Human Diseased Articular Cartilage Contains a Mesenchymal Stem Cell-Like Population of Chondroprogenitors with Strong Immunomodulatory Responses

Abstract

Background: osteoarthritic human articular cartilage (AC)-derived cartilage cells (CCs) with same-donor bone marrow (BMSCs) and adipose tissue (ASCs)-derived mesenchymal stem cells were compared, in terms of stemness features, and secretory and immunomodulatory responses to inflammation. Methods: proteoglycan 4 (PRG4) presence was evaluated in AC and CCs. MSCs and CCs (n = 8) were cultured (P1 to P4) and characterized for clonogenicity, nanog homeobox (NANOG), and POU class 5 homeobox 1 (POU5F1) expression, immunotypification, and tri-lineage differentiation. Their basal and interleukin-1β (IL-1β)-stimulated expression of matrix metalloproteases (MMPs), tissue inhibitors (TIMPs), release of growth factors, and cytokines were analyzed, along with the immunomodulatory ability of CCs. Results: PRG4 was mainly expressed in the intact AC surface, whereas shifted to the intermediate zone in damaged cartilage and increased its expression in CCs upon culture. All cells exhibited a similar phenotype and stemness maintenance over passages. CCs showed highest chondrogenic ability, no adipogenic potential, a superior basal secretion of growth factors and cytokines, the latter further increased after inflammatory stimulation, and an immunomodulatory behavior. All stimulated cells shared an increased MMP expression without a corresponding TIMP production. Conclusion: based on the observed features, CCs obtained from pathological joints may constitute a potential tissue-specific therapeutic target or agent to improve damaged cartilage healing, especially damage caused by inflammatory/immune mediated conditions.

Intervertebral disc and endplate cell characterisation highlights annulus fibrosus cells as the most promising for tissue-specific disc degeneration therapy

“…Differences were observed only in terms of loss of proliferative potential by NPCs and EPCs and of chondrogenic potential and pluripotent stem cell gene expression (POU5F1 and NANOG) by AFCs and EPCs at P3. The surface marker layout after expansion indicated a typical MSC-like immunophenotype (Dominici et al, 2006) of IVD cells and the positivity for CD44, CD73, CD90, CD105 and CD166 was consistent with what has already been observed in IVD cells (Blanco et al, 2010;Brisby et al, 2013;Liu et al, 2011;Wang et al, 2014). Furthermore, although they have an MSC-like immunophenotype, IVD cells do not share a perivascular origin given the lack of CD146 expression.…”

“…Furthermore, although they have an MSC-like immunophenotype, IVD cells do not share a perivascular origin given the lack of CD146 expression. To reinforce the MSClike phenotype of the analysed disc cells, CD151 also showed a high expression already previously observed in articular chondroprogenitor cells (De Luca et al, 2019) and MSCs (Lee et al, 2009).…”

“…Flow cytometry analyses were conducted on 2.5 × 10 5 expanded cells (P3) incubated with anti-human primary monoclonal antibodies: CD34-biotinylated, CD44-FITC, CD45-FITC, CD105-biotinylated, CD166-FITC (all from Ancell Corporation, Bayport, MN, USA), CD90-FITC and CD73-PE (Miltenyi Biotec) and CD151 (R&D Systems) (De Luca et al, 2019). Cells stained with biotinylated antibodies were incubated with streptavidin-PE (Ancell Corporation), whereas samples stained with anti-CD151 primary antibody were incubated with a FITC-conjugated goat antimouse secondary antibody (Ancell Corporation).…”

“…CFU ability has been also reported as a parameter associated with a good clinical outcome for the treatment of IVD degeneration (Pettine et al, 2016;Pettine et al, 2015;Pettine et al, 2017). A recent study of osteoarthritic human articular cartilage-derived cells has observed an enhanced clonogenic ability and sustained expression of stemness markers in cells throughout in vitro expansion, suggesting that this heterogeneous cell population conserves (or even expands) a chondrogenic progenitor pool of cells over time (De Luca et al, 2019). Therefore, it would 158 www.ecmjournal.org P De Luca et al…”

Biological potential alterations of migratory chondrogenic progenitor cells during knee osteoarthritic progression

“…In addition, CPCs are capable of migrating to injured sites after cartilage trauma [7] or diseased osteoarthritic cartilages [8]. Furthermore, CPCs have recently attracted interest due to their immunoregulatory properties [9,10] and phagocytic capacity [11], which have been suggested as valuable potentials for cell-based therapies [6,[12][13][14]. However, numerous studies from independent teams brought inconclusive information in understanding CPCs activity at different phase of knee OA progression.…”