Effects of ASC Secretome on Human Osteochondral Explants

I.R.C.C.S Ospedale Galeazzi-Sant'Ambrogio36 enrolled

Overview

Since the need of finding effective disease-modifying anti-osteoarthritis (OA) treatments is still unmet, with this study the investigators aim to gather further evidences of the therapeutic potential of Mesenchymal Stem/stromal Cell (MSC) secretome in order to pave the way to its future use as a cell-free biological product. In detail, the investigators predict to validate the promising results obtained in vitro, ex vivo on osteochondral explants, an OA model more representative of the physiological situation.

Study Type

OBSERVATIONAL

Enrollment

Conditions

Osteoarthritis

Interventions

ASC secretomeBIOLOGICAL

The osteochondral explants isolated from arthroplasty patients will be induced to an OA phenotype and treated with ASC secretome (either complete conditioned medium or extracellular vesicles) in order to investigate its therapeutic potential.

Eligibility

Sex: ALLMin age: 18 Years

Medical Language ↔ Plain English

Inclusion Criteria: * males and females * age≥18 years old * patients hospitalized in IRCCS (Istituto di Ricovero e Cura a Carattere Scientifico) Istituto Ortopedico Galeazzi, undergoing arthroplasty Exclusion Criteria: * age\<18 years old * patients unable to sign the Informed Consent * positivity to serological test (HIV-Human Immunodeficiency Virus, HCV -Hepatitis C Virus, HBV -Hepatitis B Virus and TPHA -Treponema Pallidum Hemagglutination Assay)

Locations (1)

IRCCS Istituto Ortopedico Galeazzi

Milan, MI, Italy

Outcomes

Primary Outcomes

Validation of the cell-free approach based on the use of ASC secretome on an ex vivo OA model by evaluation of cell viability, histological features and gene/protein expression of cartilage and bone biomarkers

Biochemical and functional characterization of untreated osteochondral explants (to set up the model) and explants treated with inflammatory cytokines (to simulate osteoarthritic pathology) by evaluation of several parameters. The multiple measurements that will concur to describe the phenotype of the specimens are the following: * cell viability by AlamarBlue (Arbitrary Fluorescence Units). * histological assessments (qualitative differences between specimens). * analysis of gene expression by RT-PCR (real-time polymerase chain reaction, relative quantification by 2-ΔΔCT method). * analysis of protein expression by Western Blot (relative quantification by densitometry) and Luminex (pg/ml). Evaluation of the effects of ASC secretome (either complete conditioned medium or its subcomponents) on osteochondral biopsies treated with inflammatory cytokines through the biochemical and functional analyses described above.

Time frame: 3 years

Data from ClinicalTrials.gov

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