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. 2016 Sep 9:12:6-12.
doi: 10.1016/j.btre.2016.09.001. eCollection 2016 Dec.

Evaluation of methods for pore generation and their influence on physio-chemical properties of a protein based hydrogel

Nicholas Bodenberger  1 , Dennis Kubiczek  1 , Irina Abrosimova  1 , Annika Scharm  1 , Franziska Kipper  1 , Paul Walther  1 , Frank Rosenau  1
Affiliations

Evaluation of methods for pore generation and their influence on physio-chemical properties of a protein based hydrogel

Nicholas Bodenberger et al. Biotechnol Rep (Amst). .

Abstract

Different methods to create and manipulate pore sizes in hydrogel fabrication are available, but systematic studies are normally conducted with hydrogels made of synthetic chemical compounds as backbones. In this study, a hydrogel made of natural and abundant protein in combination with different, well-available techniques was used to produce different architectures within the hydrogel matrix. Pore sizes and distribution are compared and resulting hydrogel properties like swelling ratio, resistance towards external stimuli and enzymatic degradation were investigated. Porous hydrogels were functionalized and two cancer cell lines were successfully adhered onto the material. With simple methods, pores with a radius between 10 and 80 μm and channels of 25 μm radius with a length of several hundreds of μm could be created and analyzed with laser scanning confocal microscopy and electron microscopy respectively. Furthermore, the influence of different methods on swelling ratio, enzymatic degradation and pH and temperature resistance was observed.

Keywords: Biopolymers; Cell culture; Degradation; Hydrogel properties; Pore size.

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Figures

Fig. 1
Fig. 1
Cryo scanning electron microscopy of untreated BSA Hydrogel at 10,000× (scale bar 5 μm) which was frozen under high pressure to maintain its native form.
Fig. 2
Fig. 2
Influence of pore formation and pore size for a freeze-drying approach with protein based hydrogels frozen at different temperatures. (A) Size and distribution of pores within the protein hydrogels matrix for hydrogels frozen in liquid nitrogen and at −20 °C (B)(C) Hydrogels were frozen and in liquid nitrogen (B) and at −20 °C (C) followed by subsequent freeze-drying, stained with rhodamine B and investigated via confocal laser scanning microscopy. Scale bar 50 μm.
Fig. 3
Fig. 3
Size and distribution of pores in protein hydrogels treated with a salt-leaching procedure. (A) Size distribution of pores within the protein hydrogels matrix. The used NaCl crystals were grinded with a pestle and mortar for a specific times: (B) 10 min. (C) 2 min. (D) 1 min. (E) not grinded. Protein hydrogels were stained with rhodamine B and investigated via confocal laser scanning microscopy. Scale bar 50 μm.
Fig. 4
Fig. 4
Size and distribution of pores in protein hydrogels treated with a chalk-leaching procedure. (A) Size distribution of pores within the protein hydrogels matrix for hydrogels which were mixed with untreated CaCO3 (B) and CaCO3 which was grinded with a pestle and mortar for 10 min. (C). Hydrogels were stained with rhodamine B and investigated via confocal laser scanning microscopy. Scale bar 50 μm.
Fig. 5
Fig. 5
Structure of protein hydrogel which was treated with a gradient freezing method: protein hydrogels were placed on a block of dried ice at 37 °C for 10 min, stained with rhodamine B in PBS for 30 min and visualized with confocal laser scanning microscopy at 514 nm. Scale bar 50 μm.
Fig. 6
Fig. 6
Representative enzymatic degradation pattern for macroporous hydrogels treated with a particle leaching approach and incubated with 300 U of trypsin (pH 7.4) and pepsin (pH 2.0) over a period of 12 h.
Fig. 7
Fig. 7
Growth and adhesion of cells onto a functionalized protein hydrogel. 2*105 cells were seeded onto a hydrogel surface on native (left side) and functionalized (right side) hydrogels. To functionalize hydrogels, a cell-adhesive RGD peptide was co-polymerized during hydrogel formation. After 24 h, the cells were stained with 5 μl of 300 U phalloidin-rhodamine B in 200 μl PBS and cellular adhesion of MCF7 and A549 cells was observed at 514 nm at a confocal laser scanning microscopy.
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References

    1. Park H., Guo X., Temenoff J.S., Tabata Y., Caplan A.I., Kasper F.K., Mikos A.G., Box P.O. Effect of swelling ratio of injectable hydrogel composites on chondrogenic differentiation of encapsulated rabbit marrow mesenchymal stem cells in vitro. Biomacromolecules. 2010;10:541–546. - PMC - PubMed
    1. Raic A., Rödling L., Kalbacher H., Lee-Thedieck C. Biomimetic macroporous PEG hydrogels as 3D scaffolds for the multiplication of human hematopoietic stem and progenitor cells. Biomaterials. 2014;35:929–940. - PubMed
    1. Li X.J., Valadez A.V., Zuo P., Nie Z. Microfluidic 3D cell culture: potential application for tissue-based bioassays. Bioanalysis. 2012;4:1509–1525. - PMC - PubMed
    1. Sasaki J.-I., Asoh T.-A., Matsumoto T., Egusa H., Sohmura T., Alsberg E., Akashi M., Yatani H. Fabrication of three-dimensional cell constructs using temperature-responsive hydrogel. Tissue Eng. Part A. 2010;16:2497–2504. - PubMed
    1. Hikmet Geckil Feng Xu Xiaohui Zhang SangJun Moon U.D. Engineering hydrogels as extracellular matrix mimics. Nanomedicine. 2011;5:469–484. - PMC - PubMed