Versatile lysine dendrigrafts and polyethylene glycol hydrogels with inherent biological properties: in vitro cell behavior modulation and in vivo biocompatibility
Mariana Carrancá
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Laboratory for Materials Engineering and Science, CNRS INSA, Villeurbanne, France
Search for more papers by this authorLouise Griveau
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Laboratory for Materials Engineering and Science, CNRS INSA, Villeurbanne, France
Search for more papers by this authorNoëlle Remoué
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorChloé Lorion
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorPierre Weiss
INSERM, Laboratory of Osteo-Articlular and Dental Engineering, Nantes, France
Search for more papers by this authorValérie Orea
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorDominique Sigaudo-Roussel
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorDaniel Ferri-Angulo
Laboratory for Materials Engineering and Science, CNRS INSA, Villeurbanne, France
Search for more papers by this authorRomain Debret
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorCorresponding Author
Jérôme Sohier
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Laboratory for Materials Engineering and Science, CNRS INSA, Villeurbanne, France
Correspondence
Jérôme Sohier, CNRS INSA, UMR 5510, Laboratory for Materials Engineering and Science, Bat. B. Pascal, 7 Avenue Jean Capelle 69621, Villeurbanne Cedex, France.
Email: [email protected]
Search for more papers by this authorMariana Carrancá
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Laboratory for Materials Engineering and Science, CNRS INSA, Villeurbanne, France
Search for more papers by this authorLouise Griveau
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Laboratory for Materials Engineering and Science, CNRS INSA, Villeurbanne, France
Search for more papers by this authorNoëlle Remoué
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorChloé Lorion
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorPierre Weiss
INSERM, Laboratory of Osteo-Articlular and Dental Engineering, Nantes, France
Search for more papers by this authorValérie Orea
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorDominique Sigaudo-Roussel
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorDaniel Ferri-Angulo
Laboratory for Materials Engineering and Science, CNRS INSA, Villeurbanne, France
Search for more papers by this authorRomain Debret
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Search for more papers by this authorCorresponding Author
Jérôme Sohier
Laboratory of Tissue Biology and Therapeutic Engineering, IBCP, CNRS Université, Lyon, France
Laboratory for Materials Engineering and Science, CNRS INSA, Villeurbanne, France
Correspondence
Jérôme Sohier, CNRS INSA, UMR 5510, Laboratory for Materials Engineering and Science, Bat. B. Pascal, 7 Avenue Jean Capelle 69621, Villeurbanne Cedex, France.
Email: [email protected]
Search for more papers by this authorPresent address: Clément Faye, GLPBiocontrol, Bat CAP ALPHA, Clapiers, France
Funding information: Agence Nationale de la Recherche, Grant/Award Number: TECSAN 016-01; Consejo Nacional de Ciencia y Tecnología; Région Auvergne-Rhône-Alpes, Grant/Award Number: grant 17 002601 ARC 2016
Abstract
Poly(ethylene glycol) (PEG) hydrogels have been extensively used as scaffolds for tissue engineering applications, owing to their biocompatibility, chemical versatility, and tunable mechanical properties. However, their bio-inert properties require them to be associated with additional functional moieties to interact with cells. To circumvent this need, we propose here to reticulate PEG molecules with poly(L-lysine) dendrigrafts (DGL) to provide intrinsic cell functionalities to PEG-based hydrogels. The physico-chemical characteristics of the resulting hydrogels were studied in regard of the concentration of each component. With increasing amounts of DGL, the cross-linking time and swelling ratio could be decreased, conversely to mechanical properties, which could be tailored from 7.7 ± 0.7 to 90 ± 28.8 kPa. Furthermore, fibroblasts adhesion, viability, and morphology on hydrogels were then assessed. While cell adhesion significantly increased with the concentration of DGL, cell viability was dependant of the ratio of DGL and PEG. Cell morphology and proliferation; however, appeared mainly related to the overall hydrogel rigidity. To allow cell infiltration and cell growth in 3D, the hydrogels were rendered porous. The biocompatibility of resulting hydrogels of different compositions and porosities was evaluated by 3 week subcutaneous implantations in mice. Hydrogels allowed an extensive cellular infiltration with a mild foreign body reaction, histological evidence of hydrogel degradation, and neovascularization.
