Enzymatic formation of modular cell-instructive fibrin analogs for tissue engineering

Ehrbar, Martin; Rizzi, Simone C.; Hlushchuk, Ruslan; Djonov, Valentin; Zisch, Andreas H.; Hubbell, Jeffrey A.; Weber, Franz E.; Lütolf, Matthias P.

doi:10.1016/j.biomaterials.2007.03.027

Cited by 206 publications

(244 citation statements)

References 37 publications

Supporting

Mentioning

240

Contrasting

Order By: Relevance

“…This result suggested that a cleavage between the Ala-46 and the Cys-47 occurred upon the electrochemical desorption of Fg. Additionally, in this 2.7 kDa fragment we found the α-chain polymerization site EA Gly-Pro-Arg-Val (36)(37)(38)(39). Finally, we confirmed that the peak at 1536.7 Da is the FPA (Supporting Information Table S3 and Figure S5).…”

Section: Eqcm: Readsorption Of Electrochemically Desorbed Fg Revealedsupporting

confidence: 72%

“…It is important to mention that interfacial pH changes affect the structure and function of other blood coagulation factors not studied in this work. As most of enzymes, thrombin and factor XIII show pH-dependent activity, [37,38] and we recently demonstrated that the activity of factor XIII is indeed changed close to the electrode surface. [39] We thus consider the electrochemical degradation of Fg and the consequent loss of Fg clottability to be an important mechanism explaining the reduced coagulation at the cathode, but not the only one.…”

Section: Possible Mechanisms Leading To the Electrochemical Desorptiomentioning

confidence: 94%

See 1 more Smart Citation

Coagulation at the Blood–Electrode Interface: The Role of Electrochemical Desorption and Degradation of Fibrinogen

et al. 2014

View full text Add to dashboard Cite

The influence of electrochemistry on the coagulation of blood on metal surfaces was demonstrated several decades ago. In particular, the application of cathodic currents resulted in reduced surface thrombogenicity, but no molecular mechanism has been so far proposed to explain this observation. In this article we used for the first time the quartz crystal microbalance with dissipation monitoring technique coupled with an electrochemical setup (EQCM-D) to study thrombosis at the blood-electrode interface. We confirmed the reduced thrombus deposition at the cathode, and we subsequently studied the effect of cathodic currents on adsorbed fibrinogen (Fg). Using EQCM and mass spectrometry, we found that upon applying currents Fg desorbed from the electrode and was electrochemically degraded. In particular, we show that the flexible N-terminus of the -chain, containing an important polymerization site, was cleaved from the protein, thus affecting its clottability. Our work proposes a molecular mechanism that at least partially explains how cathodic currents reduce thrombosis at the blood-electrode interface and is a relevant contribution to the rational development of medical devices with reduced thrombus formation on their surface. ABSTRACTThe influence of electrochemistry on the coagulation of blood on metal surfaces was demonstrated several decades ago. In particular, the application of cathodic currents resulted in reduced surface thrombogenicity, but no molecular mechanism has been so far proposed to explain this observation. In this article we used for the first time the quartz crystal microbalance with dissipation monitoring technique coupled with an electrochemical setup (EQCM-D) to study thrombosis at the blood-electrode interface. We confirmed the reduced thrombus deposition at the cathode, and we subsequently studied the effect of cathodic currents on adsorbed fibrinogen (Fg). Using EQCM and mass spectrometry, we found that upon applying currents Fg desorbed from the electrode and was electrochemically degraded. In particular, we show that the flexible N-terminus of the α-chain, containing an important polymerization site, was cleaved from the protein, thus affecting its clottability. Our work proposes a molecular mechanism that at least partially explains how cathodic currents reduce thrombosis at the blood-electrode interface and is a relevant contribution to the rational development of medical devices with reduced thrombus formation on their surface.2

show abstract

Section: Eqcm: Readsorption Of Electrochemically Desorbed Fg Revealedsupporting

confidence: 72%

Section: Possible Mechanisms Leading To the Electrochemical Desorptiomentioning

confidence: 94%

Coagulation at the Blood–Electrode Interface: The Role of Electrochemical Desorption and Degradation of Fibrinogen

et al. 2014

View full text Add to dashboard Cite

show abstract

“…To investigate this function, we incorporated Fg β15-66 (2) into a multifunctional poly(ethylene glycol) (PEG) matrix designed to mimic fibrin's main functions. The fibrin-mimetic matrix backbone comprised two eight-arm PEG peptide conjugates (at a concentration of 1.75%) crosslinkable by the fibrin stabilizing factor, transglutaminase factor XIIIa (16,17). PEG was conjugated with a peptide containing a factor XIIIa substrate sequence derived from α 2 -plasmin inhibitor (α 2 PI 1-8 , NQEQVSPL) (18) or with a lysine donor peptide containing a substrate sequence for matrix metalloproteases (MMPs) and plasmin (VPMSMRGG) (19), allowing degradation of the matrix in situ.…”

