Gelatin-based biomaterials with tailorable mechanical properties as promising matrices for soft-tissue replacement


Gelatin-based biomaterials with tailorable mechanical properties as promising matrices for soft-tissue replacement

Ullm, S.; Tondera, C.; Gebauer, T.; Neffe, A. T.; Lendlein, A.; Pietzsch, J.

Objectives
Gelatin-based hydrogels are promising degradable materials for soft tissue regeneration. Our approach aims at biopolymer-based polymer networks with tailorable elastic properties and degradation behavior due to different degrees of crosslinking with lysine diisocyanate ethyl ester. Two gelatin-based hydrogel films were compared regarding their influence on vitality, adhesion and proinflammatory activation of both endothelial cells and human macrophages, to investigate representative cells being responsible for tissue integration and degradation of the material.
Materials & methods
10 wt% gelatin solutions were crosslinked with three- (G10LNCO3) or eight-fold (G10LNCO8) excess of isocyanate groups, resulting in hydrogels with tailored Young’s moduli (13 and 55 kPa), swelling (1200 and 350 vol%) and degradation time [1]. For experiments on cell vitality, human leukemia HL-60 cells differentiated to macrophages (MΦ), and primary human aortic endothelial cells (HAEC) were incubated with material eluates for 24 and 48 h. Furthermore, these cells were seeded directly on swollen hydrogels for adhesion assays at 2 or 4 h, as well as for microscopic studies on their infiltration ability into the hydrogels after 48 h and 7 days. In order to characterize the ability of hydrogels to induce proinflammatory effects in cells, expression of COX-2 and the receptor for advanced glycation endproducts was quantified by western blotting after 48 h.
Results
MΦ showed a higher vitality and HAEC showed a lower vitality after incubation with material eluates, which can only be related to fragments formed by partial degradation. Interestingly, the strong difference in degradation rate, with G10LNCO3 showing a mass loss of 60 wt% within 6 days, while G10LNCO8 showed only 5–10 wt% mass loss, did not have an influence on the vitality. The adhesion ability of MΦ to swollen hydrogels was significantly decreased to 30% (p<0.05, ANOVA) for G10LNCO3 and to 38% for G10LNCO8, respectively, when compared with adhesion on normal cell culture plastic. By contrast, HAEC in part showed enhanced adhesion to the materials (116% for G10LNCO3, n.s.; 145% for G10LNCO8, p<0.05). This suggests that gelatin offers adhesion sequences for HAEC, but not for MΦ. A higher degree of crosslinking resulted in higher adhesion of both cell types. Additionally, both cell types infiltrated the materials within 4 days, which highlights the degradability of the material, putatively supported by cell-mediated mechanisms. Direct contact with the materials resulted in an increment of COX-2 expression in both cell lines, with a higher degree of hydrogel crosslinking leading to elevated COX-2 expression. Receptor for advanced glycation endproducts synthesis remained unaffected in both MΦ and HAEC.
Conclusion
The hydrogels provide both a surface preventing adhesion of macrophages and supporting adhesion of endothelial cells, which might lead to good tissue integration. However, the materials or their degradation products induced proinflammatory effects on MΦ depending on the degree of crosslinking. In further studies, these hydrogel films will be studied in animal models concerning their initial interaction with the organism after implantation and their degradation.
Financial & competing interests disclosure
This work is part of a research initiative within the Helmholtz-Portfoliotheme “Technologie und Medizin - Multimodale Bildgebung zur Aufklaerung des In vivo -Verhaltens von polymeren Biomaterialien”.
Reference
1 Tronci G, Neffe AT, Piercea BF, Lendlein A. An entropy–elastic gelatin-based hydrogel system. J. Mater. Chem. 20, 8875–8884

  • Abstract in refereed journal
    Regenerative Medicine 8(2013), S201
  • Poster
    World Conference on Regenerative Medicine, 23.-25.10.2013, Leipzig, Deutschland

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Publ.-Id: 19759