CONFLICTS OF INTEREST
The authors declare no conflict of interest.
Supporting Information
| Filename | Description |
|---|---|
| jbma37083-sup-0001-FigureS1.zipapplication/x-zip-compressed, 1.4 MB | Supplementary Figure 1 Subcutaneous implantation of dense DGL/PEG hydrogel (2/19 mM DGL/PEG). Masson's trichrome staining of the full explants and close-ups highlighting the hydrogel (#), the fibrous capsule (*), macrophages (+). |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1Tibbitt MW, Anseth KS. Hydrogels as extracellular matrix mimics for 3D cell culture. Biotechol Bioeng. 2009; 103: 655-663.
- 2Jia X, Kiick KL. Hybrid multicomponent hydrogels for tissue engineering. Macromol Biosci. 2009; 9: 140-156.
- 3Park S, Park K. Engineered polymeric hydrogels for 3D tissue models. Polymers. 2016; 8: 23.
- 4Figueiredo L, Pace R, D'Arros C, et al. Assessing glucose and oxygen diffusion in hydrogels for the rational design of 3D stem cell scaffolds in regenerative medicine. J Tissue Eng Regen Med. 2018; 12: 1238-1246.
- 5Van Vlierberghe S, Dubruel P, Schacht E. Biopolymer-based hydrogels as scaffolds for tissue engineering applications: a review. Biomacromolecules [Internet. 2011; 12: 1387-1408. Available from. https://doi.org/10.1021/bm200083n.
- 6Franz S, Rammelt S, Scharnweber D, Simon JC. Immune responses to implants—a review of the implications for the design of immunomodulatory biomaterials. Biomaterials. 2011; 32: 6692-6709. Available from:. https://doi.org/10.1016/j.biomaterials.2011.05.078.
- 7Zhu J. Bioactive modification of poly(ethylene glycol) hydrogels for tissue engineering. Biomaterials. 2010; 31: 4639-4656.
- 8Spicer CD. Hydrogel scaffolds for tissue engineering: the importance of polymer choice. Polym Chem Royal Society of Chemistry. 2020; 11: 184-219.
- 9Lin C, Anseth KS. PEG hydrogels for the controlled release of biomolecules in regenerative medicine. Pharm Res. 2009; 26: 631-643.
- 10Sargeant TD, Desai AP, Banerjee S, Agawu A, Stopek JB. An in situ forming collagen—PEG hydrogel for tissue regeneration. Acta Materialia Inc. 2012; 8: 124-132. https://doi.org/10.1016/j.actbio.2011.07.028.
- 11Barros D, Conde-sousa E, Gonçalves AM, et al. Engineering hydrogels with affinity-bound laminin as 3D neural stem cell culture systems. Biomater Sci Royal Soc Chem. 2019; 7: 5338-5349.
- 12Hern DL, Hubbell JA. Incorporation of adhesion peptides into nonadhesive hydrogels useful for tissue resurfacing. J Biomed Mater Res. 1998; 39: 266-276.
10.1002/(SICI)1097-4636(199802)39:2<266::AID-JBM14>3.0.CO;2-B CAS PubMed Web of Science® Google Scholar
- 13Ouyang L, Dan Y, Shao Z, et al. MMP—sensitive PEG hydrogel modified with RGD promotes bFGF, VEGF and EPC - mediated angiogenesis. Exp Ther Med. 2019; 18: 2933-2941.
- 14Burdick JA, Anseth KS. Photoencapsulation of osteoblasts in injectable RGD-modified PEG hydrogels for bone tissue engineering. 2002; 23: 4315-4323.
- 15Nemir S, West L. J. Synthetic materials in the study of cell response to substrate rigidity. Ann Biomed Eng. 2010; 38: 2-20.
- 16Papavasiliou G, Sokic S, Turturro M. Synthetic PEG Hydrogels as Extracellular Matrix Mimics for Tissue Engineering Applications, Biotechnology - Molecular Studies and Novel Applications for Improved Quality of Human Life, Reda Helmy Sammour, IntechOpen. https://doi.org/10.5772/31695. Available from: https://www.intechopen.com/books/biotechnology-molecular-studies-and-novel-applications-for-improved-quality-of-human-life/synthetic-peg-hydrogels-as-extracellular-matrix-mimics-for-tissue-engineering-applications.
