Promotion of cutaneous diabetic wound healing by subcutaneous administration of Wharton's jelly mesenchymal stem cells derived from umbilical cord.

Wound healing is a major problem in diabetic patients, and current treatments have been confronted with limited success. The present study examined the benefit of Wharton's jelly mesenchymal stem cells (WJ-MSCs) derived from the human umbilical cord (UC) in wound healing in diabetic rats. Thirty days after inducing diabetes, a circular excision was created in the skin of rats, and the treatments were performed for 21 days. Two groups were studied, which included the Control group and WJ-MSCs group. The studied groups were sampled on the 7th, 14th, and 21st days after wounding. Histological ultrasound imaging of dermis and epidermis in the wound area for thickness and density measurement and skin elasticity were evaluated. Our results on post-wounding days 7, 14, and 21 showed that the wound closure, thickness, and density of new epidermis and dermis, as well as skin elasticity in the healed wound, were significantly higher in the WJ-MSCs group compared to the Control group. Subcutaneous administration of WJ-MSCs in diabetic wounds can effectively accelerate healing. Based on this, these cells can be used along with other treatment methods in the healing of different types of chronic wounds.

Cytotoxic and Apoptotic Effects of Celecoxib and Topotecan on AGS and HEK 293 Cell Lines.

PURPOSE: This study is aimed to assess the anti-cancer effects of Celecoxib and topotecan against Human Gastric cancer cell line (AGS) in comparison to the control in an in-vitro study. METHODS: In this experimental study, Celecoxib and topotecan was prepared at concentrations of 500, 250, 125, 62.5, 31.2, 15.6 and 7.8 mg/ml. The effect of celecoxib and topotecan separately and in mixed form were investigated on AGS and normal HEK cells. To investigate the cell survival, MTT method was used to study the pathway of apoptosis using flowcytometry and Caspase kits based on colorimetric. Finally, one-way ANOVA and t-test were used to analyze the data. RESULTS: The results of this study indicated that Celecoxib was cytotoxic against AGS and HEK cell lines. The topotecan indicated a significant cytotoxicity against AGS cells and was not toxic against HEK cell line. Our results indicated that Celecoxib and topotecan have synergist effects against AGS and HEK cell lines and were more effective than separate celecoxib or topotecan. CONCLUSION: The mixture of clecoxib and topotecan was more effective than celecoxib and topotecan in separate form. Our results indicated that use mixed forms of treatments can cause excellent therapeutic effects and can cause less side effects.

Biological Characteristics and Optical Reflectance Spectroscopy of Human Placenta Derived Mesenchymal Stem Cells for Application in Regenerative Medicine.

Introduction: The efficiency of stem cell isolation, culture, and biological characterization techniques for treatment is facing serious challenges. The purpose of this study was to provide a protocol for isolation and culture of three types of mesenchymal stem cells (MSCs) derived from the human placenta, amniotic membrane, and umbilical cord with high efficiency used for cell therapy. Methods: During this experimental laboratory study, 10 complete placenta samples were prepared from cesarean section mothers. The protocol for isolation and culture of mesenchymal cells from the placenta tissue, umbilical cord, and amniotic membrane was enzymatically optimized. The morphological features of mesenchymal cells were investigated using an inverted microscope and their biological features were measured using flow cytometry. The differentiation potential of the cells was evaluated by measuring their differentiation capacity into osteocytes and adipocytes. The absorption and reflectance features of the cells were recorded by optical spectroscopy. Finally, the data were statistically analyzed. Results: The expression of CD44, CD73, CD90 and CD29 markers in human placenta tissue-derived cells was significant. CD14, CD34 and CD45 markers were not expressed or were slightly expressed. These cells were highly viable and successfully differentiated into osteocytes and adipocytes. MSCs absorbed more light than visible light by showing light absorption peaks at wavelengths of about 435 and 550 nm. Conclusion: The protocol used in this study for isolation and culture of human placenta tissue-derived MSCs had significant efficiency for the production of MSCs for use in cell therapy and tissue engineering.

Anti-proliferative effects of Ziziphus spina-christi and Phlomis russeliana leaf extracts on HEK293 and MCF-7 Cell Lines and Evaluation of Bax and Bcl-2 Genes Expression Level in MCF-7 Cells.

