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

Detection Techniques ◽

Non Invasive ◽

Msu 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.

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 ◽

Optical Diffraction Tomography ◽

Time Lapse Imaging ◽

Msu 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.

Evaluation of Diff Quik Staining Method for Detection of Mono Sodium Urate Crystals in Synovial Fluid Using Light and Polarizing Microscopy: A Cross-Sectional Study

Bangladesh Journal of Medical Microbiology ◽

10.3329/bjmm.v12i2.51693 ◽

2018 ◽

Vol 12 (2) ◽

pp. 10-13

Author(s):

Ina Rahim ◽

Jamal Pasha Chowdhury

Keyword(s):

Synovial Fluid ◽

Light Microscopy ◽

Sensitivity And Specificity ◽

Cross Sectional Study ◽

Sectional Study ◽

Monosodium Urate ◽

Polarizing Microscopy ◽

Cross Sectional ◽

Msu Crystals ◽

Gout is one of the oldest diseases of the medical history that results from the deposition of monosodium urate crystals in joint structures and in periarticular sites in the form of tophi. The evaluation of synovial fluid is integral for the diagnosis of gout and other arthritis of microcrystals. The objective of this cross-sectional study was to evaluate the analysis of synovial fluid by polarizing and light microscopy using wet film and Diff Quik stained films and evaluating usefulness of Diff Quik stain in identifying monosodium urate (MSU) crystals on permanent mounted slides. It was conducted on 100 clinically suspected gout patients in the Department of Clinical Pathology, Bangabandhu Sheikh Mujib Medical University, Dhaka from May 2008 to April 2009. Polarizing Microscopy detected the presence of MSU crystals in 35.71% cases from wet films and 36.73% cases from Diff Quik stained films. Light Microscopy detected crystals in 28 (28.57%) cases from Diff Quik stained samples and in 31.63% cases from wet film samples. Considering wet film polarizing microscopy as gold standard, the sensitivity and specificity of wet film light microscopy was 88.6% and 100.0% respectively, whereas sensitivity and specificity were 80.0% and 100.0% respectively in Diff Quik light microscopy.In Diff Quik polarizing microscopy, sensitivity and specificity were100.0% and 98.4% respectively. The sensitivity and specificity was highest in Diff Quik stained films examined by polarizing microscopy Bangladesh J Med Microbiol 2018; 12 (2): 10-13

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

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-eular.2316 ◽

2021 ◽

Vol 80 (Suppl 1) ◽

pp. 278.1-278

Author(s):

Y. Huang ◽

Y. Liu ◽

Q. Huang ◽

W. Deng ◽

T. W. Li

Keyword(s):

Cell Migration ◽

Annexin V ◽

Cell Counting ◽

Ros Production ◽

Monosodium Urate ◽

Migration Ability ◽

Migration Assay ◽

Msu Crystals ◽

Background:Ligament destruction is a frequent complication of gout and is strongly associated with tophi. Ligament fibroblasts are important cellular mediators of ligament remodeling. None of study has paid attention to the effects of monosodium urate (MSU) crystals on ligament fibroblasts.Objectives:The study aims to investigate the effects and mechanism of MSU crystals on ligament fibroblasts.Methods:MSU crystals were added to human ligament fibroblasts(HLFs) cultures or primary ligament cells cultures. Cell counting kit-8 (CCK-8) assay, cell migration assay, Annexin V-FITC/PI assay were conducted. Reactive Oxygen Species(ROS) was tested by ROS Assay Kit.Results:The higher concentrations of MSU crystals (0.5-1mg/mL) reduced the viability of HLFs or primary ligament cells after 24 h as assessed by CCK8 assays, with a further reduction in viability observed at the 48 h time point. When observed under light microscopy, HLFs cultured with MSU crystals (0.5mg/mL) appeared unhealthy with fewer cells present. The cell migration ability of HLFs was decreased significantly on MSU crystals (0.5mg/mL). According to the result of Annexin V-FITC/PI assay, the survival rate of HLFs on MSU crystals (0.5mg/mL) was lower than that of 0.25mg/ml and 0 mg/ml at 72h. ROS assay results showed that the production of ROS increased as the concentrations of MSU crystals increased.Conclusion:MSU crystals inhibit human ligament cells viability through the increase of ROS production. It may contribute to disordered ligament remodeling in gout patients with ligament destruction.References:[1]Ashika Chhana, et al. Monosodium urate crystals reduce osteocyte viability and indirectly promote a shift in osteocyte function towards a proinflammatory and proresorptive state. Arthritis Res Ther. 2018, 20(1): 208.Figure 1.MSU crystals reduce human ligament fibroblasts and primary human ligament cells viability over time. A: CCK-8 assay; B: Observation of HLFs morphology; C: Annexin V-FITC/PI assay.Disclosure of Interests:None declared

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 ◽

Predominant Symptom ◽

Essential Components ◽

Unique Approach ◽

Ayurvedic Treatment ◽

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.