes in 1X PBS (0.14 M NaCl), and decreased ( P < 0.05) for all 6 electrodes in 10X PBS (1.4 M NaCl). This relationship corresponds to similar studies where streaming potentials were directly measured on cartilage. EAG coefficients, obtained after trypsin degradation, were reduced ( P < 0.05) in 6 of 8, and 7 of 8 electrodes, during simulated standing and walking, respectively. Trypsin degradation was confirmed by direct cartilage assessments. Streaming potentials, measured by directly contacting cartilage, indicated lower cartilage stiffness ( P < 10−5). Unconfined compression data revealed reduced Em, representing proteoglycan matrix stiffness ( P = 0.005), no change in Ef, representing collagen network stiffness ( P = 0.15), and no change in permeability ( P = 0.24). Trypsin depleted proteoglycan as observed by both dimethylmethylene blue assay ( P = 0.0005) and safranin-O stained histological sections. Conclusion These data show that non-invasive EAG detects streaming potentials produced by cartilage during joint compression and has potential to become a diagnostic tool capable of detecting early cartilage degeneration.

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Exosomes derived from miR-126-3p-overexpressing synovial fibroblasts suppress chondrocyte inflammation and cartilage degradation in a rat model of osteoarthritis

Cell Death Discovery ◽

10.1038/s41420-021-00418-y ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Yan Zhou ◽

Jianghua Ming ◽

Yaming Li ◽

Bochun Li ◽

Ming Deng ◽

...

Keyword(s):

Synovial Fluid ◽

Apoptotic Cell ◽

Synovial Fibroblasts ◽

Cartilage Degeneration ◽

Cartilage Degradation ◽

Exosomal Mirnas ◽

Exosomal Mirna ◽

And Control ◽

And Cartilage

AbstractMicroRNAs (miRNAs) encapsulated within exosomes can serve as essential regulators of intercellular communication and represent promising biomarkers of several aging-associated disorders. However, the relationship between exosomal miRNAs and osteoarthritis (OA)-related chondrocytes and synovial fibroblasts (SFCs) remain to be clarified. Herein, we profiled synovial fluid-derived exosomal miRNAs and explored the effects of exosomal miRNAs derived from SFCs on chondrocyte inflammation, proliferation, and survival, and further assessed their impact on cartilage degeneration in a surgically-induced rat OA model. We identified 19 miRNAs within synovial fluid-derived exosomes that were differentially expressed when comparing OA and control patients. We then employed a microarray-based approach to confirm that exosomal miRNA-126-3p expression was significantly reduced in OA patient-derived synovial fluid exosomes. At a functional level, miRNA-126-3p mimic treatment was sufficient to promote rat chondrocyte migration and proliferation while also suppressing apoptosis and IL-1β, IL-6, and TNF-α expression. SFC-miRNA-126-3p-Exos were able to suppress apoptotic cell death and associated inflammation in chondrocytes. Our in vivo results revealed that rat SFC-derived exosomal miRNA-126-3p was sufficient to suppress the formation of osteophytes, prevent cartilage degeneration, and exert anti-apoptotic and anti-inflammatory effects on articular cartilage. Overall, our findings indicate that SFC exosome‐delivered miRNA-126-3p can constrain chondrocyte inflammation and cartilage degeneration. As such, SFC-miRNA-126-3p-Exos may be of therapeutic value for the treatment of patients suffering from OA.

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Treadmill Exercise after Controlled Abnormal Joint Movement Inhibits Cartilage Degeneration and Synovitis

Life ◽

10.3390/life11040303 ◽

2021 ◽

Vol 11 (4) ◽

pp. 303

Author(s):

Yuichiro Oka ◽

Kenij Murata ◽

Kaichi Ozone ◽

Takuma Kano ◽

Yuki Minegishi ◽

...

Keyword(s):

