Extracorporeal Shock Waves Increase Markers of Cellular Proliferation in Bronchial Epithelium and in Primary Bronchial Fibroblasts of COPD Patients.

Abstract

Chronic obstructive pulmonary disease (COPD) is due to structural changes and narrowing of small airways and parenchymal destruction (loss of the alveolar attachment as a result of pulmonary emphysema), which all lead to airflow limitation. Extracorporeal shock waves (ESW) increase cell proliferation and differentiation of connective tissue fibroblasts. To date no studies are available on ESW treatment of human bronchial fibroblasts and epithelial cells from COPD and control subjects. We obtained primary bronchial fibroblasts from bronchial biopsies of 3 patients with mild/moderate COPD and 3 control smokers with normal lung function. 16HBE cells were also studied. Cells were treated with a piezoelectric shock wave generator at low energy (0.3 mJ/mm2, 500 pulses). After treatment, viability was evaluated and cells were recultured and followed up for 4, 24, 48, and 72 h. Cell growth (WST-1 test) was assessed, and proliferation markers were analyzed by qRT-PCR in cell lysates and by ELISA tests in cell supernatants and cell lysates. After ESW treatment, we observed a significant increase of cell proliferation in all cell types. C-Kit (CD117) mRNA was significantly increased in 16HBE cells at 4 h. Protein levels were significantly increased for c-Kit (CD117) at 4 h in 16HBE (p < 0.0001) and at 24 h in COPD-fibroblasts (p = 0.037); for PCNA at 4 h in 16HBE (p = 0.046); for Thy1 (CD90) at 24 and 72 h in CS-fibroblasts (p = 0.031 and p = 0.041); for TGFβ1 at 72 h in CS-fibroblasts (p = 0.038); for procollagen-1 at 4 h in COPD-fibroblasts (p = 0.020); and for NF-κB-p65 at 4 and 24 h in 16HBE (p = 0.015 and p = 0.0002). In the peripheral lung tissue of a representative COPD patient, alveolar type II epithelial cells (TTF-1+) coexpressing c-Kit (CD117) and PCNA were occasionally observed. These data show an increase of cell proliferation induced by a low dosage of extracorporeal shock waves in 16HBE cells and primary bronchial fibroblasts of COPD and control smoking subjects.

• Authors: Antonino Di Stefano, Roberto Frairia, Fabio L M Ricciardolo, Isabella Gnemmi, Antonella Marino Gammazza, Alessio Piraino, Francesco Cappello, Bruno Balbi, Maria Graziella Catalano
• Journal: Canadian respiratory journal (Can Respir J) Vol. 2020 Pg. 1524716 ( 2020) ISSN: 1916-7245 [Electronic] Egypt
• PMID: 32831979 (Publication Type: Journal Article)
• Copyright: Copyright © 2020 Antonino Di Stefano et al.

Chemical References

• Collagen Type I
• Proliferating Cell Nuclear Antigen
• RNA, Messenger
• TGFB1 protein, human
• Transcription Factor RelA
• Transforming Growth Factor beta1
• Proto-Oncogene Proteins c-kit

Topics

• Aged
• Bronchi (cytology)
• Case-Control Studies
• Cell Differentiation (radiation effects)
• Cell Line
• Cell Proliferation (radiation effects)
• Collagen Type I (genetics, metabolism, radiation effects)
• Epithelial Cells (radiation effects)
• Extracorporeal Shockwave Therapy
• Fibroblasts (radiation effects)
• Humans
• Male
• Middle Aged
• Primary Cell Culture
• Proliferating Cell Nuclear Antigen (genetics, metabolism, radiation effects)
• Proto-Oncogene Proteins c-kit (genetics, metabolism, radiation effects)
• Pulmonary Disease, Chronic Obstructive (metabolism, physiopathology)
• RNA, Messenger (metabolism, radiation effects)
• Smokers
• Transcription Factor RelA (genetics, metabolism, radiation effects)
• Transforming Growth Factor beta1 (genetics, metabolism, radiation effects)