mahmoud orazizadeh

Diabetic nephropathy (DN) is a critical complication of diabetes mellitus. This study evaluates whether administration of conditioned medium from kidney tubular cells (KTCs‑CM) has the ability to be efficacious as an alternative to cell‑based therapy for DN.

CM of rabbit kidney tubular cells (RK13; KTCs) has been collected and after centrifugation, filtered with 0.2 filters. Four groups of rats have been utilized, including control, DN, DN treated with CM, and sham group. After diabetes induction by streptozotocin (50 mg/kg body weight) in rats, 0.8 ml of the CM was injected to each rat three times per day for 3 consecutive days. Then, 24‑h urine protein, blood urea nitrogen (BUN), and serum creatinine (Scr) have been measured through detection kits. The histopathological effects of CM on kidneys were evaluated by periodic acid–Schiff staining and the expression of microRNAs (miRNAs) 29a and 377 by using the real‑time polymerase chain reaction. The expression of aquapurin‑1 (AQP1) protein was also examined by Western blotting.

Intravenous injections of KTCs‑CM significantly reduced the urine volume, protein 24‑h, BUN, and Scr, decreased the miRNA‑377, and increased miRNA‑29a and AQP1 in DN treated with CM rats.

KTCs‑CM may have the potential to prevent kidney injury from diabetes by regulating the microRNAs related to DN and improving the expression of AQP1.

Type 1 diabetes mellitus is a common autoimmune and multifactorial disorder. Researchers have been interested in making a favorable islet-like tissue model for the treatment of diabetes. The main objective of this study was to determine the effects of the spleen extracellular matrix (S-ECM) on the function of the MIN6 cell line (a β-cell model). 

In this experimental research, Wistar rat spleens were decellularized by sodium dodecyl sulfate (SDS) and Triton X-100. S-ECM was characterized by histological assessments, scanning electron microscopy, determination of residua DNA, and examination of the mechanical tensile property. Then, MIN6 cells were seeded on S-ECM scaffold. Glucose-stimulated insulin secretion and mRNA expression of insulin-related genes were examined to confirm the function of the cells. 

The main components of S-ECM such as collagen and glycosaminoglycan remained after decellularization. Furthermore, very low residual DNA and appropriate mechanical behavior of S-ECM provided an ideal extracellular microenvironment for the MIN6 cells. GSIS results showed that the seeded cells in S-ECM secreted more insulin than the traditional two-dimensional (2D) culture. The expression of specific insulin-related genes such as PDX-1, insulin, Maf-A, and Glut-2 in the recellularized scaffold was more significant than in the 2D traditional cultured cells. Also, MTT assay results showed that S-ECM were no cytotoxic effects on the MIN6 cells. 

These results collectively have evidenced that S-ECM is a suitable scaffold for stabilizing artificial pancreatic islands.

The main objective of this study is to determine the myogenic effects of skeletal muscle extracellular matrix, vascular endothelial growth factor and human umbilical vein endothelial cells on adipose-derived stem cells to achieve a 3-dimensional engineered vascular-muscle structure.

The present experimental research was designed based on two main groups, i.e. monocultureof adipose tissue-derived stem cells (ADSCs) and co-culture of ADSCs and human umbilical vein endothelial cells ( HUVECs) in a ratio of 1:1. Skeletal muscle tissue was isolated, decellularized and its surface was electrospun using polycaprolactone/gelatin parallel nanofibers and then matrix topography was evaluated through H&E, trichrome staining and SEM. The expression of MyHC2 gene and tropomyosin protein were examined through real-time reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence, respectively. Finally, the morphology of mesenchymal and endothelial cells and their relationship with each other and with the engineered scaffold were examined by scanning electron microscopy (SEM).

According to H&E and Masson’s Trichrome staining, muscle tissue was completely decellularized. SEM showed parallel Polycaprolactone (PCL)/gelatin nanofibers with an average diameter of about 300 nm. The immunofluorescence proved that tropomyosin was positive in the ADSCs monoculture and the ADSCs/HUVECs coculture in horse serum (HS) and HS/VEGF groups. There was a significant difference in the expression of the MyHC2 gene between the ADSCs and ADSCs/HUVECs culture groups (P<0.05) and between the 2D and 3D models in HS/ VEGF differentiation groups (P<001). Moreover, a significant increase existed between the HS/VEGF group and other groups in terms of endothelial cells growth and proliferation as well as their relationship with differentiated myoblasts (P<0.05).

