POS0133 MONOSODIUM URATE CRYSTALS REDUCE HUMAN LIGAMENT CELLS VIABILITY THROUGH INCREASE OF ROS PRODUCTION

Background:The prevalence of peripheral neuropathy in patients with gout almostly reaches 25%[1]. Demyelination caused by Schwann cell (SCs) injury and apoptosis is the major pathological feature of peripheral[2]. None of study has focused on the effects of monosodium urate (MSU) crystals on SCs.Objectives:To assess the effect of MSU crystals on SCs.Methods:Mouse-derived Schwann cells (RSC96) are stimulated with different concentrations of MSU crystals (0mg/ml,0.25mg/ml,0.5mg/ml) and time (24h,48h,72h). The migration ability of Schwann cells is evaluated by acratch assay, the proliferation level is assessed by the cell counting kit-8 (CCK-8) assay, and the apoptosis rate is detected by flow cytometry.Results:The acratch assay showed that the migration ability of SCs was worsened, CCK-8 assay suggested that proliferation of SCs was reduced in a dose-dependent manner (P<0.05). According to the result of flow cytometry, the survival rate of SCs at 0.5mg/ml(78.60%±2.26%) was lower than that 0.25mg/ml(87.50%±0.95%)and 0mg/ml (98.80%±0.26%)(p<0.05) at 24h. When the stimulation time increased to 72h, the survival rate at 0.5mg/ml(47.90%±11.70%) dropped significantly, which was significantly different from the other two groups(p<0.05).Conclusion:MSU crystals can cause damage to Schwann cells. It may help to explain the reason of peripheral neuropathy in gout patients.References:[1]López-López, C.O., et al., Peripheral neuropathies in rheumatic diseases: More diverse and frequent than expected. A cross-sectional study. International journal of rheumatic diseases, 2020. 23(2): p. 226-232.[2]Liu, Y., S. Shao and H. Guo, Schwann cells apoptosis is induced by high glucose in diabetic peripheral neuropathy. Life sciences, 2020. 248: p. 117459.Figure 1.Flow cytometry assays of RSC96 on MSU crystals at 72hDisclosure of Interests:None declared.

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Detection of intracellular monosodium urate crystals in gout synovial fluid using optical diffraction tomography

Scientific Reports ◽

10.1038/s41598-021-89337-7 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Sangwoo Park ◽

Lucy Eunju Lee ◽

Hanna Kim ◽

Ji Eun Kim ◽

Seung Jun Lee ◽

...

Keyword(s):

Three Dimensional ◽

Time Lapse ◽

Optical Diffraction ◽

Diffraction Tomography ◽

Monosodium Urate ◽

Monosodium Urate Crystals ◽

Optical Diffraction Tomography ◽

Time Lapse Imaging ◽

Msu Crystals ◽

Urate Crystals

AbstractOptical diffraction tomography (ODT) enables imaging of unlabeled intracellular components by measuring the three-dimensional (3D) refractive index (RI). We aimed to detect intracellular monosodium urate (MSU) crystals in synovial leukocytes derived from gout patients using ODT. The 3D RI values of the synthetic MSU crystals, measured by ODT, ranged between 1.383 and 1.440. After adding synthetic MSU crystals to a macrophage, RI tomograms were reconstructed using ODT, and the reconstructed RI tomograms discerned intracellular and extracellular MSU crystals. We observed unlabeled synthetic MSU crystal entry into the cytoplasm of a macrophage through time-lapse imaging. Furthermore, using gout patient-derived synovial leukocytes, we successfully obtained RI tomogram images of intracellular MSU crystals. The 3D RI identification of MSU crystals was verified with birefringence through polarization-sensitive ODT measurements. Together, our results provide evidence that this novel ODT can identify birefringent MSU crystals in synovial leukocytes of patients with gout.

