Therapeutic Potential of Bone Marrow Derived Mesenchymal Stem Cells in Modulating Astroglyosis of Surgical Induced Experimental Spinal Cord Injury

Background: Spinal cord injury (SCI) unsuccessful regeneration was due to glial scar development. It was a major obstacle to axonal restoration. Safe therapeutic intervention by the use of bone marrow derived stem cells (BMMSCs) transplantation applied in the present study could reduce spinal disability. Material and methods: Forty male albino rats were divided into four groups: GI: negative control (n = 10 rats);GII: positive control after SCI (n = 10 rats);GIII: SCI + BM - MSCs intravenous injected and GIV: SCI + BM - MSCs intra lesion injected (n = 10 rats in each group). The samples were taken from spinal cord tissues around the region of injury and were subjected to histological, immunohistochemical assessment. RNA extraction and real time PCR for detection of nerve regeneration and astrocyte response to the injury were also performed. Results: Clinical improvement occurred by the enhancement in the Basso, Beattie and Bresnahan (BBB) score after SCI. Histological examinations showed positive regenerative responses in GIV compared to GIII. Conclusion: BM-MSCs transplantation has a promising role in enhancing the microenvironment for nerve regeneration through stumbling the glial scaring formation and inflammatory response after chronic spinal cord injury especially by using intra-lesion route injection.

Molecular detection of monocyte chemotactic protein-1 polymorphism in spontaneous bacterial peritonitis patients

AIM: To investigate the association of the functional monocyte chemotactic protein-1(MCP-1) promoter polymorphism(A-2518G) with spontaneous bacterial peritonitis(SBP).METHODS: Fifty patients with post-hepatitis C liver cirrhosis and ascites were categorized into two groups; group Ⅰ included 25 patients with SBP and group Ⅱ included 25 patients free from SBP. In addition, a group of 20 healthy volunteers were included. We assessed the MCP-1 gene polymorphism and gene expression as well as interleukin(IL)-10 levels in both blood and ascitic fluid. RESULTS: A significant MCP-1 gene polymorphism was detected in groups Ⅰ and Ⅱ(P = 0.001 and 0.02 respectively). Group Ⅰ was associated with a significantly higher frequency of AG genotype [control 8(40%) vs SBP 19(76.0%), P < 0.001], and group Ⅱ was associated with a significantly higher frequency of GG genotype when compared to healthy volunteers [control 1(5%) vs cirrhotic 16(64%), P < 0.001]. Accordingly, the frequency of G allele was significantly higher in both groups(Ⅰ and Ⅱ) [control 10(25%) vs SBP 27(54%), P < 0.001 and vs cirrhotic 37(74.0%), P < 0.001, respectively]. The total blood and ascetic fluid levels of IL-10 and MCP-1 gene expression were significantly higher in group Ⅰ than in group Ⅱ. Group Ⅰ showed significant reductions in the levels of MCP-1 gene expression and IL-10 in the whole blood and ascetic fluid after therapy. CONCLUSION: MCP-1 GG genotype and G allele may predispose HCV infected patients to a more progressive disease course, while AG genotype may increase the susceptibility to SBP. Patients carrying these genotypes should be under supervision to prevent or restrict further complications.

Assessment of heme oxygenase-1 （HO-1） activity in the cavernous tissues of sildenafil citrate-treated rats

Aim： To assess heine oxygenase-1 （HO-1） activity in the cavemous tissue of sildenafil citrate-treated rats. Methods： One hundred and ninety-two Sprague-Dawley male rats, divided into four equal groups, were investigated. Group 1, the control group, received regular animal chow; group 2 received sildenafil citrate by intragastric tube; group 3 received sildenafil and HO inhibitor （zinc protoporphyrin, ZnPP）; and group 4 received sildenafil and nitric oxide synthase （NOS） inhibitor L-nitroarginine methyl ester （L-NAME）. Twelve rats from each group were killed after 0.5 h, 1 h, 2 h and 3 h of drug administration. Then HO-1 activity, cGMP levels and NOS enzymatic activity in the cavernous tissues were estimated. Results： In cavemous tissue, HO-1 activity, NOS enzymatic activity and cGMP concentration increased significantly in sildenafil-treated rats compared to other groups throughout the experiment. Rats receiving either HO or NOS inhibitors showed a significant decrease in these parameters. HO- 1 cavemous tissue activity and NOS enzymatic activity demonstrated a positive significant correlation with cGMP levels （r = 0.646, r = 0.612 respectively; P 〈 0.001）. Conclusion： The actions of PDE5 inhibitor sildenafil citrate in the cavernous tissue are partly mediated through the interdependent relationship between both HO-1 and NOS activities.

Does vitamin C have the ability to augment the therapeutic effect of bone marrow-derived mesenchymal stem cells on spinal cord injury？

Methylprednisolone（MP） is currently the only drug confirmed to exhibit a neuroprotective effect on acute spinal cord injury（SCI）. Vitamin C（VC） is a natural water-soluble antioxidant that exerts neuroprotective effects through eliminating free radical damage to nerve cells. Bone marrow mesenchymal stem cells（BMMSCs）, as multipotent stem cells, are promising candidates in SCI repair. To evaluate the therapeutic effects of MP, VC and BMMSCs on traumatic SCI, 80 adult male rats were randomly divided into seven groups： control, SCI（SCI induction by weight-drop method）, MP（SCI induction, followed by administration of 30 mg/kg MP via the tail vein, once every other 6 hours, for five times）, VC（SCI induction, followed by intraperitoneal administration of 100 mg/kg VC once a day, for 28 days）, MP ＋ VC（SCI induction, followed by administration of MP and VC as the former）, BMMSCs（SCI induction, followed by injection of 3 × 10~6 BMMSCs at the injury site）, and BMMSCs ＋ VC（SCI induction, followed by BMMSCs injection and VC administration as the former）. Locomotor recovery was assessed using the Basso Mouse Scale. Injured spinal cord tissue was evaluated using hematoxylin-eosin staining and immunohistochemical staining. Expression of transforming growth factor-beta, tumor necrosis factor-alpha, and matrix metalloproteinase-2 genes was determined using real-time quantitative PCR. BMMSCs intervention better promoted recovery of nerve function of rats with SCI, mitigated nerve cell damage, and decreased expression of transforming growth factor-beta, tumor necrosis factor-alpha, and matrix metalloproteinase-2 genes than MP and/or VC. More importantly, BMMSCs in combination with VC induced more obvious improvements. These results suggest that VC can enhance the neuroprotective effects of BMMSCs against SCI.

Low-density lipoprotein receptor genetic polymorphism in chronic hepatitis C virus Egyptian patients affects treatment response

AIM: To correlate a genetic polymorphism of the low-density lipoprotein(LDL) receptor with antiviral responses in Egyptian chronic hepatitis C virus(HCV) patients.METHODS: Our study included 657 HCV-infected patients with genotype 4 who received interferonbased combination therapy. Patients were divided into two groups based on their response to therapy: 356 were responders, and 301 were non-responders. Patients were compared to 160 healthy controls. All patients and controls underwent a thorough physical examination, measurement of body mass index(BMI) and the following laboratory tests: serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, albumin, total bilirubin, direct bilirubin, prothrombin time, prothrombin concentration, INR, complete blood count, serum creatinine, fasting blood sugar, HCV antibody, and hepatitis B surface antigen. All HCV patients were further subjected to the following laboratory tests: HCV-RNA using quantitative polymerase chain reaction(PCR), antinuclear antibodies, thyroid-stimulating hormone, an LDL receptor(LDLR) genotype study of LDLR exon8 c.1171G>A and exon-10 c.1413G>A using real-time PCR-based assays, abdominal ultrasonography, ultrasonographic-guided liver biopsy, and histopathological examination of liver biopsies. Correlations of LDL receptor polymorphisms with HAI, METAVIR score, presence of steatosis, and BMI were performed in all cases.RESULTS: There were no statistically significant differences in response rates between the different types of interferon used or LDLR exon10 c.1413G>A. However, there was a significant difference in the frequency of the LDL receptor exon8 c.1171G>A genotype between cases(AA: 25.9%, GA: 22.2%, GG: 51.9%) and controls(AA: 3.8%, GA: 53.1% and GG: 43.1%)(P < 0.001). There was a statistically significant difference in the frequency of the LDLR exon 8C:1171 G>A polymorphism between responders(AA: 3.6%, GA: 15.2%, GG: 81.2%) and nonresponders(AA: 52.2%, GA: 30.6%, GG: 17.2%)(P < 0.001). The G allele of LDL receptor exon8 c.1171G>A predominated in cases and controls over the A allele, and a statistically significant association with response to interferon was observed. The frequency of the LDLR exon8 c.1171G>A allele in non-responders was: A: 67.4% and G: 32.6 vs A: 11.2% and G: 88.8% in responders(P < 0.001). Therefore, carriers of the A allele exhibited a 16.4 times greater risk for nonresponse. There was a significant association between LDL receptors exon8 c.1171G>A and HAI(P < 0.011). There was a significant association between LDL receptors exon8 c.1171G>A and BMI. The mean BMI level was highest in patients carrying the AA genotype(28.7 ± 4.7 kg/m2) followed by the GA genotype(28.1 ± 4.8 kg/m2). The lowest BMI was the GG genotype(26.6 ± 4.3 kg/m2)(P < 0.001). The only significant associations were found between LDL receptors exon8 c.1171G>A and METAVIR score or steatosis(P < 0.001).CONCLUSION: LDL receptor gene polymorphisms play a role in the treatment response of HCV and the modulation of disease progression in Egyptiansinfected with chronic HCV.

