Integration of a miniaturized DMMB assay with high-throughput screening for identifying regulators of proteoglycan metabolism was written by Sun, Yi;Tsui, Yuen-kee;Yu, Mengqi;Lyu, Minmin;Cheung, Kenneth;Kao, Richard;Leung, Victor. And the article was included in Scientific Reports in 2022.Reference of 61-73-4 This article mentions the following:
Defective biosynthesis or function of proteoglycans causes pathol. conditions in a variety of tissue systems. Osteoarthritis (OA) is a prevalent degenerative joint disorder characterized by progressive cartilage destruction caused by imbalanced proteoglycan synthesis and degradation Identifying agents that regulate proteoglycan metabolism may benefit the development of OA-modifying therapeutics. High-throughput screening (HTS) of chem. libraries has paved the way for achieving this goal. However, the implementation and adaptation of HTS assays based on proteoglycan measurement remain underexploited. Using primary porcine chondrocytes as a model, we report a miniaturized dimethyl-methylene blue (DMMB) assay, which is commonly used to quant. evaluate sulfated glycosaminoglycan (GAG) content, with an optimized detection range and reproducibility and its integration with HTS. Treatment with TGF-β1 and IL1-α, known as pos. and neg. proteoglycan regulators, resp., supported the assay specificity. A pre-test of chem. screening of 960 compounds identified both stimulators (4.48%) and inhibitors (6.04%) of GAG production Fluorophore-assisted carbohydrate electrophoresis validated the activity of selected hits on chondroitin sulfate expression in an alginate culture system. Our findings support the implementation of this simple colorimetric assay in HTS to discover modifiers of OA or other diseases related to dysregulated proteoglycan metabolism In the experiment, the researchers used many compounds, for example, 3,7-Bis(dimethylamino)phenothiazin-5-ium chloride (cas: 61-73-4Reference of 61-73-4).
3,7-Bis(dimethylamino)phenothiazin-5-ium chloride (cas: 61-73-4) belongs to organic chlorides. Organic chlorides are compounds containing a carbon-chlorine bond, which are widely used in the oil field as a wax dissolver. Alkyl chlorides are versatile building blocks in organic chemistry. While alkyl bromides and iodides are more reactive, alkyl chlorides tend to be less expensive and more readily available.Reference of 61-73-4
Referemce:
Chloride – Wikipedia,
Chlorides – an overview | ScienceDirect Topics