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Raman spectroscopy of monosodium urate crystals: A novel tool for non-invasive gout diagnosis

10.32920/ryerson.14639202 ◽

2021 ◽

Author(s):

Faizan Bilwani

Keyword(s):

Raman Spectroscopy ◽

Sensitivity And Specificity ◽

Diagnostic Tool ◽

False Positives ◽

Monosodium Urate ◽

Monosodium Urate Crystals ◽

Detection Techniques ◽

Non Invasive ◽

Msu Crystals ◽

Urate Crystals

Gout is the most common form of inflammatory arthritis. It results in the deposition of monosodium urate crystals (MSU) at the surface of a joint or the articular cartilage. The gold standard for gout diagnosis is synovial fluid (SF) analysis which requires aspiration of the fluid and subsequent analysis by polarized light microscopy (PLM). This has poor reproducibility, is invasive and requires trained personal to perform the analysis. Raman spectroscopy (RS) has the potential to be a non-invasive diagnostic tool that can detect the presence of MSU crystals. The purpose of the research was to determine whether Raman spectroscopy applied onto the surface of a joint could detect MSU crystals through the skin. Two clinical studies were conducted, entitled the Pilot study (Pilot) and the sons of gout study (SOG). Pilot (n=20) considered ten clinically diagnosed gout sufferers and ten clinically diagnosed Osteoarthritis (OA) patients that acted as control. SOG (n=25) considered nine asymptomatic patients where gout was confirmed by clinicians at time of experiment by ultrasound, and 16 Non-Gouty patients, which did not show any signs of gout. An algorithm was implemented in Matlab® 2016 that removed background florescence, performed denoising and identified the presence or absence of MSU peaks. The comparisons were made against Raman peaks that are known to relate to MSU according to the literature. Three peak combinations, entitled C1, C2 and C3 were evaluated based on their resulting sensitivities and specificities for both studies. C1 was chosen as it provided the highest sensitivity for both studies. Pilot was found to have a sensitivity and specificity of 0.8 and 0.7, respectively. SOG had a sensitivity and specificity of 1 and 0.5, respectively. The results indicated that RS diagnosis is able to achieve good to high sensitivity comparable to other gout detection techniques but a moderate to good specificity. The results also show that RS is fully capable of detecting MSU crystals in-vivo, but results in a high number of false positives, 2 for Pilot and 7 for SOG. However, the false positives in Pilot may be attributed to the control subjects suffering from osteoarthritis, which can be a precursor of gout, and the false positives in the SOG study may be a result of ultrasound (US) being used as the confirmatory diagnostic technique that RS is being compared to. US is known to have sensitivity as low as 0.22 and is operator-dependent. Larger population studies are needed to confirm the ability of RS as a diagnostic tool for detecting gout.

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In Vivo Transmigrated Human Neutrophils Are Highly Primed for Intracellular Radical Production Induced by Monosodium Urate Crystals

International Journal of Molecular Sciences ◽

10.3390/ijms21113750 ◽

2020 ◽

Vol 21 (11) ◽

pp. 3750 ◽

Cited By ~ 1

Author(s):

Lisa Davidsson ◽

Agnes Dahlstrand Rudin ◽

Felix Peter Sanchez Klose ◽

Alicia Buck ◽

Lena Björkman ◽

...

Keyword(s):

Neutrophil Extracellular Trap ◽

Human Neutrophils ◽

Ros Production ◽

Monosodium Urate ◽

Intracellular Compartments ◽

Blood Neutrophils ◽

Msu Crystals ◽

Urate Crystals

Gout is an inflammatory disease caused by monosodium urate (MSU) crystals. The role of neutrophils in gout is less clear, although several studies have shown neutrophil extracellular trap (NET) formation in acutely inflamed joints of gout patients. MSU crystals are known to induce the production of reactive oxygen species (ROS) and NET formation in neutrophils isolated from blood, but there is inconclusive knowledge on the localization of ROS production as well as whether the ROS are required for NET formation. In this report we demonstrate that MSU crystals activate human neutrophils to produce ROS exclusively in intracellular compartments. Additionally, in vivo transmigrated neutrophils derived from experimental skin chambers displayed markedly increased ROS production as compared to resting blood neutrophils. We also confirmed that MSU stimulation potently induced NET formation, but this response was not primed in in vivo transmigrated neutrophils. In line with this we found that MSU-triggered NET formation was independent of ROS production and proceeded normally in neutrophils from patients with dysfunctional respiratory burst (chronic granulomatous disease (CGD) and complete myeloperoxidase (MPO) deficiency). Our data indicate that in vivo transmigrated neutrophils are markedly primed for oxidative responses to MSU crystals and that MSU triggered NET formation is independent of ROS production.

