Time-sequential fibroblast-to-myofibroblast transition in elastin-variable 3D hydrogel environments by collagen networks

Biomaterials Research

Research Article

Daeho Kim¹  ChaeHo Shin²  Yoon Seo Lee³  Jin Gyeong Son³  Sun Young Lee³  Nhuan T. Do⁴  Se-Hwa Kim⁴  Tae Geol Lee⁵ 

[1] Bruker Nano Surface & Metrology, Bruker Korea, 13493, Seongnam, Republic of Korea;Interdisciplinary Materials Measurement Institute, Korea Research Institute of Standards and Science, 267 Gajeong-Ro, Yuseong-Gu, 34113, Daejeon, Republic of Korea;Nanoconvergence Measurement, University of Science and Technology, 217 Gajeong-Ro, Yuseong-Gu, 34113, Daejeon, Republic of Korea;Safety Measurement Institute, Korea Research Institute of Standards and Science, 267 Gajeong-Ro, Yuseong-Gu, 34113, Daejeon, Republic of Korea;Safety Measurement Institute, Korea Research Institute of Standards and Science, 267 Gajeong-Ro, Yuseong-Gu, 34113, Daejeon, Republic of Korea;BioMedical Measurement, University of Science and Technology, 217 Gajeong-Ro, Yuseong-Gu, 34113, Daejeon, Republic of Korea;Safety Measurement Institute, Korea Research Institute of Standards and Science, 267 Gajeong-Ro, Yuseong-Gu, 34113, Daejeon, Republic of Korea;Nanoconvergence Measurement, University of Science and Technology, 217 Gajeong-Ro, Yuseong-Gu, 34113, Daejeon, Republic of Korea;

关键词: Elastin;    Collagen;    Fibroblast-to-myofibroblast transition;    Calcium signaling;   

DOI  :  10.1186/s40824-023-00439-x

 received in 2023-07-10, accepted in 2023-09-25,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundFibrosis plays an important role in both normal physiological and pathological phenomena as fibroblasts differentiate to myofibroblasts. The activation of fibroblasts is determined through interactions with the surrounding extracellular matrix (ECM). However, how this fibroblast-to-myofibroblast transition (FMT) is regulated and affected by elastin concentration in a three-dimensional (3D) microenvironment has not been investigated.MethodsWe developed an insoluble elastin-gradient 3D hydrogel system for long-lasting cell culture and studied the molecular mechanisms of the FMT in embedded cells by nanoflow LC–MS/MS analysis along with validation through real-time PCR and immunofluorescence staining.ResultsBy optimizing pH and temperature, four 3D hydrogels containing fibroblasts were successfully fabricated having elastin concentrations of 0, 20, 50, and 80% in collagen. At the low elastin level (20%), fibroblast proliferation was significantly increased compared to others, and in particular, the FMT was clearly observed in this condition. Moreover, through mass spectrometry of the hydrogel environment, it was confirmed that differentiation proceeded in two stages. In the early stage, calcium-dependent proteins including calmodulin and S100A4 were highly associated. On the other hand, in the late stage after several passages of cells, distinct markers of myofibroblasts were presented such as morphological changes, increased production of ECM, and increased α-SMA expression. We also demonstrated that the low level of elastin concentration induced some cancer-associated fibroblast (CAF) markers, including PDGFR-β, and fibrosis-related disease markers, including THY-1.ConclusionUsing our developed 3D elastin-gradient hydrogel system, we evaluated the effect of different elastin concentrations on the FMT. The FMT was induced even at a low concentration of elastin with increasing CAF level via calcium signaling. With this system, we were able to analyze varying protein expressions in the overall FMT process over several cellular passages. Our results suggest that the elastin-gradient system employing nonlinear optics imaging provides a good platform to study activated fibroblasts interacting with the microenvironment, where the ECM plays a pivotal role.Graphical Abstract

【 授权许可】

CC BY   
© The Korean Society for Biomaterials 2023

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