Treadmill Exercise ◽

Cruciate Ligament ◽

Cartilage Degeneration ◽

Research Society ◽

Therapeutic Effects ◽

Joint Movement ◽

Knee Oa ◽

Anterior Cruciate ◽

Safranin O ◽

Walking Group

Cartilage degeneration is the main pathological component of knee osteoarthritis (OA), but no effective treatment for its control exists. Although exercise can inhibit OA, the abnormal joint movement with knee OA must be managed to perform exercise. Our aims were to determine how controlling abnormal joint movement and treadmill exercise can suppress cartilage degeneration, to analyze the tissues surrounding articular cartilage, and to clarify the effect of treatment. Twelve-week-old ICR mice (n = 24) underwent anterior cruciate ligament transection (ACL-T) surgery on their right knees and were divided into three groups as follows: ACL-T, animals in the walking group subjected to ACL-T; controlled abnormal joint movement (CAJM), and CAJM with exercise (CAJM + Ex) (n = 8/group). Walking-group animals were subjected to treadmill exercise 6 weeks after surgery, including walking for 18 m/min, 30 min/day, 3 days/week for 8 weeks. Safranin-O staining, hematoxylin-eosin staining, and immunohistochemical staining were performed. The OARSI (Osteoarthritis research Society international) score was lower in the CAJM group than in the ACL-T group and was even lower in the CAJM + Ex group. The CAJM group had a lower meniscal injury score than the ACL-T group, and the CAJM + Ex group demonstrated a less severe synovitis than the ACL-T and CAJM groups. The observed difference in the perichondrium tissue damage score depending on the intervention method suggests different therapeutic effects, that normalizing joint motion can solve local problems in the knee joint, and that the anti-inflammatory effect of treadmill exercise can suppress cartilage degeneration.

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Selective STAT3 Inhibitor Alantolactone Ameliorates Osteoarthritis via Regulating Chondrocyte Autophagy and Cartilage Homeostasis

Frontiers in Pharmacology ◽

10.3389/fphar.2021.730312 ◽

2021 ◽

Vol 12 ◽

Author(s):

Wenbin Pei ◽

Xiaojian Huang ◽

Bowei Ni ◽

Rui Zhang ◽

Guangyi Niu ◽

...

Keyword(s):

Western Blot ◽

Inflammatory Responses ◽

Cartilage Degeneration ◽

Cartilage Destruction ◽

Signaling Molecules ◽

Inflammatory Factors ◽

Signal Pathways ◽

Autophagic Flux ◽

Safranin O

Osteoarthritis (OA), which is identified by chronic pain, impacts the quality of life. Cartilage degradation and inflammation are the most relevant aspects involved in its development. Signal transducer and activator of transcription 3(STAT3), a member of the STATs protein family, is associated with inflammation. Alantolactone (ALT), a sesquiterpene lactone compound, can selectively suppress the phosphorylation of STAT3. However, the pharmacological effect of ALT on OA is still imprecise. In this study, IL-1β (10 ng/ml) was applied to cartilage chondrocytes, which were treated with different concentrations of Alantolactone for 24 h. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2(COX2), matrix metalloproteinases (MMPs) and thrombospondin motifs-5 (ADAMTS5) were detected by western blot. Protein expression of Collagen Ⅱ was observed by western blot, safranin O staining and immunofluorescence. Manifestation of autophagy related proteins such as autophagy-related gene-5 (ATG5), P62, LC3Ⅱ/Ⅰ and PI3K/AKT/mTOR-related signaling molecules were measured by western blot and autophagic flux monitored by confocal microscopy. Expression of STAT3 and NF-κB-related signaling molecules were evaluated by western blot and immunofluorescence. In vivo, 2 mg/kg ALT or equal bulk of vehicle was engaged in the destabilization of medial meniscus (DMM) mouse models by intra-articular injection, the degree of cartilage destruction was classified by Safranin O/Fast green staining. Our findings reported that the enhance of inflammatory factors containing iNOS, COX2, MMPs and ADAMTS5 induced by IL-1β could be ameliorated by ALT. Additionally, the diminish of Collagen Ⅱ and autophagy which was stimulated by IL-1β could be alleviated by ALT. Mechanistically, STAT3, NF-κB and PI3K/AKT/mTOR signal pathways might be involved in the effect of ALT on IL-1β-induced mouse chondrocytes. In vivo, ALT protected cartilage in the DMM mouse model. Overall, this study illustrated that ALT attenuated IL-1β-induced inflammatory responses, relieved cartilage degeneration and promoted impaired autophagy via restraining of STAT3 and NF-κB signal pathways, implying its auspicious therapeutical effect for OA.

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247 SUCCESSFUL ARTIFICIAL INSEMINATION IN THE CORN SNAKE (ELAPHE GUTATTA), USING FRESH AND COOLED SEMEN

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab247 ◽

2007 ◽

Vol 19 (1) ◽

pp. 240 ◽

Cited By ~ 1

Author(s):

J. K. Mattson ◽

A. T. DeVries ◽

S. M. McGuire ◽

J. Krebs ◽

E. E. Louis ◽

...

