Artificial cell membrane binding thrombin constructs drive in situ fibrin hydrogel formation

Robert C. Deller; Thomas Richardson; Rebecca Richardson; Laura Bevan; Ioannis Zampetakis; Fabrizio Scarpa; Adam W. Perriman

doi:10.1038/s41467-019-09763-0

journal article Open Access Apr 23, 2019

Artificial cell membrane binding thrombin constructs drive in situ fibrin hydrogel formation

Robert C. Deller Thomas Richardson

Rebecca Richardson

Laura Bevan Ioannis Zampetakis Fabrizio Scarpa

Adam W. Perriman

Nature Communications Vol. 10 No. 1 · Springer Science and Business Media LLC

View at Publisher Save 10.1038/s41467-019-09763-0

Abstract

AbstractCell membrane re-engineering is emerging as a powerful tool for the development of next generation cell therapies, as it allows the user to augment therapeutic cells to provide additional functionalities, such as homing, adhesion or hypoxia resistance. To date, however, there are few examples where the plasma membrane is re-engineered to display active enzymes that promote extracellular matrix protein assembly. Here, we report on a self-contained matrix-forming system where the membrane of human mesenchymal stem cells is modified to display a novel thrombin construct, giving rise to spontaneous fibrin hydrogel nucleation and growth at near human plasma concentrations of fibrinogen. The cell membrane modification process is realised through the synthesis of a membrane-binding supercationic thrombin-polymer surfactant complex. Significantly, the resulting robust cellular fibrin hydrogel constructs can be differentiated down osteogenic and adipogenic lineages, giving rise to self-supporting monoliths that exhibit Young’s moduli that reflect their respective extracellular matrix compositions.

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References

53

[1]

Karp, J. M. & Leng Teo, G. S. Mesenchymal stem cell homing: the devil is in the details. Cell Stem Cell 4, 206–216 (2009). 10.1016/j.stem.2009.02.001

[2]

Discher, D. E., Janmey, P. & Wang, Y. L. Tissue cells feel and respond to the stiffness of their substrate. Science 310, 1139–1143 (2005). 10.1126/science.1116995

[3]

Discher, D. E., Mooney, D. J. & Zandstra, P. W. Growth factors, matrices, and forces combine and control stem cells. Science 324, 1673–1677 (2009). 10.1126/science.1171643

[4]

Kim, T. G., Shin, H. & Lim, D. W. Biomimetic Scaffolds for Tissue Engineering. Adv. Funct. Mater. 22, 2446–2468 (2012). 10.1002/adfm.201103083

[5]

Shin, H., Jo, S. & Mikos, A. G. Biomimetic materials for tissue engineering. Biomaterials 24, 4353–4364 (2003). 10.1016/s0142-9612(03)00339-9

[6]

Drury, J. L. & Mooney, D. J. Hydrogels for tissue engineering: scaffold design variables and applications. Biomaterials 24, 4337–4351 (2003). 10.1016/s0142-9612(03)00340-5

[7]

Ahmed, T. A., Dare, E. V. & Hincke, M. Fibrin: a versatile scaffold for tissue engineering applications. Tissue Eng. Part B Rev. 14, 199–215 (2008). 10.1089/ten.teb.2007.0435

[8]

Tennent, G. A. et al. Human plasma fibrinogen is synthesized in the liver. Blood 109, 1971–1974 (2007). 10.1182/blood-2006-08-040956

[9]

Kim, I. L., Khetan, S., Baker, B. M., Chen, C. S. & Burdick, J. A. Fibrous hyaluronic acid hydrogels that direct MSC chondrogenesis through mechanical and adhesive cues. Biomaterials 34, 5571–5580 (2013). 10.1016/j.biomaterials.2013.04.004

[10]

Gailit, J. et al. Human fibroblasts bind directly to fibrinogen at RGD sites through integrin alpha(v)beta3. Exp. Cell Res. 232, 118–126 (1997). 10.1006/excr.1997.3512

[11]

Martino, M. M., Briquez, P. S., Ranga, A., Lutolf, M. P. & Hubbell, J. A. Heparin-binding domain of fibrin(ogen) binds growth factors and promotes tissue repair when incorporated within a synthetic matrix. Proc. Natl Acad. Sci. USA 110, 4563–4568 (2013). 10.1073/pnas.1221602110

[12]

El-Sherbiny, I. M. & Yacoub, M. H. Hydrogel scaffolds for tissue engineering: progress and challenges. Glob. Cardiol. Sci. Pr. 2013, 316–342 (2013).

