Osteochondral defect repair using bilayered hydrogels encapsulating both chondrogenically and osteogenically pre-differentiated mesenchymal stem cells.

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Osteochondral defect repair using bilayered hydrogels encapsulating both chondrogenically and osteogenically pre-differentiated mesenchymal stem cells in a rabbit model J. Lam, S. Lu, E.J. Lee, J.E. Trachtenberg, V.V. Meretoja, R.L. Dahlin, J.J.J.P. van den Beucken, Y. Tabata, M.E. Wong, J.A. Jansen, A.G. Mikos, F.K. Kasper Osteoarthritis and Cartilage Volume 22, Issue 9, Pages 1291-1300 (September 2014) DOI: 10.1016/j.joca.2014.06.035 Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Histological sections showing representative osteochondral tissue repair at 12 weeks postoperatively for the MSC/MSC group. Sections were stained with (a) Safranin-O/Fast Green, (b) H&E, and (c) van Gieson's Picrofuchsin (scale bars: 1000 μm). Images from this sample (a–c), which received a score of 2 for cartilage morphology, showed thin fibrocartilage formation filling the chondral defect space and ossifying chondrocytes in the subchondral defect space. (d) The newly formed cartilage in the osteochondral defect mostly comprised fibrocartilage at the joint surface and hypertrophic cartilage in the subchondral region (scale bar: 500 μm). (e) One other joint from the MSC/MSC group displayed bone-like tissue formation in the chondral defect region, as can be seen by the presence of osteocytes in the H&E section (scale bar: 500 μm). Osteoarthritis and Cartilage 2014 22, 1291-1300DOI: (10.1016/j.joca.2014.06.035) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Histological sections showing representative osteochondral tissue repair at 12 weeks postoperatively for the MSC/OS group. Sections were stained with (a) Safranin-O/Fast Green, (b) H&E, and (c) van Gieson's Picrofuchsin (scale bars: 1000 μm). (d) This sample, which received a score of 2 for cartilage morphology, displayed thicker cartilage formation, which mainly comprised a mixture of fibro- and hyaline cartilage (scale bar: 500 μm). This sample also showed the development of a relatively discernable osteochondral interface. (e) An example of regional hypertrophy in the chondral portion of the defect from another sample displayed enlarged chondrocytes and differential matrix staining when compared to the surrounding cartilage (scale bar: 500 μm). Osteoarthritis and Cartilage 2014 22, 1291-1300DOI: (10.1016/j.joca.2014.06.035) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Histological sections showing representative osteochondral tissue repair at 12 weeks postoperatively for the CG14/OS group. Sections were stained with (a) Safranin-O/Fast Green, (b and d) H&E, and (c) van Gieson's Picrofuchsin (scale bars: 1000 μm). (d) This sample, which received a score of 2 for cartilage morphology, showed primarily fibrocartilage tissue repair with some fibrous tissue formation at the joint surface (scale bar: 500 μm). (e) One example of joint surface disruption in another sample is shown in the H&E section (scale bar: 500 μm). Osteoarthritis and Cartilage 2014 22, 1291-1300DOI: (10.1016/j.joca.2014.06.035) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Histological sections showing representative osteochondral tissue repair at 12 weeks postoperatively for the CG7/OS group. Sections were stained with (a) Safranin-O/Fast Green, (b) H&E, and (c) van Gieson's Picrofuchsin (scale bars: 1000 μm). (d) This sample, which received a score of 4 for cartilage morphology, displayed high quality articular cartilage with columnar arrangement of chondrocytes (scale bars: 1000 μm). (e) This sample also displayed compact and trabecular bone formation in the subchondral defect regions of the osteochondral defects (scale bars: 1000 μm). Osteoarthritis and Cartilage 2014 22, 1291-1300DOI: (10.1016/j.joca.2014.06.035) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Histological score distribution for the new (a) subchondral bone morphology and the new (b) cartilage tissue morphology. Osteoarthritis and Cartilage 2014 22, 1291-1300DOI: (10.1016/j.joca.2014.06.035) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions

Supplementary Fig. 1 : Representative gross morphology of the osteochondral defects after 12 weeks from the (a) MSC/MSC group, (b) the MSC/OS group, (c) the CG7/OS group, and (d) the CG14/OS group. Defects are indicated with a white circle. It should be noted that the white circles are only indicative and do not reflect the actual dimensions of the defect. (e) H&E section showing the partially degraded hydrogel particulates encapsulated in fibrous tissue, as indicated by arrows, typically found remaining in the subchondral defect region of samples. This particular sample is from the MSC/MSC group. Osteoarthritis and Cartilage 2014 22, 1291-1300DOI: (10.1016/j.joca.2014.06.035) Copyright © 2014 Osteoarthritis Research Society International Terms and Conditions