Tehran University Medical Journal

Stem cell therapy for intervertebral disc regeneration: review article

Mahdieh Ghiasi

Intervertebral disks (IVD) acts as shock absorber between each of the vertebrae in the spinal column by keeping the vertebrae separated when the shock caused by the action. They also serve to protect the nerves that run down the middle of the spine and intervertebral disks. The disks are made of fibrocartilaginous material. The outside of the disk is made of a strong material called the annulus fibrosus. Inside this protective covering is a jelly-like substance known as mucoprotein gel. This interior is known as the nucleus pulposus. The nucleus pulposus consists of large vacuolated notochord cells, small chondrocyte-like cells, collagen fibrils, and aggrecan, a proteoglycan that aggregates by binding to hyaluronan. Attached to each aggrecan molecule are glycosaminoglycan (GAG) chains of chondroitin sulfate and keratan sulfate. Intervertebral disks degeneration is frequently associated with low back and neck pain, which accounts as a disability. Despite the known outcomes of the Intervertebral disks degeneration cascade, the treatment of IVD degeneration is limited in that available conservative and surgical treatments do not reverse the pathology or restore the IVD tissue. Regenerative medicine for IVD degeneration, by injection of Intervertebral disks cells, chondrocytes or stem cells, has been extensively studied in the past decade in various animal models of induced IVD degeneration, and has progressed to clinical trials in the treatment of various spinal disease. Despite preliminary results showing positive effects of cell-injection strategies for IVD regeneration, detailed basic research on Intervertebral disks cells and their niche demonstrates that transplanted cells are unable to survive and adapt in the avascular niche of the IVD. For this therapeutic strategy to succeed, the indications for its use and the patients who would benefit need to be better defined. To surmount these obstacles, the solution will be identified only by focused research, both in the laboratory and in the clinic. In present paper, the potential utilization of different adult stem cells for intervertebral disc regeneration has been reported. Bone marrow mesenchymal stem cells, adipose tissue derived stem cells, synovial stem cells and committed IVD cells have been studied for this purpose either in vitro or in vivo.

Effects of cyclophosphamide on proliferation and viability of malignant HN5 cell line and comparing with embryonic fibroblast cells

Elham Hoveizi

Background: One of the major causes of death in the world is cancer and therefore any study in the field of cancer biology is of great importance. Head and neck cancers represent approximately 2-5% of neoplasms which is higher in some countries. The most appropriate therapy for various cancers is identifying effective and efficient ways that contribute to initiation of apoptosis. Cyclophosphamide is an alkylating agent that stops the replication of DNA and then, it stops the cell proliferation and viability. Therefore, cyclophosphamide is used to treat various types of cancer. In this study we evaluate the cytotoxic effects of cyclophosphamide on viability of (head and neck cancer cells) HN5 cell line and compare it with fibroblast cells as noncancerous cells.

Methods: This experimental study was done in cell and developmental laboratory in faculty of science, Shahid Chamran University of Ahvaz in Spring of 2016. HN5 cell line and embryonic fibroblast cells were grown in DMEM supplemented with 10% fetal bovine serum (FBS), penicillin/streptomycin (100 U/ml, 100 µg/ml) at 37 °C, then the effects of different concentrations of cyclophosphamide on cell viability was evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay. 4',6-diamidino-2-phenylindole (DAPI) staining was performed to determine the proportion of apoptotic cells by manually counting pyknotic nuclei. According to standard procedures from day 13 embryos of outbred strains naval medical research institute (NMRI), fibroblast cells were isolated. In this study HN5 cell line and fibroblasts were exposed to cytostatics for 72 hours.

Results: Various concentrations of cyclophosphamide were effective in cytotoxicity of HN5 cancer and fibroblast cells. A significant cytotoxicity was observed with the examined concentration of 1 µg/ml of cyclophosphamide with 50% in 3th day and P< 0.001. Interestingly, at low concentrations, cyclophosphamide was more toxic than at higher concentrations.

