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. 2012 Mar 12;13(3):706-13.
doi: 10.1021/bm201596w. Epub 2012 Feb 22.

Biomimetic poly(ethylene glycol)-based hydrogels as scaffolds for inducing endothelial adhesion and capillary-like network formation

Junmin Zhu  1 , Ping He, Lin Lin, Derek R Jones, Roger E Marchant
Affiliations

Biomimetic poly(ethylene glycol)-based hydrogels as scaffolds for inducing endothelial adhesion and capillary-like network formation

Junmin Zhu et al. Biomacromolecules. .

Abstract

The extracellular matrix (ECM) is an attractive model for designing synthetic scaffolds with a desirable environment for tissue engineering. Here, we report on the synthesis of ECM-mimetic poly(ethylene glycol) (PEG) hydrogels for inducing endothelial cell (EC) adhesion and capillary-like network formation. A collagen type I-derived peptide GPQGIAGQ (GIA)-containing PEGDA (GIA-PEGDA) was synthesized with the collagenase-sensitive GIA sequence attached in the middle of the PEGDA chain, which was then copolymerized with RGD capped-PEG monoacrylate (RGD-PEGMA) to form biomimetic hydrogels. The hydrogels degraded in vitro with the rate dependent on the concentration of collagenase and also supported the adhesion of human umbilical vein ECs (HUVECs). Biomimetic RGD/GIA-PEGDA hydrogels with incorporation of 1% RGD-PEGDA into GIA-PEGDA hydrogels induced capillary-like organization when HUVECs were seeded on the hydrogel surface, while RGD/PEGDA and GIA-PEGDA hydrogels did not. These results indicate that both cell adhesion and biodegradability of scaffolds play important roles in the formation of capillary-like networks.

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Figures

Figure 1
Figure 1
(A) Schematic of the natural ECM structure and its interaction with cells. FN, fibronectin; LN, laminin; HA, hyaluronic acid; GAG, glycosaminoglycan; PG, proteoglycan. (B) Model of ECM-mimetic PEG hydrogel with incorporation of enzyme-sensitive peptides (ESPs) and cell adhesive peptides (CAPs).
Figure 2
Figure 2
(A) Collagenase-sensitive sequences, GPQG↓IAGQ (GIA) and GPQG↓LLGA (GLL) in the collagenase-sensitive domain of collagen type I (↓ indicating the cleavage site). (B) MALDI mass spectrum of GPQGIAGQ-Dap (GIA-Dap) prepared by SPPS.
Figure 3
Figure 3
(A) Synthesis of GIA-PEGDA and RGD-PEGMA macromers. (B) MALDI mass spectra of GIA-PEGDA with a maximum peak at 7,996. (C) MALDI mass spectrum of RGD-PEGMA with a maximum peak at 4,178.
Figure 4
Figure 4
Degradation of GIA-PEGDA (20%, w/v) hydrogels in the presence of collagenase in PBS at 37°C.
Figure 5
Figure 5
Phase contrast images of HUVECs (5x104 cells/cm2) seeded on PEG hydrogels. (A, B) PEGDA hydrogels for 4 and 24 h, respectively. (C, D) 1% (w/v) RGD/PEGDA hydrogels for 4 and 24 h, respectively (Scale bar = 100 μm). (E) Quantitative comparison of cell coverage area on PEGDA and 1% (w/v) RGD/PEGDA hydrogels.
Figure 6
Figure 6
Phase contrast images of HUVECs 24 h after seeded on 1% (w/v) RGD/PEGDA hydrogels with various seeding densities. (A) 2.5x104 cells/cm2; (B) 5.0x104 cells/cm2; (C) 7.5x104 cells/cm2; (D) 1.0x105 cells/cm2 (Scale bar = 100 μm). (E) Quantitative comparison of cell coverage area on the hydrogel surface.
Figure 7
Figure 7
Phase contrast images of HUVECs seeded on collagenase-sensitive hydrogels. (A, B) GIA-PEGDA hydrogels for 4 and 24 h, respectively. (C, D, E) 1% (w/v) RGD/GIA-PEGDA hydrogels for 4, 12 and 24 h, respectively. (F) Zooming in the boxed area in (E). (Scale bar = 100 μm).
Figure 8
Figure 8
Quantitative comparison of cell coverage area on GIA-PEGDA and 1% (w/v) RGD/GIA-PEGDA hydrogels.
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