Tehran University Medical Journal

The mammalian cochlea is a highly complex structure which contains several cells, including sensory receptor or hair cells. The main function of the cochlear hair cells is to convert the mechanical vibrations of the sound into electrical signals, then these signals travel to the brain along the auditory nerve. Auditory hair cells in some amphibians, reptiles, fish, and birds can regenerate or replace by new cells, but irreversible damage to the mammalian hair cells are not being replaced through differentiation of the internal epithelial cells in the inner ear. Indeed, mammalian auditory hair cells do not spontaneously repair or regenerate after development. Sometimes, functions of damaged hair cells may be restored, but in most cases, there is no such possibility and permanent hearing loss occurs. Several factors such as chronic ear infections, genetic disorders, drug abuse, acoustic trauma and aging can damage the cochlea, resulting in permanent hearing loss. More than 250 million people in the world have disabling hearing impairment. Deafness is caused by damage to sensory hair cells or spiral ganglion neurons. Although hearing aids and cochlear implants were used for improvement of hearing loss, but they do not restore normal hearing. In addition, application of new biological approaches to induce auditory hair cell regeneration provides more comprehensive treatment for hearing loss. Cell therapy is considered a promising way in the treatment of several diseases such as Parkinson, diabetes and cardiac diseases. According to recent research, cell therapy can be useful in hair cell regeneration. Cell therapy is effective in hearing loss when stem cell differentiates into hair cells with appropriate morphology, electrical activity and capacity for suitable innervations with inner ear tissues. In fact, stem cell-derived neurons need to project neural processes toward the sensory hair cells and the cochlear nucleus neurons. In this regard, studies focus on methods in which hair cells can be provided from exogenous and endogenous stem cells. Here, we review cell therapy approaches in repair damaged cochlear hair cells, as well as imitations and problems of its clinical application.

Background: Aphanizomenon flos-aquae (AFA) is a type of blue-green algae and contains a source of biological compounds. These microalgae have many beneficial health effects. Recently, fucoidan, known sulfated polysaccharide component of AFA algae, has been claimed to stimulate stem-cell mobilization in animal models. Stem cells play an essential role in tissue repair process. In this study, we use excisional full thickness wound model to investigate the effectiveness of trademark AFA extract on skin wound repair process.
Methods: In this experimental study, 21 adult male Wistar rats (weighing 200-250 g) were used and under general anesthesia (intraperitoneally with a ketamine/xylazine solution), two round excisional wounds were created under sterile conditions by a 6 mm punch on the dorsum (paravertebral area) of all rats. Animals were randomly assigned into 3 groups. In groups 1 and 2 (SE-200, SE-400), StemEnhance© (StemTech Health Sciences Inc. British Columbia, Canada) were given respectively 200 or 400 mg/kg by oral gavage once daily and in group 3 (Sham), distilled water (DW) was given to all subjects. Post-wounding gavage of StemEnhance or DW started from 1st day and continued to 7th day. The wound surface area was monitored daily by digital camera and assessed by Image Tool™ software, version 3.5 (UTHSCSA, San Antonio, TX, USA). At 9th day post-wounding animals were sacrificed and repaired tissues were harvested by and assessed by a 8 mm punch. Repaired skin areas were processed for hematoxylin and eosin (H&E). Histopathological parameters of healing including inflammatory cell infiltration, angiogenesis, and fibroblast count were assessed by pathologist. Our study was conducted in the Physiology Department of Medical School, Bushehr University of Medical Sciences, Iran, from October 2016 to March 2016.
Results: Macroscopic imaging of wound area revealed that there was statistically significant difference in wound area reduction between SE-200 group and sham group on day 6 post wounding (P=0.032). Moreover, histological findings showed that the number of neutrophils, macrophages, fibroblasts, and microvessel density decreased in both StemEnhace-treated groups. There were no significant differences between two treatment groups.
Conclusion: According to the obtained results it seems that the extract of Aphanizomenon flos-aquae algae positively affects wound healing process by ameliorating inflammatory response in early healing phases.

