Lack of anti-inflammatory and anti-catabolic effects on basal inflamed osteoarthritic chondrocytes or synoviocytes by adipose stem cell-conditioned medium

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Presentation transcript:

Lack of anti-inflammatory and anti-catabolic effects on basal inflamed osteoarthritic chondrocytes or synoviocytes by adipose stem cell-conditioned medium C. Manferdini, M. Maumus, E. Gabusi, F. Paolella, F. Grassi, C. Jorgensen, S. Fleury-Cappellesso, D. Noël, G. Lisignoli Osteoarthritis and Cartilage Volume 23, Issue 11, Pages 2045-2057 (November 2015) DOI: 10.1016/j.joca.2015.03.025 Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Effects of different ratios of ASC-CM (DMEM:CM 75:25, 50:50, 25:75) both on chondrocytes (n = 5) and synoviocytes (n = 5). A. Gene expression of IL6 and CXCL8/IL8 tested on chondrocytes or synoviocytes treated with three different ratio of ASC-CM. B. ADAMTS5 and TIMP1 gene expression of chondrocytes or synoviocytes treated with different ratios of ASC-CM. Data show fold increase mRNA expression vs basal chondrocytes or synoviocytes (median with interquartile range). Significant differences vs basal *P < 0.05. Osteoarthritis and Cartilage 2015 23, 2045-2057DOI: (10.1016/j.joca.2015.03.025) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 ASC-CM effects on IL6 and CXCL8/IL8 chemokines released by chondrocytes (n = 8) and synoviocytes (n = 12). A. IL6 and CXCL8/IL8 (pg/ml) detected in chondrocytes or synoviocytes monoculture alone or treated with ASC-CM. Significant differences *P < 0.001. B. IL6 and CXCL8/IL8 (pg/ml) detected in chondrocytes micromass alone or treated with ASC-CM. Data are expressed as median with interquartile range. Significant differences * P = 0.0175. C. ASC-CM effects on CCL2/MCP-1,CCL3/MIP-1α,CCL5/RANTES and CXCL1/GROα chemokines released by chondrocytes (n = 8) and synoviocytes (n = 12). CCL2/MCP-1,CCL3/MIP-1α,CCL5/RANTES and CXCL1/GROα (pg/ml) detected in chondrocytes or synoviocytes alone or treated with ASC-CM. Data are expressed as median with interquartile range. Significant differences *P < 0.001. Osteoarthritis and Cartilage 2015 23, 2045-2057DOI: (10.1016/j.joca.2015.03.025) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 ASC-CM effects on IL6 and CXCL8/IL8 chemokines released by chondrocytes (n = 8) and synoviocytes (n = 12). A. IL6 and CXCL8/IL8 (pg/ml) detected in chondrocytes or synoviocytes monoculture alone or treated with ASC-CM. Significant differences *P < 0.001. B. IL6 and CXCL8/IL8 (pg/ml) detected in chondrocytes micromass alone or treated with ASC-CM. Data are expressed as median with interquartile range. Significant differences * P = 0.0175. C. ASC-CM effects on CCL2/MCP-1,CCL3/MIP-1α,CCL5/RANTES and CXCL1/GROα chemokines released by chondrocytes (n = 8) and synoviocytes (n = 12). CCL2/MCP-1,CCL3/MIP-1α,CCL5/RANTES and CXCL1/GROα (pg/ml) detected in chondrocytes or synoviocytes alone or treated with ASC-CM. Data are expressed as median with interquartile range. Significant differences *P < 0.001. Osteoarthritis and Cartilage 2015 23, 2045-2057DOI: (10.1016/j.joca.2015.03.025) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 ASC-CM effects on ADAMTS4, ADAMTS5, TIMP1, TIMP3 and MMP13 gene expression and protein release of chondrocytes (n = 8). A. Gene expression of ADAMTS4, ADAMTS5, TIMP1, TIMP3 and MMP13 was tested on chondrocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Data are expressed as median with interquartile range of fold basal release. B. Protein release of ADAMTS4, ADAMTS5, and MMP13 (pg/ml) was tested on chondrocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Significant differences: ADAMTS4 P = 0.0286, ADAMTS5 P = 0.0286, MMP13 P = 0.048. C. Protein release of TIMP1 (ng/ml) and TIMP3 (pg/ml) was tested on chondrocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Data are expressed as median with interquartile range. Significant differences: TIMP1 (both left and right side) *P = 0.048. Osteoarthritis and Cartilage 2015 23, 2045-2057DOI: (10.1016/j.joca.2015.03.025) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 