Accumulation of mitochondrial DNA with 4977-bp deletion in knee cartilage – an association with idiopathic osteoarthritis  Ming-Chau Chang, M.D., Shih-Chieh Hung, M.D., Ph.D., Winby York-Kwan Chen, M.D., Tzu-Ling Chen, B.Sc., Cheng-Feng Lee, M.S., Hsin-Chen Lee, Ph.D., Kai-Li Wang, B.Sc., Chung-Chih Chiou, B.Sc., Yau-Huei Wei, Ph.D.  Osteoarthritis and Cartilage  Volume 13, Issue 11, Pages 1004-1011 (November 2005) DOI: 10.1016/j.joca.2005.06.011 Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 1 An illustration of the application of the histologic-histochemical grading system to the osteoarthrotic knee cartilage of specimen no. 12-1 (54 y/o female), 15-1 (76 y/o man), 3-1 (78 o/y female), 13-1 (54 8/0 female), and 6-1 (72 y/o man). The photomicrographs A, B, C, D, E indicate the cartilage specimens stained with HE (4X in A, B, C, E, 10X in D, upper photograph) and with Safranin-O–fast green–iron hemoxylin (40X in A, B, C, D, 20X in E, lower photograph). The Safranin-O-fast stain recording scores of A, B, C, D and E were normal (0), slight reduction (1), moderate reduction (2), severe reduction (3) and no dye noted (4). They demonstrate the histological appearance and histochemical appearance on which the grading was based. Osteoarthritis and Cartilage 2005 13, 1004-1011DOI: (10.1016/j.joca.2005.06.011) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 1 An illustration of the application of the histologic-histochemical grading system to the osteoarthrotic knee cartilage of specimen no. 12-1 (54 y/o female), 15-1 (76 y/o man), 3-1 (78 o/y female), 13-1 (54 8/0 female), and 6-1 (72 y/o man). The photomicrographs A, B, C, D, E indicate the cartilage specimens stained with HE (4X in A, B, C, E, 10X in D, upper photograph) and with Safranin-O–fast green–iron hemoxylin (40X in A, B, C, D, 20X in E, lower photograph). The Safranin-O-fast stain recording scores of A, B, C, D and E were normal (0), slight reduction (1), moderate reduction (2), severe reduction (3) and no dye noted (4). They demonstrate the histological appearance and histochemical appearance on which the grading was based. Osteoarthritis and Cartilage 2005 13, 1004-1011DOI: (10.1016/j.joca.2005.06.011) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions

Fig. 2 Gel chromatograms of the PCR products amplified from mtDNA with 4977-bp deletion (upper panel, 524-bp) and total mtDNA (lower panel, 533-bp) in the semi-quantitative analysis of 4977-bp-deleted mtDNA. The PCR products of serially diluted DNA samples were separated on 1.5% agarose gels followed by staining with 0.01% ethidium bromide. The two typical gel chromatograms revealed 0.012% of 4977-bp-deleted mtDNA (A) and detectable but not distinct level of 4977-bp deletion of mtDNA (B) in two bone tissue samples, respectively. M, 100-bp size marker; P, positive control. Osteoarthritis and Cartilage 2005 13, 1004-1011DOI: (10.1016/j.joca.2005.06.011) Copyright © 2005 OsteoArthritis Research Society International Terms and Conditions