Altered functional loading causes differential effects in the subchondral bone and condylar cartilage in the temporomandibular joint from young mice

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Altered functional loading causes differential effects in the subchondral bone and condylar cartilage in the temporomandibular joint from young mice J. Chen, D.D.S., Ph.D., K.P. Sorensen, B.A., T. Gupta, D.M.D., T. Kilts, B.A., M. Young, Ph.D., S. Wadhwa, D.D.S., Ph.D. Osteoarthritis and Cartilage Volume 17, Issue 3, Pages 354-361 (March 2009) DOI: 10.1016/j.joca.2008.05.021 Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Body weight of female CD-1 mice exposed to different masticatory conditions. Points are the mean and s.e.m. for n=58 for normal loading group, n=9 incisor trimming, n=9 soft diet, n=59 for altered functional loading (incisor trimming in combination with soft diet administration) and n=30 for the return to normal loading (Re-Normal). aSignificant difference between altered functional loading and normal loading (P<0.05). bSignificant difference between altered functional loading and return to normal loading (P<0.05). Osteoarthritis and Cartilage 2009 17, 354-361DOI: (10.1016/j.joca.2008.05.021) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 21-day-old female CD-1 mice were exposed to different masticatory conditions. Real time PCR analysis for Prg4, Pthrp, Sox9, Col II, Ihh, Col X, Vegf, Opn, Oc, Opg, Runx2, Col I and Rankl gene expression from the mandibular condylar head from (A) 4 weeks of normal loading, soft diet, incisor trimming or soft diet and incisor trimming (altered functional loading), and (B) 6 weeks of normal loading, altered functional loading or return to normal loading conditions (Re-Normal). Points are the mean and SEM for n=5 for the 4-week and n=6 for the 6-week loading groups. aSignificant difference between altered functional loading and normal loading (P <0.05). bSignificant difference between altered functional loading and return to normal loading (P<0.05). Osteoarthritis and Cartilage 2009 17, 354-361DOI: (10.1016/j.joca.2008.05.021) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Measurements of mandibles from 21-day-old female CD-1 mice subjected to normal loading (2, 4 or 6 weeks), altered functional loading (2, 4 or 6 weeks) and return to normal loading (Re-Normal) (6weeks). (A and B) Gross mandibular and condylar measurements of the whole mandible and the distal condylar end of the mandible. (C–F) Statistical results of the measurements of cartilage width (C), condyle head length (D), mandible length (E) and cartilage thickness (F). Points are the mean and s.e.m. for n=9 for the normal loading group, n=9 for the altered functional loading group and n=9 for the return to normal loading group. aSignificant difference between altered functional loading and normal loading (P<0.05). bSignificant difference between altered functional loading and return to normal loading (P<0.05). Osteoarthritis and Cartilage 2009 17, 354-361DOI: (10.1016/j.joca.2008.05.021) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Representatives of the Safranin O staining (red) of the condylar cartilage. The slides were counter stained with Fast Green. (A) 4-Week normal loading; (B) 4-week altered functional loading; (C) 6-week normal loading; (D) 6-week altered functional loading; and (E) return to normal loading (4-week altered loading followed by 2-week normal loading). Osteoarthritis and Cartilage 2009 17, 354-361DOI: (10.1016/j.joca.2008.05.021) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Micro-CT analysis of the subchondral bone. Representative images of mid-sagittal cross sections from the mandibular condylar head of 21-day-old CD-1 mice subjected to 4 weeks Normal Loading (A) or Altered functional loading (B). (C–G) Micro-CT analysis of bone volume fraction (C), total volume (D), bone volume (E), trabecular thickness (F) and trabecular spacing (G) from the mandibular condylar subchondral bone of 21-day-old CD-1 mice subjected to 0 weeks (n=4), 2 weeks (n=11), 4 weeks (n=10), or 6 weeks (n=19) of normal loading, altered functional loading or return to normal loading TMJ conditions. aSignificant difference between altered loading and normal loading (P<0.05). bSignificant difference between altered loading and return to normal loading (P<0.05). Osteoarthritis and Cartilage 2009 17, 354-361DOI: (10.1016/j.joca.2008.05.021) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Immunohistochemistry of Col II (A – normal loading, B – altered functional loading) and negative control (C) performed on sagittal sections of the TMJ area from female CD-1 49-day-old mice. (D) Semiquantitative analysis of COL II positive area normalized by total mandibular condylar cartilage area from 49-day-old CD-1 mice exposed to 4 weeks of normal loading or altered loading. Points are the mean and SEM for n=11 for the normal loading group and n=11 for the altered loading group. aSignificant difference between altered loading and normal loading (P<0.05). Osteoarthritis and Cartilage 2009 17, 354-361DOI: (10.1016/j.joca.2008.05.021) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions