Decalcification of calcified tissues induced by inorganic polyphosphate in chondrogenic ATDC5 cells in the presence of insulin

Purpose: Inorganic polyphosphate (PolyP), a polymer of orthophosphate, strongly promotes mineralized tissue formation. This study explored the conditions necessary for PolyP to induce calcified deposits in cartilage and assessed the role of insulin in modulating PolyP-induced tissue calcification. Methods: Murine chondrogenic ATDC5 cells were cultured under growth, mineralization, or PolyP-induced calcification conditions, with or without insulin. Calcified nodules were stained with Alizarin Red S, and conditioned media were analyzed for pH and lactate concentration using a pH meter and a lactate assay kit-WST. Results: PolyP treatment of ATDC5 cells led to calcified deposits by day 5, both with and without insulin. However, in the presence of insulin, these deposits were nearly fully decalcified by day 14. Conditioned media with insulin had a lower pH and a higher lactate concentration compared to those without insulin, with lactate levels sufficient to demineralize the PolyP-induced calcified deposits. Conclusion: These data suggest that treatment of ATDC5 chondrogenic cells with PolyP accelerates the formation of mineralized tissue. However, PolyP-induced calcified nodules undergo demineralization owing to lactate production by cells in the presence of insulin.