A Novel In Vitro Disease Model for Systemic Sclerosis Using Polyisocyanide Hydrogels

Abstract

Systemic sclerosis (SSc) is a rare autoimmune disease with limited treatment options that is characterized by fibrosis in various organs. To screen the effectiveness of new therapies, there is an urgent need for reliable in vitro models. Key is that diseased cells' characteristics are maintained, which is challenging in currently used setups. In this study, an in vitro 3D culture system is described using the biocompatible polyisocyanide (PIC‐RGD) hydrogel and SSc patient‐derived fibroblasts from affected (lesional cells) and from healthy‐skin (healthy cells). In contrast to the standard collagen‐coated 2D cultures, the cells in the 3D PIC‐RGD gels maintain the native phenotype and functionality of the primary cells. The functionality of the model is studied in the presence of the fibrosis stimulator transforming growth factor β1 (TGFβ1) and the suppressor tumor necrosis factor (TNFα). In this study, it is observed that lesional cells have a stronger fibrotic character with increased contraction, proliferation, and expression of collagen, and myofibroblast markers α‐smooth muscle actin and fibroblast activation protein. The high tunability of the hydrogel, which can maintain the native functionality of fibroblasts in in vitro cultures, delivers a crucial step in developing these materials into an effective tool for personalized medicine approaches of SSc patients.

Conductive Polyisocyanide Hydrogels Inhibit Fibrosis and Promote Myogenesis

“…The fraction of viable cells was calculated from the stained images. The intensity of the green (alive) and red (dead) channels was used using eq , % o f .25em v i a b l e .25em c e l l s = i n t e n s i t y .25em l i v e .25em c e l l s i n t e n s i t y ( normall normali normalv normale + normald normale normala normald ) c e l l s × 100 % …”

“…The fraction of viable cells was calculated from the stained images. The intensity of the green (alive) and red (dead) channels was used using eq 3 ( 35 , 36 ) …”

Repeated Application and Removal of Polyisocyanopeptide Hydrogel Wound Dressings in a Splinted Full-Thickness Wound Model

“…Triethylene glycol-grafted PIC polymers (Radboud University, Nijmegen, the Netherlands) were synthesised and prepared as described in detail previously [ 9 , 10 , 15 , 18 , 32 ]. Synthesis of DTPA-functionalized PIC polymers (purchased from Noviocell BV, Oss, The Netherlands) and radioactive labelling with 111 In was performed as described previously [ 10 ].…”

An Improved Understanding of the Pathophysiology of Pelvic Organ Prolapse: A 3D In Vitro Model under Static and Mechanical Loading Conditions

“…The highly biomimetic PIC hydrogel is an ideal material to investigate the differences between 3D cultured POP and non‐POP fibroblasts because it is able to maintain the native phenotype and functionality of these fibroblasts. [ 25 ] Moreover, the PIC hydrogel has the same fibrous architecture and mechanical properties, including strain‐stiffening behavior in a biologically relevant stress regime, as native ECM components. [ 26 ] In the mechanical loading conditions, we mimic intermittent and continuous loading exerted on the pelvic floor by exposing the 3D in vitro model to two different straining protocols.…”