Keyword(s):

Artificial Insemination ◽

Isoamyl Alcohol ◽

Invasive Technique ◽

Non Invasive ◽

Microcentrifuge Tube ◽

Tuberculin Syringe ◽

Potential Damage ◽

Corn Snake ◽

Phosphate Buffered Saline ◽

Parentage Test

The purpose of this investigation was to develop a non-invasive technique to artificially inseminate snakes using the corn snake, Elaphe gutatta, as the model representative for this taxon. Semen was collected by first applying pressure to the lower abdomen in a continuous distal motion toward the cloaca to remove any feces or urates. The cloaca was then gently washed using phosphate-buffered saline, and a more localized pressure was applied to each side of the vent to evert the hemipenes and, subsequently, the ejaculate. The semen was collected using a sterile transfer pipette and placed into 70 to 90 �L of medium (TL-HEPES solution; Cambrex Bio Science, Inc., Baltimore, MD, USA04–616F) in a sterile microcentrifuge tube, and then analyzed for overall motility, rate of forward progression (RFP, 0–5), and concentration. Based on a previously reported procedure, 10 females were inseminated with either fresh (n = 5) or cooled semen (n = 5; refrigerated for 3 days) one week after recovering from a hibernation period required to stimulate reproduction in this species. The overall sperm motility and concentration for females inseminated with fresh or cooled semen was 92%, 9.6 million sperm mL-1; and 85%, 6.1 million sperm mL-1, respectively. Immediately prior to insemination, the same method for expressing feces and urates in the males was applied to the females. The insemination dose (50 �L semen per oviduct) was drawn into a 1-mL latex- and silicone-free tuberculin syringe (Norm-Ject; VWR, Batavia, IL, USA) that was connected to a feeding/dosing needle (EJAY International, Issaquah, WA, USA) with a ball tip to prevent any potential damage during the insemination. The tip of the needle was then moved around the inner tissue of the vent to relax the cloaca, and the insertion continued until resistance was found indicating the vicinity of the oviducts. The extended semen was carefully deposited on both sides, and then the needle was slowly withdrawn. The offspring were tested for parentage to verify the success of the insemination. Blood was collected from the dorsal aorta posterior to the cloaca and stored in 10 mM Tris at 4�C. The DNA was extracted using a phenyl : chloroform : isoamyl alcohol (PCI) extraction method. Eight microsatellite loci were used for the paternity exclusion analysis: Eob�1, Eob�3, Eob�10, Eob�13, Eob�16, Eob�34, Eob�366, and Eob�373 (IDT, Coraville, IA, USA). All males and females in the collection were tested, and parental candidates were excluded if 2 or more allele mismatches occurred. From the total number of females inseminated, 3 females laid 51 eggs. Two females inseminated with cooled semen laid 36 eggs, of which 5 eggs were hatched, and the remaining were either unfertilized (n = 25) or non-viable (n = 6). All 5 hatched eggs were laid by one of the females. The third female inseminated with fresh semen laid 15 eggs, resulting in 5 hatching and 10 unfertilized eggs. The parentage test validated the AI a success as the alleles correlated between the adults and the offspring. In conclusion, artificial insemination was successful using both freshly collected and cooled (3 days) semen. Further studies are underway to improve the success rate in order to maximize the efficiency of this technology, and thus assist in the genetic preservation of endangered snake species.

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HDAC4 mutant represses chondrocyte hypertrophy by locating in the nucleus and attenuates disease progression of posttraumatic osteoarthritis

BMC Musculoskeletal Disorders ◽

10.1186/s12891-021-04947-6 ◽

2022 ◽

Vol 23 (1) ◽

Author(s):

Xiaodong Gu ◽

Fei Li ◽

Yangyang Gao ◽

Xianda Che ◽

Pengcui Li

Keyword(s):