Background: Although cervical malignancy rate had grown up in recent years, primary cervical lymphoma is so rare. It must be high index of suspicious for primary cervical lymphoma diagnosis in patient with malignancy-like signs and symptoms for early detection. Primary cervical lymphoma has no standard treatment or follow-up protocol; so the management still is in doubt and based on previous case reports. In the other hand, the precise prognosis of patient is undetermined. In the present study, a case of primary cervical lymphoma is presented which was misdiagnosed at first. The patient accurate diagnosis was made at last due to multidisciplinary team working.
Case presentation: A 51-year-old woman, gravida 2, para 2, presented with complaint of abnormal vaginal bleeding and discharge, with no abnormal finding in cervical cytology and sonography, so uneventfully a diagnostic error had happened in the assessment of her. After several months and multiple different treatment, the patient referred to the Oncology Department of Obstetrics and Gynecology Center, Ghaem Hospital, Mashhad, Iran in May 2017. Re-assessment was performed by biopsy and imaging, and the final pathologic diagnosis of diffuse large B-cell non-Hodgkin's lymphomas was confirmed.
Conclusion: Primary cervical lymphoma is an uncommon malignancy; the diagnosis could be missed simply by low suspicious due to low accuracy of Pap smear and imaging in this situation. So an accurate evaluation and pelvic examination, high suspicious and close communication between clinician and pathologist are needed. By timely diagnosis of patient in early stage and appropriate approach, the prognosis could be excellent most of the time.

Background: Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin's lymphoma (NHL), which is the most common type of lymphoma. NHL comprises a group of clinically and biologically diverse diseases, which range from indolent to aggressive clinical courses. Despite treatment advances in the last three decades with the use of combination immunotherapy, a significant fraction of patients relapses or are refractory to these treatments. Actually, there is no standard method for detection of recurrence. The purpose of this study was to evaluate the level of this enzyme in the patients with recurrent DLBCL compared to healthy controls.
Methods: In this randomized double-blind, placebo-controlled trial, the serum level of pyruvate dehydrogenase (PDH) enzyme in total of 26 patients with DLBCL recurrence in compare with 26 healthy individuals in the Sayed Al-Shohada Hospital, Isfahan University of Medical Sciences in Iran from September 2016 to September 2018, were assessed. The clinical data including age criteria, the Eastern Cooperative Oncology Group (ECOG) score rating, total bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), prothrombin time and international normalized ratio (PT/INR), CT-scan, serum creatinine, platelet count, the absolute number of neutrophils, and the interval until the last treatment were gathered. After obtaining informed consent, blood samples were taken. and the PDH enzyme was measured in case and control groups.
Results: Fifty-three percent of patients were male and the mean age of participants in case and control groups was 37.2±17.3 and 34.8±8.9, respectively. Subsequently, the PDH levels were measured according to the enzyme protocol. The levels of enzyme in patients with relapse were significantly lower than normal ones (P=0.0003). The PDH serum level was also evaluated by age and sex, which did not show any significant differences (P=0.86). 
Conclusion: In patients with relapsing B-cell lymphoma, the pyruvate dehydrogenase (PDH) enzyme serum levels were significantly lower than healthy subjects, but this difference was not related to age and gender. In the case of further studies and comparisons beyond this study, this enzyme could be a good candidate, used as an alternative diagnosis tool, in patients with recurrent lymphoma.

