title: Prostanoid pattern and iNOS expression during chondrogenic differentiation of human mesenchymal stem cells.
abstract: Availability of human chondrocytes is a major limiting factor regarding drug discovery projects and tissue replacement therapies. As an alternative human mesenchymal stem cells (hMSCs) from bone marrow are taken into consideration as they can differentiate along the chondrogenic lineage. However, it remains to be shown whether they could form a valid model for primary chondrocytes with regards to inflammatory mediator production, like nitric oxide (NO) and prostanoids. We therefore investigated the production of NO and prostanoids in hMSCs over the course of chondrogenic differentiation and in response to IL-1beta using primary OA chondrocytes as reference. Chondrogenic differentiation was monitored over 28 days using collagen I, collagen II, and collagen X expression levels. Expression levels of inducible nitric oxide synthase (iNOS), levels of NO, and prostanoids were assessed using PCR, Griess assay, and GC/MS/MS, respectively. The hMSCs collagen expression profile during course of differentiation was consistent with a chondrocytic phenotype. Contrary to undifferentiated cells, differentiated hMSCs expressed iNOS and produced NO following stimulation with IL-1beta. Moreover, this induction of iNOS expression was corticosteroid insensitive. The spectrum of prostanoid production in differentiated hMSCs showed similarities to that of OA chondrocytes, with PGE2 as predominant product. We provide the first detailed characterization of NO and prostanoid production in hMSCs in the course of chondrogenic differentiation. Our results suggest that differentiated hMSCs form a valid model for chondrocytes concerning inflammatory mediator production. Furthermore, we propose that IL-1beta stimulation, leading to corticosteroid-insensitive NO synthesis, can be used as a sensitive marker of chondrogenesis.
authors.affiliation: ALTANA Pharma AG, Byk Gulden Street 2, 78467 Konstanz, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Mais
authors.forename: Thomas
authors.initials: T
authors.lastname: Klein
authors.forename: Volker
authors.initials: V
authors.lastname: Ullrich
authors.forename: Christian
authors.initials: C
authors.lastname: Schudt
authors.forename: Gereon
authors.initials: G
authors.lastname: Lauer
mesh_terms: D000293:Adolescent; D000328:Adult; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000201:enzymology*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007375:Interleukin-1 / Q000494:pharmacology; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000201:enzymology*; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis*; D011453:Prostaglandins / Q000096:biosynthesis*
chemical_list: D007375:Interleukin-1; D011453:Prostaglandins; D052247:Nitric Oxide Synthase Type II