Abstract:Calcium pyrophosphate deposition disease, previously known as pseudogout, is a type of chronic and painful joint arthropathy. Accurate identification of calcium pyrophosphate dihydrate (CPPD) single crystals is crucial for determining...
“…1 c). The crystals of CPPDs were prepared as previously described 15 (see Supplementary Page S2 online). Figure 1 Crystal characteristics.…”
Section: Resultsmentioning
confidence: 99%
“…This problem was initially indicated back in 1986 by Schumacher et al 11 and several other studies proposed for quality control of synovial fluid analyses 12 , 13 . Thus far, various strategies have been developed to improve the sensitivity of MSUM and CPPD detection, including the use of Fv antibodies 14 , fluorescent probes 15 , nanostructured TiO 2 material 16 , X-ray dark field radiography 17 , optical diffraction tomography 18 , lens-free polarized microscopy 19 and dual energy computed tomography 20 . These methods provide high sensitivity for gouty crystal detection, but they require specific reagents and sophisticated instruments.…”
Conventional polarized light microscopy has been widely used to detect gouty crystals, but its limited sensitivity increases the risk of misidentification. In this study, a number of methods were investigated to improve the sensitivity of polarized light microscopy for the detection of monosodium urate monohydrate (MSUM) and calcium pyrophosphate dihydrate (CPPD) crystals. We found that coating glass slides with poly-L-lysine, a positively charged polymer, improved the attachment of crystals to the glass surface, resulting in clearer crystal images compared to non-coated slides. Additionally, the sensitivity of detection was further enhanced by selective dissolution, in which 40% v/v formalin phosphate buffer was employed to dissolve MSUM crystals but not CPPD while 10% ethylenediamine tetraacetic acid (EDTA) was employed to dissolved CPPD but not MSUM. The other possible interferences were dissolved in both EDTA and formalin solution. These methods were successfully applied to detect gouty crystals in biological milieu, including spiked porcine synovial fluid and inflamed rat subcutaneous air pouch tissues.
“…1 c). The crystals of CPPDs were prepared as previously described 15 (see Supplementary Page S2 online). Figure 1 Crystal characteristics.…”
Section: Resultsmentioning
confidence: 99%
“…This problem was initially indicated back in 1986 by Schumacher et al 11 and several other studies proposed for quality control of synovial fluid analyses 12 , 13 . Thus far, various strategies have been developed to improve the sensitivity of MSUM and CPPD detection, including the use of Fv antibodies 14 , fluorescent probes 15 , nanostructured TiO 2 material 16 , X-ray dark field radiography 17 , optical diffraction tomography 18 , lens-free polarized microscopy 19 and dual energy computed tomography 20 . These methods provide high sensitivity for gouty crystal detection, but they require specific reagents and sophisticated instruments.…”
Conventional polarized light microscopy has been widely used to detect gouty crystals, but its limited sensitivity increases the risk of misidentification. In this study, a number of methods were investigated to improve the sensitivity of polarized light microscopy for the detection of monosodium urate monohydrate (MSUM) and calcium pyrophosphate dihydrate (CPPD) crystals. We found that coating glass slides with poly-L-lysine, a positively charged polymer, improved the attachment of crystals to the glass surface, resulting in clearer crystal images compared to non-coated slides. Additionally, the sensitivity of detection was further enhanced by selective dissolution, in which 40% v/v formalin phosphate buffer was employed to dissolve MSUM crystals but not CPPD while 10% ethylenediamine tetraacetic acid (EDTA) was employed to dissolved CPPD but not MSUM. The other possible interferences were dissolved in both EDTA and formalin solution. These methods were successfully applied to detect gouty crystals in biological milieu, including spiked porcine synovial fluid and inflamed rat subcutaneous air pouch tissues.