Conclusion: Totally cyclophosphamide had low toxicity effects on both of the cell lines but the toxicity effects of cyclophosphamide on HN5 were significantly greater than fibroblast cells. These results indicate that cyclophosphamide can be a potential anticancer agent.

Cardiac systolic function in cirrhotic patients’ candidate of liver trans-plantation compared with control group

Babak Geraiely

Background: We assessed different systolic cardiac indices to describe left and right ventricular dysfunction in cirrhotic patients before liver transplantation.

Methods: In this case-control study, eighty-one consecutive individuals with the confirmed hepatic cirrhosis and candidate for liver transplantation in the Imam Khomeini Hospital between March 2008 and March 2010 were selected. Thirty-two age and gender cross-matched healthy volunteers were also selected as the control group. A detailed two-dimensional and Doppler echocardiography was obtained in all patients and controls performed by the same operator on the day of admission.

Results: Dimensions of both left and right atriums as well as left ventricular end-diastolic volume and basal right ventricular dimension in the cirrhotic group were significantly higher than control group. Left ventricular end-systolic dimensions as well as aortic annulus diameter were not different between the two study groups. Left ventricular outflow tract velocity time integral, isovolumic pre-ejection time, isovolumic relaxation time, stroke volume, left ventricular ejection fraction, IVCT+IVRT+ET, systolic velocity of tricuspid annulus, systolic velocity of basal segment of RV free wall, systolic velocity of basal segment of septal wall, peak strain of septal margin (base), peak strain of septal margin (midpoint), peak strain of lateral margin (midpoint), strain rate of septal margin (base), strain rate of septal margin (midpoint), strain rate of lateral margin (base), strain rate of lateral margin (midpoint), Tei index (left and right ventricles), systolic time interval and tricuspid annular plane systolic excursion were higher in cirrhotic group, significantly, (P< 0.05). Left ventricular ejection time and systolic velocity of mid segment of lateral wall were lower in cirrhotic group, significantly, (P< 0.05).

Conclusion: In this study, the effects of liver on heart were volume overload, hyperdynamic state and systolic dysfunction in cirrhotic patients. These effects were due to chamber enlargement and we cannot use the most of cardiac indices for evaluation systolic function in cirrhotic patients. So, we suggest that systolic time interval and Tei index are useful indices in evaluation of systolic function in cirrhotic patients.

Comparative study of cytomegalovirus antibody and viral load in schizophrenia and bipolar patients

Jila Yavarian

Background: Schizophrenia (SC) and bipolar disorder (BD) are two chronic psychiatric illnesses with worldwide distribution. People could be involved at any age, particularly in early adolescence. Main symptoms of SC are non- affective symptoms such as auditory hallucination and illogical thinking. In contrast, BD represents affective symptoms such as depression and mania. Although the main cause of these mood disorders has been remained elusive, there are some potential contributing factors that could be considered in the pathogenesis of mentioned illnesses including, genetic and environmental factors. Cytomegalovirus (CMV) is one of the probable contributing factors in SC and BD. CMV is a prototype of herpesviridae family which may infect different cell types such as endothelial and differentiated hematopoietic cells. CMV infections in immunocompromised patients as well as congenitally infected children represent CNS complication such as microcephaly and hearing loss. This virus has capability to impair the limbic structures in brain.

Methods: This descriptive study was designed to evaluate the role of CMV in these illnesses. We investigated the level of serum IgG antibody and the presence of CMV DNA in serum and peripheral blood mononuclear cells (PBMCs) samples of 46 SC and BD patients admitted to Iran Psychiatry Hospital Tehran, Iran from 2014 to 2015 as well as 46 healthy control groups at Tehran University of Medical Sciences. First, the level of CMV IgG antibody was evaluated in serum samples, by enzyme-linked immunosorbent assay (ELISA). Then, DNA extraction conducted by using the high pure viral nucleic acid kit (Roche, Germany). Serologically positive sera along with PBMC samples were tested by Real-time PCR, to investigate the presence of CMV DNA.

Results: Results indicated higher levels of CMV IgG antibody in psychiatric patients, compared with a healthy control group. Afterward, we did not observe the presence of CMV DNA in either case or control groups.