Background: Most studies show that 9 to 24% of people who are in in vitro fertilization (IVF) cycles are women who respond poorly to ovarian stimulation. Women with poor ovarian response (POR) are a group of infertile patients whose ovarian reserve, ovarian response to medication, and the quality of ovum are declining. Therefore, the number of female cycles, the number of fetuses from the oocyte and the rate of pregnancy in these women is reduced. The purpose of this study was to investigate the expression of three adrenoceptor receptor genes in the cumulus cells of women with poor ovarian response in culture medium.
Methods: This case-control study was conducted in two groups: study (POR) and control (oocyte donor's women) groups. POR diagnosis was performed by ESHRE Bologna criteria. After puncture of the follicles, cumulus-oocyte complex was collected and the cumulus cells (CCs) were isolated by enzyme and are counted with Neobar lamella and then were added in the culture medium. After completing the culture, RNA was extracted from cumulus cells and the RNA concentration was read by the Spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). Then cDNA synthesized and primers designed for ADR-α1, 2 and ADR-B2 or gene expression by real-time polymerase chain reaction (PCR) technique. The research was done in Reproductive Health Research Center of Imam Khomeini Hospital, Tehran University of Medical Sciences, Tehran, Iran, from April to December 2017.
Results: Comparison of the results of ADR-α1, 2 gene expressions in cumulus cells showed a significant decrease, but ADR-B2 was not significant in two groups. Correlation coefficients also showed that there are relationship between three adrenoceptors and their effects on each other.
Conclusion: Our results showed that the decreased expressions of ADR-α1, 2 probably related to activation of the sympathetic system and release of the more neurotransmitter that lead to down-regulation of ADR-α1, 2 in the cell membrane of cumulus in culture medium.

Chick embryos are a great historical research model in basic and applied sciences. Along with other animal models, avian and specifically chicken embryo has been attended, as well. Avian fertilized eggs as a natural bioreactor are an efficient tool for producing recombinant proteins and vaccines manufacturing. Due to the limitations of birds' eggs for viral replication, avian stem cells culture technologies access to safe methods as well as large-scale production of a variety of human and animal vaccines. Chicken pluripotent stem cells present the unique property of self-renewal and the ability to generate differentiated progeny in all embryonic lineages such as ectoderm, mesoderm, and endoderm in vitro. For the first time, chicken embryonic stem cells (cESCs) derived from the blastodermal cells of stage X embryos in vitro. Chicken ESC provides a great model of early embryo and they are useful for gene manipulation, virus proliferation, and the generation of transgenic birds. In addition to blastodermal cells, pluripotent cell lines can be produced by reprogramming of chicken fibroblasts into induced pluripotent stem cells (iPSCs) with transcription factors such as OCT4, NANOG, SOX2, KLF4, LIN28, and C-MYC that are well known to contribute to the reprogramming of somatic cells into an iPSCs. Similar to chicken ESCs, iPSCs have properties of unlimited self-renewal in vitro and the capacity for differentiation to all three embryonic germ layers. Chicken iPSCs have been a useful tool for the production of transgenic birds and viral vaccines. Despite the benefits and multiple applications of chicken pluripotent stem cells, the propagation of these cells is limited and some important challenges should be eliminated before their use in vaccine manufacturing. It is necessary to define the appropriate culture conditions for chicken pluripotent stem cells. For example, the presence of endogenous viruses in the avian species should be evaluated for human vaccine production. Currently, primary chicken fibroblast cells are still mainly used for vaccine production. This review covers the resources to achieve chicken derived cell lines for vaccine manufacturing.

Background: Non-viral Nano carriers such as liposomes and cationic polymers based on engineered properties are regarded in gene delivery field. Although these carriers do not have weaknesses of viral vectors, but they are less efficient than viruses and they still need to be improved as favorable gene delivery carriers. Amongst non-viral carriers, cationic liposomes have been proposed for clinical applications, but limitations such as low nucleic acid transfer and endosome escape and conduction of plasmid to the nucleus have challenged their use in clinical trials. Therefore, the combination of liposomes and cationic polymers for nucleic acid transfer has been considered because this approach makes it possible to use the desirable properties of liposomes and polymers so that it is even suggested for the gene treatment of some diseases such as Parkinson's. In this study, a combination of liposomes and cationic polymers were used for the preparation of lipopolyplexes. This approach allows simultaneous utilizing of the desirable properties of liposomes and polymers.
Methods: This interventional-experimental study was conducted in the medical faculty of Mashhad University of Medical Sciences from April 2017 to February 2018. In this study, PEI-based lipopolyplex with a molecular weight of 25 and 10 kDa and a liposome-to-polymer ratio of 1:1 were combined with plasmid containing the GFP (Green Fluorescent Protein) marker. The physicochemical properties of the synthesized carriers such as size, cytotoxicity and gene transferability in human prostate cancer (PC3) cells were evaluated.