10.5772/31695 Google Scholar
- 17Rizzi SC, Hubbell JA. Recombinant protein-co-PEG networks as cell-adhesive and Proteolytically degradable hydrogel matrixes. Part I: development and physicochemical characteristics. Biomacromolecules. 2005; 6: 1226-1238.
- 18Wathier M, Jung PJ, Carnahan MA, Kim T, Grinstaff MW. Dendritic Macromers as in situ polymerizing biomaterials for securing cataract incisions. J Am Chem Soc. 2004; 126: 12744-12745.
- 19Wathier M, Johnson CS, Kim T, Grinstaff MW. Hydrogels formed by multiple peptide ligation reactions to fasten corneal transplants. Bioconjugate Chem. 2006; 17: 873-876.
- 20Kaga S, Arslan M, Sanyal R, Sanyal A. Dendrimers and Dendrons as versatile building blocks for the fabrication of functional hydrogels. Molecules. 2016; 21(4): 497. https://doi.org/10.3390/molecules21040497.
- 21Oliveira JM, Salgado AJ, Sousa N, Mano JF, Reis RL. Dendrimers and derivatives as a potential therapeutic tool in regenerative medicine strategies—a review. Prog Polym Sci [internet]. Elsevier. 2010; 35: 1163-1194.
- 22N. Desai P, Yuan Q, Yang H. Synthesis and characterization of Photocurable Polyamidoamine dendrimer hydrogels as a versatile platform for tissue engineering and drug delivery. Biomacromolecules 2011; 11: 666–73.
- 23Navath RS, Menjoge AR, Dai H, Romero R, Kannan S, Kannan RM. Injectable PAMAM dendrimer - PEG hydrogels for the treatment of genital infections: formulation and in vitro and in vivo evaluation. Mol Pharmaceutics. 2011; 8: 1209-1223.
- 24Unal B, Hedden RC. Gelation and swelling behavior of end-linked hydrogels prepared from linear poly (ethylene glycol) and poly (amidoamine) dendrimers. Polymer. 2006; 47: 8173-8182.
- 25Labieniec-Watala M, Watala C. PAMAM dendrimers: destined for success or doomed to fail? Plain and modified PAMAM dendrimers in the context of biomedical applications. J pharm Sci [internet]. Elsevier Masson SAS. 2015; 104: 2-14. https://doi.org/10.1002/jps.24222.
- 26Romestand B, Rolland J, Commeyras A, Desvignes I, Pascal R, Vandenabeele-trambouze O. Dendrigraft poly- L -lysine: a non-immunogenic synthetic carrier for antibody production. Biomacromolecules. 2010; 11: 1169-1173.
- 27Francoia J, Vial L. Everything you always wanted to know about poly-l-lysine Dendrigrafts (but were afraid to ask). Chem—Eur J. 2018; 24: 1-10.
- 28Huang R, Liu S, Shao K, et al. Evaluation and mechanism studies of PEGylated dendrigraft poly-L-lysines as novel gene delivery vectors. Nanotechnology. 2010; 21:265101.
- 29Lorion C, Faye C, Maret B, et al. Biosynthetic support based on dendritic poly(L-lysine) improves human skin fibroblasts attachment. J Biomater Sci, Polym Ed [Internet. 2014; 25: 136-149.
- 30Collet H, Souaid E, Cottet H, et al. An expeditious multigram-scale synthesis of lysine dendrigraft (DGL) polymers by aqueous n-carboxyanhydride polycondensation. Chem—A Eur J. 2010; 16: 2309-2316.
- 31Maret B, Crépet A, Faye C, Garrelly L, Ladavière C. Molar-mass analysis of dendrigraft poly(L-lysine) (DGL) polyelectrolytes by SEC-MALLS: the “cornerstone” refractive index increment. Macromol Chem Phys. 2015; 216: 95-105.
- 32Schneider CA, Rasband WS, Eliceiri KW. NIH image to ImageJ: 25 years of image analysis. Nat Methods. Nature Publishing Group. 2012; 9: 671-675.