To investigate the effects of Phlomis russeliana and Ziziphus spina-christi leaf extracts on apoptosis in breast cancer MCF-7 cells. Cell lines were divided into a control group and the groups exposed to 0.001, 0.01, 0.1, 1, and 10 mg/ml of Ziziphus spina-christi and Phlomis russeliana leaf extracts. Cell viability was quantified by the MTT assay. The expression of Bax and Bcl-2 genes was evaluated by Real-time PCR analysis. Statistical analysis was performed using ANOVA. HEK293 cell viability significantly increased in the groups exposed to 0.001, 0.01, and 0.1 mg/ml of Z.christi leaf extract and decreased in the group exposed to 10 mg/ml of P.russeliana leaf extract. MCF-7 cells viability significantly decreased in the groups exposed to 0.001, 0.01, 0.1, 1 and 10 mg/ml of Z.christi leaf extract and increased in the groups exposed to 0.001 and 0.01 mg/ml of P.russeliana leaf extract. The exposure of MCF-7 cells to 1 and 10 mg/ml of P.russeliana leaf extract also led to a significant decrease in cell viability. The cytotoxic effect of Z.christi was higher than P.russeliana leaf extracts on MCF7 cells.  1 mg/ml of Z.christi leaf extracts also significantly increased the expression level of Bax and Bcl-2 genes in MCF7 cells. Bcl-2 gene expression significantly increased in the group exposed to 10 mg/ml of P.ruseliana leaf extract.Despite P.russeliana leaf extract, lower Z.christi leaf extract concentrations inhibited MCF-7 cells proliferation. Ziziphus spina-christi and phlomis russeliana leaf extracts mechanism of action has occurred through the Bax-independent apoptotic pathway on MCF-7 cells.

Characterization of an Enzyme-Catalyzed Crosslinkable Hydrogel as a Wound Dressing in Skin Tissue Engineering.

Introduction: Wound healing can have a very important impact on the patients' quality of life. For its treatment, wound dressings have vital and effective uses. Indeed, the use of a proper wound dressing can improve the healing process and duration. Recently, wound dressings with unique properties have been prepared using natural hydrogels. In addition to the general wound characteristics, new generations of wound dressings, such as those lasting longer on the wound, can have specific properties such as transferring allogeneic cells to enhance the healing effect and speed up the healing process. The present study aimed to prepare a gelatin-based hydrogel and to characterize it for therapeutic purposes. Methods: In this experimental-laboratory study, a gelatin hydrogel was made using a microbial transglutaminase (mTG) enzyme. The prepared hydrogel was evaluated in terms of appearance, physical, and chemical properties. To investigate the biological properties of the hydrogel, cells were cultured on it and the toxicity of the hydrogel for the cells was investigated. The location of the cells on the hydrogel was imaged via an electron microscope. The absorption and reflectance characteristics of the hydrogel were recorded by optical spectroscopy. Data were collected and statistical analysis was performed. Results: The results showed that the mTG gelatin hydrogel had a uniform pore size and good physical, chemical, and mechanical properties for use in wound healing. Cell experiments showed evident cell proliferation and high viability. The results also revealed that the cells grew vigorously and adhered tightly to the hydrogel. Conclusion: The preparation of a gelatin hydrogel under GMP conditions can be considered in the healing of diabetic wounds and burns.

Supplementation of freezing media with alpha lipoic acid preserves the structural and functional characteristics of sperm against cryodamage in infertile men with asthenoteratozoospermia.

The aim of this study was to examine the effects of alpha lipoic acid (ALA) supplementation during semen cryopreservation on the sperm quality, chromatin integrity, oxidative stress, and expression level of BAX, BCL2, HSP70 and iNOS genes in semen samples obtained from infertile men with asthenoteratozoospermia. METHODS: Twenty freshly ejaculated semen samples were cryopreserved with sperm freezing medium supplemented with 0.00, 0.02, 0.05, 0.1, 0.5, and 1 mmol/mL of ALA. The samples were analyzed according to the WHO guidelines before and after freezing. Sperm ROS production level, DNA fragmentation and cryo-capacitation were assessed using flow cytometry, TUNEL assay and chlortetracycline (CTC) test, respectively. Expression level of stress protein (HSP70), pro-apoptotic Bax, anti-apoptotic Bcl-2, and iNOS genes was assessed by real-time PCR assay. RESULTS: The effective concentrations of ALA (0.02 and 0.5 mM) significantly improved the motility, viability and morphology of the frozen-thawed sperms compared to the control group treated with 0.00 mM of ALA. During cryopreservation, treatment of semen with 0.02 mM of ALA, as the optimal concentration, significantly decreased DNA fragmentation and oxidative stress level (P < 0.05), protected the acrosome integrity, and led to insignificant reduction in BAX gene expression level and significant increase in expression level of BCL2, HSP70, and iNOS genes compared with control group. CONCLUSION: Our findings revealed that the adding ALA to semen samples obtained from infertile men with asthenoteratozoospermia plays a significant protective role against cryodamage by preserving the sperm functional parameters.