Articular Cartilage ◽

Histone Deacetylase ◽

Cartilage Damage ◽

Adenovirus Vector ◽

Cartilage Degeneration ◽

Sprague Dawley ◽

Chondrocyte Hypertrophy ◽

Sprague Dawley Rats ◽

Histone Deacetylase 4 ◽

Safranin O

Abstract Background The aim of this study was to evaluate whether histone deacetylase 4 S246/467/632A mutant (m-HDAC4) has enhanced function at histone deacetylase 4 (HDAC4) to attenuate cartilage degeneration in a rat model of osteoarthritis (OA). Methods Chondrocytes were infected with Ad-m-HDAC4-GFP or Ad-HDAC4-GFP for 24 h, incubated with interleukin-1β (IL-1β 10 ng/mL) for 24 h, and then measured by RT-qPCR. Male Sprague-Dawley rats (n = 48) were randomly divided into four groups and transduced with different vectors: ACLT/Ad-GFP, ACLT/Ad-HDAC4-GFP, ACLT/Ad-m-HDAC4-GFP, and sham/Ad-GFP. All rats received intra-articular injections 48 h after the operation and every 3 weeks thereafter. Cartilage damage was assessed using radiography and Safranin O staining and quantified using the OARSI score. The hypertrophic and anabolic molecules were detected by immunohistochemistry and RT-qPCR. Results M-HDAC4 decreased the expression levels of Runx-2, Mmp-13, and Col 10a1, but increased the levels of Col 2a1 and ACAN more effectively than HDAC4 in the IL-1β-induced chondrocyte OA model; upregulation of HDAC4 and m-HDAC4 in the rat OA model suppressed Runx-2 and MMP-13 production, and enhanced Col 2a1 and ACAN synthesis. Stronger Safranin O staining was detected in rats treated with m-HDAC4 than in those treated with HDAC4. The resulting OARSI scores were lower in the Ad-m-HDAC4 group (5.80 ± 0.45) than in the Ad-HDAC4 group (9.67 ± 1.83, P = 0.045). The OARSI scores were highest in rat knees that underwent ACLT treated with Ad-GFP control adenovirus vector (14.93 ± 2.14, P = 0.019 compared with Ad-HDAC4 group; P = 0.003 compared with Ad-m-HDAC4 group). Lower Runx-2 and MMP-13 production, and stronger Col 2a1 and ACAN synthesis were detected in rats treated with m-HDAC4 than in those treated with HDAC4. Conclusions M-HDAC4 repressed chondrocyte hypertrophy and induced chondrocyte anabolism in the nucleus. M-HDAC4 was more effective in attenuating articular cartilage damage than HDAC4.

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The Role of Nanoparticles-Mediated Ligustrazine in the Treatment of Knee Osteoarthritis and Its Effect on Matrix Metalloproteinases and Upstream NF-κB Signaling Pathway in Knee Osteoarthritis

Journal of Nanoscience and Nanotechnology ◽

10.1166/jnn.2021.18647 ◽

2021 ◽

Vol 21 (2) ◽

pp. 1372-1377

Author(s):

Lei Ji ◽

Weihua Li ◽

Wenbin Qian ◽

Shuang Feng ◽

Hui Shi ◽

...

Keyword(s):

Chinese Medicine ◽

Traditional Chinese Medicine ◽

Synovial Fluid ◽

Knee Osteoarthritis ◽

Signaling Pathway ◽

Raw Materials ◽

Cartilage Degeneration ◽

Effective Components ◽

Traditional Chinese Medicine Preparation ◽

Effective Component

Knee osteoarthritis (KOA) is a joint degenerative arthropathy, characterized by cartilage degeneration of knee joint. Ligustrazine is an effective component of traditional Chinese medicine chuanqiong. It is reported that ligustrazine is used as a kind of anti-inflammatory medicine in folk prescription, especially in the treatment of knee osteoarthritis. The study is aimed to study the therapeutic effect of ligustrazine mediated by nanoparticle on knee osteoarthritis and its impact on MMPs and upstream NF-κB signaling pathway in synovial fluid. Nanoparticle-mediated system is a kind of nano traditional Chinese medicine preparation, which is made by taking nanoparticle and combining with the effective components, effective parts, raw materials, and their compounds in a certain way. We found that the combination of nanoparticle and ligustrazine can improve its bioavailability and targeting, reduce the adverse reactions in the treatment of knee osteoarthritis. The ligustrazine mediated by nanoparticle can effectively alleviate knee osteoarthritis by reducing the level of MMPs in synovial fluid and the expression of NF-κB in upstream NF-κB signaling pathway.

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Hyaluronan concentration and size distribution in human knee synovial fluid: variations with age and cartilage degeneration

Arthritis Research & Therapy ◽

10.1186/s13075-016-0922-4 ◽

2016 ◽

Vol 18 (1) ◽

Cited By ~ 41

Author(s):

Michele M. Temple-Wong ◽

Shuwen Ren ◽

Phu Quach ◽

Bradley C. Hansen ◽

Albert C. Chen ◽

...