Co-culture of ADSCs/HUVECs on the engineered cell-free muscle scaffold and the dual effects of VEGF can lead to formation of a favorable engineered vascular-muscular tissue. These engineered structures can be used as an acceptable tool for tissue implantation in muscle injuries and regeneration, especially in challenging injuries such as volumetric muscle loss, which also require vascular repair.

One of the severe complications and well-known sources of end stage renal disease (ESRD) from diabetes mellitus is diabetic nephropathy (DN). Exosomes secreted from diverse cells are one of the novel encouraging therapies for chronic renal injuries. In this study, we assess whether extracted exosomes from kidney tubular cells (KTCs) could prevent early stage DN in vivo.

In this experimental, exosomes from conditioned medium of rabbit KTCs (RK13) were purified by ultracentrifuge procedures. The exosomes were assessed in terms of morphology and size, and particular biomarkers were evaluated by transmission electron microscopy (TEM), scanning electron microscopy (SEM), Western blot, atomic force microscopy (AFM) and Zetasizer Nano analysis. The rats were divided into four groups: DN, control, DN treated with exosomes and sham. First, diabetes was induced in the rats by intraperitoneial (i.p.) administration of streptozotocin (STZ, 50 mg/kg body weight). Then, the exosomes were injected each week into their tail vein for six weeks. We measured 24-hour urine protein, blood urea nitrogen (BUN), and serum creatinine (Scr) levels with detection kits. The histopathological effects of the exosomes on kidneys were evaluated by periodic acid-Schiff (PAS) staining and expressions of miRNA-29a and miRNA-377 by quantitative real-time polymerase chain reaction (qRT-PCR).

The KTC-Exos were approximately 50-150 nm and had a spherical morphology. They expressed the CD9 and CD63 specific markers. Intravenous injections of KTC-Exos potentially reduced urine volume (P<0.0001), and 24- hour protein (P<0.01), BUN (P<0.001) and Scr (P<0.0001) levels. There was a decrease in miRNA-377 (P<0.01) and increase in miRNA-29a (P<0.001) in the diabetic rats. KTC-Exos ameliorated the renal histopathology with regulatory changes in microRNAs (miRNA) expressions.

KTC-Exos plays a role in attenuation of kidney injury from diabetes by regulating the miRNAs associated with DN.

This study was designed to fabricate a suitable permanent scaffold for the normal aligned myotube formation and improve the process of myogenic differentiation of selected stem cells.

In this experimental study, an engineered scaffold composed of decellularized human amniotic membrane (DHAM) and electrospun fibers of poly(ε-caprolactone) (PCL) was fabricated and characterized. PCL nanofibers were superimposed on DHAM (PCL-DHAM) in two different patterns, including randomized fibers (Random) and aligned fibers (Aligned). Adipose derived stem cells (ADSCs) were isolated from adult Wistar rats and cultured on designed scaffold and induced to myotube differentiation. Using an MTT assay, the vitality of cells was determined. Then, myogenic cell differentiation was assessed using scan electron microscopy (SEM), immunofluorescence assay, and reverse transcription-polymerase chain reaction (RT-PCR).

The mechanical properties of engineered PCL-DHAM composite improved significantly compared to DHAM as a control. The engineered PCL-DHAM promoted cell growth and high expression of myosin, Mhc2 and myogenin and thus enhanced the myotube formation.

These findings revealed that bio-composite membrane prepared from PCL nanofibers and DHAM, may represent a promising biomaterial as a desirable scaffold for applying in the bioengineered muscle repair.

Wounds have a bad prognostic nature and excessive discharges whose regular wound dressings are ineffective. Hydrogels are the best candidates for dressing such wounds due to their high water content and ability to exchange substances. Accordingly, the purpose of this study was to make a novel hydrogel wound dressing following the integration of various findings on wound healing and the use of regenerative medicine.

Various compounds were fabricated by glycerol/chitosan/polyvinyl alcohol (PVA) and then characterized to obtain the optimal composition using several techniques, including a water vapor passage test, scanning electron microscopy, water absorption, tensile strength, biodegradability, Fourier transform infrared spectroscopy, and antibacterial test.

