Snake venom-defined fibrin architecture dictates fibroblast survival and differentiation

Abstract

Fibrin is the provisional matrix formed after injury, setting the trajectory for the subsequent stages of wound healing. It is commonly used as a wound sealant and a natural hydrogel for three-dimensional (3D) biophysical studies. However, the traditional thrombin-driven fibrin systems are poorly controlled. Therefore, the precise roles of fibrin’s biophysical properties on fibroblast functions, which underlie healing outcomes, are unknown. Here, we establish a snake venom-controlled fibrin system with precisely and independently tuned architectural and mechanical properties. Employing this defined system, we show that fibrin architecture influences fibroblast survival, spreading phenotype, and differentiation. A fine fibrin architecture is a key prerequisite for fibroblast differentiation, while a coarse architecture induces cell loss and disengages fibroblast’s sensitivity towards TGF-β1. Our results demonstrate that snake venom-controlled fibrin can precisely control fibroblast differentiation. Applying these biophysical principles to fibrin sealants has translational significance in regenerative medicine and tissue engineering.

Hybrid hydrogel-extracellular matrix scaffolds identify distinct ligand and mechanical signatures in cardiac aging

“…While dECM-based biomaterials can provide faithful recapitulation of native ECM composition to some degree, preserving mechanical and architectural aspects of the native tissue remain a challenge. This is crucial because ECM architecture has been shown to regulate cellular phenotypes and organ physiology in recent studies 7,35,44,45 , while ECM mechanics have become increasingly appreciated in the field of cell biology over the past two decades [46][47][48] . Although some dECM-based biomaterial systems can provide tunable stiffness, albeit within a limited range and typically much softer than the original tissue stiffness, the tunability depends on other ECM properties.…”

“…5a-c). Pro-fibrotic CFs increase crosslinking of ECM, which not only directly alters cellular behavior through architecture-mediated mechanisms 7,35 , but also exacerbates myocardial stiffening 3 . In addition, these samples showed significantly downregulated MMPs 2 and 3, which are responsible for normal tissue ECM turnover and degradation, respectively 36 (Fig.…”

“…In healthy tissue, CFs largely remain in a quiescent state, but external stimuli, including biochemical, structural, and mechanical cues [5][6][7] , are able to activate quiescent CFs, leading to their differentiation into a proto-myofibroblast phenotype and subsequently into a mature myofibroblast phenotype when these stimuli are impactful and persistent 6,8 . The process of CF activation and proper myofibroblast maturation are essential for ECM deposition and the maintenance of matrix homeostasis but can also lead to fibrosis and result in functional consequences 9 .…”