| Popis: |
Idiopathic pulmonary fibrosis (IPF) is a devastating lung disease associated with heterogeneous and abnormal build-up of extracellular matrix (ECM) proteins and increased fibroblast activity. We hypothesize that the alteration in material properties of lung tissue is one of the driving forces in the disease, where ECM stiffening has a fundamental impact on cellular responses. Our aim is to find a correlation between structural alterations of the ECM and cellular mechanisms associated with disease pathology such as increased proliferation, ECM synthesis and mechanotransduction. Thinly sliced distal tissue (350µm x 1cm2) from healthy (n=4) respective IPF (n=4) subjects was decellularized using a detergent-based protocol to produce acellular scaffolds. The scaffolds were characterized with histology, SEM, mass spectrometry, measurements of DNA content, tissue density and tensile strength. Next, the scaffolds were mounted in a holder, seeded with primary human fibroblasts from healthy donors and cultured for 9 days. Metabolic activity was measured after 1, 3, 6, and 9 days, with collection of histology samples at day 1 and 9. DNA content and tensile strength were measured after 9 days. The IPF-derived scaffolds displayed denser morphology with higher matrix stiffness in comparison to healthy tissue scaffolds. Mass spectrometric characterization of scaffolds separated IPF and healthy scaffolds into two cluster groups. Repopulated scaffolds displayed increased fibroblast proliferation and tissue stiffness over time. Further evaluations are performed to clarify the regulation of mechanotransduction and its association to pathological characteristics of the IPF matrisome. |