The findings revealed the optimal dressing ratio. Better antibacterial activity was found for the silver nanoparticle (AgNP) dressing.

Our new fabricated dressing, glycerol/chitosan/PVA hydrogel loaded with AgNPs, exhibited satisfactory wound healing properties.

Researchers have been interested in the creation of a favorable cellular model for use in vascular-muscle tissue engineering. The main objective of this study is to determine the myogenic effects of vascular endothelial growth factor (VEGF) and human umbilical vein endothelial cells (HUVECs) on adipose-derived stem cells (ADSCs) to achieve an in vitro vascular-muscle cellular model.

The present experimental research was conducted on two primary groups, namely ADSCs monoculture and ADSCs/HUVECs co-culture that were divided into control, horse serum (HS), and HS/VEGF differentiation subgroups. HUVECs were co-cultured by ADSC in a ratio of 1:1. The myogenic differentiation was evaluated using the reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence in different experimental groups. The interaction between ADSCs and HUVECs, as well as the role of ADSCs conditional medium, was investigated for endothelial tube formation assay.

Immunofluorescence staining indicated that Tropomyosin was positive in ADSCs and ADSCs and HUVECs co-culture groups on HS and HS/VEGF culture medium. Furthermore, the MyHC2 gene expression significantly increased in HS and HS/VEGF groups in comparison with the control group (P<0.001). More importantly, there was a significant difference in the mRNA expression of this gene between ADSCs and ADSCs and HUVECs co-culture groups on HS/VEGF culture medium (P<0.05). Current data revealed that the co-culture of ADSCs and HUVECs could develop endothelial network formation in the VEGF-loaded group. Also, the ADSCs-conditioned medium improved the viability and formation of the endothelial tube in the HS and VEGF groups, respectively.

It was concluded that ADSCs/HUVECs co-culture and dual effects of VEGF can lead to the formation of differentiated myoblasts in proximity to endothelial network formations. These in vitro cellular models could be potentially used in vascular-muscle tissue engineering implanted into organ defects where muscle tissue and vascular regeneration were required.

This research studied the effects of glycyrrhizic acid (GA) on apoptosis induced with by titanium dioxide (NTiO2) in the liver of rats.

It is widely accepted that the contamination resulting from nanoparticles (NPs) is an emerging dangerous issue. Metal oxide nanoparticles have high environmental stability and cause toxicity in the food chain. Thus, the present study investigated the anti-apoptotic effects of glycyrrhizic acid (GA) on the hepatotoxicity generated by titanium dioxide (NTiO2) NPs in the liver tissue.

Thirty-two male Wistar rats were randomly divided into four groups. NTiO2-treated rats were given 300 mg / kg of NTiO2 solution via gavage for 14 days; GA-treated were administered 100 mg/kg GA for 14 days; protection group was pre-treated with GA before NTiO2 administration for 7 days. Then, apoptotic index was evaluated through immunolocalization of Bax and Bcl-2 and TUNEL assay.

we found that HSCORE of Bax expression and apoptotic index experienced a significant increase with NTiO2 (P <0.001), while Bcl-2 expression significantly diminished in NTiO2-treated rats (P <0.001). The results revealed that the increased Bax expression and apoptotic index were reversed by GA and enhanced the activities of Bcl2.

The results revealed that GA effectively attenuated apoptosis against NTiO2 in rats.

Fabrication of an antibiotic-loaded scaffold with controlled release properties for wound dressing is one of tissue engineering challenges. The aim of this study was to evaluate the wound-healing effectiveness of 500-µm thick polycaprolactone (PCL) nanofibrous mat containing silver sulfadiazine (SSD) as an antibacterial agent. In this experimental study, an electrospun membrane of PCL nanofibrous mat containing 0.3% weight SSD with 500 µm thickness, was prepared. Morphological and thermomechanical characteristics of nanofibers were evaluated. Drug content and drug release properties as well as the surface hydrophobicity of the nanofibrous membrane were determined. Antimicrobial properties and cellular viability of the scaffold were also examined. A full thickness wound of 400 mm2 was created in rats, to evaluate the wound-healing effects of PCL/SSD blend in comparison with PCL and vaseline gas used as the control group. SSD at a concentration of 0.3% improved physicochemical properties of PCL. This concentration of SSD did not inhibit the attachment of human dermal fibroblasts (HDFs) to nanofibers in vitro, but showed antibacterial activity against Gram-positive Staphylococcus aureus (ST) and Gram-negative Pseudomonas aeruginosa (PS). Overall, results showed that SSD improves characteristics of PCL nanofibrous film and improves wound-healing process in one-week earlier compared to control. Cytotoxicity of SSD in fabricated nanofibrous mat is a critical challenge in designing an effective wound dressing that neutralizes cellular toxicity and improves antimicrobial activity. The PCL/SSD nanofibrous membrane with 500- µm thickness and 0.3% (w/v) SSD showed applicable characteristics as a wound dressing and it accelerated wound healing process in vivo.