Background: Lung cancer is a disease with high mortality rate that conventional drug treatments have not been successful in controlling it. The activity of iron chelators in various studies has been considered by scientists as a new treatment strategy. The primary objective of this study was to synthesize a novel Fe3O4 thiosemicarbazone complex and investigate its anti-proliferative activity against A549 cells of lung cancer.
Methods: This experimental study was carried out in Islamic Azad University of Rasht Branch, from September of 2018 to September 2019. First thiosemicarbazone (PTSC) was synthesized by the method of the condensation reaction of amine and aldehyde groups. Also, the Fe3O4 nanoparticulates were synthesized using the co-precipitation method in the presence of glutamic acid. Then, Fe3O4@Glu complex functionalized with bio-reactive PTSC moiety. Besides, morphological characteristics of Fe3O4@Glu/PTSC complex were determined by scanning electron microscope (SEM) images. The cell viability was detected in 62.5, 125, 250, 500, and 1000 µg/ml for treated cells with Fe3O4@Glu/PTSC complex via MTT assay. Changes of NDRG1 gene expression the level in treated cells were investigated via qRT-PCR analysis. Therefore, total RNA was extracted after culturing the cells and cDNA of NDRG1 and GAPDH genes as the study and control gene was obtained, respectively. Ultimately, the level of NDRG1 gene expression was compared with level of GAPDH mRNA expression via the 2^{– ΔΔCt} method.

Background: Clopidogrel is an effective anti-platelet drug that is commonly prescribed for patients with atherosclerotic coronary and peripheral vascular diseases, especially after angioplasty and coronary artery bypass graft surgery. Even though this drug seems safe, there are case reports of important side effects with its use.
Case Presentation: The patient was 67 years old male admitted with a diagnosis of inferior myocardial infarction who underwent angioplasty on RCA with a drug-eluting stent and was discharged with an uneventful course from hospital with medical treatment, including Clopidogrel. The patient came back to the clinic 30 days later with icter, nausea and loss of appetite. The lab data showed: Bilirubin (total= 5, direct=4), ALT=220, AST=100, Gama GT=830, Alk.phop=445. Virologic and imaging studies were normal. With a possible diagnosis of drug-induced hepatic injury all possible Prescribed drugs were held stepwise but bilirubin and the patient’s symptoms did not relieve until Clopidogrel has been changed. After 10 days bilirubin and his symptom relieved.

Background: Granular cell tumor (Abrikossoff’s tumor) is a rare and slow-growing tumor of the soft tissue. Originated from the Schwann cells, it is often a benign tumor, but it can be malignant in 1-3% of the cases. Malignant cases can cause significant morbidity and mortality. It may develop in many anatomic locations, especially in the head and neck region, and also in skin and subcutaneous tissue.

Background: The application of mesenchymal stem cells in the healing of chronic wounds is one of the most challenging issues in cell therapy. The present study investigated the efficacy of intradermal injection of umbilical cord Wharton's Jelly-derived mesenchymal stem cells in diabetic wound healing using ultrasound imaging in an animal model.
Methods: During this experimental laboratory study that was performed in the Skin and Stem Cell Research Center, Tehran University of Medical Sciences between October 2017 and October 2016, mesenchymal stem cells were isolated from umbilical cord Wharton's jelly of 10 neonates. The cells were passage. The differentiation potential of cells to osteocyte and adipose cells was evaluated. The expression of specific markers of mesenchymal stem cells was evaluated using flow cytometry. The viability and quality of cells were evaluated before transplantation. The diabetes model was developed by intraperitoneal injection of streptozotocin in 42 male Wistar rats. The animals were randomly divided into two groups: normal saline injection (control) and cell injection. Cell transplantation was performed intradermally. Skin thickness and density were assessed using ultrasound imaging on days 7, 14 and 21. Finally, the data were analyzed using a t-test and analysis of variance.

