Microstructural remodeling of articular cartilage following defect repair by osteochondral autograft transfer  C.B. Raub, S.C. Hsu, E.F. Chan, R. Shirazi, A.C. Chen, E. Chnari, E.J. Semler, R.L. Sah  Osteoarthritis and Cartilage  Volume 21, Issue 6, Pages 860-868 (June 2013) DOI: 10.1016/j.joca.2013.03.014 Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Safranin-O histology and qPLM maps of osteochondral sections from non-operated knees. (A, B) Safranin-O, (C, D) PI, (E, F) orientation (α) and (G, H) retardance (Γ) maps for representative samples of contralateral non-operated MFC (A, C, E, G, I–L) and lateral trochlea (LT) (B, D, F, H, M–P), with boxed areas in A–H shown in insets in I–L and M–P. ICRS and qPLM scores, and qPLM scores for superficial zone (SZ), middle zone (MZ) and deep zone (DZ) are indicated. ICRS subcategory A = surface, B = matrix, C = cell distribution, D = cell population viability, E = subchondral bone, and F = cartilage mineralization. Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Effect of OATS grafts on qPLM parameters at 6 and 12 months. PI (A–C), α (D–F), and Γ (G–I) of superficial (SZ, A, D, G), middle (MZ, B, E, H) and deep (DZ, C, F, I) zones of articular cartilage of the contralateral non-operated (Non-Op) or osteochondral autograft (OATS) study groups, at 6 and 12 months post-implantation. OATS repair regions were analyzed at the proximal (P) and distal (D) host (h), as well as the proximal interface (P), central region (C), and distal interface (D) of the graft (g). Data are mean ± SD (n = 4 for OATS; n = 6 for Non-Op). * indicates difference vs time-matched Non-Op (P < 0.05). Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Co-localization of qPLM parameters with histological indices of graft deterioration. (A–C) Safranin-O, (D–F) PI, (G–I) α, and (J–L) Γ maps for representative samples of good OATS repair (A, D, H, J), moderate OATS repair (B, E, H, K), and poor OATS repair (C, F, I, L) at 6 months post-operatively. ICRS scores, qPLM overall scores, and qPLM zonal values are indicated. ICRS subcategory A = surface, B = matrix, C = cell distribution, D = cell population viability, E = subchondral bone, and F = cartilage mineralization. PI, α, and Г averages in regions from proximal (P), central (C), and distal (D) graft superficial (SZ), middle (MZ) and deep zones (DZ) are tabulated. Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Correlation of qPLM scores with histopathological indices of osteochondral deterioration. Weighted (open markers) and zone-weighted (filled markers) qPLM scores vs (A) summed ICRS score and (B) summed Modified O'Driscoll (MOD) score, qPLM DZ subscore vs (C) ICRS cell distribution subscore and (D) MOD cell morphology subscore, and qPLM DZ subscore vs (E) ICRS cartilage mineralization subscore and (F) MOD structural characteristics subscore. Data are from n = 8 OATS and n = 12 contralateral non-operated samples. Values of Spearman's correlation coefficient, ρ, are indicated. Data marker types, defined in the legend, indicate OATS and Non-Op samples for weighted and unweighted scores. Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Overlay of ellipse representation of qPLM scores with H&E sections of non-operated, good OATS repair, and poor OATS repair samples. Overlays are shown for (A) non-operated MFC, (B, C) a good OATS repair, and (D–F) a poor OATS repair. qPLM parameters PI, α, and Γ were averaged over local areas, both in the host (h) and graft (g) regions, separated by dotted line in repairs (B, D), and used to calculate ellipse aspect ratio, orientation, and area, respectively (see cartoon). Boxes areas in (B, D) are shown in detail in insets (C, E, F) to show chondrocyte clusters (arrows) near the host-graft cleft. Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10. 1016/j. joca Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10. 