supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo...

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Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj Tripathi, and Ashok Kumar Interface Volume 8(57):540-554 April 6, 2011 ©2011 by The Royal Society

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Page 1: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage

tissue engineering

by Sumrita Bhat, Anuj Tripathi, and Ashok Kumar

InterfaceVolume 8(57):540-554

April 6, 2011

©2011 by The Royal Society

Page 2: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

Schematic diagram showing the procedure for isolation of chondrocytes from goat knee joint and FDA-stained cells (viable) at the time of seeding on the scaffolds.

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 3: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

Scanning electron microscopy (SEM) images showing microstructure of chitosan–agarose–gelatin CAG gels at (a) 80× and (b) 200× magnification.

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 4: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

FTIR spectroscopy analysis of the CAG blend showing a prominent peak for glycosidic linkage.

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 5: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

Cyclic swelling and de-swelling kinetics of CAG cryogels 4.5% (filled triangles), 5% (open squares).

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 6: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

Degree of degradation of chitosan–agarose cryogels 4.5% (filled squares), 5% (filled triangles), after eight weeks of incubation with 0.1 M PBS under aseptic conditions at 37°C.

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 7: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

SEM images of cryogel samples after eight week of degradation showing cracks in the pore walls at a magnification of 200×.

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 8: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

Unconfined compression stress–strain curve of chitosan–agarose cryogels 4.5% (filled squares) and 5% (open squares).

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 9: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

SEM images of (a) chondrocytes adhesion to the surface of the gel after 24 h at the magnification of 2500×, (b,c,d) Chondrocytes synthesizing extracellular matrix (ECM) after 4 days at a

magnification of 10 000×, (e) dividing chondrocytes at a magnification...

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 10: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

Cell viability and proliferation of chondrocytes in CAG cryogels as evaluated by MTT assay.

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 11: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

SEM images of (a) implanted scaffolds after two weeks of implantation showing cells attached to the scaffold surface (b) after four weeks of implantation and (c) after six weeks of implantation

showing degraded cryogel matrix and deposition of ECM. (d) Cont...

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society

Page 12: Supermacroprous chitosan–agarose–gelatin cryogels: in vitro characterization and in vivo assessment for cartilage tissue engineering by Sumrita Bhat, Anuj

Histological examination of the implanted constructs stained by H&E (a) after two weeks showing the integration of scaffolds with the native tissue and exhibiting the process of neo-

vascularization (b) after four weeks showing the disintegration of the cryo...

Sumrita Bhat et al. J. R. Soc. Interface 2011;8:540-554

©2011 by The Royal Society