Impact of Isolation Procedures on the Development of a Preclinical Synovial Fibroblasts/Macrophages in an In Vitro Model of Osteoarthritis

There is a lack ofin vitromodels able to properly represent osteoarthritis (OA) synovial tissue (ST). We aimed to characterize OA ST and to investigate whether a mechanical or enzymatic digestion procedures influence synovial cell functional heterogeneity in vitro. Procedures using mechanical nondigested fragments (NDF), synovial digested fragments (SDF), and filtrated synovial digested cells (SDC) were compared. An immunophenotypic profile was performed to distinguish synovial fibroblasts (CD55, CD73, CD90, CD106), macrophages (CD14, CD68), M1-like (CD80, CD86), and M2-like (CD163, CD206) synovial macrophages. Pro-inflammatory (interleukin 6 IL6), tumor necrosis factor alpha (TNFα), chemokine C-C motif ligand 3 (CCL3/MIP1α), C-X- motif chemokine ligand 10 (CXCL10/IP10) and anti-inflammatory (interleukin 10 (IL10)), transforming growth factor beta 1 (TGFβ1), C-C motif chemokine ligand 18 (CCL18) cytokines were evaluated. CD68 and CD163 markers were higher in NDF and SDF compared to the SDC procedure, while CD80, CD86, and CD206 were higher only in NDF compared to the SDC procedure. Synovial fibroblast markers showed similar percentages. TNFα, CCL3/MIP1α, CXCL10/IP10, and CCL18 were higher in NDF compared to SDC, but not compared to SDF. IL10 and TGFβ1 were higher in NDF than SDC at the molecular level, while IL6 did not show differences among procedures. We demonstrated that NDF isolation procedures better preserved the heterogeneity of specific OA synovial populations (fibroblasts, macrophages), fostering their use for testing new cell therapies or drugs for OA, reducing or avoiding the use of animal models.

Medienart:

E-Artikel

Erscheinungsjahr:

2020

Erschienen:

2020

Enthalten in:

Zur Gesamtaufnahme - volume:9

Enthalten in:

Biology - 9(2020), 12 vom: 10. Dez.

Sprache:

Englisch

Beteiligte Personen:

Manferdini, Cristina [VerfasserIn]
Saleh, Yasmin [VerfasserIn]
Dolzani, Paolo [VerfasserIn]
Gabusi, Elena [VerfasserIn]
Trucco, Diego [VerfasserIn]
Filardo, Giuseppe [VerfasserIn]
Lisignoli, Gina [VerfasserIn]

Links:

Volltext

Themen:

Isolation methods
Journal Article
Osteoarthritis
Synovial cells
Synovial fibroblasts
Synovial macrophages

Anmerkungen:

Date Revised 29.12.2020

published: Electronic

Citation Status PubMed-not-MEDLINE

doi:

10.3390/biology9120459

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM31887105X