BLINDED PLACEBO STUDY OF BILATERAL OSTEOARTHRITIS TREATMENT USING ADIPOSE DERIVED MESENCHYMAL STEM CELLS
Abstract
Mesenchymal stem cell therapies attract a lot of attention and also controversy in veterinary medicine. In the present study, mesenchymal stem cell therapy, using autologous cells from adipose tissue was evaluated for the treatment of osteoarthritis in the knee. Ten dogs with bilateral osteoarthritis in the knees were included in the study. After collection of adipose tissue and expansion of cells, 2 to 3 millions of mesenchymal stem cells were injected into one knee while sterile phosphate buffer (same as used for resuspension of cells) was injected into the other knee as a placebo. Dog owners were blind in regard which knee received cells and which placebo. Dogs were clinically evaluated before treatment and 3, 6 and 12 months after the treatment, and synovial fluid was collected and evaluated before treatment and after 12 months. Radiographs of both knees were taken before treatment and 6 and 12 months after the treatment. In 9 out of 10 dogs enrolled in the study, there was a significant improvement of lameness after 1 year. Radiographs did not show improvement in the cartilage condition 1 year after the treatment. However, in 7 out of 10 treated joints the osteoarthritis did not progress while in all 10 of placebo treated joints there was a significant worsening of the osteoarthritis. Analysis of the synovial fluid before and 1 year after the treatment showed no statistically significant differences between mesenchymal stem cells treated and placebo treated knees. Results of this study suggest beneficial effects of mesenchymal stem cells treatment in osteoarthritis in dogs and confirm that mesenchymal stem cells treatment is a viable option for managing this debilitating disease in dogs.
Key words: dog; osteoarthritis; knee; mesenchymal stem cells
SLEPA PLACEBO RAZISKAVA ZDRAVLJENJA VNETJA KOLENSKEGA SKLEPA Z MEZENHIMSKIMI MATIČNIMI CELICAMI
Zdravljenje z mezenhimskimi matiènimi celicami v zadnjih letih priteguje veliko pozornosti in tudi razprav v veterinarski medicini. V predstavljeni raziskavi smo preverjali uèinkovitost zdravljenja z matiènimi celicami pri ventju kolenskih sklepov pri psih. V raziskavo je bilo vkljuèenih 10 psov z vnetjem obeh kolenskih sklepov. Psom je bil odvzet košèek mašèobnega tkiva, iz katerega smo osamili in pripravili mezenhimske matiène celice. Dva do tri milijone celic v sterilnem fosfatnem pufru smo nato injicirali v eno obolelo koleno, medtem ko smo v drugo obolelo koleno injicirali samo fosfatni pufer kot placebo. Lastniki psov niso vedeli, v katero koleno so psi prejeli celice in v katero koleno placebo. Psi so bili klinièno pregledani 3, 6 in 12 mesecev po zdravljenju. Sinovialno tekoèino smo iz sklepa odvzeli ob vnosu matiènih celic ter 12 mesecev po zdravljenju. Ob zaèetku zdravljenja ter 6 in 12 mesecev po zdravljenju smo naredili tudi rentgenske posnetke obeh kolenskih sklepov. Od 10 zdravljenih psov smo po enem letu pri devetih opazili znaèilno izboljšanje šepanja po enem letu. Rentgenske slike po enem letu sicer niso pokazale izboljšanja sklepne površine, vendar pa pri 7 od 10 zdravljenih sklepov ni prišlo do napredovanja bolezni, medtem ko je pri vseh 10 nezdravljenih sklepih prišlo do znaèilnega poslabšanja sklepne površine (veèje število oziroma velikost izrastkov). Analiza sinovialne tekoèine ob zaèetku zdravljenja ter eno leto po zdravljenju ni pokazala statistièno znaèilnih razlik v preiskovanih parametrih, kar kae, da v sklepih ni prišlo do vnetnih sprememb kot posledica zdravljenja z mezenhimskimi matiènimi celicami. Rezultati te raziskave kaejo pomembne uèinke mezenhimskih matiènih celic pri zdravljenju vnetja sklepov pri psih in potrjujejo, da je takšno zdravljenje primeren naèin zdravljenja te nevarne kroniène bolezni psov.
Kljuène besede: pes; vnetje sklepov; koleno; mezenhimske matiène celice
References
(1) Rychel JK. Diagnosis and treatment of osteoarthritis. Top Companion Anim Med 2010; 25(1): 20–5.
(2) Sanderson RO, Beata C, Flipo RM, Genevois JP, Macias C, Tacke S et al. Systematic review of the management of canine osteoarthritis. Vet Rec 2009; 164(14): 418-24.
(3) Johnston SA. Osteoarthritis. Joint anatomy, physiology, and pathobiology. Vet Clin North Am Small Anim Pract. 1997; 27(4): 699–723.
(4) Fortier LA, Travis AJ. Stem cells in veterinary medicine. Stem Cell Res Ther 2011; 2(1): e9 (6 pp.) http://stemcellres.biomedcentral.com/articles/10.1186/scrt50
(5) Koh S, Piedrahita JA. From “ES-like†cells to induced pluripotent stem cells: a historical perspective in domestic animals. Theriogenology 2014; 81(1):103–11.
(6) Sylvester KG, Longaker MT. Stem cells: review and update. Arch Surg 2004; 139(1): 93–9.
(7) Whitworth DJ, Banks TA. Stem cell therapies for treating osteoarthritis: prescient or premature? Vet J 2014; 202(3): 416–24.
