title: Pirfenidone and nintedanib modulate properties of fibroblasts and myofibroblasts in idiopathic pulmonary fibrosis.
abstract: BACKGROUND
  Idiopathic pulmonary fibrosis (IPF) is an incurable lung disease with a poor prognosis. Fibroblasts and myofibroblasts are the key cells in the fibrotic process. Recently two drugs, pirfenidone and nintedanib, were approved for clinical use as they are able to slow down the disease progression. The mechanisms by which these two drugs act in in vitro cell systems are not known. The aim of this study was therefore to examine the effects of pirfenidone and nintedanib on fibroblasts and myofibroblasts structure and function established from patients with or without IPF.
  
  
  METHODS
  Stromal cells were collected and cultured from control lung (n = 4) or IPF (n = 7). The cells were treated with pirfenidone and/or nintedanib and the effect of treatment was evaluated by measuring cell proliferation, alpha smooth muscle actin (α-SMA) and fibronectin expression by Western analysis and/or immunoelectron microscopy, ultrastructural properties by transmission electron microscopy and functional properties by collagen gel contraction and invasion assays.
  
  
  RESULTS
  Both pirfenidone and nintedanib reduced in vitro proliferation of fibroblastic cells in a dose dependent manner. The number of cells from control lung was reduced to 47 % (p = 0.04) and of IPF cells to 42 % (p = 0.04) by 1 mM pirfenidone and correspondingly to 67 % (p = 0.04) and 68 % (p = 0.04), by 1 μM nintedanib. If both drugs were used together, a further reduced proliferation was observed. Both pirfenidone and nintedanib were able to reduce the amount of α-SMA and the myofibroblastic appearance although the level of reduction was cell line dependent. In functional assays, the effect of both drugs was also variable.
  
  
  CONCLUSIONS
  We conclude that the ultrastructure and function of fibroblasts and myofibroblasts are affected by pirfenidone and nintedanib. Combination of the drugs reduced cell proliferation more than either of them individually. Human lung derived cell culture systems represent a potential platform for screening and testing drugs for fibrotic diseases.
authors: Department of Anatomy and Cell Biology, Cancer and Translational Medicine Research Unit, University of Oulu, Aapistie 7 A, FIN-90 220, Oulu, Finland. siri.lehtonen@oulu.fi.|Department of Internal Medicine, Respiratory Research Unit and Medical Research Center, Oulu University Hospital, Aapistie 5 A, FIN-90220, Oulu, Finland. siri.lehtonen@oulu.fi.
authors: Siri T
authors: ST
authors: Lehtonen
authors: Department of Internal Medicine, Respiratory Research Unit and Medical Research Center, Oulu University Hospital, Aapistie 5 A, FIN-90220, Oulu, Finland. anniveijo@windowslive.com.
authors: Anniina
authors: A
authors: Veijola
authors: Department of Internal Medicine, Respiratory Research Unit and Medical Research Center, Oulu University Hospital, Aapistie 5 A, FIN-90220, Oulu, Finland. Henna.Karvonen@oulu.fi.|Laboratory of Tissue Repair and Regeneration, Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, 150 College Street, Toronto, ON, M5S 3E2, Canada. Henna.Karvonen@oulu.fi.
authors: Henna
authors: H
authors: Karvonen
authors: Department of Pathology, Oulu University Hospital, P.O. Box 50, FIN90029, Oulu, Finland. elisa.lappi-blanco@ppshp.fi.|Department of Pathology, Cancer and Translational Medicine Research Unit, University of Oulu, Aapistie 5 B, FIN-90220, Oulu, Finland. elisa.lappi-blanco@ppshp.fi.
authors: Elisa
authors: E
authors: Lappi-Blanco
authors: Department of Pathology, Oulu University Hospital, P.O. Box 50, FIN90029, Oulu, Finland. raija.sormunen@oulu.fi.|Department of Pathology, Cancer and Translational Medicine Research Unit, University of Oulu, Aapistie 5 B, FIN-90220, Oulu, Finland. raija.sormunen@oulu.fi.|Biocenter Oulu, University of Oulu, Aapistie 5 A, FIN-90220, Oulu, Finland. raija.sormunen@oulu.fi.
authors: Raija
authors: R
authors: Sormunen
authors: Department of Anatomy and Cell Biology, Cancer and Translational Medicine Research Unit, University of Oulu, Aapistie 7 A, FIN-90 220, Oulu, Finland. o3kosa01@students.oamk.fi.|Department of Internal Medicine, Respiratory Research Unit and Medical Research Center, Oulu University Hospital, Aapistie 5 A, FIN-90220, Oulu, Finland. o3kosa01@students.oamk.fi.
authors: Saara
authors: S
authors: Korpela
authors: Department of Medical Epidemiology & Biostatistics, Karolinska Institutet, SE-17177, Stockholm, Sweden. Ulrika.Zagai@ki.se.
authors: Ulrika
authors: U
authors: Zagai
authors: Respiratory Medicine Unit, Department of Medicine Solna and Centre for Molecular Medicine, Karolinska Institutet, SE-17177, Stockholm, Sweden. Magnus.Skold@ki.se.
authors: Magnus C
authors: MC
authors: Sköld
authors: Department of Internal Medicine, Respiratory Research Unit and Medical Research Center, Oulu University Hospital, Aapistie 5 A, FIN-90220, Oulu, Finland. Riitta.Kaarteenaho@kuh.fi.|Research Unit of Internal Medicine, Respiratory Research Unit and Medical Research Center, University of Oulu, Aapistie 5 A, FIN-90220, Oulu, Finland. Riitta.Kaarteenaho@kuh.fi.|Unit of Medicine and Clinical Research, Pulmonary Division, University of Eastern Finland, Kuopio, Finland. Riitta.Kaarteenaho@kuh.fi.|Center for Medicine and Clinical Research, Division of Respiratory Medicine, Kuopio University Hospital, Kuopio, Finland. Riitta.Kaarteenaho@kuh.fi.
authors: Riitta
authors: R
authors: Kaarteenaho
mesh_terms: D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004359:Drug Therapy, Combination; D005347:Fibroblasts / Q000187:drug effects* / Q000473:pathology*; D006801:Humans; D054990:Idiopathic Pulmonary Fibrosis / Q000188:drug therapy* / Q000473:pathology*; D007211:Indoles / Q000008:administration & dosage*; D058628:Myofibroblasts / Q000187:drug effects / Q000473:pathology; D011728:Pyridones / Q000008:administration & dosage*; D016896:Treatment Outcome
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D007211:Indoles; D011728:Pyridones; C093844:pirfenidone; C530716:nintedanib