1016/j. joca Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Figure S1 Determination of boundaries between cartilage zones from α depth profiles for non-operated MFC (A–C, G–I) and LT (D–F, J–L) sites. (A, D) Representative depth-averaged α curve, showing zonal boundaries (round blue markers), and (B, E) depth-averaged PI curve of a single ROI. (C, F) Zonal boundary positions averaged from normal (non-operated) samples, for regions along an 11 mm span along the articular surface. From logistic fits to α ROI data, zonal (G, J) boundary values and (H, K) thickness, as well as (I, L) % of total thickness. Data shown as mean ± SD, n = 12 Non-Op condyles. Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Figure S2 Quantification of PI, α, and Γ from non-operated goat MFC. (A, C, E) Colormaps (qPLM) with ROI boundaries (black dotted line) and (B, D, F) ROI quantification, subdivided by zone, for PI (A, B), α (C, D), and Γ (E, F). Graph data markers and bars represent individual ROI mean ± SD (across all depth levels within a zone); lines represent non-operated control group zonal mean ± SE (n = 6 condyles/Non-Op group). Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Figure S3 Effect of joint site, time post-implantation and zone on qPLM parameters of articular cartilage of non-operated goat knees. (A) PI, (B) α, and (C) Γ for articular cartilage from the non-operated (contralateral) knee, medial femoral condyle (MFC) and lateral trochlea (LT) at 6 and 12 months post-implantation, for superficial (SZ), middle (MZ) and deep (DZ) zones. Data are mean ± SD (n = 6/group). * Indicates difference vs. time- or site-matched group (P < 0.05). Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Figure S4 Quantification of PI, α, and Γ from a good OATS repair of goat MFC. (A, C, E) Colormaps (qPLM) with ROI boundaries (black dotted line) and (B, D, F) ROI quantification, subdivided by zone, for PI (A, B), α (C, D), and Γ (E, F). Graph data markers and bars represent individual ROI mean ± SD (across all depth levels within a zone); lines represent non-operated control group zonal mean ± SE (n = 6 condyles/Non-Op group). Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Figure S5 Quantification of PI, α, and Γ from moderate OATS repair of goat MFC. (A, C, E) Colormaps (qPLM) with ROI boundaries (black dotted line) and (B, D, F) ROI quantification, subdivided by zone, for PI (A, B), α (C, D), and Γ (E, F). Graph data markers and bars represent individual ROI mean ± SD (across all depth levels within a zone); lines represent non-operated control group zonal mean ± SE (n = 6 condyles/Non-Op group). Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Figure S6 Quantification of PI, α, and Γ of a poor OATS repair of goat medial femoral condyle. (A, C, E) Colormaps (qPLM) with ROI boundaries (black dotted line) and (B, D, F) ROI quantification, subdivided by zone, for PI (A, B), α (C, D), and Γ (E, F). Graph data markers and bars represent individual ROI mean ± SD (across all depth levels within a zone); lines represent non-operated control group zonal mean ± SE (n = 6 condyles/Non-Op group). Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Figure S7 Site-matched group PI, α, and Γ from OATS graft-implanted goat medial femoral condyles, 6 and 12 months post-implantation. PI (A, B), α (C, D), and Γ (E, F) at 6 months (A, C, E) and 12 months (B, D, E) post-operatively averaged over matched sites at the proximal interface (P1, P2), central implant (C1–C5) and distal interface (D1, D2) for superficial (SZ), middle (MZ) and deep (DZ) zones. Graph data markers represent OATS group mean ± SE (n = 4 condyles). Horizontal lines represent non-operated control group zonal mean ± SE (n = 6 condyles/Non-Op group). Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions

Figure S8 Effect of implant site, time-point and zone on PI and α values. Effect on PI vs. α means (polar coordinates) of OATS at (A) 6 and (B) 12 months post-operatively by implant site (proximal interface, P; central, C; distal interface, D) and zone (superficial, SZ; middle, MZ; deep, DZ), plotted with zone and time-matched controls. Data markers represent average values for n = 4 condyles/experimental group and n = 6 condyles/non-operated group. Osteoarthritis and Cartilage 2013 21, 860-868DOI: (10.1016/j.joca.2013.03.014) Copyright © 2013 Osteoarthritis Research Society International Terms and Conditions