Proteomic and genomic biomarkers for Non-Small Cell Lung Cancer: Peroxiredoxin, Haptoglobin, and Alpha-1 antitrypsin.

BACKGROUND: The development of lung cancer is a multifactorial process that involves the environmental and genetic factors. The mortality rate of this cancer is higher than breast, colorectal, and prostate cancers. In this study, we try to analyze the proteome of patients with Non-Small Cell Lung Cancer (NSCLC) and compare it with the healthy samples. METHODS: This study has compared 30 lung tissue samples from patients with NSCLC and 30 healthy samples using proteomics and RT-PCR. Hence, tissue samples were obtained from the surgical ward in sterile conditions, and then, protein extraction applied to them. At the next stage, two-dimensional electrophoresis and mass spectrometry LCMS/MS were performed for protein isolation and sequencing, respectively. RESULTS: The proteome analysis identified more than 40 differences in proteomic pattern of normal lung tissues compared to lung tissues with NSCLC. Peroxiredoxin, Haptoglobin, and Alpha-1 antitrypsin proteins were identified. Molecularly, it has also been shown that the two main proteins of Peroxiredoxin-2 and Alpha-1 antitrypsin were upregulated, and the expression of Haptoglobin protein was downregulated in cancer tissue. CONCLUSION: The results of this study showed that there are some differences in term of protein content between the normal and cancerous lung tissues. Further studies are needed to evaluate these proteins that investigate whether these proteins can candidate as biomarkers to use in the early diagnosis of patients with NSCLC.

Evaluation of the efficacy of intralesional Glucantime plus niosomal zinc sulphate in comparison with intralesional Glucantime plus cryotherapy in the treatment of acute cutaneous leishmaniasis, a randomized clinical trial.

Current treatment modalities in cutaneous leishmaniasis have low efficacy and high toxicity as well as high rate of resistance to treatment. In this study, for the first time we decided to evaluate efficacy of intralesional Glucantime plus niosomal zinc sulphate in comparison with intralesional Glucantime plus cryotherapy in the treatment of acute cutaneous leishmaniasis. This is a case-control study on 64 patients with cutaneous leishmaniasis in Kerman-Iran. Patients were categorized in 2 groups A and B whom were treated with weekly intralesional meglumine antimonite plus twice daily niosomal topical zinc sulphate versus weekly intralesional Glucantime plus every other week cryotherapy, respectively. We assessed the efficacy of treatment modalities (as partial and complete response) and their adverse effects by measuring size of the lesions every 2 weeks up to maximum of 12 weeks and 3 months after the end of the treatment. Partial response rate was 16.6% and 12.9% in group A and B, respectively (P = 0.784). Complete response rate was 73.3% and 80.6% in group A and B, respectively (P = 0.784). Complete response rate was achieved in 4.73 ± 0.29 weeks and 4.69 ± 0.28 weeks in group A and B, respectively (P = 0.925). Partial response rate was achieved in 2.92 ± 0.23 weeks and 2.65 ± 0.18 weeks, respectively (P = 0.365). Combination of niosomal zinc sulphate with intralesional Glucantime has equal efficacy versus combination of cryotherapy plus intralesional Glucantime in the treatment of acute cutaneous leishmaniasis. So, it can be used in cases that have resistance to first-line treatments.

Multi-Gene Expression in Anthracosis of the Lungs as One of the Risk Factors for Non-Small Cell Lung Cancer

Background: Anthracosis of the lung occurs due to the deposition of carbon and silica in the mucosa and submucosa, manifested as black lesions. The association of anthracosis with lung cancer has remained to be clearly elucidated The current study aimed to assess the P16, CDH1 and LUNX genes expression level to evaluate the association of anthracotic lesions in the lungs with the occurrence of non-small cell lung cancer. Methods: Forty biopsy samples were taken from the center and 40 from the margins of black anthracotic lesions in the lungs; RNA was extracted from the samples and cDNA was synthesized. Real-time reverse-transcription polymerase chain reaction (RT-PCR) was performed to assess the expression of P16, CDH1 and LUNX genes. All steps were performed in triplicate. Results: A significant reduction in P16 gene expression was noted at the center compared to the margins of the lesions (P<0.001). expression level of CDH1 at the center of lesions was significantly lower than margins (P<0.001). However, LUNX gene had significantly higher expressionlevel at the center compared to margins (P<0.001). Conclusion: Decreased expression of P16 and CDH1 and increased expression of LUNX tumor genes were noted at the center of anthracotic lesions. Significant increase in expression of LUNX gene in NSCLC indicates an association between anthracosis and NSCLC, according to which, anthracotic patients may carry a high risk for NSCLC.