- 33Ma PX, Choi PDJ. Biodegradable polymer scaffolds with well-defined interconnected spherical pore network. Tissue Eng. 2001; 7: 23-33.
- 34Anseth KS, Bowman CN, Brannon-Peppas L. Mechanical properties of hydrogels and their experimental determination. Biomaterials. 1996; 17: 1647-1657.
- 35Coussot G, Nicol E, Commeryras A, Desvignes I, Pascal R, Vandenabeele-trambouze O. Colorimetric quantification of amino groups in linear and dendritic structures. Polym Int. 2009; 58: 511-518.
- 36Wang Y, Zhao Q, Zhang H, Yang S, Jia X. A novel poly(amido amine)-dendrimer-based hydrogel as a mimic for the extracellular matrix. Adv Mater. 2014; 26: 4163-4167.
- 37Gitsov I, Zhu C. Amphiphilic hydrogels constructed by poly(ethylene glycol) and shape-persistent dendritic fragments. Macromolecules. 2002; 35: 8418-8427.
- 38Quinton MP, Philpott CWA. Role for anionic sites in epithelial architecture: effects of cationic polymers on cell membrane structure. J Cell Biol. 1978; 56: 787-796.
10.1083/jcb.56.3.787 Google Scholar
- 39Tang M, Dong H, Li Y, Ren T. Harnessing the PEG-cleavable strategy to balance cytotoxicity, intracellular release and the therapeutic effect of dendrigraft poly-L-lysine for cancer gene therapy. J Mater Chem B. 2016; 4: 1284-1295.
- 40Jiang G, Huang AH, Cai Y, Tanase M, Sheetz MP. Rigidity sensing at the leading edge through avb3 Integrins and RPTPa. Biophys J. 2006; 90: 1804-1809.
- 41Yeung T, Georges PC, Flanagan LA, et al. Effects of substrate stiffness on cell morphology, cytoskeletal structure, and adhesion. Cell Motil Cytoskeleton. 2005; 60: 24-34.
- 42Ghosh K, Pan Z, Guan E, et al. Cell adaptation to a physiologically relevant ECM mimic with different viscoelastic properties. Biomaterials. 2007; 28: 671-679.
- 43Lo CM, Wang HB, Dembo M, Wang YL. Cell movement is guided by the rigidity of the substrate. Biophys J. 2000; 79: 144-152.
- 44Solon J, Levental I, Sengupta K, Georges PC, Janmey PA. Fibroblast adaptation and stiffness matching to soft elastic substrates. Biophys J. 2007; 93: 4453-4461.
- 45Kono T, Tanii T, Furukawa M, et al. Cell cycle analysis of human dermal fibroblasts cultured on or in hydrated type I collagen lattices. Arch Dermatol Res. 1990; 282: 258-262.
- 46Rhudy RW, McPherson JM. Influence of the extracellular matrix on the proliferative response of human skin fibroblasts to serum and purified platelet-derived growth factor. J Cell Physiol. 1988; 137: 185-191.
- 47Sarber R, Hull B, Merrill C, Sorrano T, Bell E. Regulation of proliferation of fibroblasts of low and high population doubling levels grown in collagen lattices. Mech Ageing Dev. 1981; 17: 107-117.
- 48Lynn AD, Kyriakides TR, Bryant SJ. Characterization of the in vitro macrophage response and in vivo host response to poly(ethylene glycol)-based hydrogels. J Biomed Mater Res—Part A. 2010; 93: 941-953.
- 49Griffon DJ, Sedighi MR, Schaeffer DV, Eurell JA, Jonhnson AL. Chitosan scaffolds: interconnective pore size and cartilage engineering. Acta Biomater. 2006; 2: 313-320.
- 50Rouwkema J, Rivron NC, Van BCA. Vascularization in tissue engineering. Trends Biotechnol. 2008; 26: 434-441.
- 51Annabi N, Nichol JW, Zhong X, et al. Controlling the porosity and microarchitecture of hydrogels for tissue engineering. Tissue Eng Part B Rev. 2010; 16: 371-383.
Citing Literature
