The beneficial effect of delayed compressive loading on tissue-engineered cartilage constructs cultured with TGF-β3 E.G. Lima, M.Phil., M.S., L. Bian,

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The beneficial effect of delayed compressive loading on tissue-engineered cartilage constructs cultured with TGF-β3 E.G. Lima, M.Phil., M.S., L. Bian, M.S., K.W. Ng, M.S., R.L. Mauck, Ph.D., B.A. Byers, Ph.D., R.S. Tuan, Ph.D., G.A. Ateshian, Ph.D., C.T. Hung, Ph.D. Osteoarthritis and Cartilage Volume 15, Issue 9, Pages 1025-1033 (September 2007) DOI: 10.1016/j.joca.2007.03.008 Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Schematic of TGF-β3 time-course and loading time-course for each study. Loading was initiated at days indicated by arrow; TGF-β3 was supplemented during periods indicated by hatch marks. Osteoarthritis and Cartilage 2007 15, 1025-1033DOI: (10.1016/j.joca.2007.03.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 (a) Loading profile adjusted for system compliance delivered by bioreactor over time in culture. Dotted line shows increasing tare strain as a result of increasing tissue thickness with time. Solid lines show decreasing applied dynamic strain as a result of tissue stiffening over time. (b) Representative load vs time curve of tissue-engineered constructs on day 42. A load of zero would have indicated platen lift-off. Inset represents full curve. Osteoarthritis and Cartilage 2007 15, 1025-1033DOI: (10.1016/j.joca.2007.03.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Study 1: The effect of temporal application of TGF-β3 to a chemically defined medium: (a) EY, (b) G∗ at 1Hz, (c) GAG, and (d) collagen. *P<0.005 for TGF continued vs TGF discontinued (n=4). Osteoarthritis and Cartilage 2007 15, 1025-1033DOI: (10.1016/j.joca.2007.03.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Study 2: The effect of dynamic deformational loading applied concurrently with exposure to TGF-β3: (a) EY, (b) G∗ at 1Hz, (c) GAG, and (d) collagen. *P<0.005 for FS vs CDL (n=5). Osteoarthritis and Cartilage 2007 15, 1025-1033DOI: (10.1016/j.joca.2007.03.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Study 3: The effect of dynamic deformational loading initiated after the discontinuation of TGF-β3: (a) EY, (b) G∗ at 1Hz, (c) GAG, and (d) collagen. *P<0.005 for FS vs DDL (n=8). Osteoarthritis and Cartilage 2007 15, 1025-1033DOI: (10.1016/j.joca.2007.03.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 (1) Safranin O staining for GAG, (2) Picrosirius Red staining for collagen, (3) hematoxylin and eosin staining for visualization of local multiplication of cell nuclei (mag. 40×), and (4) immunohistochemical staining for type II collagen. All slides taken from Study 3 on either day 0 or day 42 with either free-swelling (FS) or dynamic compressive loading (DL) groups. Osteoarthritis and Cartilage 2007 15, 1025-1033DOI: (10.1016/j.joca.2007.03.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions