Therapeutic Effects of Human Mesenchymal Stem Cells on Traumatic Brain Injury in Rats: Secretion of Neurotrophic Factors and Inhibition of Apoptosis

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1 Therapeutic Effects of Human Mesenchymal Stem Cells on Traumatic Brain Injury in Rats: Secretion of Neurotrophic Factors and Inhibition of Apoptosis Hwa-Jung Kim, Jae-Ho Lee, and Se-Hyuk Kim Presentation title / topic OR Presenter's name 1

2 Cell death Biochemistry, Genetics and Molecular Biology» "Apoptosis and Medicine", book edited by Tobias M. Ntuli, ISBN , Published: August 16, 2012 under CC BY 3.0 license. The Author(s). Cell Death and Cancer, Novel Therapeutic Strategies By Silvina Grasso, M. Piedad Menéndez-Gutiérrez, et al DOI: /

3 Cell death Pathways of apoptotic and non-apoptotic death in tumour cells, Hitoshi Okada & Tak W. Mak Nature Reviews Cancer 4, (August 2004) doi: /nrc1412 3

4 Traumatic brain injury Austria Incidence: 303/100,000/year Mortality rate: 11/100,000/year Occurrence highest in: in male teens female octogenarians leading cause of death in people aged year globally Epidemiology of traumatic brain injury in Austria Walter Mauritz et al. Wien Klin Wochenschr. 2014; 126(1-2): Published online 2013 Nov 19. doi: /s T he impact of traumatic brain injuries: a global perspective. Hyder AA et al. NeuroRehabilitation. 2007;22(5):

5 Symptoms of TBI Mild TBI Fatigue Headaches Visual disturbances, Sensitivity to light and sounds Memory loss Poor attention/concentration Sleep disturbances Dizziness/loss of balance Irritability-emotional disturbances Feelings of depression Seizure Nausea Loss of smell last access on

6 Symptoms of TBI Severe TBI not understanding the spoken word (receptive aphasia) difficulty speaking and being understood (expressive aphasia) partial or total loss of vision weakness of eye muscles and double vision (diplopia) loss or diminished sense of smell (anosmia) Convulsions Physical paralysis/spasticity Chronic pain Control of bowel and bladder Loss of stamina Appetite changes Regulation of body temperature last access on

7 Materials and Methods 6-8 weeks old Sprague Dawley rats Group 1: Sham control group (craniotomy, without TBI) Group 2: Placebo group (TBI with saline injection) Group 3: Treatment group (TBI with hmsc injection) neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

8 Materials and Methods First set of animals (8 per subgroup) Neurological assessment ELISA Immunohistochemistry Sacrifice on day 2,8,15 and 29 (total n=96) Second set of animals (4 per subgroup) Westernblot Sacrifice on day 2 and 8 (total n=24) neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

9 Experimental design Mononuclear cells from human bone marrow (posterior iliac crest) Differentiation in culture to human mesenchymal stem cells (hmsc) Intravenous application 24h after TBI neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

10 Traumatic brain injury Fixation in stereotaxic frame Drilling an 8mm hole Adjacent to central suture, between lambda and bregma Impacting right cortex with pneumatic piston 2mm of compression, Velocity of 3,2 m/sec neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu Picture: last access date

11 Methods Westernblot Immunoprecipitation Immunofluorescent staining Cleaved caspase 3 (apoptosis) Neurological severity score/ Behaviour neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

12 Modified neurological Severity Score Human marrow stromal cell therapy for stroke in rat. Y Li et al. Neurology August 27, 2002 vol. 59 no neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

13 Rotarod motor test neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

14 Results neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

15 Results Role for RIP1 in mediating necroptosis in experimental intracerebral hemorrhage model both in vivo and in vitro. Shen H, Liu C et al. Cell Death Dis Mar 2;8(3):e2641. doi: /cddis

16 Results neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

17 Results neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

18 Results Role for RIP1 in mediating necroptosis in experimental intracerebral hemorrhage model both in vivo and in vitro. Shen H, Liu C et al. Cell Death Dis Mar 2;8(3):e2641. doi: /cddis

19 Results neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

20 Results neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

21 Results neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu

22 Discussion hmsc reach the lesional area Surrogate effect on tissue regeneration not sufficient Activation of neurotrophic factors Amelioration of neurological function due to paracrine effect neurotrophic factors and inhibition of apoptosis. Kim HJ, Lee JH, Kim SH. J Neurotrauma Jan;27(1): doi: /neu