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1.
Fig. 2.

Fig. 2. From: Injectable preformed scaffolds with shape-memory properties.

SEM cross-sectional image of 1% (wt/vol) MA-alginate cryogel showing interconnected macroporous network (A) and nanoporous structure of conventional hydrogels (B). Two-dimensional micro-CT images of alginate cryogel (C) and conventional hydrogel (D), and reconstructed three-dimensional micro-CT image of alginate cryogel (E). Photographs of cylindrical MA-alginate hydrogels, demonstrating the effect of Ca2+ treatment on cryogels and conventional hydrogels (F).

Sidi A. Bencherif, et al. Proc Natl Acad Sci U S A. 2012 Nov 27;109(48):19590-19595.
2.
Fig. 5.

Fig. 5. From: Injectable preformed scaffolds with shape-memory properties.

Adhesion of MSC cells in MA-alginate cryogels. Phase-contrast image showing elongated cells in RGD-containing cryogels (A). Confocal micrographs of seeded cells after 5 d of culture are shown in a typical RGD-modified cryogel (B), unmodified cryogel (C), and RGE-modified cryogel (D). Live/dead cell assay of MSCs (E) (1 d incubation postinjection) and confocal image showing cells (F) (6 d incubation postinjection) in RGD-modified MA-alginate cryogels. Cryogels were injected through a 16-gauge needle before imaging (E and F). (Scale bars: A–C, E, F, 20 μm; D, 10 μm.)

Sidi A. Bencherif, et al. Proc Natl Acad Sci U S A. 2012 Nov 27;109(48):19590-19595.
3.
Fig. 3.

Fig. 3. From: Injectable preformed scaffolds with shape-memory properties.

Comparison of physical properties of conventional nanoporous gels versus cryogels. Both gel types were fabricated using 1% (wt/vol) MA-alginate. Young’s moduli for alginate cryogels and conventional hydrogels (A). Stress vs. strain curves for macroporous and nanoporous rhodamine-labeled alginate hydrogels subjected to compression tests (B). Evaluation of pore connectivity (C) and weight swelling ratio (D) of macroporous and conventional alginate hydrogels. Swelling ratios were determined for gels in the absence (−) and presence (+) of CaCl2. Values in A, C, and D represent mean and SD (n = 4).

Sidi A. Bencherif, et al. Proc Natl Acad Sci U S A. 2012 Nov 27;109(48):19590-19595.
4.
Fig. 1.

Fig. 1. From: Injectable preformed scaffolds with shape-memory properties.

(A) Overview of the cryogelation process: alginate is chemically modified to allow radical polymerization (1); MA-alginate is added to APS/TEMED initiator system before incubation at −20 °C to allow ice crystal formation (2); the process of cryogelation takes place via the following steps: phase separation with ice crystal formation, cross-linking, and polymerization followed by thawing of ice crystals (porogens) to form an interconnected porous cryogel network (3); and conventional needle–syringe injection of preformed cryogels (4). (B) Photographs showing placement of a cryogel in 1-mL syringe (before injection) and gel recovery (after injection) via a conventional 16-gauge needle. (C) Rhodamine-labeled 1% MA-alginate gels with various sizes and shapes can be easily prepared by cryogenic polymerization. Fluorescent square-shaped gels suspended in 0.2 mL of PBS were syringe injected with a complete geometric restoration as illustrated in the microscopy image before/after injection. (D) Photographs showing cryogels prepared with different geometric shapes (disk, pentagon, heart, and star) retained their original shapes after syringe injection.

Sidi A. Bencherif, et al. Proc Natl Acad Sci U S A. 2012 Nov 27;109(48):19590-19595.
5.
Fig. 6.

Fig. 6. From: Injectable preformed scaffolds with shape-memory properties.

Injectable preseeded scaffolds promote in situ localization and retention of bioluminescent reporter cells. Photographs showing alginate cryogel scaffolds (white) and rhodamine-labeled alginate scaffolds (pink) (A). Bioluminescent cells were seeded on 1% RGD-modified MA-alginate cryogels at a concentration of 200 × 103 cells/scaffold, and cultured for 1 d. Optical imaging depicts distribution of bioluminescent cells (B). SEM image shows cells (pseudocolored blue) homogeneously adherent to the gel (C). Real-time fluorescence in vivo imaging showing injected Rhod-labeled cryogels. Cells injected s.c. via Rhod-labeled cryogels (1), Rhod-labeled RGD-cryogels (2), or as a free-floating cell suspension (B) in BALB/c mice at day 0 (D). Noninvasive bioluminescence in vivo imaging of transplanted cells in BALB/c mice at day 2 (E) and day 15 (F) postinjection. The same mice were used in each of these images and were arranged in the same relative positions before imaging (D–F).

Sidi A. Bencherif, et al. Proc Natl Acad Sci U S A. 2012 Nov 27;109(48):19590-19595.
6.
Fig. 4.

Fig. 4. From: Injectable preformed scaffolds with shape-memory properties.

Minimally invasive s.c. injection of alginate cryogels into the backs of C57BL/6J mice (A). Hydrogel localization after s.c. injection of preformed rhodamine-labeled 1% MA-alginate cryogels (4 × 4 × 1 mm) in the subcutis of a mouse after 3 d (B). Histological analysis (H&E stain) of explanted alginate cryogel at day 3. The arrows indicate the interface between the s.c. connective tissue (Lower Left) and the cryogel matrix (Upper Right). (Magnification: 10×.) (C). Photographs showing merged phase-contrast and fluorescence of s.c. injected rhodamine-labeled alginate macroporous scaffold with restoration of geometry after placement (D). Three days postinjection, the dashed lines indicate original square-shaped geometry of recovered scaffolds and the arrows show the spread of released BSA into the surrounding tissue (E). In vivo release profiles of cross-linked (chemically anchored) or encapsulated (physically entrapped) rhodamine-labeled BSA containing injected cryogels (F). Values represent mean and SD (n = 4).

Sidi A. Bencherif, et al. Proc Natl Acad Sci U S A. 2012 Nov 27;109(48):19590-19595.

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