We proposed a novel differentiation method for the efficient differentiation of adipose-derived mesenchymal stem cells (ADMSCs) into functional insulin-producing cells (IPCs) based on MafA overexpression. In this experimental study, a eukaryotic expression vector containing MafA [MafA/pcDNA3.1(+)] was constructed and purified. ADMSCs were differentiated into IPCs. ADMSCs were assigned in two groups including control (C), and the MafA overexpressed (MafA+) groups. The ADMSCs were transfected by MafA/pcDNA 3.1(+) at day 10 of the differentiation. Differentiated cells were analyzed for the expression of multiple β cell specific genes (Nkx2.2, Ngn3, Isl-1, Pdx1, MafA, Nkx6.1, and Insulin) using real-time polymerase chain reaction (PCR). The insulin secretion potency of the differentiated cells in response to glucose exposure was also determined using an enzyme-linked immunosorbent assay (ELISA) method and Dithizone (DTZ) staining. The IPCs from the control manipulated group, and un-differentiated ADMSCs group were transplanted to streptozotocin (STZ)-diabetic rats. Rats were monitored for blood glucose and insulin concentration. The results revealed that ADMSCs were successfully differentiated into IPCs through the 14 day differentiation protocol. The expression of β-cell specific genes in MafA+ IPCs was higher than in control cells. Glucose-induced insulin secretion after the exposure of IPCs to glucose was higher in MafA+ group than the control group. The STZ- diabetic rats showed an ability to secrete insulin and apparent hyperglycemic condition adjustment after transplantation of the control IPCs. The mean insulin concentration of diabetic rats that were transplanted by manipulated IPCs was significantly higher than ADMSCs-transplanted rats; however, no effect was observed in the concentration of blood glucose. The overexpression of MafA can be used as a novel promising approach for the efficient production of IPCs from ADMSCs in vitro. However, the future therapeutic use of the MafA+ IPCs in diabetic animals needs further investigations.

Asthenozoospermia (astheno) is a common male infertility disorder associated with low sperm motility. The progressive movement of sperm is an important factor in the fertilization rate, and it requires a high level of adenosine triphosphate (ATP). This experimental study aimed to identify the role of cytochrome c oxidase 6B2 (COX6B2) as an important functional subunit of Cytochrome-c Oxidase in sperm motility. According to the World Health Organization (WHO) criteria, Semen samples were collected from 14 asthenozoospermia and 16 normospermia individuals that were referring to the Infertility Research and Treatment Center of Khuzestan, Iran in October 2016- May 2017. The sperm from two groups was isolated via the Percoll density gradient centrifugation to prepare healthy, motile sperm for COX6B2 immunofluorescent staining and real-time polymerase chain reaction (PCR). In addition, apoptosis assessment was carried out simultaneously to compare apoptosis and the COX6B2 expression level. To analyze the data, descriptive statistics including frequency and mean and analytical statistics including Fisher’s exact test and two independent samples were used. COX6B2 was detected in midpiece of the sperm by immunofluorescence assays. In addition, the percentage of COX6B2 positive sperm in the astheno samples was almost half that of the normal group (49.0 ± 15.8 to 28.7 ± 14.1, P = 0.641). Real-time PCR definitely reconfirmed the immunofluorescent staining result. A decrease in apoptosis was shown in as the no samples compared with the normal group (19.1 ± 0.4 to 9 ± 0.2, P = 0.04). The expression of COX6B2 in the sperm midpiece represents the OXPHOS pathway and functionality of mitochondria in sperm. This study introduced COX6B2 staining as a potential functional test for the recognition of competent mitochondria in sperm and it could be assigned as a biomarker in male-factor patients