Conclusion: According to the CNS impairment mediated by CMV infection, this virus has been supposed to play an important role in pathogenesis of mental disorders such as SC and BD. We suggest further investigation to be conducted, particularly on other samples such as cerebrospinal fluid.

Association between 497A>G polymorphism in FCGR2A gene with recurrent miscarriage in infertile women

Khadijeh Onsory

Background: The results indicated that the immunologic and genetic factors play a key role in the susceptibility to this syndrome compared to other risk factors. Immunoglobulin G, representing approximately 80% of Immunoglobulins in humans and the only way that IgG2 can be passed from mother to fetus blood circulation is binding to Fcgamma receptor (FcγR) classes which have been coded by Fcgamma receptor (FcγRIIA) gene. Any changes in the FcγRIIA gene structure such as mutations or polymorphisms can be considered as risk factors on the incidence of abortion through causing the inflammation or decreasing fetus safety. This receptor is the only which can have an interaction with IgG2 antibody and the Therefore, the current study was carried out to assess the association between R/H131 polymorphism in the FcγRIIA gene and susceptibility to recurrent abortions in Iranian women.

Methods: For this reason, a case-control study was confirmed to compare the frequency of FCGR2A gene R/H131 polymorphism in 150 women with recurrent miscarriage history having normal karyotype and 150 healthy women with no abortion history as control which were collected in March 2014 up to September 2015, from Royan Institute for Reproductive, Tehran, Iran. The genomic DNA was extracted from peripheral blood leukocytes and genotyping was performed using amplification refractory mutation system-polymerase chain reaction (ARMS-PCR).

Results: The frequency of AA, AG, and GG genotypes in case and control groups were 31.3%, 54.7%, 14% and 27.3%, 49.2%, 23.5% respectively. According to the findings, the presence of the risk allele was not associated with increased risk of recurrent miscarriage compared with individuals lacking the risk allele and it statistically was significant (P= 0.11). No significant association was found between the age of participants and risk of abortion in Iranian studied population (P= 0.083).

Conclusion: The results of present study do not support the previous findings of an association between R/H131 polymorphism in FCGR2A gene and recurrent miscarriage.

Determination the bacterial etiologies for sepsis in premature newborns admitted in neonatal intensive care unit

Samileh Noorbakhsh

Background: Bacterial sepsis is a main cause of mortality and morbidity especially in preterm newborns. The aim of this study was to search the bacterial etiologies of neonatal sepsis in NICU admitted preterm neonates.

Methods: A descriptive cross-sectional study had done in NICU of Ali Asghar Hospital, Tehran, Iran from March 2007 to March 2009. Seventy septicemic preterm newborns (<37 weeks) were studied. At admission day, for blood culture, 1-2 ml of venous blood was drawn after swabbing the venipuncture site with alcohol. After centrifugation of blood samples, deposits were cultured on sheep blood agar and incubated in a candle jar at 37 °C for 48 h and followed by subcultured. Isolates were identified using standard techniques (Nima pouyesh, Iran). Type of isolated bacterial organisms determined. Its correlation with gestational age, birth weight, premature rupture of membranes (PROM) and other variables determined we used the nonparametric two independent sample test, Mann-Whitney U test. Chi-square values (CI 95%, P< 0.05) were calculated for all categorical variables. P-value less than 0.05 considered statistically significant.

Results: Of 70 studied septicemic preterm cases, 17 (10.6%) cases had positive blood culture. Overall gram-negative organisms were more frequent than gram-positive organisms, Klebsiella (K.) pneumoniae, Escherichia (E.) coli and Staphylococcus (S.) aureus organisms were the 3 common causes of bacterial sepsis in studied cases. Early onset sepsis produced by K. pneumonia (40%), E. coli (20%) and S. aureus (20%). K. pneumonia, E. coli, S. aureus had equal incidence in late onset sepsis (26.8%). K. pneumonia was more frequent in early onset sepsis (P= 0.05), and in low birth weight (< 1500 g) neonates (P= 0.005, and PROM (P= 0.05).