Background: Low-level lasers are used for various medical applications including wound healing and hair loss treatment. Cell Therapy using skin stem cells could be a novel approach to hair transplantation. However, there is no study on the effect of low-level laser on the hair follicle stem cells. So, in this study, we investigated the effect of low level laser irradiation on viability and ROS production in the hair follicle stem cells.
Methods: This study was performed in the cell culture laboratory of Medical Laser Research Center, Yara Institute in 2020 (June 2020 to February 2020). The hair follicle was isolated from the Safe Donor Area (SDA) using the 4mm punch method. In the laboratory, after separating the follicular units, the bulb region of each follicle was isolated via mechanical and enzymatic methods and cultured in FBS+F12-DMEM. Afterward, the stem cells were characterized via flow cytometry. The effect of low-level laser (685 nm) with different doses (1-20 J/cm²) was investigated on cell proliferation, viability and ROS production.

Background: Mesenchymal stem cells are non-hematopoietic stromal cells that are used in the treatment of many chronic and autoimmune diseases by modulating the immune system. Due to the limitations of using autologous mesenchymal stem cells, the use of allogeneic stem cells is a promising therapeutic approach in the treatment of immunological disorders. This study aimed to investigate the ability of allogeneic mesenchymal stem cells to induce Programmed death-ligand 1(PD-L1) expression on the surface of splenic lymphocytes and the role of this molecule in the mesenchymal stem cell-treated cells tolerogenicity. 
Methods: This study was conducted from February 2019 to December 2020 in the department of Immunology of Mazandaran University of medical sciences. Mesenchymal stromal cells were isolated from the femur and tibia of C57 mice. C57 bone marrow-derived mesenchymal stem cells were co-cultured with allogeneic BALB/c splenic cells. After 72 hours, the expression of PD-L1 on the surface of splenic lymphocytes was evaluated by flow cytometry. Interferon-gamma (IFN-γ) and Interleukin-10 (IL-10) cytokine assay were done in the cell culture supernatant. Mesenchymal stem cell-treated BALB/c lymphocytes were then exposed to allogeneic C57 splenocyte as stimuli in the mixed lymphocyte reaction (MLR) and the rate of proliferation was assessed by CFSE.

Background: Primary intraosseous squamous cell carcinoma (PIOC) of the mouth is a very rare but well-known carcinoma. It is locally invasive and its prognosis is quite poor. It may originate from the walls of an odontogenic cyst or de novo from the remnant epithelium at this region. Because the early symptoms of such malignancies are sometimes similar to those of inflammatory or periodontal diseases, early diagnosis is the most important step toward appropriate treatment.
Case Presentation: The case was a 46-year-old female with a sudden luxation of two posterior mandibular molars referred to a general dentist in May 2019. With diagnosis of periodontitis, the teeth were extracted without finding the cause of the tooth luxation. After four months, because of pain, swelling and non-healing socket of extracted teeth she referred to the department of Oral medicine in Mashhad Faculty of Dentistry. The patient was healthy with no systemic disease, drug history or familial history of cancer and no history of trauma, habits, tobacco, alcohol, and smoking. ln clinical examination, a nodular swelling with extensive ulcer and rolled borders along with hyperkeratosis and erythema on the posterior part of the right mandibular ridge was observed. The Extraoral examination revealed a tender mobile lymph node in the right submandibular region. After radiographic evaluation, with an initial diagnosis of SCC originated from the socket of extracted mandibular molars, incisional biopsy was performed, and histopathological analysis of the specimen revealed a primary intraosseous squamous cell carcinoma (grade II) and then necessary treatments (surgery, radiotherapy, chemotherapy) were performed.