Curcumin (Cur) has been found to be very efficacious against many different types of cancer cells. However, the major disadvantage associated with the use of Cur is its low systemic bioavailability. Our present work investigated the toxic effect of encapsulation of Cur in PLGA (poly lactic-coglycolic acid) nanospheres (NCur) on PC3 human cancer prostate cell. In the present study, we have investigated the effects of NCur on growth, autophagia, and apoptosis in PC3 cells, respectively, by MTT assay, fluorescence microscopy, and Flow cytometry. MTT assays revealed that the NCur at the concentration of 25 μg/mL for 48 h were able to exert a more pronounced effect on the PC3 cells as compared to free Cur. Apoptotic index was significantly increased in NCur-treated cells compared to free Cur. The percentage of autophagic cells (LC3- II positive cells) was also significantly increased in NCur treatment in comparison to free Cur. These data indicate that the NCur has considerable cytotoxic activity more than Cur on P3 cell lines, which is mediated by induction of both apoptotic and autophagic processes. Thus, NCur has high potential as an adjuvant therapy for clinical application in prostate cancer.

In the present study, we assessed the effects of β-carotene on Titanium oxide Nanoparticle (TNP) induced mouse Spermatozoon Stem Cells (SSCs) apoptosis, at molecular level.
After isolation from cryptorchid mouse testis and characterization, spermatogonial stem cells were divided into four groups. In the control group, spermatogonial cells were cultured in α-MEM supplemented with 2% BSA (Bovine Serum Albumin). β-Carotene (BC) group was composed of control culture condition supplemented with 1 µg/ml β-carotene. TNP group comprised control culture condition supplemented with 1 µg/ml titanium oxide (TiO2). Ultimately the last group contained control culture condition supplemented with both 1 µg/ml BC and 1 µg/ml TiO2for three days. After that, spermatozoon viability was evaluated by MTT (3-[4,5-dimethylthiazolyl-2]-2,5-diphenyltetrazolium bromide) assay, apoptotic and necrotic indices with Annexin V/PI kit and gene expression of CASP3 and MAPK14 using qRT-PCR method.
TiO2 could significantly decrease viability of the cultured spermatozoon in TNP group compared to the control group. In BC group, we determined increased frequency of live spermatozoon compared to TNP or control group. Expression of apoptotic related genes significantly increased in TNP group. Spermatozoon induced by titanium oxide might be useful in clinical procedures. Measurement of apoptosis index using Annexin V/PI method also showed significant increase in apoptotic index of germ cells in TiO2 treated spermatozoon (P Expression of apoptotic related genes in cultured spermatozoon could efficiently be decreased by β-carotene treatment. Application of BC had a potential protective effect in preventing apoptosis in germ cells and might be useful in clinic.

Curcumin has been found to be very efficacious against many different types of diseases. However, the major disadvantage associated with the use of curcumin is its low systemic bioavailability. In the present study the protective effects of curcumin-loaded poly lactic-co-glycolic acid nanoparticles (nanocurcumin) against mono-iodoacetate-induced osteoarthritis in rats was investigated. Mono-iodoacetate was injected into right knee joints to induce osteoarthritis. In experimental groups, 14 days after injection of mono-iodoacetate, curcumin (200 mg kg-1) and nanocurcumin (200 mg kg-1) were gavaged, respectively, for two weeks. Then the rats were sacrificed and the right knee joints were removed and fixated in 10% formalin for histological assessments. Cellularity and matrix staining were significantly increased in articular cartilage of curcumin-treated animals compared to mono-iodoacetate group (p

The organotypic co-culture is a well-known technique to examine cellular interactions and their roles in stem cell proliferation and differentiation. This study aims to evaluate the effects of dermal fibroblasts (DFs) on epidermal differentiation of adipose-derived stem cells (ASCs) using a three-dimensional (3D) organotypic coculture technique.
In this experimental research study, rat DFs and ASCs were isolated and cultured separately on electrospun polycaprolactone (PCL) matrices. The PCL matrices seeded by ASCs were superimposed on to the matrices seeded by DFs in order to create a 3D organotypic co-culture. In the control groups, PCL matrices seeded by ASCs were placed on matrices devoid of DFs. After 10 days, we assessed the expressions of keratinocyte-related genes by real-time reverse transcriptase- polymerase chain reaction (RT-PCR) and expression of pan-cytokeratin protein by immunofluorescence in the differentiated keratinocyte-like cells from coculture and control groups. Keratinocyte-like cell morphologies were also observed by scanning electron microscopy (SEM).
The early, intermediate, and terminal differentiation keratinocyte markers-Cytokeratin14, Filaggrin, and Involucrin significantly expressed in the co-culture groups compared to the control ones (P The 3D organotypic co-culture bilayered construct that consisted of DFs and ASCs was an effective technique for epidermal differentiation of ASCs. This co-culture might be useful for epidermal differentiation of stem cells for future applications in skin regeneration.

