BACKGROUND: Recently, cells derived from synovial mesenchymal stem cells (MSCs) have been regarded as a potential source of cells to induce repair of articular cartilage. To investigate more effective methods for promoting chondrogenesis, we examined the effects of osteogenic protein (OP)-1 with or without transforming growth factor-beta (TGFbeta1) on chondrogenesis of human MSCs in vitro. METHODS: MSCs were isolated from the synovial membrane of patients with rheumatoid arthritis undergoing knee replacement surgery. After expansion of the cells, pellet cultures were performed in chondrogenic medium with OP-1 100-200 ng/ml, TGFbeta1 10 ng/ml, or both agents for 3 or 6 weeks. Chondrogenesis was evaluated histologically with safranin O staining, reverse transcription polymerase chain reaction for aggrecan and type II collagen mRNA, and quantification of glycosaminoglycan (GAG) content using a dimethylmethylene blue dye-binding assay. GAG content was normalized by DNA content measured using Hoechst 33258 dye. RESULTS: At 3 weeks of culture, mRNAs for type II collagen and aggrecan were expressed by MSCs treated with either TGFbeta1 or OP-1; however, substantial matrix production was not induced. At 6 weeks, OP-1 increased GAG accumulation dose-dependently in the presence or absence of TGFbeta1, and the GAG content was the highest after combined treatment with 200 ng OP-1 and TGFbeta1. Histological staining for safranin O was poor after treatment with OP-1 or TGFbeta1 alone and slightly increased after combined treatment with TGFbeta1 and OP-1 at 3 weeks. At 6 weeks, OP-1 increased the intensity of staining dose-dependently in the presence or absence of TGFbeta1. However, the histological appearance of the cells treated with OP-1 alone was similar to that of hypertrophic chondrocytes, which was different from that of cells with combined treatment with OP-1 and TGFbeta1. CONCLUSIONS: A high dose of OP-1 was useful for enhancing chondrogenesis from synovium-derived MSCs in combined treatment with TGFbeta1.
BACKGROUND: Recently, cells derived from synovial mesenchymal stem cells (MSCs) have been regarded as a potential source of cells to induce repair of articular cartilage. To investigate more effective methods for promoting chondrogenesis, we examined the effects of osteogenic protein (OP)-1 with or without transforming growth factor-beta (TGFbeta1) on chondrogenesis of human MSCs in vitro. METHODS: MSCs were isolated from the synovial membrane of patients with rheumatoid arthritis undergoing knee replacement surgery. After expansion of the cells, pellet cultures were performed in chondrogenic medium with OP-1 100-200 ng/ml, TGFbeta1 10 ng/ml, or both agents for 3 or 6 weeks. Chondrogenesis was evaluated histologically with safranin O staining, reverse transcription polymerase chain reaction for aggrecan and type II collagen mRNA, and quantification of glycosaminoglycan (GAG) content using a dimethylmethylene blue dye-binding assay. GAG content was normalized by DNA content measured using Hoechst 33258 dye. RESULTS: At 3 weeks of culture, mRNAs for type II collagen and aggrecan were expressed by MSCs treated with either TGFbeta1 or OP-1; however, substantial matrix production was not induced. At 6 weeks, OP-1 increased GAG accumulation dose-dependently in the presence or absence of TGFbeta1, and the GAG content was the highest after combined treatment with 200 ng OP-1 and TGFbeta1. Histological staining for safranin O was poor after treatment with OP-1 or TGFbeta1 alone and slightly increased after combined treatment with TGFbeta1 and OP-1 at 3 weeks. At 6 weeks, OP-1 increased the intensity of staining dose-dependently in the presence or absence of TGFbeta1. However, the histological appearance of the cells treated with OP-1 alone was similar to that of hypertrophic chondrocytes, which was different from that of cells with combined treatment with OP-1 and TGFbeta1. CONCLUSIONS: A high dose of OP-1 was useful for enhancing chondrogenesis from synovium-derived MSCs in combined treatment with TGFbeta1.
Authors: Jorge Paz Rodriguez; Michael P Murphy; Soonjun Hong; Marialaura Madrigal; Keith L March; Boris Minev; Robert J Harman; Chien-Shing Chen; Ruben Berrocal Timmons; Annette M Marleau; Neil H Riordan Journal: Int Arch Med Date: 2012-02-08
Authors: A Martin-Pena; R M Porter; G Plumton; T M McCarrel; A J Morton; M V Guijarro; S C Ghivizzani; B Sharma; G D Palmer Journal: Eur Cell Mater Date: 2018-10-12 Impact factor: 3.942
Authors: Lisa A Fortier; Joseph U Barker; Eric J Strauss; Taralyn M McCarrel; Brian J Cole Journal: Clin Orthop Relat Res Date: 2011-10 Impact factor: 4.176
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