Time of phenotype onset critical for modelling skeletal effects of T2DM

or 25mM (HG) of glucose.The mRNA expression of several ECM components synthetized by these cells was measured using RT-qPCR.Then BMAd ECMs in LG and HG were devitalized by hypotonic shock for RAMAN spectroscopy characterization and to test osteogenic differentiation of BMSC using mRNA expression.Among tested genes, fibronectin is down-regulated in BMAds compared to BMSCs (-2fold, pb 0.05) and OBs (-5fold, pb 0.05).Western-blot analysis confirms this difference.The devitalization protocol was validated through the absence of nucleic acids and triglycerides while proteins could be detected using a silver nitratestained gel.For a first donor, intensities of Raman signal of type I collagen at 1653cm -1 and 1667cm -1 , and, proline and hydroxyproline are increased in BMAd ECM obtained in HG compared to LG condition.Following seeding on BMAd ECM, BMSC adhesion and proliferation were not modified.Osteogenic differentiation on BMAd ECM obtained in HG shows altered expression of osteogenic markers (such as Runx2, Alkaline Phosphatase and Osteocalcin) and fibronectin compared to differentiation on ECM in LG or on plastic.In summary, BMAds produce a specific ECM which could be modified under HG chronic exposure.Such BMAd ECM could interfere with osteoblast differentiation and mineralization.

Abstract licence: CC BY-NC

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