The goal of the study described here, was to investigate the potential of umbilical cord derived mesenchymal stem cell (UC-MSCs) into hepatocyte like cells in a sequential 2D and 3D differentiation protocols as alternative therapy.
Mesenchymal stem cells (MSCs) were isolated from the umbilical cord (UC) and CD markers were analyzed by flow cytometry. For hepatic differentiation of UC-MSCs, cells were induced with a sequential 4-step protocol in 3D and 2D culture system. Urea concentration and albumin secretion into the culture medium was quantified by ELISA. Gene expression levels of AFP, ALB, and CK18 were determined by RT-PCR. Data were statistically analyzed by the SPSS software. The difference between the mean was considered significant when p Growth factor dependent morphological changes from elongated fibroblast-like cells to round epithelial cell morphology were observed in 2D culture. Cell proliferation analysis showed round-shaped morphology with clear cytoplasm and nucleus on the alginate scaffold in 3D culture. The mean valuses of albumin production and urea secretion were significantly higher in the 3D Culture system when compared with the 2D culture (p = 0.005 vs p = 0.001), respectively. Treatment of cells with TSA in the final step of differentiation induced an increased expression of CK18 and a decreased expression of αFP in both the 3D and 2D cultures (p = 0.026), but led to a decreased albumin gene expression, and an increased expression in the 2D culture (p = 0.001).
Findings of the present study indicated that sequential exposure of UC-MSCs with growth factors in 3D culture improves hepatic differentiation.

Due to increasing demand for liver tissue engineering, three-dimensional (3D) liver cells culture techniques have been proposed. Therefore, the aim of the present study was to examine the cells isolation and expansion of umbilical cord derived mesenchymal stem cells and in vitro 2D and 3D hepatocyte differentiation. Also functional characteristics of hepatocytes were analyzed. The study performed in several phases. In the first umbilical cord derived mesenchymal stem cells obtained and isolated, thereafter cellss expanded. Determination of Immunophenotype using Flowcytometry performed by DAKO – Galaxy Hepatic differentiation UC-MSCs was performed by four step sequential method using FGF-4, ITS, HGF, dexamethasone, glucagon, OSM and TSA. Urea production was quantified by ELISA. Section of tissue constructs stained with hematoxyllin and eosin for histological examination. MSCs isolated from umbilical cord expressed mesenchymal surface antigen such as CD73, but were negative against CD31. Several cell clusters mainly between the round cells were observed in alginate scaffold after 3d differentiation. Urea production was increased time- dependable and was significantly higher in the experimental group of 3D culture (P=0.001). Tissue construct of 3D culture revealed multicellular tissue with several euchromatin cell plates. The finding of the present study indicated that four step differentiation of umbilical cord derived mesenchymal stem cells within hydrogel scaffold induced functionally and morphologically characteristics of hepatocytes such as urea production and cell plates.

Zinc oxide nanoparticles (ZnO-NPs) are increasingly used in sunscreens, biosensors, food additives, pigments, manufacture of rubber products, and electronic materials. There are several studies about the effects of NPs on dermal fibroblast or keratinocytes, but very little attention has been directed towards adipose-derived mesenchymal stem cells (ASCs). A previous study has revealed that ZnO-NPs restricted the migration capability of ASCs. However, the potential toxicity of these NPs on ASCs is not well understood. This study intends to evaluate the effects of ZnO-NPs on subcutaneous ASCs. In this experimental study, In order to assess toxicity, we exposed rat ASCs to ZnO-NPs at concentrations of 10, 50, and 100 μg/ml for 48 hours. Toxicity was evaluated by cell morphology changes, cell viability assay, as well as apoptosis and necrosis detection. ZnO-NPs concentration dependently reduced the survival rates of ASCs as revealed by the trypan blue exclusion and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium- bromide (MTT) tests. ZnO-NPs, at concentrations of 10 and 50 μg/ml, induced a significant increase in apoptotic indices as shown by the annexin V test. The concentration of 10 μg/ml of ZnO-NPs was more toxic. Lower concentrations of ZnO-NPs have toxic and apoptotic effects on subcutaneous ASCs. We recommend that ZnO-NPs be used with caution if there is a dermatological problem.