“…Therefore, the measurement of ATP hydrolysis by monitoring its hydrolyzed product, inorganic pyrophosphate (PPi), would be a simpler and more accurate strategy for quantifying ENPP1 activity, without complications due to chemical derivatization and allosteric modulation. For this purpose, sensitive and real-time detection of PPi can be achieved by using Kyoto Green (xanthene dipicolylamine Zn(II) complex), a molecular fluorescent sensor extensively employed for the detection of PPi under physiological conditions and in synovial fluids 7 , 23 – 26 . However, to our knowledge, Kyoto Green has not been leveraged for screening of ENPP1 inhibitors.…”
Ecto-nucleotide pyrophosphatases/phosphodiesterases 1 (ENPP1) is a key enzyme in purinergic signaling pathways responsible for cell-to-cell communications and regulation of several fundamental pathophysiological processes. In this study, Kyoto Green, a rapid chemical sensor of pyrophosphate, was employed to screen for effective ENPP1 inhibitors among five representative flavonoids (quercetin, myricetin, morin, kaempferol, and quercetin-3-glucoside), five nucleosides (adenosine, guanosine, inosine, uridine, and cytidine), and five deoxynucleosides (2′- and 3′-deoxyadenosine, 2′-deoxyguanosine, 2′-deoxyinosine, and 2′-deoxyuridine). Conventional colorimetric, fluorescence, and bioluminescence assays revealed that ENPP1 was effectively inhibited by quercetin (Ki ~ 4 nM) and myricetin (Ki ~ 32 nM) when ATP was used as a substrate at pH 7.4. In silico analysis indicated that the presence of a chromone scaffold, particularly one containing a hydroxyl group at the 3′ position on the B ring, may promote binding to the active site pocket of ENPP1 and enhance inhibition. This study demonstrated that the naturally derived quercetin and myricetin could effectively inhibit ENPP1 enzymatic activity and may offer health benefits in arthritis management.
Biomimicking the chemical, mechanical, and topographical properties of bone on an implant model is crucial to obtain rapid and effective osteointegration, especially for the large‐area fractures of the skeletal system. Titanium‐based biomaterials are more frequently preferred in clinical use in such cases and coating these materials with oxide layers having chemical/nanotopographic properties to enhance osteointegration and implantation success rates has been studied for a long time. The objective of this study is to examine the high and rapid mineralization potential of anodized aluminum oxide (AAO) coated and atomic layer deposition (ALD)‐alumina coated titanium substrates on large deformation areas with difficult spontaneous healing. AAO‐coated titanium (AAO@Ti) substrates were fabricated via anodization technique in different electrolytes and their osteogenic potential was analyzed by comparing them to the bare titanium surface as a control. In order to investigate the effect of the ionic characters gained by the surfaces through anodization, the oxidized nanotopographic substrates were additionally coated with an ultrathin alumina layer via ALD (ALD@AAO@Ti), which is a sensitive and conformal coating vapor deposition technique. Besides, a bare titanium sample was also coated with pure alumina by ALD (ALD@Ti) to investigate the effect of nanoscale surface morphology. XPS analysis after ALD coating showed that the ionic character of each surface fabricated by anodization was successfully suppressed. In vitro studies demonstrated that, among the substrates investigated, the mineralization capacity of MG‐63 osteosarcoma cells were highest when incubated on ALD‐treated and bare AAO@Ti samples that were anodized in phosphoric acid (H3PO4_AAO@Ti and ALD@H3PO4_AAO@Ti). Mineralization on these substrates also increased consistently beginning from day 2 to day 21. Moreover, immunocytochemistry for osteopontin (OPN) demonstrated the highest expression for ALD@H3PO4_AAO@Ti, followed by the H3PO4_AAO@Ti sample. Consequently, it was observed that, although ALD treatment improves cellular characteristics on all samples, effective mineralization requires more than a simple ALD coating or the presence of a nanostructured topography. Overall, ALD@H3PO4_AAO@Ti substrates can be considered as an implant alternative with its enhanced osteogenic differentiation potential and rapid mineralization capacity.
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