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Raman spectroscopy of monosodium urate crystals: A novel tool for non-invasive gout diagnosis

10.32920/ryerson.14639202.v1 ◽

2021 ◽

Author(s):

Faizan Bilwani

Keyword(s):

Raman Spectroscopy ◽

Sensitivity And Specificity ◽

Diagnostic Tool ◽

False Positives ◽

Monosodium Urate ◽

Monosodium Urate Crystals ◽

Detection Techniques ◽

Non Invasive ◽

Msu Crystals ◽

Urate Crystals

Gout is the most common form of inflammatory arthritis. It results in the deposition of monosodium urate crystals (MSU) at the surface of a joint or the articular cartilage. The gold standard for gout diagnosis is synovial fluid (SF) analysis which requires aspiration of the fluid and subsequent analysis by polarized light microscopy (PLM). This has poor reproducibility, is invasive and requires trained personal to perform the analysis. Raman spectroscopy (RS) has the potential to be a non-invasive diagnostic tool that can detect the presence of MSU crystals. The purpose of the research was to determine whether Raman spectroscopy applied onto the surface of a joint could detect MSU crystals through the skin. Two clinical studies were conducted, entitled the Pilot study (Pilot) and the sons of gout study (SOG). Pilot (n=20) considered ten clinically diagnosed gout sufferers and ten clinically diagnosed Osteoarthritis (OA) patients that acted as control. SOG (n=25) considered nine asymptomatic patients where gout was confirmed by clinicians at time of experiment by ultrasound, and 16 Non-Gouty patients, which did not show any signs of gout. An algorithm was implemented in Matlab® 2016 that removed background florescence, performed denoising and identified the presence or absence of MSU peaks. The comparisons were made against Raman peaks that are known to relate to MSU according to the literature. Three peak combinations, entitled C1, C2 and C3 were evaluated based on their resulting sensitivities and specificities for both studies. C1 was chosen as it provided the highest sensitivity for both studies. Pilot was found to have a sensitivity and specificity of 0.8 and 0.7, respectively. SOG had a sensitivity and specificity of 1 and 0.5, respectively. The results indicated that RS diagnosis is able to achieve good to high sensitivity comparable to other gout detection techniques but a moderate to good specificity. The results also show that RS is fully capable of detecting MSU crystals in-vivo, but results in a high number of false positives, 2 for Pilot and 7 for SOG. However, the false positives in Pilot may be attributed to the control subjects suffering from osteoarthritis, which can be a precursor of gout, and the false positives in the SOG study may be a result of ultrasound (US) being used as the confirmatory diagnostic technique that RS is being compared to. US is known to have sensitivity as low as 0.22 and is operator-dependent. Larger population studies are needed to confirm the ability of RS as a diagnostic tool for detecting gout.

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Management of Vatarakta w.s.r. to Gout through Panchakarma

Journal of Ayurveda and Integrated Medical Sciences (JAIMS) ◽

10.21760/jaims.v3i4.13298 ◽

2018 ◽

Vol 3 (4) ◽

Author(s):

Dr. Chandramohan Arya ◽

Dr.Sanjay Gupta

Keyword(s):

Metabolic Disorders ◽

Molecular Level ◽

Modern Science ◽

Monosodium Urate ◽

Monosodium Urate Crystals ◽

Predominant Symptom ◽

Essential Components ◽

Unique Approach ◽

Ayurvedic Treatment ◽

Urate Crystals

Vatarakta is more distressing and common metabolic disorder prevalent in present era. It is Vatapradhana Tridoshaja Vatavyadhi where Rakta is main Dushya. Vata is a predominant Dosha in Vatarakta, when it is excessively aggravated with vitiated Rakta is called Vatarakta. Vatarakta is correlated with gout in modern science. Gout is an abnormality of purine metabolism causes hyperuricemia and deposition of monosodium urate crystals in joints. Pain is predominant symptom of gout, which disturbs day-today life of the patients. The Panchakarma is not only a important component of Ayurvedic treatment but it is also forms the fundamental basis of Ayurveda therapy. The different procedures like Swedana, Vamana, Virechana, Basti, Sirovirechana, Raktamokshana focus on the purification which detoxifies the human body, i.e. the correction of the metabolism at molecular level. These detoxification procedures are essential components of the management in various metabolic disorders. So Panchakarma is an unique approach in the management of Vatarakta.

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