Secreted Factors and EV-miRNAs Orchestrate the Healing Capacity of Adipose Mesenchymal Stem Cells for the Treatment of Knee Osteoarthritis.

Enrico Ragni,Laura De Girolamo,Paola De Luca,Alessandra Colombini,Carlotta Perucca Orfei,Marco Viganò

doi:10.3390/ijms21051582

Enrico Ragni, Laura De Girolamo + Show 4 more

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https://doi.org/10.3390/ijms21051582

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Abstract

Mesenchymal stem cells (MSCs) derived from adipose tissue and used either as expanded cells or minimally manipulated cell preparations showed positive clinical outcomes in regenerative medicine approaches based on tissue restoration and inflammation control, like in osteoarthritis (OA). Recently, MSCs’ healing capacity has been ascribed to the large array of soluble factors, including soluble cytokines/chemokines and miRNAs conveyed within extracellular vesicles (EVs). Therefore, in this study, 200 secreted cytokines, chemokines and growth factors via ELISA, together with EV-embedded miRNAs via high-throughput techniques, were scored in adipose-derived MSCs (ASCs) cultivated under inflammatory conditions, mimicking OA synovial fluid. Both factors (through most abundantly expressed TIMP1, TIMP2, PLG and CTSS) and miRNAs (miR-24-3p, miR-222-3p and miR-193b-3p) suggested a strong capacity for ASCs to reduce matrix degradation activities, as those activated in OA cartilage, and switch synovial macrophages, often characterized by an M1 inflammatory polarization, towards an M2 phenotype. Moreover, the crucial importance of selecting the target tissue is discussed, showing how a focused search may greatly improve potency prediction and explain clinical outcomes. In conclusion, herein presented data shed light about the way ASCs regulate cell homeostasis and regenerative pathways in an OA-resembling environment, therefore suggesting a rationale for the use of MSC-enriched clinical products, such as stromal vascular fraction and microfragmented adipose tissue, in joint pathologies.

Highlights

Osteoarthritis (OA) is a heterogeneous disease with a wide range of underlying pathways and involves structural alterations mainly in the articular cartilage, subchondral bone and synovial membrane [1]
The aim of this study was to identify secreted cytokines, chemokines and growth factors and extracellular vesicles (EVs)-shuttled miRNAs from adipose-derived MSCs (ASCs) treated with inflammatory mediators at concentrations resembling those in the OA synovial fluid based on literature reports [33,34,35,36,37,38,39,40,41] in order to clarify how the ASCs regulate cell homeostasis and regenerative pathways in OA joints
ASCs were characterized by flow cytometry for both Mesenchymal stem cells (MSCs) and hemato-endothelial markers in the absence and presence of OA-resembling cytokines TNFα (5 pg/mL) + IL1β (10 pg/mL) + IFNγ

Summary

Introduction

Osteoarthritis (OA) is a heterogeneous disease with a wide range of underlying pathways and involves structural alterations mainly in the articular cartilage, subchondral bone and synovial membrane [1]. Total knee replacement showed greater pain relief and functional improvement than nonsurgical treatments [7], up to 25% of patients still complain of pain and disability after surgery [8]. In this perspective, new biological-based products aimed to prevent the progression of OA and restoring tissue homeostasis have been introduced. New biological-based products aimed to prevent the progression of OA and restoring tissue homeostasis have been introduced They are mostly based on mesenchymal stem cells (MSCs) whose pro-regenerative properties are well known. In addition to expanded MSCs, MSC-enriched products prepared at the point of care under minimal manipulation are available, both from bone marrow (bone marrow aspirate concentrate—BMAC)

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