Conclusion: Three causes for sepsis in premature newborns were determined: K. pneumonia, E. coli and S. aureus, it is so important for initial antibiotic treatment in admission day. Low birth weight, prematurity, and, PROM were the common risk factors for sepsis in cases. By preventing of low birth weight, low gestational age, and PROM the risk of sepsis could be decreased. We recommend empiric antibiotic in septic preterm newborns which can cover: K. pneumonia, E. coli and S. aureus in our center.

Clinical, psychological and demographic parameters of body pain in multiple sclerosis

Ghasem Salehpoor

Background: Body pain in multiple sclerosis (MS) is a common phenomenon that can create or exacerbate by different parameters of clinical, psychological and demographic. The aim of this study was to investigate the relationship between parameters of clinical (fatigue, clinical course, body mass index and duration), psychological (depression, anxiety and stress) and demographic (age, gender, marital status and education) characters with multiple sclerosis patient’s body pain.

Methods: This cross-sectional study has been performed in the Multiple Sclerosis Society of Guilan Province and Imam Reza Specialized and Sub-specialized Clinic, Rasht City, Iran during June to February 2010. In this study 162 patients with MS were selected by consecutive sampling. We used the clinical and demographic variables inventory, body pain subscale of the health survey questionnaire, depression, anxiety and stress scale and fatigue severity scale along with identical analog-spring balance. The data were analyzed by Pearson correlation coefficient and point bi-serial, one-way analysis of variance, Gabriel test and stepwise multiple regression.

Results: The findings showed that patients who scored 3 or higher in relapses experienced significantly more body pain than patients who scored 1-2 times of relapses (P= 0.031). In the meantime, significant differences were not found between the two groups of patients with a score of 3 or higher in relapses and non-relapse and between non-relapse patients and with a score 1-2 times of relapses in terms of body pain. Also, significant differences were not found in different groups of hospitalization in terms of body pain. However, anxiety and fatigue together could explain significantly 25% of the shared variance of body pain (F= 26.29, P≤ 0.0009).

Conclusion: This study showed the effect of psychological and clinical factors on body pain exacerbation in MS patients. Therefore, it is necessary for clinicians to consider identifying these factors and the relationships of the factors with increasing pain in patients with MS.

The interactive effects of two forced and voluntary exercise training method and Nanocurcumin supplement on doxorubicin-induced hepatotoxicity in aging induced by D-galactose

Valiollah Dabidi Roshan

Background: Doxorubicin (DOX) is a commonly used chemotherapeutic agent that causes hepatotoxicity via depletion of anti-oxidants and activation of apoptosis. Present study was aimed to investigate the interactive effects of two forced treadmill running and voluntary wheel running exercise training method and Nanocurcumin supplement on hepatic damage, in aging model subjects.

Methods: This experimental research was performed in animals and exercise physiology laboratory of Faculty of Physical Education and Sport Sciences, University of Mazandaran, Iran, in April, 2014. The statistical population was eighty Wistar male rats that, received a daily injection of D-galactose solution for nine weeks (100 mg/kg body weight per day, i.p.) and then, they randomly assigned to 10 groups. The forced treadmill running protocol was progressively between 25 to 54 min/day at the intensity of 15 to 20 m/min for 5 days per week for six weeks and voluntary wheel running exercise was six weeks. DOX was administrated for 15 days (1 mg/mL/kg body weight per day, i.p.). Nanocurcumin supplement was administrated for 14 days (100 mg/kg body weight per day. orally). Superoxide dismutase and apoptosis inducing factor levels were measured by enzyme-linked immunosorbent assay (ELISA) method.

Results: Implementation of two forced treadmill running and voluntary wheel running exercise with Nanocurcumin supplement, respectively led to insignificant decrease and increase in superoxide dismutase levels in comparison with the implementation of this exercise methods alone (P= 0.955 and P= 1.000, respectively). Apoptosis Inducing Factor levels following these two training method with Nanocurcumin supplement, has insignificant decrease in comparison with the implementation of this exercise methods alone (P= 1.000 and P= 1.000, respectively).