Zinc oxide nanoparticles (ZNP) are increasingly used in sunscreens, biosensors, food additives and pigments. In this study the effects of ZNP on liver of rats was investigated. Experimental groups received 5, 50 and 300 mg/kg ZNP respectively for 14 days. Control group received only distilled water. ALT, AST and ALP were considered as biomarkers to indicate hepatotoxicity. Lipid peroxidation (MDA), SOD and GPx were detected for assessment of oxidative stress in liver tissue. Histological studies and TUNEL assay were also done. Plasma concentration of zinc (Zn) was significantly increased in 5 mg/kg ZNP-treated rats. Liver concentration of Zn was significantly increased in the 300 mg/kg ZNP-treated animals. Weight of liver was markedly increased in both 5 and 300 mg/kg doses of ZNP. ZNP at the doses of 5 mg/kg induced a significant increase in oxidative stress through the increase in MDA content and a significant decrease in SOD and GPx enzymes activity in the liver tissue. Administration of ZNP at 5 mg/kg induced a significant elevation in plasma AST, ALT and ALP. Histological studies showed that treatment with 5 mg/kg of ZNP caused hepatocytes swelling, which was accompanied by congestion of RBC and accumulation of inflammatory cells. Apoptotic index was also significantly increased in this group. ZNP at the dose of 300 mg/kg had poor hepatotoxicity effect. It is concluded that lower doses of ZNP has more hepatotoxic effects on rats, and recommended to use it with caution if there is a hepatological problem.

In recent years tissue engineering developed to replace or repair damaged tissues using cell and biomaterial. One of the most important fields of tissue engineering is simulation of in vivo micro environment of body tissues. This study aimed to develop culture chamber and cell biomaterial sheet engineering for the reconstruction of cartilage tissue. Stainless steel culture chamber was designed with mechanical factor affecting cartilage. Shear exerted on the wall of the chamber was predicted with computational fluid dynamic modeling with Fluent. The meshes were created with Gambit software.After isolation of chondrocytes from cartilage, cells mixed with natural biomaterial and hydrogel tissue construct cultured in chamber. Sections of resulted cell-biomaterial construct were examined with histological methods. The designed chamber mimicked synovial joint with perfusion flow. Maximum wall shear was predicted with fluent was 2.407× 10-3 Pa. Chondrocyte-scaffold was created as a thin sheet. Histological examination of chondrocyte biomaterial revealed morphology of native cartilage tissue with round cluster chondrocyte profile. Isogenic group of constract proliferated and expanded. According to simulation of in vivo environment of natural cartilage joint and simplification of tissue constructs production from chondrocyte and hydrogel scaffold, the novel strategy described here has great advantages to the improvement of cartilage tissue engineering.

In Iranian traditional medicine, Punica granatum, cabbage, and plantago have been used to prompt wound healing. The effects of cabbage, Punica granatum, and Plantago in wound healing have been investigated to some extent, but no experimental study has been performed to assess the impact of an ointment containing all these three substances on wound healing.. The aim of the present study was to evaluate the effect of an ointment prepared from a mixture of cabbage, Punica granatum, and plantago on local wound healing in Wistar rats.. This experimental study was performed on 21 male Wistar rats. Firstly, a full thickness wound was created on rats flank, near the spine by a bistoury blade and forceps. Then they were randomly divided into three groups of seven rats as the control group, a positive control group (the treatment group with phenytoin), and the third one received a treatment with studied ointment. The wounds were evaluated on days three, six, nine, and 12. Results were compared by SPSS software using ANOVA test.. The average percentage of wound healing in different groups on days 3, 6, 9 and 12 were as follows; 21.3%, 37.87%, 67.39% and 77.17% in control group; 24.98%, 34.21%, 70.74% and 88.55% in positive control group; and 32.35%, 61.27%, 94.53%, and 99.91%, in case group. A significant difference was observed between the control and case groups (P < 0.05)..

