Ömer BEŞALTI, Pınar CAN, Ferda PINARLI ALPASLAN, Sevil ATALAY VURAL

Ratlarda Deneysel Omurilik Yaralanmalarinda Allojenik Mezenkimal Kök Hücre Transplantasyonunun Etkilerinin Klinik, Histopatolojik ve Elektrofizyolojik Değerlendirmesi

Bu çalışmanın amacı, allojenik kemik iliği kaynaklı MKH'lerin (KK-MKH) transplantasyonunun sıçanlarda deneysel olarak oluşturulan şiddetli omurilik hasarının (OH) akut evresindeki etkilerini farklı aralıklarla araştırmaktır. Bu çalışmada yetmiş yedi adet Sprague-Dawley sıçan kullanıldı. Kemik iliği, bir günlük sıçanların femur ve tibialarından elde edildi, daha sonra standart yöntemlerde olduğu gibi işlendi. OH, T9-T10 laminektomiden sonra bir anevrizma klipsiyle oluşturuldu. Kök hücre veya kök hücre sulandırıcısının intraparenkimal transplantasyonu hasar oluşturulduktan 6 saat sonra yapıldı. KK-MKH'lerin etkileri postoperatif 1, 3, 7 ve 21. günlerde klinik, elektrofizyolojik ve histopatolojik incelemelerle değerlendirildi. Klinik muayenede, hematüri oranı tedavi grubunda kontrol grubuna göre anlamlı derecede düşüktü (P= 0.003). 21. gün tedavi grubunun sağkalım oranı kontrol grubundan anlamlı olarak yüksekti. Tedavi grubunda 3. ve 7. günlerde somatosensoriyel uyarılmış potansiyellerde (SUP) belirgin bir ilerleme görüldü. Histopatolojik değişiklikler kontrol ve tedavi gruplarında tüm aralıklarda benzerdi. Sonuç olarak, omurilik hasarının akut fazında KK-MKH'lerin intraparenkimal transplantasyonunun hematüri oranı, SUP skorları ve sağkalım oranı ile ilgili bazı umut verici etkiler sağladığı görülmüştür.

Clinical, Histopathological and Electrophysiological Evaluation of the Effects of Allogeneic Mesenchymal Stem Cell Transplantation in Experimental Induced Spinal Cord Injury in Rats

The aim of this study was to investigate the effects of allogeneic bone marrow-derived MSCs (BMMSCs)transplantation in acute stage of experimentally generated severe spinal cord injury (SCI) inrats at different intervals. Seventy seven Sprague-Dawley rats were used in this study. The bonemarrow was harvested from femurs and tibias of the one day-old rats, then processed using thestandard methods. SCI was generated with an aneurysm clip after T9-T10 laminectomy.Intraparenchymal transplantation of the stem cell or stem cell diluent was perfomed at 6th hourafter injury. The effects of the BM-MSCs were evaluated by clinical, electrophysiological andhistopathological examinations on the 1st, 3rd, 7th and 21st day postoperatively. In clinicalexamination, hematuria rate was significantly low in treatment comparing to control (P=0.003). Thesurvival rate of the 21th day treatment group was significantly higher than the control group.Significant improvement in somatosensory evoked potentials (SEP) was observed on the 3rd and7th day in treatment group. The histopathological changes were similar in the control and treatmentgroups at all intervals. In conclusion, intraparenchymal transplantation of BM-MSCs in the acutephase of the SCI provided some benefical effects regarding to hematuria rate, SEP scores andsurvival rate. The results of this study found as promising for doing further, more detailed studieson the treatments of spinal cord injury by the MSCs.