Conclusion: Our findings suggest that, however implementation of these training methods with Nanocurcumin supplement, partly mitigates the side effects of doxorubicin, but this level of intervention is not sufficient to protect against doxorubicin-induced hepatotoxicity in aging model rats.

Mental retardation due to chromosomal translocation in an Iranian consanguineous family: report of three cases

Mohammad Miryounesi

Background: Mental retardation is defined as impaired mental capacity and ability to comply with environmental and social conditions. Chromosomal abnormalities are the most important causes of mental retardation. Carriers of balanced chromosomal translocation are phenotypically normal, although they may be at risk of infertility, recurrent miscarriage or giving birth to mentally retarded children. These abnormalities are caused because chromosomes participated in the reciprocal translocations produce quadrivalents at meiosis. These quadrivalents segregate and lead to several different meiotic outcomes, just two of which are normal or balanced.

Case Presentation: A consanguineous family with three mentally retarded daughters at the ages of 24, 18 and 10 years was referred to Comprehensive Medical Genetics Centre, Shiraz, Iran in 2015. Family history showed a case of unexplained infant death as well as a spontaneous abortion. Three survived siblings had hypotonia and severe developmental delay during infantile period. In addition, they suffer from primary amenorrhea. Two siblings have vesicoureteral reflux (VUR). Cytogenetic analysis of two patients showed 46,XX,t(6;12)(q23;q22),der(9)t(8;9)(q24;p24) with partial monosomy of chromosome 9 and partial trisomy of 8q24 segment, while the other patient had 46,XX,der(12)t(6;12)(q23;q22) with partial monosomy of 12q22qter and partial trisomy of 6q23qter segment. Their mother had two balanced chromosomal translocations (46, XX, t(6,12)(q21;q22), t(8,9)(q24;p24)).

Conclusion: The above presented case is another example for the rare occurrence of double balanced chromosomal translocations in a phenotypically normal person. Although the most important causes of mental retardation in consanguineous marriages are autosomal recessive disorders, the role of chromosomal aberrations in mental retardation in these families must not be neglected. In other words, cytogenetic studies should be performed as a first line test in either situation.

Choriocarcinoma after caesarian section: case report

Amir Hossein Jafarian

Background: The most common symptom of patients with gestational trophoblastic neoplasia is abnormal vaginal bleeding. Despite repeated visits of patients with postpartum choriocarcinoma and abnormal postpartum hemorrhage, delayed diagnosis leads to advanced disease with widespread metastasis. Therefore, occurrence of choriocarcinoma with variable patterns in different diagnosis of late onset postpartum hemorrhage should be considered. Early diagnosis of choriocarcinoma after term pregnancy is important that resulted in decrease of maternal morbidity. Therefor late onset postpartum hemorrhage should have an awareness. The aim of this study was to report a case of choriocarcinoma after caesarian section.

Case Presentation: A 33-years-old woman one month after antecedent caesarian section in her second pregnancy admitted with abnormal vaginal bleeding. Based on raised titer concentration of β-hCG was 187000 u, with clinical suspicious of choriocarcinoma she was referred to oncology department of Ghaem Hospital, Mashhad University of Medical Sciences, Iran, in 2016. Vaginal exam revealed an enlarged uterus about 10 weeks of pregnancy. Transvaginal sonography showed an intracavitary heterogeneous mass with irregular surface in fundus without myometrium invasion. Extra pelvic metastasis excluded via vaginal exam, pulmonary X-ray and, ultrasonography and computed tomography scan. Due to early stage of gestational trophoblastic neoplasia and diagnosis of this condition, single agent chemotherapy (methotrexate) was recommended, but because of unresponsive disease, subsequently, she was treated with combination chemotherapy (etoposide, methotrexate, and actinomycin, followed by cyclophosphamide and vincristine) that led to remarkable response. After three courses of therapy, normal level of β-hCG was observed and now the patient is free of disease and under-serial follow-up visit for choriocarcinoma.

Conclusion: Diagnosis of choriocarcinoma should be considered in any postpartum woman with abnormal vaginal bleeding.