Background: Skin cancer is the most prevalent type of cancer and melanoma is the deadliest kind of skin cancer in the world. Due to enhanced induction of apoptosis and ROS levels, low-level lasers can be utilized to destroy skin cancer cells. Lasers are used to treat some skin lesions. Vitamin A is beneficial in the prevention and treatment of skin cancer. Vitamin A inhibits the pathway of cancer signals in the skin and suppresses tumor growth. In this study, the combined effect of low-level laser radiation (LLL) and vitamin A on cellular factors of skin melanoma cancer cells was investigated.
Methods: An in-vitro interventional laboratory study was performed in the cell culture laboratory of Medical Laser Research Center, Yara Institute in 2020-2021 (July 2020 to July 2021). First, A375 skin cancer cells were cultured in DMEM with 10% FBS. After preparation and culture of A375 cell lines, different concentrations of vitamin A (1, 5, 50, 100 μM) and LLL energy doses (1, 2, 5, 10 J/cm²) as treatments were done. Combination research of these treatments was performed to eliminate skin melanoma cancer cells. The rate of viability was determined using the MTT test, and the rate of apoptosis was determined using flow cytometry.

Background: Connexon is a membrane structural protein in the gap junctions. These cellular connections are responsible for transporting ions and messenger molecules to the oocyte. This study aimed to investigate the role of beta-2-adrenoceptors in the process of follicle growth based on the expression of the two connexins 37/43 in the gap junctions that have a primary role in the mitotic resumption and oocyte maturation.
Methods: This case-control study was conducted from April 2019 to November 2020 at the Reproductive Health Research Center of Tehran Imam Khomeini Hospital was performed on women with the poor ovarian response (POR) and control (women donate eggs) groups. Both groups had entry criteria with a body mass index under 28 m²/kg and 20-45 years old. Exit criteria were including no drug use except ovarian stimulants and no illness. The diagnosis was made agreeing to the Bologna model criterion. Ovulation stimulation cycle was performed, and then after the puncture, cumulus cells were isolated by enzyme and were freezing in -80 centigrade until the time of inserting into the cell culture medium. Isoproterenol (agonist) and propranolol (antagonist) at a concentration of 100 nM were added to the culture medium as the beta-2 adrenoceptors selective drugs. After culture, RNA extraction was performed and the concentration was read by Nanodrop, and then cDNA was synthesized. Gene expression was determined by real-time PCR.

Background: Cerebral palsy is a movement disorder syndrome in early childhood. Signs and symptoms vary among people and over time but include poor coordination, stiff muscles, and weak muscles. Some affected children can achieve near-normal adult lives with appropriate treatment. In recent years, transplantation of human mesenchymal stem cells (hMSC) has become a promising therapeutic strategy for CP. Every year, a lot of costs are spent on the treatment and management of this disease. The purpose of this study is to investigate the safety and effectiveness of this method on CP.

Methods: This article is a systematic review. At first, a search strategy was written and performed in Scopus, PubMed, and Google Scholar databases(The search was conducted from March 14 to March 28, 2021), and the inclusion and exclusion criteria were determined. Study inclusion criteria: review studies, cohort studies, clinical trial studies (randomized and non-randomized), control case, and retrospective , exclusion criteria: non-English articles and studying on animals. After removing duplicate articles, two authors independently reviewed the studies according to the inclusion and exclusion criteria. Disagreements among the authors were resolved through discussion.
Findings: In total, 9236 articles were found in the initial search, after reading the titles, of 37 articles, 21 articles were selected in the abstract stage and 18 articles remained in the full-text stage. We finally found 18 articles that showed that using stem cell technology as a scientific method could improve sick patients’ quality of life and movement defects.
Conclusion: According to the available evidence and limited studies, stem cell technology can be safeand cost-effective in improving CP patients, but there is insufficient evidence. On the other hand, there are many studies confirming the effectiveness of these cells in the treatment of movement impairment. In conclusion, stem cells may have a very promising future. Finally, stem cell technology combined with innovative biotechnologies may soon bring promising results to patients.