The Influence of IL-10 and TNFα on Chondrogenesis of Human Mesenchymal Stromal Cells in Three-Dimensional Cultures

Abstract

Chondrogenic differentiated mesenchymal stromal cells (MSCs) are a promising cell source for articular cartilage repair. This study was undertaken to determine the effectiveness of two three-dimensional (3D) culture systems for chondrogenic MSC differentiation in comparison to primary chondrocytes and to assess the effect of Interleukin (IL)-10 and Tumor Necrosis Factor (TNF)α on chondrogenesis by MSCs in 3D high-density (H-D) culture. MSCs were isolated from femur spongiosa, characterized using a set of typical markers and introduced in scaffold-free H-D cultures or non-woven polyglycolic acid (PGA) scaffolds for chondrogenic differentiation. H-D cultures were stimulated with recombinant IL-10, TNFα, TNFα + IL-10 or remained untreated. Gene and protein expression of type II collagen, aggrecan, sox9 and TNFα were examined. MSCs expressed typical cell surface markers and revealed multipotency. Chondrogenic differentiated cells expressed cartilage-specific markers in both culture systems but to a lower extent when compared with articular chondrocytes. Chondrogenesis was more pronounced in PGA compared with H-D culture. IL-10 and/or TNFα did not impair the chondrogenic differentiation of MSCs. Moreover, in most of the investigated samples, despite not reaching significance level, IL-10 had a stimulatory effect on the type II collagen, aggrecan and TNFα expression when compared with the respective controls.

Chondrogenic potential of IL-10 in mechanically injured cartilage and cellularized collagen ACI grafts

“…Interpretations based on type 1 collagen staining were limited by the scaffolds nature itself (type 1/ 3 collagen composition), but there appeared a trend for less type 1 collagen especially in the co-treatment group. Similar findings have been shown by Schulze-Tanzil et al, who demonstrated an IL-10 induced COL2A1 expression in mesenchymal stem cells, that were embedded in a polyglycolic acid scaffolds 29 . Interestingly, HA biosynthesis was much more pronounced compared to aggrecan.…”

“…Interestingly, joint fluid analyses revealed relatively low in vivo concentrations ranging from 1 pg/ml e 1 ng/ml after traumatic cartilage defects or OA 2,28 . Thematically related studies that directly applied IL-10 used dosages ranging from 10 to 300 ng/ml, but there is no study we are aware of that investigated dosages lower than 1 ng/ml in cartilage yet 13,15,29,30 . In addition, we could demonstrate that rising dosages of IL-10 led to reduced mRNA expression of IL-10 indicating an autocrine feedback mechanism in chondrocytes ( Fig.…”

Testing the potency of anti‐TNF‐α and anti‐IL‐1β drugs using spheroid cultures of human osteoarthritic chondrocytes and donor‐matched chondrogenically differentiated mesenchymal stem cells

“…MSCs obtained from RA and OA patients displayed similar chondrogenic potential as MSCs isolated from healthy individuals 47 Also, MSCs isolated from bone marrow of OA patients had capacity to produce hyaline cartilage suitable for tissue repair. [66][67][68] Therefore we see no limitations in using these kinds of cells for in vitro testing of anti-inflammatory drugs.…”

Key transcription factors in the differentiation of mesenchymal stem cells

“…The transfection resulted in a significant increase in the chondrogenic differentiation of MSCs (Park et al, 2011). TNF-α has been also found to upregulate Sox9 gene expression (Jagielski et al, 2014). Adenovirus-mediated BMP2 and Sox9 expression in mice embryonic MSCs have shown to effectively enhance chondrogenic differentiation in vitro (Liao et al, 2014).…”