Section: Resultsmentioning

confidence: 99%

Heparin-binding domain of fibrin(ogen) binds growth factors and promotes tissue repair when incorporated within a synthetic matrix

Martino

Briquez

Ranga

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

417

366

View full text Add to dashboard Cite

By binding growth factors (GFs), the ECM tightly regulates their activity. We recently reported that the heparin-binding domain II of fibronectin acts as a promiscuous high-affinity GF-binding domain. Here we hypothesized that fibrin, the provisional ECM during tissue repair, also could be highly promiscuous in its GF-binding capacity. Using multiple affinity-based assays, we found that fibrin(ogen) and its heparin-binding domain bind several GFs from the PDGF/VEGF and FGF families and some GFs from the TGF-β and neurotrophin families. Overall, we identified 15 unique binding interactions. The GF binding ability of fibrinogen caused prolonged retention of many of the identified GFs within fibrin. Thus, based on the promiscuous and high-affinity interactions in fibrin, GF binding may be one of fibrin’s main physiological functions, and these interactions may potentially play an important and ubiquitous role during tissue repair. To prove this role in a gain-of-function model, we incorporated the heparin-binding domain of fibrin into a synthetic fibrin-mimetic matrix. In vivo, the multifunctional synthetic matrix could fully mimic the effect of fibrin in a diabetic mouse model of impaired wound healing, demonstrating the benefits of generating a hybrid biomaterial consisting of a synthetic polymeric scaffold and recombinant bioactive ECM domains. The reproduction of GF–ECM interactions with a fibrin-mimetic matrix could be clinically useful, and has the significant benefit of a more straightforward regulatory path associated with chemical synthesis rather than human sourcing.

show abstract

“…Numerous methods have been developed to produce PEG hydrogels through covalent crosslinking of PEG prepolymer; free radical polymerization of PEG acrylates, Michael-type addition, enzymatic reaction, and radiation. [32][33][34][35][36][37][38][39] Although PEG displays many desirable features as a biomaterial, the inability for cells to degrade, remodel, and attach to PEG make it difficult to implement as a hydrogel for tissue engineering, where such characteristics are critical. The lack of bioactivity of PEG has spurred the development of functionalized PEG polymers, where peptide sequences are attached to PEG acrylate groups or polymerized to 4-arm PEG using Michael-type addition.…”

Section: Figmentioning

confidence: 99%

Synthesis and Characterization of Tunable Poly(Ethylene Glycol): Gelatin Methacrylate Composite Hydrogels

Hutson

Nichol

Aubin

et al. 2011

Tissue Engineering Part A

278

290

View full text Add to dashboard Cite

Poly(ethylene glycol) (PEG) hydrogels are popular for cell culture and tissue-engineering applications because they are nontoxic and exhibit favorable hydration and nutrient transport properties. However, cells cannot adhere to, remodel, proliferate within, or degrade PEG hydrogels. Methacrylated gelatin (GelMA), derived from denatured collagen, yields an enzymatically degradable, photocrosslinkable hydrogel that cells can degrade, adhere to and spread within. To combine the desirable features of each of these materials we synthesized PEGGelMA composite hydrogels, hypothesizing that copolymerization would enable adjustable cell binding, mechanical, and degradation properties. The addition of GelMA to PEG resulted in a composite hydrogel that exhibited tunable mechanical and biological profiles. Adding GelMA (5%-15% w/v) to PEG (5% and 10% w/v) proportionally increased fibroblast surface binding and spreading as compared to PEG hydrogels ( p < 0.05). Encapsulated fibroblasts were also able to form 3D cellular networks 7 days after photoencapsulation only within composite hydrogels as compared to PEG alone. Additionally, PEG-GelMA hydrogels displayed tunable enzymatic degradation and stiffness profiles. PEG-GelMA composite hydrogels show great promise as tunable, cell-responsive hydrogels for 3D cell culture and regenerative medicine applications.

show abstract

Enzymatic formation of modular cell-instructive fibrin analogs for tissue engineering

Cited by 206 publications

References 37 publications

Coagulation at the Blood–Electrode Interface: The Role of Electrochemical Desorption and Degradation of Fibrinogen

Coagulation at the Blood–Electrode Interface: The Role of Electrochemical Desorption and Degradation of Fibrinogen

Heparin-binding domain of fibrin(ogen) binds growth factors and promotes tissue repair when incorporated within a synthetic matrix

Synthesis and Characterization of Tunable Poly(Ethylene Glycol): Gelatin Methacrylate Composite Hydrogels

Contact Info

Product

Resources

About