PDF

___

Webb AA, Ngan S, Fowler JD. Spinal cord injury II: Prognostic indicators, standards of care, and clinical trials. Can Vet J 2010; 51: 598-604.
Donovan J, Kirshblum S. Clinical trials in traumatic spinal cord ınjury. Neurotherapeutics 2018; 15: 654-668.
Granger N, Blamires H, Franklin RJM, Jeffery ND. Autologous olfactory mucosal cell transplants in clinical spinal cord injury: a randomized double-blinded trial in a canine translational model. Brain 2012; 135: 3227-3237.
Park EH, White GA, Tieber LM. Mechanisms of injury and emergency care of acute spinal cord injury in dogs and cats. J Vet Emerg Crit Care 2012; 22: 160-178.
Besalti O, Aktas Z, Can P, et al. The use of autologous neurogenically-induced bone marrow-derived mesenchymal stem cells for the treatment of paraplegic dogs without nociception due to spinal trauma. J Vet Med Sci 2016; 78: 1465-1473.
Olby NJ, Muguet-Chanoit AC, Lim JH, et al. A placebocontrolled, prospective, randomized clinical trial of polyethylene glycol and methylprednisolone sodium succinate in dogs with intervertebral disk herniation. J Vet Intern Med 2016; 30: 206-214.
Hurlbert RJ. Methylprednisolone for acute spinal cord injury: an inappropriate standard of care. J Neurosurg 2000; 93: 1-7.
Miękisiak G, Łątka D, Jarmużek P, et al. Steroids in acute spinal cord ınjury: All but gone within 5 years. World Neurosurg 2019; 122: 467-471.
Ramer LM, Au E, Richter MW et al. Peripheral olfactory ensheathing cells reduce scar and cavity formation and promote regeneration after spinal cord injury. J Comp Neurol 2004; 473: 1-15.
Biernaskie J, Sparling JS, Liu J, et al. Skin-derived precursors generate myelinating Schwann cells that promote remyelination and functional recovery after contusion spinal cord injury. J Neurosci 2007; 27: 9545- 9559.
Karimi-Abdolrezaee S, Efekharpour E, Wang J, Schut D, Fehlings MG. Synergistic effects of transplanted adult neural stem/progenitor cells, chondrotinase, and growth factors promote functional repair and plasticity of the chronically injured spinal cord. J Neurosci 2010; 30: 1657- 1676.
Lammertse DP, Jones LA, Charlifue SB, et al. Autologous incubated macrophage therapy in acute, complete spinal cord injury: results of the phase 2 randomized controlled multicenter trial. Spinal Cord 2012; 50: 661-671.
Yazdani So, Pedram M, Hafizi M, et al. A comparison between neurally induced bone marrow derived mesenchymal stem cells and olfactory ensheathing glial cells to repair spinal cord injuries in rat. Tissue Cell 2012; 44: 205-213.
Liu J, Chen P, Wang Q, et al. Meta analysis of olfactory ensheathing cell transplantation promoting functional recovery of motor nerves in rats with complete spinal cord transection. Neural Regen Res 2014; 9: 1850-1858.
Besalti O, Can P, Akpinar E et al. Intraspinal transplantation of autologous neurogenically-induced bone marrow-derived mesenchymal stem cells in the treatment of paraplegic dogs without deep pain perception secondary to intervertebral disk disease. Turkish Neurosurgery 2015; 25: 625-632.
Gazdic M, Volarevic V, Harrell CR et al. Stem cells therapy for spinal cord ınjury. Int J Mol Sci 2018; 19: 1039.
Keirstead HS, Nistor G, Bernal G et al. Human embryonic stem cell derived oligodendrocyte progenitor cell transplants remyelinate and restore locomotion after spinal cord injury. J. Neurosci 2005; 25: 4694–4705.
Lee KH, Suh-Kim H, Choi JS et al. Human mesenchymal stem cell transplantation promotes functional recovery following acute spinal cord injury in rats. Acta Neurobiol Exp 2007; 67: 13-22.
Ide C, Nakai Y, Nakano N et al. Bone marrow stromal cell transplantation for treatment of sub-acute spinal cord injury in the rat. Brain Res 2010; 21: 32-47.
Park WB, Kim SY, Lee SH et al. The effect of mesenchymal stem cell transplantation on the recovery of bladder and hindlimb function after spinal cord contusion in rats. BMC Neuroscience 2010; 11: 119-130.
Nakajima H, Uchida K, Guerrero AR et al. Transplantation of mesenchymal stem cells promotes an alternative pathway of macrophage activation and functional recovery after spinal cord injury. J Neurotrauma 2012; 29: 1614- 1625.
Tan JW, Wang KY, Liao GJ, Chen FM, Mu MZ. Neuroprotective effect of methylprednisolone combined with placenta-derived mesenchymal stem cell in rabbit model of spinal cord injury. Int J Clin Exp Pathol 2015; 8: 8976-8982.
Wang A, Brown EG, Lankford L et al. Placental mesenchymal stromal cells rescue ambulation in ovine myelomeningocele. Stem Cells Transl Med 2015; 4: 659- 669.
Chung HJ, Chung WH, Lee JH et al. Expression of neurotrophic factors in injured spinal cord after transplantation of human-umbilical cord blood stem cells in rats. J Vet Sci 2016; 17: 97-102.
Yaghoobi K, Kaka G, Mansouri K et al. Lavandulaangustifolia extract ımproves the result of human umbilical mesenchymal wharton's jelly stem cell transplantation after contusive spinal cord ınjury in wistar rats. Stem Cells Int 2016; 2016: 5328689.
Carrade DD, Affolter VK, Outerbridge CA et al. Intradermal injections of equine allogeneic umbilical cordderived mesenchymal stem cells are well tolerated and do not elicit immediate or delayed hypersensitivity reactions. Cytotherapy 2011; 13: 1180-1192.
Sekiya I, Larson BL, Smith JR et al. Expansion of human adult stem cells from bone marrow stroma: Conditions that maximize the yields of early progenitors and evaluate their quality. Stem Cells 2002; 20: 530-541.
Şahin F, Saydam G, Omay S. Kök hücre plastisitesi ve klinik pratikte kök hücre tedavisi. Türk hematoloji-onkoloji dergisi 2005; 1: 48-56.
Basso DM, Beattie MS, Bresnahan JC. A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma 1995; 12: 1-21.
Şirin YS, Keleş H, Beşaltı Ö, Vural SA. Deneysel spinal kordtravmalarında atp-mgcl2 ve metilprednizolonun karşılaştırılması. J Clin Anal Med 2012; 3: 442-447.
Rivlin AS, Tator CH. Effect of duration of acute spinal cord compression in a new acute cord injury model in the rat. Surg Neurol 1978; 10: 38-43.
Caliskan M, Simsek S, Vural SA, Besalti O. Comparison of etanercept, etomidate and erythropoietin and their combinations in experimentally-ınduced spinal cord ınjury. Turkish Neurosurgery 2016; 26: 930-936.
Wu S, Cui G, Shao H, et al. The cotransplantation of olfactory ensheathing cells with bone marrow mesenchymal stem cells exerts antiapoptotic effects in adult rats after spinal cord ınjury. Stem Cells International 2015; 2015: 516215.
Aras Y, Sabancı Pa, Kabatas S, et al. The effects of adipose tissue-derived mesenchymal stem cell transplantation during the acute and subacute phases following spinal cord injury. Turkish Neurosurgery 2016; 26: 127-129.
Wang Y, Zhang S, Luo M, Li Y. Hyperbaric oxygen therapy improves local microenvironment after spinal cord injury. Neural Regeneration Research 2014; 9: 2182-2188.