1. An in vivo cross-linkable hyaluronan gel with inherent anti-inflammatory properties reduces OA cartilage destruction in female mice subjected to cruciate ligament transection 
C. Aulin, P. Lundbäck, K. Palmblad, L. Klareskog, H. Erlandsson Harris 
Osteoarthritis and Cartilage 
Volume 25, Issue 1, Pages (January 2017)
DOI: /j.joca
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

2. Fig. 1
Disease progression. OA was induced by destabilization of the knee joint through ACL transection and disease progression was monitored over 8 weeks. Each time point includes n = 5 animals. (a) Histological examination of Safranin-O stained knee joints revealing a rapid progression of cartilage damage. OARSI histology scoring showed high scores already at week 2, with cartilage degradation remaining or progressing until termination week 8. (b) Histology of total joints demonstrated destruction of the AC and chondrogenesis in the medial joint capsule (arrowheads). Osteophytes were detected mainly in the medial compartment (arrows). (c–f) demonstrate the different stages of destruction including fibrillation of the surface cartilage (arrow) (c), areas with loss of proteoglycans seen as weaker or total loss of the red Safranin-O staining (d), erosion of surface cartilage down to the calcified cartilage below the tidemark (dotted line) (e) and total erosion down to subchondral bone (B) (g). Serum samples from OA mice at different time points were analyzed for proinflammatory cytokines (IL-β, TNF, KC, MCP-1 and IFN-γ). Levels were <10 pg/ml in all cases, indicating no or low systemic inflammation. (h–j) Local inflammation was assessed by immunohistochemical analysis of the MHC class II expression. In areas with small cartilage fibrillation, no or low staining of MHC-II was observed (h), whereas in menisci (i), and fibrous tissue (j) strong expression was detected, evident at all time points evaluated. Here representative images are shown. Error bars represent 95% confidence interval. MFC – medial femoral condyle, LFC – lateral femoral condyle, MTP – medial tibia plateau, LTP – lateral tibia plateau. Scale bars 100 μm.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

3. Fig. 2
HA gel therapy rescues cartilage destruction. (a) Frontal sections from the central joint area were scored according to OARSI histopathology initiative. Treatment with HA gel injection rescued the cartilage destruction when compared to PBS injections shown as a lower score. (b) In PBS treated animals total erosion of AC down to subchondral bone could be observed in some cases. HA gel treated animals also showed destruction, but less severe, mainly appearing as loss of Safranin-O (arrow) and surface irregularity (arrowheads). Three individual experiments were performed, with the data pooled together. Each treatment included n = 5 or n = 3 animals. Error bars represent 95% confidence interval. Scale bars 100 μm.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

4. Fig. 3
Lipid peroxidation product expressed in hypertrophic chondrocytes of AC and the GP. OA features an increase in lipid peroxidation and lipid peroxidation products such as the aldehyde 4-HNE. Immunohistochemistry of OA knee joints revealed that 4-HNE was present in the AC at all time points, mainly in hypertrophic chondrocytes present in the surface layer of the AC. 4-HNE was also detected in hypertrophic chondrocytes in the GP (inset). Scale bars 100 μm.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

5. Fig. 4
Aldehyde quenching by gel component results in cartilage rescue. The aldehyde-reactive carbazate-modified component was injected intra-articularly to investigate its potential aldehyde quenching effect (n = 5). (a) The overall OARSI histological score was assessed, and a significant decrease in score was observed (P = 0.046), demonstrating an ameliorating effect compared to PBS treatment. (b) Histomorphometric analysis of 4-HNE stained sections showed no significant decrease compared to PBS or HA gel treated animals (n = 5). (c) Inflammation was assessed by immunohistochemistry of MHC class II in the articular cartilage. A similar expression in MHC class II was detected in all treatment groups (n = 5). (d) Comparison of consecutive sections stained with Safranin-O, 4-HNE and MHC class II revealed that 4-HNE staining did not overlap with Safranin-O depletion, which was the case for MHC class II. Error bars represent 95% confidence interval. Scale bars 100 μm.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions

6. Fig. 5
Decreased apoptosis with carbazate-modified component and HA gel treatment. TUNEL staining was performed to assess the degree of apoptosis in the articular cartilage (n = 5). Histomorphometry of AC showed a significant decrease in apoptosis in the aldehyde quenching gel component group compared to PBS treatment. Scale bars 100 μm.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2016 Osteoarthritis Research Society International Terms and Conditions