engleski

Biological effects of three newly synthesized compounds on the cell cultures from the patients affected by Dupuytren’ s contracture

Dupuytren’ s contracture (DC) is a palmar fibromatosis having an obscure etiology. It is characterized by nodular fibroblastic proliferation and accumulation of myofibroblasts. So far, the only successful treatment was surgical excision or incision of diseased fascia, often accompanied by high recurrence rates. Experimental chemotherapy, such as 5-fluorouracil (5-FU), proved to be useful in acting locally in the palm by inhibiting myofibroblasts proliferation. 5-FU was shown to inhibit both, proliferation and myofibroblast differentiation in DC’ s cell cultures. We have therefore established primary cell cultures as a model for studying the effects of 3 newly synthesized chemical compounds on proliferation and differentiation of Dupuytren’ s fibroblasts/myofibrobalsts. Subsequently, cells were characterized by several proliferation assays and the cell cycle distribution was determined using flow cytometry.We showed cells from the affected part (D) of fascia to proliferate faster then the cells from non-affected part (ND). Also, the percentage of cells in the S-phase of the cell cycle was higher in the D than in the ND cells. The presence of myofibroblasts in culture was ascertained by immunocytochemistry and Western blot method, as we assed the quantity of α -smooth actin as a molecular marker for differentiated fibroblasts. We have further tested cytotoxic and antiproliferatve effects of 3 compounds (a N-sulfonylpyrimidine derivative, an amidino substituted benzimidazo(1, 2-a)quinoline derivative and an amido-substituted dithieno-kinolone derivative) by MTT-test and showed them all to have a marked antiproliferative effect on established cell lines. Their mechanisms of action were further studied in parallel experiments by cell cycle analyses, immunocitochemistry and Western blots. Obtained results implicated that tested compounds exerted different but rather specific effects related to the progression through the cell cycle and differentiation of fibroblasts into myofibroblasts.

Dupuytren's contracture; antiproliferative effect; primary cell cultures

nije evidentirano