Inflammatory effects of autologous, genetically modified autologous, allogeneic, and xenogeneic mesenchymal stem cells after intra-articular injection in horses

J. H. Pig; A. Ishihara; M. L. Wellman; D. S. Russell; A. L. Bertone

doi:10.3415/VCOT-13-01-0008

Veterinary and Comparative Orthopaedics and Traumatology, Table of Contents

Vet Comp Orthop Traumatol 2013; 26(06): 453-460
DOI: 10.3415/VCOT-13-01-0008

Original Research

Schattauer GmbH

Inflammatory effects of autologous, genetically modified autologous, allogeneic, and xenogeneic mesenchymal stem cells after intra-articular injection in horses

Authors

J. H. Pig

¹Comparative Orthopedics Research Laboratory, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Ohio State University, Columbus, Ohio, USA
A. Ishihara

¹Comparative Orthopedics Research Laboratory, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Ohio State University, Columbus, Ohio, USA
M. L. Wellman

²Comparative Orthopedics Research Laboratory, Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH, USA
D. S. Russell

²Comparative Orthopedics Research Laboratory, Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH, USA
A. L. Bertone

¹Comparative Orthopedics Research Laboratory, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Ohio State University, Columbus, Ohio, USA

Abstract

Summary

Objectives: To compare the clinical and inflammatory joint responses to intra-articular injection of bone marrow-derived mesenchymal stem cells (MSC) including autologous, genetically modified autologous, allogeneic, or xenogeneic cells in horses.

Methods: Six five-year-old Thoroughbred mares had one fetlock joint injected with Gey's balanced salt solution as the vehicle control. Each fetlock joint of each horse was subsequently injected with 15 million MSC from the described MSC groups, and were assessed for 28 days for clinical and inflammatory parameters representing synovitis, joint swelling, and pain.

Results: There were not any significant differences between autologous and genetically modified autologous MSC for synovial fluid total nucleated cell count, total protein, interleukin (IL)-6, IL-10, fetlock circumference, oedema score, pain-free range-of-motion, and soluble gene products that were detected for at least two days. Allogeneic and xenogeneic MSC produced a greater increase in peak of inflammation at 24 hours than either autologous MSC group.

Clinical significance: Genetically engineered MSC can act as vehicles to deliver gene products to the joint; further investigation into the therapeutic potential of this cell therapy is warranted. Intra-articular MSC injection resulted in a moderate acute inflammatory joint response that was greater for allogeneic and xenogeneic MSC than autologous MSC. Clinical management of this response may minimize this effect.

