Substrate properties influence calcification in valvular interstitial cell culture.

摘要: BACKGROUND AND AIM OF THE STUDY Valvular calcification is an active, cell-mediated process that results in significant morbidity and mortality. In standard culture, valvular interstitial cells (VICs) elicit as a result of myofibroblast activation, this limits their use characterization studies. The study aim was to identify culturing substrates would suppress atypical VIC calcification, investigate culture representing more physiological system. METHODS Several platforms were selected compare contrast the influence biochemical mechanical properties on calcification. Substrates investigated included: tissue polystyrene (TCPS), TCPS coated with either fibronectin or fibrin, elastic poly(ethylene glycol) (PEG) hydrogel, also fibrin coupled surface. Experiments repeated profibrotic growth factor transforming factor-beta 1 (TGF-beta1). characterized by calcific nodule formation, alkaline phosphatase activity calcium accumulation. Gene protein expression alpha smooth muscle actin (aSMA) core binding factor-1 (CBFa-1) analyzed qRT-PCR immunostaining. RESULTS Unmodified had innate ability promote markers studied. addition TGF-beta1 enhanced levels all osteoblastic When surfaces modified fibronectin, for repressed, but alphaSMA - marker myofibroblastic unchanged. Meanwhile, fibrin-modified over unmodified substrates. On soft PEG hydrogels, regardless surface chemistry, while remained unaffected. CONCLUSION Collectively, are highly linked microenvironment. Both, environment has effect spontaneous VICs, may have profound molecular properties, related understanding disease vivo.