GC-07. Production of injectable implant mesenchymal stem cells for treatment of osteoarthritis
Hanna A. Zhernasechanka1, Yanina I. Isaikina1, Dzmitry V. Bukach2
1 Belarusian Research Center for Pediatric Oncology, Hematology and Immunology, Minsk, Republic of Belarus
2 Republican Scientific and Practical Center of Traumatology and Orthopedics, Minsk, Republic of Belarus
Contact: Dr. Hanna A. Zhernasechanka, phone: +375 (29) 399-37-65, e-mail: sapphire.anna@gmail.com
Summary
Due to the restricted capacity of resident chondrocytes to regenerate injury of cartilage, stem cell-based therapies have been proposed as a novel therapeutic approach for cartilage repair. The aim of this work was to develop the method for injection of MSCs in the treatment of osteoarthritis.
Materials and methods
MSCs have been derived from bone marrow and expanded in vitro in DMEM with 10% FBS-MSC. Chondrogenic pre-differentiation of MSCs in 2D were obtained by culturing in DMEM with 10 ng/ml TGFβ3, 100 ng/ml IGF, 100 nM dexamethasone (Life Technologies, USA), 1% antibiotic for 7 days. The injectable implant was obtained by mixing predifferentiated MSCs with 1% hyaluronic acid in a ratio of 1:1. Chondrogenesis of MSCs was confirmed by immunofluorescence staining, PCR. The clinical efficacy of injectable implants was assessed by VAS (Visual Analogue Scale) and WOMAC (Western Ontario McMaster Universities OA Index).
Results
The study included twelve patients with osteoarthritis at the 2-3rd severity stage. The cells had high 98 (97.5; 99.5)% viability, phenotype of MSCs (more than 95% of cells had CD90, CD105, and CD73). Chondrogenesis after supplementation with the growth factors was proven by detection of the type II collagen and aggrecan synthesis by MSCs, as shown by immunofluorescence. A change in morphology MSCs from spindle-shaped to “cobblestone-like” was detached. We have shown an increased expression of COL2 by 10 times (p=0.002), COL11 by 3 times (p=0.03), along with 5-fold decrease of COL1 expression (p=0.03). A total of 12 clinical preparations of chondrogenic pre-differentiated MSCs with 1% hyaluronic acid were obtained, at the average number of 31.0 (25.0; 37.5)×106 cells in a mean volume of 0.95 (0.65; 1.05) mL. The cellular implants were injected to the patients intra-articularly. After a follow-up periods of 6 and 12 months, clinical effects were evaluated by means of VAS and WOMAC assessment scales. By the VAS scale, distinct changes were shown at the end of follow-up periods, i.e., from 5 cm (4; 6) to 2.5 cm (2; 3.5), and to 2 cm (1; 4), respectively (p <0.05). The data of WOMAC evaluation score showed improvement during the follow-up periods from 56.5 (37; 63) to 31 (22; 41) points, and to 31 (9; 48) respectively (p <0.05). Systemic or local complications were not observed in the patients after injections of implanted cells into the knee joint.
Conclusion
The results of this study confirm the efficacy of intra-articular injections of chondrogenically predifferentiated allogeneic MSCs in hyaluronic acid when treating patients with osteoarthritis.
Keywords
Mesenchymal stem cells, cell-based therapy, osteoarthritis.