(8) Minteer DM, Marra KG, Rubin JP. Adipose stem cells: biology, safety, regulation, and regenerative potential. Clin Plast Surg 2015; 42(2): 169–79.
(9) Arnhold S, Wenisch S. Adipose tissue derived mesenchymal stem cells for musculoskeletal repair in veterinary medicine. Am J Stem Cells. 2015; 4(1): e1-12. https://www.ncbi.nlm.nih. gov/pmc/articles/PMC4396154/
(10) Black LL, Gaynor J, Adams C, et al. Effect of intraarticular injection of autologous adipose-derived mesenchymal stem and regenerative cells on clinical signs of chronic osteoarthritis of the elbow joint in dogs. Vet Ther 2008; 9(3): 192–200.
(11) Black LL, Gaynor J, Gahring D, et al. Effect of adipose-derived mesenchymal stem and regenerative cells on lameness in dogs with chronic osteoarthritis of the coxofemoral joints: a randomized, double-blinded, multicenter, controlled trial. Vet Ther 2007; 8(4): 272–84.
(12) Nixon AJ, Dahlgren LA, Haupt JL, Yeager AE, Ward DL. Effect of adipose-derived nucleated cell fractions on tendon repair in horses with collagenase-induced tendinitis. Am J Vet Res 2008; 69(7): 928–37.
(13) Patruno M, Martinello T. Treatments of the injured tendon in veterinary medicine: from scaffolds to adult stem cells. Histol Histopathol 2014; 29(4): 417–22.
(14) Vilar JM, Batista M, Morales M, et al. Assessment of the effect of intraarticular injection of autologous adipose-derived mesenchymal stem cells in osteoarthritic dogs using a double blinded force platform analysis. BMC Vet Res 2014; 10: e143 (7 pp.) http://bmcvetres.biomedcentral. com/articles/10.1186/1746-6148-10-143
(15) Brunnberg L, Johnson KA. Diagnosing lameness in dogs. Oxford : Blackwell Science, 2001: uporab. str. ?
(16) Desando G, Cavallo C, Sartoni F, et al. Intra-articular delivery of adipose derived stromal cells attenuates osteoarthritis progression in an experimental rabbit model. Arthritis Res Ther 2013; 15(1): R22 (15 pp.) http://arthritis-research.biomedcentral.com/ articles/10.1186/ar4156
(17) Hatsushika D, Muneta T, Horie M, Koga H, Tsuji K, Sekiya I. Intraarticular injection of synovial stem cells promotes meniscal regeneration in a rabbit massive meniscal defect model. J Orthop Res 2013; 31(9): 1354–9.
(18) Horie M, Choi H, Lee RH, et al. Intra-articular injection of human mesenchymal stem cells (MSCs) promote rat meniscal regeneration by being activated to express Indian hedgehog that enhances expression of type II collagen. Osteoarthritis Cartilage 2012; 20(10): 1197–207.
(19) Murphy JM, Fink DJ, Hunziker EB, Barry FP. Stem cell therapy in a caprine model of osteoarthritis. Arthritis Rheum 2003; 48(12): 3464–74.
(20) ter Huurne M, Schelbergen R, Blattes R, et al. Antiinflammatory and chondroprotective effects of intraarticular injection of adipose-derived stem cells in experimental osteoarthritis. Arthritis Rheum 2012; 64(11): 3604–13.
(21) Diekman BO, Wu CL, Louer CR, et al. Intra-articular delivery of purified mesenchymal stem cells from C57BL/6 or MRL/MpJ superhealer mice prevents posttraumatic arthritis. Cell Transplant 2013; 22(8): 1395–408.
(22) Frisbie DD, Kisiday JD, Kawcak CE, Werpy NM, McIlwraith CW. Evaluation of adipose-derived stromal vascular fraction or bone marrow-derived mesenchymal stem cells for treatment of osteoarthritis. J Orthop Res 2009; 27(12): 1675–80.
(23) Lee DH, Sonn CH, Han SB, Oh Y, Lee KM, Lee SH. Synovial fluid CD34(-) CD44(+) CD90(+) mesenchymal stem cell levels are associated with the severity of primary knee osteoarthritis. Osteoarthritis Cartilage 2012; 20(2): 106–9.
(24) Schelbergen RF, van Dalen S, ter Huurne M, et al. Treatment efficacy of adipose-derived stem cells in experimental osteoarthritis is driven by high synovial activation and reflected by S100A8/A9 serum levels. Osteoarthritis Cartilage 2014; 22(8): 1158–66.
(25) van Buul GM, Villafuertes E, Bos PK, et al. Mesenchymal stem cells secrete factors that inhibit inflammatory processes in short-term osteoarthritic synovium and cartilage explant culture. Osteoarthritis Cartilage 2012; 20(10): 1186–96.
(26) Vilar JM, Morales M, Santana A, et al. Controlled, blinded force platform analysis of the effect of intraarticular injection of autologous adipose-derived mesenchymal stem cells associated to PRGF-Endoret in osteoarthritic dogs. BMC Vet Res 2013; 9: e131 (6 pp.) http://bmcvetres.biomedcentral.com/articles/10.1186/1746-6148-9-131
(27) Wilke MM, Nydam DV, Nixon AJ. Enhanced early chondrogenesis in articular defects following arthroscopic mesenchymal stem cell implantation in an equine model. J Orthop Res 2007; 25(7): 913–25.
(28) Brunnberg L. Myeloperoxidase, a highly sensitive and specific pain indicator in osteoarthritis. In: Hill’s European Symposium on Osteoarthritis and Joint Health. Genova, 2005: 20–5.