Keywords

Mesenchymal stem cell - bone marrow - horse - inflammation

Full Text

References

References
1 Agung M, Ochi M, Yanada S. et al. Mobilization of bone marrow-derived mesenchymal stem cells into the injured tissues after intra-articular injection and their contribution to tissue regeneration. Knee Surg Sports Traumatol Arthrosc 2006; 14: 1307-1314.
2 Lee KB, Hui JH, Song IC. et al. Injectable mesenchymal stem cell therapy for large cartilage defects--a porcine model. Stem Cells 2007; 25: 2964-2971.
3 Murphy JM, Fink DJ, Hunziker EB. et al. Stem cell therapy in a caprine model of osteoarthritis. Arthritis Rheum 2003; 48: 3464-3474.
4 Godwin EE, Young NJ, Dudhia J. et al. Implantation of bone marrow-derived mesenchymal stem cells demonstrates improved outcome in horses with overstrain injury of the superficial digital flexor tendon. Equine Vet J 2012; 44: 25-32.
5 McIlwraith CW, Frisbie DD, Rodkey WG. et al. Evaluation of intra-articular mesenchymal stem cells to augment healing of microfractured chondral defects. Arthroscopy 2011; 27: 1552-1561.
6 Arnhold SJ, Goletz I, Klein H. et al. Isolation and characterization of bone marrow-derived equine mesenchymal stem cells. Am J Vet Res 2007; 68: 1095-1105.
7 Murray SJ, Santangelo KS, Bertone AL. Evaluation of early cellular influences of bone morphogenetic proteins 12 and 2 on equine superficial digital flexor tenocytes and bone marrow-derived mesenchymal stem cells in vitro. Am J Vet Res 2010; 71: 103-114.
8 Berg L, Koch T, Heerkens T. et al. Chondrogenic potential of mesenchymal stromal cells derived from equine bone marrow and umbilical cord blood. Vet Comp Orthop Traumatol 2009; 22: 363-370.
9 Guest DJ, Ousey JC, Smith MRW. Defining the expression of marker genes in equine mesenchymal stromal cells. Stem Cells Cloning 2008; 1: 1-9.
10 Herten M, Grassman JP, Sager M. et al. Bone marrow concentrate for autologous transplantation in minipigs. Characterization and osteogenic potential of mesenchymal stem cells. Vet Comp Orthop Traumatol 2013; 26: 34-41.
11 Gafni Y, Turgeman G, Liebergal M. et al. Stem cells as vehicles for orthopedic gene therapy. Gene Ther 2004; 11: 417-426.
12 Zachos TA, Shields KM, Bertone AL. Gene-mediated osteogenic differentiation of stem cells by bone morphogenetic proteins-2 or -6. J Orthop Res 2006; 24: 1279-1291.
13 Ishihara A, Zekas LJ, Litsky AS. et al. Dermal fibroblast-mediated BMP2 therapy to accelerate bone healing in an equine osteotomy model. J Orthop Res 2010; 28: 403-411.
14 Zachos T, Diggs A, Weisbrode S. et al. Mesenchymal stem cell-mediated gene delivery of bone morphogenetic protein-2 in an articular fracture model. Mol Ther 2007; 15: 1543-1550.
15 Menendez MI, Clark DJ, Carlton M. et al. Direct delayed human adenoviral BMP-2 or BMP-6 gene therapy for bone and cartilage regeneration in a pony osteochondral model. Osteoarthritis Cartilage 2011; 19: 1066-1075.
16 Ishihara A, Bartlett JS, Bertone AL. Inflammation and immune response of intra-articular serotype 2 adeno-associated virus or adenovirus vectors in a large animal model. Arthritis. 2012 2012: 735472.
17 Carrade DD, Owens SD, Galuppo LD. et al. Clinicopathologic findings following intra-articular injection of autologous and allogeneic placentally derived equine mesenchymal stem cells in horses. Cytotherapy 2011; 13: 419-430.
18 Frisbie DD, Kisiday JD, Kawcak CE. et al. Evaluation of adipose-derived stromal vascular fraction or bone marrow-derived mesenchymal stem cells for treatment of osteoarthritis. J Orthop Res 2009; 27: 1675-1680.
19 Watts AE, Nixon AJ. Distribution and homing of stem cells after intra-articular injection to normal and arthritic joints. Proceedings of the 2011 ACVS Veterinary Symposium. 2011. November 3-5 Chicago IL, USA: 40 E50.
20 Ishihara A, Helbig H, Sanchez-Hodge R. et al. A novel gravitational marrow separator to isolate bone marrow mesenchymal stem cells. Proceedings of the 2012 Annual Meeting of the Orthopedic Research Society. 2012. February 4-7 San Francisco CA, USA: 568.
21 Bonewald LF, Harris SE, Rosser J. et al. Von Kossa staining alone is not sufficient to confirm that mineralization in vitro represents bone formation. Calc Tissue Int 2003; 72: 537-547.
22 Ishihara A, Zachos T, Bartlett J. et al. Evaluation of permissiveness and cytotoxic effects in equine chondrocytes, synovial cells and stem cells in response to infection with adenovirus 5 vectors for gene delivery. Am J Vet Res 2006; 67: 1145-1155.
23 Kay AT, Bolt DM, Ishihara A. et al. Anti-inflammatory and analgesic effects of intra-articular injection of triamcinolone acetonide, mepivacaine hydrochloride, or both on lipopolysaccharide-induced lameness in horses. Am J Vet Res 2008; 69: 1646-1654.
24 Aggarwal S, Pittenger M. Human mesenchymal stem cells modulate allogeneic immune cell responses. Blood 2005; 105: 1815-1822.
25 Guo Z, Zheng C, Chen Z. et al. Fetal BM-derived mesenchymal stem cells promote the expansion of human Th17 cells, but inhibit the production of Th1 cells. Eur J Immunol 2009; 39: 2840-2849.
26 Frisbie DD, Ghivizanni SC, Robbins PD. et al. Treatment of experimental equine osteoarthritis by in vivo delivery of the equine interleukin-1 receptor antagonist gene. Gene Ther 2002; 9: 12-20.
27 Dominici M, Le Blanc K, Mueller I. et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006; 8: 315-317.
28 Li XY, Ding J, Zheng ZH. et al. Long-term culture in vitro impairs the immunosuppressive activity of mesenchymal stem cells on T cells. Mol Med Rep 2012; 6: 1183-1189.