OBJECTIVE: Up to now several clinical trials have shown the safety and efficacy of the intra-articular injection of Adipose-derived Mesenchymal Stem/Stromal Cells (ASCs) in contrasting osteoarthritis. Since ASCs act predominantly through paracrine mechanisms, their secretome represents a promising cell-free alternative. The aim of our study was to invetigate anti-hypertrophic and anti-catabolic effects of ASC conditioned medium (ASC-CM) on TNFα-stimulated human primary articular chondrocytes (CHs). MATERIALS AND METHODS: CHs were treated with 10ng/ml TNFα and/or ASC-CM administered at a 1:5 recipient:donor cell ratio. Cell viability was assessed up to day 9. The activity, expression and/or release of hypertrophy markers (MMP-13, Collagen X and Osteocalcin), catabolic mediators (MMP-3) and cartilage-protective factors were assessed up to day 3 by enzymatic assays, qRT-PCR, Western Blot and multiplex immunoassays. RESULTS: ASC-CM blunted TNFα-induced hypertrophy, reducing the enhanced levels of MMP-13 activity (-61%), Osteocalcin (-37%) and Collagen X (-18%). In addition, also MMP-3 activity was diminished by -59%. We associated the observed reduction of MMP-3 and MMP-13 activity to the abundancy of TIMPs (Tissue Inhibitors of MMPs) in ASC secretome, rather than to a direct down-modulation of their expression and/or release. Moreover, ASC-CM contains high levels of OPG and DKK-1, other known chondroprotective factors. CONCLUSION: ASC-CM is rich in cartilage-protective factors and exerts anti-hypertrophic and anti-catabolic effects on TNFα-stimulated CHs. These evidences open the way for its possible clinical use as a cell-free approach in contrasting osteoarthritis. We are currently investigating through a differential proteomic analysis if the recognized chondroprotective effectors are enriched in the vesicular rather than the soluble component of the secretome.

Adipose-derived Stem/Stromal Cell secretome blunts TNFα-induced hypertrophy on human primary articular chondrocytes / S. Niada, C. Giannasi, S. Casati, A.T.M. Brini. ((Intervento presentato al convegno GISM tenutosi a Genova nel 2019.

Adipose-derived Stem/Stromal Cell secretome blunts TNFα-induced hypertrophy on human primary articular chondrocytes

C. Giannasi;S. Casati;A.T.M. Brini
2019-04-04

Abstract

OBJECTIVE: Up to now several clinical trials have shown the safety and efficacy of the intra-articular injection of Adipose-derived Mesenchymal Stem/Stromal Cells (ASCs) in contrasting osteoarthritis. Since ASCs act predominantly through paracrine mechanisms, their secretome represents a promising cell-free alternative. The aim of our study was to invetigate anti-hypertrophic and anti-catabolic effects of ASC conditioned medium (ASC-CM) on TNFα-stimulated human primary articular chondrocytes (CHs). MATERIALS AND METHODS: CHs were treated with 10ng/ml TNFα and/or ASC-CM administered at a 1:5 recipient:donor cell ratio. Cell viability was assessed up to day 9. The activity, expression and/or release of hypertrophy markers (MMP-13, Collagen X and Osteocalcin), catabolic mediators (MMP-3) and cartilage-protective factors were assessed up to day 3 by enzymatic assays, qRT-PCR, Western Blot and multiplex immunoassays. RESULTS: ASC-CM blunted TNFα-induced hypertrophy, reducing the enhanced levels of MMP-13 activity (-61%), Osteocalcin (-37%) and Collagen X (-18%). In addition, also MMP-3 activity was diminished by -59%. We associated the observed reduction of MMP-3 and MMP-13 activity to the abundancy of TIMPs (Tissue Inhibitors of MMPs) in ASC secretome, rather than to a direct down-modulation of their expression and/or release. Moreover, ASC-CM contains high levels of OPG and DKK-1, other known chondroprotective factors. CONCLUSION: ASC-CM is rich in cartilage-protective factors and exerts anti-hypertrophic and anti-catabolic effects on TNFα-stimulated CHs. These evidences open the way for its possible clinical use as a cell-free approach in contrasting osteoarthritis. We are currently investigating through a differential proteomic analysis if the recognized chondroprotective effectors are enriched in the vesicular rather than the soluble component of the secretome.
Settore BIO/14 - Farmacologia
Settore BIO/13 - Biologia Applicata
Adipose-derived Stem/Stromal Cell secretome blunts TNFα-induced hypertrophy on human primary articular chondrocytes / S. Niada, C. Giannasi, S. Casati, A.T.M. Brini. ((Intervento presentato al convegno GISM tenutosi a Genova nel 2019.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/647689
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