Dendritic cell co-stimulatory and co-inhibitory markers in chronic HCV: An Egyptian study

AIM:To assess co-stimulatory and co-inhibitory markers of dendritic cells(DCs)in hepatitis C virus(HCV)infected subjects with and without uremia.METHODS:Three subject groups were included in the study:group 1 involved 50 control subjects,group2 involved 50 patients with chronic HCV infection and group 3 involved 50 HCV uremic subjects undergoing hemodialysis.CD83,CD86 and CD40 as co-stimulatory markers and PD-L1 as a co-inhibitory marker were assessed in peripheral blood mononuclear cells by realtime polymerase chain reaction.Interleukin-10(IL-10)and hyaluronic acid(HA)levels were also assessed.All findings were correlated with disease activity,viral load and fibrogenesis.RESULTS:There was a significant decrease in costimulatory markers;CD83,CD86 and CD40 in groups2 and 3 vs the control group.Co-stimulatory markers were significantly higher in group 3 vs group 2.There was a significant elevation in PD-L1 in both HCV groups vs the control group.PD-L1 was significantly lower in group 3 vs group 2.There was a significant elevation in IL-10 and HA levels in groups 2 and 3,where IL-10was higher in group 3 and HA was lower in group 3 vs group 2.HA level was significantly correlated with disease activity and fibrosis grade in group 2.IL-10 was significantly correlated with fibrosis grade in group 2.There were significant negative correlations between co-stimulatory markers and viral load in groups 2 and3,except CD83 in dialysis patients.There was a significant positive correlation between PD-L1 and viral load in both HCV groups.CONCLUSION:A significant decrease in DC co-stimulatory markers and a significant increase in a DC coinhibitory marker were observed in HCV subjects and to a lesser extent in dialysis patients.

Mesenchymal stem cells transplantation attenuates experimentally induced brain injury after neonatal hypoxia by different two routes of administrations

The neonatal hypoxic-ischemic encephalopathy(HIE)is an important cause of neurological morbidity and mortality in neonates.Cell therapy is considered a promising method for treating severe neurological disorders such as this one.Stem cells have the capacity for self-renewal and differentiation into certain cell lineages.The present study was aimed to find out the most beneficial route of bone marrow-derived mesenchymal stem cells(BMSCs)administration for the attenuation of experimentally induced HIE in neonatal rats.Sixty neonatal rats were divided randomly into four groups.Group 1:control group.Group 2:rats were exposed to bilateral ligation of cephalic arteries.Group 3:rats were exposed to bilateral ligation of cephalic arteries and then underwent intravenous(IV)BMSC injection.Group 4:rats were exposed to bilateral ligation of cephalic arteries and then underwent intracerebroventricular(ICV)BMSC injection.The animals were evaluated by(a)neurobehavioral tests;(b)histopathology,i.e.,histological and immuno-histochemical studies;and(3)gene expression studies.The BMSC treated groups(3 and 4)showed improvement in neurobehavioral tests,histopathological studies,and gene expression,as compared to non-injected lesioned rats(Group 2)with better improvement in Group 4(ICV injections)than in Group 3(IV injections).