Published in:
Open Access 01-12-2013 | Research article
Effect of interleukin-1β treatment on co-cultures of human meniscus cells and bone marrow mesenchymal stromal cells
Published in: BMC Musculoskeletal Disorders | Issue 1/2013
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Background
Interleukin-1β (IL-1β) is a major mediator of local inflammation present in injured joints. In this study, we aimed at comparing the effect of IL-1β on engineered tissues from MCs, BMSCs and co-cultured MCs and BMSCs.
Methods
We compared the effect of IL-1β in 3 groups: (1) MCs, (2) BMSCs and, (3) co-cultures of MCs and BMSCs. We selected 1 to 3 ratio of MCs to BMSCs for the co-cultures. Passage two (P2) human BMSCs were obtained from two donors. Human MCs were isolated from menisci of 4 donors. Mono-cultures of MCs and BMSCs, and co-cultures of MCs and BMSCs were cultured in chondrogenic medium with TGFβ3, as cell pellets for 14 days. Thereafter, pellets were cultured for 3 more days in same medium as before with or without IL-1β (500 pg/ml). Pellets were assessed histologically, biochemically and by RT-PCR for gene expression of aggrecan, sox9, MMP-1, collagens I and II. Statistics was performed using one-way ANOVA with Tukey’s post-tests.
Results
Co-cultured pellets were the most intensely stained with safranin O and collagen II. Co-cultured pellets had the highest expression of sox9, collagen I and II. IL-1β treatment slightly reduced the GAG/DNA of co-cultured pellets but still exceeded the sum of the GAG/DNA from the proportion of MCs and BMSCs in the co-cultured pellets. After IL-1β treatment, the expression of sox9, collagen I and II in co-cultured pellets was higher compared to their expression in pure pellets. IL-1β induced MMP-1 expression in mono-cultures of MCs but not significantly in mono-cultures of BMSCs or in co-cultured pellets. IL-1β induced MMP-13 expression in mono-cultured pellets of BMSCs and in co-cultured pellets.
Conclusions
Co-cultures of MCs and BMSCs resulted in a synergistic production of cartilaginous matrix compared to mono-cultures of MCs and BMSCs. IL-1β did not abrogate the accumulated GAG matrix in co-cultures but mediated a decreased mRNA expression of aggrecan, collagen II and Sox9. These results strengthen the combinatorial use of primary MCs and BMSCs as a cell source for meniscus tissue engineering by demonstrating retention of fibrochondrogenic phenotype after exposure to IL-1β.