The potential of intra-articular injection of chondrogenicinduced bone marrow stem cells to retard the progression of osteoarthritis in a sheep model

Transcription

1 Health and Biomedicine /qfarf.2012.BMP30 The potential of intra-articular injection of chondrogenicinduced bone marrow stem cells to retard the progression of osteoarthritis in a sheep model Hamoud Hussein Alfaqeh, Ruszymah Bint haji Idrus, Nor Hamdan Mohamad Yahya Yahya, Cheng Hui Chen, Saim Aminuddin International Islamic University of Malaysia, Kuantan, MALAYSIA; Universiti Kebangsaan, Kuala Lumpur, MALAYSIA; Universiti Putra, Selangor, MALAYSIA; Ampang Puteri Specialist Hospital, Selangor, MALAYSIA fanousi08@gmail.com Background: Osteoarthritis (OA) is the most common joint disease in middle aged and older people. Despite the multiple modalities of treatment, the outcome is still poor and focuses on temporary measures to alleviate the symptoms. Objectives: We want to determine whether or not an intra-articular injection of a single dose of chondrogenic induced bone marrow mesenchymal stem cells (BMSC) can promote cartilage regeneration in surgically induced osteoarthritis in sheep. Methods: Sheep BMSCs were isolated and divided into two groups. One group was cultured in chondrogenic media containing 5 ng/ml TGFβ ng/ml IGF-1, and the other group was cultured in basal non-chondrogenic media for the duration of 3 weeks. The procedure for surgically inducing osteoarthritis was performed on the donor sheep six weeks prior to intra-articular injection to the knee joint. The injection is a single dose of BMSCs from either group, suspended in 5 ml Dulbecco's Modified Eagle Medium (DMEM) at density of 2 million cells/ml. The control groups were injected with basic cell free media. Results: Six weeks after injection, evidence of articular cartilage regeneration and meniscus repair in osteoarthritic knee joints treated with autologous BMSCs cultured in chondrogenic medium were observed. No evidence of regeneration and meniscus repair was observed for the control group and the group treated with BMSCs cultured in basal medium. Conclusions: Intra-articular injection of a single dose of BMSCs in chondrogenic culture could stimulate regeneration of articular cartilage and meniscal tissue. This abstract is available through QScience.com

2 1 Figure Legends Fig 1: Morphology of sheep bone marrow mesenchymal stem cells (BMSCs) in vitro. BMSCs cultured in FD medium supplemented with 10% FBS alone at week-1 as primary culture in passage zero (A) and at week 3 BMSCs formed dense monolayer cells and reached confluence (B) Chondrogenic induced BMSCs cultured in FD medium supplemented with 1 % FBS, 10ng/ml TGFβ-3 and 50 ng/ml IGF-1 at week-1 showing cell aggregates (C) and at week 3 cell aggregates united forming big aggregation (D). Original magnification x 40. Fig 2: Gross evaluation of OA and normal knee joints 6 weeks after intra-articular cell / medium injection. Severe OA was observed in knee joint received single dose of basal medium alone control group (A and B). Mild OA was observed in knee joint received single dose, 10x10 6 ABMSCs supplemented with TGFβ-3 and 1 % FBS group A: (C and D). In group B: Moderate OA was observed in knee joint received single dose, 10 x 10 6 ABMSCs supplemented with 10 % FBS alone (E and F). Contralateral knee joint appeared to be shiny with no OA (G and H). Arrows indicate to the OA knee joint with or without treatment. Figure 3: Histological examination of sheep femoral condyle OA 6 week s post-intraarticula injection with ABMSCs / medium. Control group: Knee joint received intra-articular injection of basal medium alone (A and-arrows). Group A: knee joint received single dos 10 x10 6 TGF-β3 induced BMSCs supplemented with 1% FBS (C & D-arrows). Group B; knee joint received intra-articular injection of 10 x 01 6 BMSCs supplemented with 10 % FBS alone (E and F-arrows). Contra-lateral knee joints (G and H). All sections were stained with H & E and Safranin-O. Magnification x 40. Arrows indicates to the OA lesions in the operated knee joints. \

3 2 Figure 1 A BMSCs +10 % FBS at week-1 (P0) C TGFβ-3 induced BMSCs at week-1 (P2) B BMSCs +10 % FBS at week-3 (P2) D TGFβ-3 induced BMSCs at week-3 (P2)

4 3 Figure 2

5 4 Figure 3