ASC-CM effects on ADAMTS4, ADAMTS5, TIMP1, TIMP3 and MMP13 gene expression and protein release of chondrocytes (n = 8). A. Gene expression of ADAMTS4, ADAMTS5, TIMP1, TIMP3 and MMP13 was tested on chondrocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Data are expressed as median with interquartile range of fold basal release. B. Protein release of ADAMTS4, ADAMTS5, and MMP13 (pg/ml) was tested on chondrocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Significant differences: ADAMTS4 P = 0.0286, ADAMTS5 P = 0.0286, MMP13 P = 0.048. C. Protein release of TIMP1 (ng/ml) and TIMP3 (pg/ml) was tested on chondrocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Data are expressed as median with interquartile range. Significant differences: TIMP1 (both left and right side) *P = 0.048. Osteoarthritis and Cartilage 2015 23, 2045-2057DOI: (10.1016/j.joca.2015.03.025) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 ASC-CM effects on ADAMTS4, ADAMTS5, TIMP1, TIMP3 and MMP13 gene expression and protein release of synoviocytes (n = 8). A. Gene expression of ADAMTS4, ADAMTS5, TIMP1, TIMP3 and MMP13 was tested on synoviocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Data are expressed as median with interquartile range of fold basal release. B. Protein release of ADAMTS4, ADAMTS5, and MMP13 (pg/ml) was tested on synoviocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Significant differences: ADAMTS4 P = 0.042, ADAMTS5 P = 0.045. C. Protein release of TIMP1 (ng/ml) and TIMP3 (pg/ml) was tested on synoviocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Data are expressed as median with interquartile range. Significant differences: TIMP1 (left side) *P = 0.0225, TIMP1 (right side). *P = 0.0177. Osteoarthritis and Cartilage 2015 23, 2045-2057DOI: (10.1016/j.joca.2015.03.025) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 ASC-CM effects on ADAMTS4, ADAMTS5, TIMP1, TIMP3 and MMP13 gene expression and protein release of synoviocytes (n = 8). A. Gene expression of ADAMTS4, ADAMTS5, TIMP1, TIMP3 and MMP13 was tested on synoviocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Data are expressed as median with interquartile range of fold basal release. B. Protein release of ADAMTS4, ADAMTS5, and MMP13 (pg/ml) was tested on synoviocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Significant differences: ADAMTS4 P = 0.042, ADAMTS5 P = 0.045. C. Protein release of TIMP1 (ng/ml) and TIMP3 (pg/ml) was tested on synoviocytes in basal conditions and after treatment with ASC-CM (left side) or co-cultured with ASC (right side). Data are expressed as median with interquartile range. Significant differences: TIMP1 (left side) *P = 0.0225, TIMP1 (right side). *P = 0.0177. Osteoarthritis and Cartilage 2015 23, 2045-2057DOI: (10.1016/j.joca.2015.03.025) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 PGE2 released by ASC-CM alone or after chondrocytes (n = 8) or synoviocytes (n = 8) treatment. A. PGE2 (pg/ml) was tested on different ratios of ASC-CM (DMEM:CM 75:25, 50:50, 25:75) B. PGE2 (pg/ml) was tested on chondrocytes or synoviocytes alone or treated with different ratios of ASC-CM. Data are expressed as median with interquartile range. Significant differences *P = 0.0036. Osteoarthritis and Cartilage 2015 23, 2045-2057DOI: (10.1016/j.joca.2015.03.025) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 ASC co-culture effects on inflamed chondrocytes and synoviocytes. A schematic model that summarizes all the ASC effects demonstrated by co-culturing ASC with inflamed chondrocytes or synoviocytes. ASC down-modulated cytokine/chemokines (IL6, IL8/CXCL8, CCL2/MCP-1, CCL3/MIP-1α, CCL5/RANTES and CXCL1/GROα), a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS4, ADAMTS5), but increased tissue metalloproteinase (TIMP1) both on chondrocytes and synoviocytes. In chondrocytes ASC down-modulated also metalloproteinase (MMP13). The down-modulation of cytokine/chemokines was mediated by PGE2. Osteoarthritis and Cartilage 2015 23, 2045-2057DOI: (10.1016/j.joca.2015.03.025) Copyright © 2015 Osteoarthritis Research Society International Terms and Conditions