Different Proliferative Capacity of Lung Fibroblasts Obtained From Control Subjects and Patients With Emphysema

Abstract

To characterize the possible role of a dysregulated proliferative capacity of pulmonary fibroblasts in insufficient tissue repair in lungs from patients with pulmonary emphysema, the authors undertook in vitro proliferative studies with pulmonary fibroblasts obtained from lung tissue of patients with emphysema. A comparison was made with fibroblasts from control subjects. The authors determined the in vitro proliferative capacity of fibroblasts at basal culture conditions and after modulation with interleukin-1beta, interferon-gamma, transforming growth factor-beta(1), and basic fibroblast growth factor. Proliferative capacity was determined by measurement of 5-bromo-2-deoxyuridine (BrdU) incorporation. BrdU incorporation by fibroblast cultures from both groups was very similar. Fibroblast cultures from control subjects, however, incorporated more BrdU after incubation with interleukin-1beta than cultures from patients with emphysema (P<.05). On the other hand, transforming growth factor-beta(1) decreased incorporation of BrdU stronger in fibroblast cultures from control subjects than from patients with emphysema (P<.05). Thus, the proliferative capacity of fibroblast cultures isolated from lung tissue of patients with pulmonary emphysema is different from that of control subjects. Although the difference is small, it may be an essential contribution to the development of pulmonary emphysema that only occurs after repeated smoke-induced injury over many years of an individual's life.

In contrast to lung fibroblasts – no signs of senescence in skin fibroblasts of patients with emphysema

“…In line with this, lung fibroblasts from patients with emphysema exhibit a reduced proliferation rate, lower number of population doublings in serial passaging (Holz et al 2004), and altered growth factor response (Holz et al 2004;Noordhoek et al 2003). In addition, we found an increased staining for senescence-associated beta-galactosidase (β-gal, (Dimri et al 1995)), as well as increased mRNA expression of the insulin-like growth factor-binding proteins (IGFBP)-3 and -rP1 in these cells (Müller et al 2006).…”

High ICAM-1 gene expression in pulmonary fibroblasts of COPD patients: a reflection of an enhanced immunological function

“…BUCKLEY et al [18] suggested that this chronic inflammation occurs because of disordered fibroblast behaviour, which leads to inappropriate survival and retention of leukocytes within inflamed tissue [38]. Thus it is not surprising that fibroblasts in COPD tissue show a functionally altered behaviour under the influence of the local cytokine microenvironment [11,18]. The differentially expressed adhesion-related genes found in GOLD stage IV COPD fibroblasts when stimulated with CD8 supernatant supports the theory of a prominent role of fibroblasts in the chronic inflammatory process observed in COPD.…”

“…Absence of mycoplasma contamination in the fibroblast cultures was confirmed with a mycoplasma detection kit (Roche Diagnostics, Almere, the Netherlands). Isolated cells were characterised as fibroblasts by morphological appearance and expression pattern of specific proteins [11]. All cells exhibited a characteristic staining pattern for vimentin, fibronectin, and the fibroblast marker prolyl-4-hydroxylase and lacked immunoreactivity for keratin.…”

“…Isolation and culture of lung fibroblasts Pulmonary fibroblast cultures were established from parenchymal lung tissue by means of an explant technique as described previously [11]. Absence of mycoplasma contamination in the fibroblast cultures was confirmed with a mycoplasma detection kit (Roche Diagnostics, Almere, the Netherlands).…”

“…The current authors have previously observed that lung fibroblasts of patients with severe COPD exhibit an altered ECM production profile compared with fibroblasts of controls pointing to an important role of fibroblasts in the defective ECM modulation observed in COPD [8,[10][11][12].…”