[13]

Aguado, B. A., Mulyasasmita, W., Su, J., Lampe, K. J. & Heilshorn, S. C. Improving viability of stem cells during syringe needle flow through the design of hydrogel cell carriers. Tissue Eng. Part A 18, 806–815 (2012). 10.1089/ten.tea.2011.0391

[14]

Abbina S., Siren E. M. J., Moon H., Kizhakkedathu J. N. Surface engineering for cell-based therapies: techniques for manipulating mammalian cell surfaces. ACS Biomater. Sci. Eng. 4, 3658–3677 (2017). 10.1021/acsbiomaterials.7b00514

[15]

Wu, P. J., Lilly, J. L., Arreaza, R. & Berron, B. J. Hydrogel patches on live cells through surface-mediated polymerization. Langmuir 33, 6778–6784 (2017). 10.1021/acs.langmuir.7b01139

[16]

Cruise, G. M., Hegre, O. D., Scharp, D. S. & Hubbell, J. A. A sensitivity study of the key parameters in the interfacial photopolymerization of poly(ethylene glycol) diacrylate upon porcine islets. Biotechnol. Bioeng. 57, 655–665 (1998). 10.1002/(sici)1097-0290(19980320)57:6<655::aid-bit3>3.0.co;2-k

[17]

Armstrong, J. P. et al. Artificial membrane-binding proteins stimulate oxygenation of stem cells during engineering of large cartilage tissue. Nat. Commun. 6, 7405 (2015). 10.1038/ncomms8405

[18]

Liska, D. J. & Suttie, J. W. Location of gamma-carboxyglutamyl residues in partially carboxylated prothrombin preparations. Biochemistry 27, 8636–8641 (1988). 10.1021/bi00423a019

[19]

Mizuochi, T., Yamashita, K., Fujikawa, K., Kisiel, W. & Kobata, A. The carbohydrate of bovine prothrombin. Occurrence of Gal beta 1 leads to 3GlcNAc grouping in asparagine-linked sugar chains. J. Biol. Chem. 254, 6419–6425 (1979). 10.1016/s0021-9258(18)50383-5

[20]

Engh, R. A. et al. Enzyme flexibility, solvent and ‘weak’ interactions characterize thrombin-ligand interactions: implications for drug design. Structure 4, 1353–1362 (1996). 10.1016/s0969-2126(96)00142-6

[21]

Supercharging Proteins Can Impart Unusual Resilience

Michael S. Lawrence, Kevin J. Phillips, David R. Liu

Journal of the American Chemical Society 2007 10.1021/ja071641y

[22]

Pesce, D., Wu, Y., Kolbe, A., Weil, T. & Herrmann, A. Enhancing cellular uptake of GFP via unfolded supercharged protein tags. Biomaterials 34, 4360–4367 (2013). 10.1016/j.biomaterials.2013.02.038

[23]

McNaughton, B. R., Cronican, J. J., Thompson, D. B. & Liu, D. R. Mammalian cell penetration, siRNA transfection, and DNA transfection by supercharged proteins. Proc. Natl Acad. Sci. USA 106, 6111–6116 (2009). 10.1073/pnas.0807883106

[24]

Lai, W. F. In vivo nucleic acid delivery with PEI and its derivatives: current status and perspectives. Expert Rev. Med Devices 8, 173–185 (2011). 10.1586/erd.10.83

[25]

Futami, J. et al. Intracellular delivery of proteins into mammalian living cells by polyethylenimine-cationization. J. Biosci. Bioeng. 99, 95–103 (2005). 10.1263/jbb.99.95

[26]

Brogan, A. P., Sharma, K. P., Perriman, A. W. & Mann, S. Enzyme activity in liquid lipase melts as a step towards solvent-free biology at 150 degrees C. Nat. Commun. 5, 5058 (2014). 10.1038/ncomms6058

[27]

Perriman, A. W. et al. Reversible dioxygen binding in solvent-free liquid myoglobin. Nat. Chem. 2, 622–626 (2010). 10.1038/nchem.700

[28]

Brogan, A. P., Sessions, R. B., Perriman, A. W. & Mann, S. Molecular dynamics simulations reveal a dielectric-responsive coronal structure in protein-polymer surfactant hybrid nanoconstructs. J. Am. Chem. Soc. 136, 16824–16831 (2014). 10.1021/ja507592b

[29]

Duong, H., Wu, B. & Tawil, B. Modulation of 3D fibrin matrix stiffness by intrinsic fibrinogen-thrombin compositions and by extrinsic cellular activity. Tissue Eng. Part A 15, 1865–1876 (2009). 10.1089/ten.tea.2008.0319

[30]

Pittenger, M. F. et al. Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–147 (1999). 10.1126/science.284.5411.143

[31]

Cory, A. H., Owen, T. C., Barltrop, J. A. & Cory, J. G. Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. Cancer Commun. 3, 207–212 (1991). 10.3727/095535491820873191

[32]

Xiao, Y., Lubin, A. A., Heeger, A. J. & Plaxco, K. W. Label-free electronic detection of thrombin in blood serum by using an aptamer-based sensor. Angew. Chem. Int. Ed. 44, 5456–5459 (2005). 10.1002/anie.200500989

[33]