Keyword(s):

Synovial Fluid ◽

Size Distribution ◽

Cartilage Degeneration ◽

Human Knee ◽

And Cartilage

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Rapid and Label-Free Isolation of Tumour Cells from the Urine of Patients with Localised Prostate Cancer Using Inertial Microfluidics

Cancers ◽

10.3390/cancers12010081 ◽

2019 ◽

Vol 12 (1) ◽

pp. 81 ◽

Cited By ~ 3

Author(s):

Alexey S. Rzhevskiy ◽

Sajad Razavi Bazaz ◽

Lin Ding ◽

Alina Kapitannikova ◽

Nima Sayyadi ◽

...

Keyword(s):

Prostate Cancer ◽

Liquid Biopsy ◽

Diagnostic Value ◽

Tumour Cells ◽

Label Free ◽

Non Invasive ◽

Diagnosis And Prognosis ◽

Potential Clinical Utility ◽

Phosphate Buffered Saline ◽

Microchannel Device

During the last decade, isolation of circulating tumour cells via blood liquid biopsy of prostate cancer (PCa) has attracted significant attention as an alternative, or substitute, to conventional diagnostic tests. However, it was previously determined that localised forms of PCa shed a small number of cancer cells into the bloodstream, and a large volume of blood is required just for a single test, which is impractical. To address this issue, urine has been used as an alternative to blood for liquid biopsy as a truly non-invasive, patient-friendly test. To this end, we developed a spiral microfluidic chip capable of isolating PCa cells from the urine of PCa patients. Potential clinical utility of the chip was demonstrated using anti-Glypican-1 (GPC-1) antibody as a model of the primary antibody in immunofluorescent assay for identification and detection of the collected tumour cells. The microchannel device was first evaluated using DU-145 cells in a diluted Dulbecco’s phosphate-buffered saline sample, where it demonstrated >85 (±6) % efficiency. The microchannel proved to be functional in at least 79% of cases for capturing GPC1+ putative tumour cells from the urine of patients with localised PCa. More importantly, a correlation was found between the amount of the captured GPC1+ cells and crucial diagnostic and prognostic parameter of localised PCa—Gleason score. Thus, the technique demonstrated promise for further assessment of its diagnostic value in PCa detection, diagnosis, and prognosis.

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Impact of Controlling Abnormal Joint Movement on the Effectiveness of Subsequent Exercise Intervention in Mouse Models of Early Knee Osteoarthritis

Cartilage ◽

10.1177/1947603519885007 ◽

2019 ◽

pp. 194760351988500

Author(s):

Yuichiro Oka ◽

Kenji Murata ◽

Takuma Kano ◽

Kaichi Ozone ◽

Kohei Arakawa ◽

...

Keyword(s):

Mechanical Stress ◽

Exercise Intervention ◽

Cruciate Ligament ◽

Cartilage Degeneration ◽

Research Society ◽

Joint Movement ◽

Knee Oa ◽

Osteoarthritis Research Society ◽

Anterior Cruciate ◽

Safranin O

Objective Moderate mechanical stress is necessary for preserving the cartilage. The clinician empirically understands that prescribing only exercise will progress osteoarthritis (OA) for knee OA patients with abnormal joint movement. When prescribing exercise for OA, we hypothesized that degeneration of articular cartilage could be further prevented by combining interventions with the viewpoint of normalizing joint movement. Design Twelve-week-old ICR mice underwent anterior cruciate ligament transection (ACL-T) surgery in their right knee and divided into 4 groups: ACL-T, controlled abnormal joint movement (CAJM), ACL-T with exercise (ACL-T/Ex), CAJM with exercise (CAJM/Ex). Animals in the walking group were subjected to treadmill exercise 6 weeks after surgery, which included walking for 18 m/min, 30 min/d, 3 d/wk for 4 weeks. Joint instability was measured by anterior drawer test, and safranin-O staining and immunohistochemical staining were performed. Results OARSI (Osteoarthritis Research Society International) score of ACL-T/Ex group showed highest among 4 groups ( P < 0.001). And CAJM/Ex group was lower than ACL-T/Ex group. Positive cell ratio of IL-1β and MMP-13 in CAJM/Ex group was lower than ACL-T/Ex group ( P < 0.05). Conclusions We found that the state of the intra-articular environment can greatly influence the effect of exercise on cartilage degeneration, even if exercise is performed under the same conditions. In the CAJM/Ex group where joint movement was normalized, abnormal mechanical stress such as shear force and compression force accompanying ACL cutting was alleviated. These findings may highlight the need to consider an intervention to correct abnormal joint movement before prescribing physical exercise in the treatment of OA.

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