Smith, J. D., Chen, A., Ernst, L. A., Waggoner, A. S. & Campbell, P. G. Immobilization of aprotinin to fibrinogen as a novel method for controlling degradation of fibrin gels. Bioconjug Chem. 18, 695–701 (2007). 10.1021/bc060265o

[34]

Rosen, E. D. & MacDougald, O. A. Adipocyte differentiation from the inside out. Nat. Rev. Mol. Cell Biol. 7, 885–896 (2006). 10.1038/nrm2066

[35]

Frith, J. & Genever, P. Transcriptional control of mesenchymal stem cell differentiation. Transfus. Med Hemother 35, 216–227 (2008). 10.1159/000127448

[36]

Loebel, C. et al. In vitro osteogenic potential of human mesenchymal stem cells is predicted by Runx2/Sox9 ratio. Tissue Eng. Part A 21, 115–123 (2015). 10.1089/ten.tea.2014.0096

[37]

Uccelli, A., Moretta, L. & Pistoia, V. Mesenchymal stem cells in health and disease. Nat. Rev. Immunol. 8, 726–736 (2008). 10.1038/nri2395

[38]

Ramirez-Zacarias, J. L., Castro-Munozledo, F. & Kuri-Harcuch, W. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with Oil red O. Histochemistry 97, 493–497 (1992). 10.1007/bf00316069

[39]

Gregory, C. A., Gunn, W. G., Peister, A. & Prockop, D. J. An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. Anal. Biochem 329, 77–84 (2004). 10.1016/j.ab.2004.02.002

[40]

Chen, J. et al. Enhanced osteogenesis of human mesenchymal stem cells by periodic heat shock in self-assembling peptide hydrogel. Tissue Eng. Part A 19, 716–728 (2013). 10.1089/ten.tea.2012.0070

[41]

Parekh, S. H. et al. Modulus-driven differentiation of marrow stromal cells in 3D scaffolds that is independent of myosin-based cytoskeletal tension. Biomaterials 32, 2256–2264 (2011). 10.1016/j.biomaterials.2010.11.065

[42]

Park, J. S. et al. The effect of matrix stiffness on the differentiation of mesenchymal stem cells in response to TGF-beta. Biomaterials 32, 3921–3930 (2011). 10.1016/j.biomaterials.2011.02.019

[43]

Chatterjee, K. et al. The effect of 3D hydrogel scaffold modulus on osteoblast differentiation and mineralization revealed by combinatorial screening. Biomaterials 31, 5051–5062 (2010). 10.1016/j.biomaterials.2010.03.024

[44]

Bialorucki, C., Subramanian, G., Elsaadany, M. & Yildirim-Ayan, E. In situ osteoblast mineralization mediates post-injection mechanical properties of osteoconductive material. J. Mech. Behav. Biomed. Mater. 38, 143–153 (2014). 10.1016/j.jmbbm.2014.06.018

[45]

Schultz, K. M., Kyburz, K. A. & Anseth, K. S. Measuring dynamic cell-material interactions and remodeling during 3D human mesenchymal stem cell migration in hydrogels. Proc. Natl Acad. Sci. USA 112, E3757–E3764 (2015). 10.1073/pnas.1511304112

[46]

Weisel, J. W. & Litvinov, R. I. Fibrin formation, structure and properties. Sub-Cell. Biochem. 82, 405–456 (2017). 10.1007/978-3-319-49674-0_13

[47]

Radosevich, M., Goubran, H. A. & Burnouf, T. Fibrin sealant: scientific rationale, production methods, properties, and current clinical use. Vox Sang. 72, 133–143 (1997). 10.1159/000461980

[48]

Sander, V., Sune, G., Jopling, C., Morera, C. & Izpisua Belmonte, J. C. Isolation and in vitro culture of primary cardiomyocytes from adult zebrafish hearts. Nat. Protoc. 8, 800–809 (2013). 10.1038/nprot.2013.041

[49]

Weyand, A. C. & Shavit, J. A. Zebrafish as a model system for the study of hemostasis and thrombosis. Curr. Opin. Hematol. 21, 418–422 (2014). 10.1097/moh.0000000000000075

[50]

Liu, Y. et al. Targeted mutagenesis of zebrafish antithrombin III triggers disseminated intravascular coagulation and thrombosis, revealing insight into function. Blood 124, 142–150 (2014). 10.1182/blood-2014-03-561027

Showing 50 of 53 references

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Published: Apr 23, 2019
Vol/Issue: 10(1)
License: View

Authors

Bristol Composites Institute (ACCIS), University of Bristol, BS8 1TR Bristol, U.K.

A

Adam W. Perriman

Cite This Article

Robert C. Deller, Thomas Richardson, Rebecca Richardson, et al. (2019). Artificial cell membrane binding thrombin constructs drive in situ fibrin hydrogel formation. Nature Communications, 10(1). https://doi.org/10.1038/s41467-019-09763-0

Artificial cell membrane binding thrombin constructs drive in situ fibrin hydrogel formation

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