The aim of this study was to evaluate the effect of exendin-4 (EX-4) on differentiation of insulin-producing cells (IPCs) from rat bone marrow-derived mesenchymal stem cells (RAT-BM-MSCs). In this experimental study, RAT-BM-MSCs were cultured and the cells characterized by flow cytometry analysis of cell surface markers. RAT-BM-MSCs were subsequently treated with induction media with or without EX-4. After induction, the presence of IPCs was demonstrated with dithizone (DTZ) staining and gene expression profiles for pancreatic cell differentiation markers (PDX-1, GLUT-2, insulin) were assessed using reverse transcription polymerase chain reaction (RT-PCR). Insulin excreted from differentiated cells was analyzed with radioimmunoassay (RIA). The two-tailed student’s t-test was used for comparison of the obtained values. The percentage of DTZ-positive cells significantly increased in EX-4 treated cells (p<0.05). Expression of the islet-associated genes PDX-1, GLUT-2 and insulin genes in EX-4 treated cells was markedly higher than in the cells exposed to differentiation media without EX-4. RIA analysis demonstrated significant release of insulin with the glucose challenge test in EX-4 treated cells compared to EX-4 untreated cells. The results of this study have demonstrated that EX-4 can enhance differentiation of IPCs from RAT-BM-MSCs.

Previous studies have shown that some cytokines have protective effects on cartilage in joint diseases. In the current study، effects of IL-4 against morphological changes and tissue degradation induced by IL-1α on bovine nasal cartilage (BNC) explants were investigated. Fresh BNC samples were prepared from a slaughterhouse under sterile conditions. BNC explants culture was treated with both IL-lα (10 ng/ml) and IL-4 (50 ng/ml) at the same time for 28 days. The morphological characteristics of explants were assessed by using histology techniques and invert microscopy. Matrix metalloproteinase-1 (MMP-1) production was assessed within different days by using Western blotting. IL-lα induced prominent cartilage morphology degradation. The pro and active form of MMP-1 band substantially increased at day 21 of culture. In the presence of both IL-lα and IL-4، chondrocytes preserved their ordinary normal phenotype with intact extracellular matrix. In addition، a significant reduction in pro-MMP-1and inhibition of active MMP-1 was seen. In conclusion، IL-4 could be regarded as a potential candidate in cartilage protecting against the degradation changes of IL-lα. It seems that the preservation effect of IL-4 is associated with significant reduction of MMP-1.

A study of the histological events under interleukin-1α (IL-lα) induction of bovine nasal cartilage (BNC) could result in useful data to better understand the mechanisms involved in tissue breakdown in joint diseases. The aim of this study was to investigate the effects of IL-lα on chondrocyte phenotype and extracellular matrix (ECM) changes in BNC explants. In this experimental study, samples were divided into two groups. Group I (control group) BNC explants were cultured only in Dulbecco’s modified Eagle’s medium (DMEM). In group II, BNC explants were treated with IL-lα (10 ng/ml) for 28 days. Then, samples were harvested on culture days 3, 7, 14, 21 and 28 and chondrocyte morphology and ECM alterations were assessed by invert microscopy and histology by hematoxylin and eosin (H&E) and Alcian blue. Cell viability was evaluated by the lactate dehydrogenase (LDH) assay test. Data were analyzed by the t test and p<0.05 was considered significant. IL-lα induced significant morphological changes in cartilage. In the presence of IL-lα, most chondrocytes transformed into a fibroblast-like morphology with a granular black point appearance. An increase in the cell: matrix ratio was observed and there were decreased numbers of chondrocytes.IL-lα induced breakdown of ECM. We observed partial degradation of ECM between days 7-14 and complete degradation occurred between days 21-28 of culture. The LDH levels increased. IL-1α induced morphological changes in chondrocytes and increased destruction of cartilage ECM. There was a parallel correlation between proteoglycan degradation and changes in chondrocyte morpholgy.

Galectin-3 (Gal-3), a β-galactoside-binding lectin, is a multifunctional lectin that involves in a number of critical biological processes.

The purpose of this study was to investigate the expression pattern of Gal-3 in mouse endometrium during estrus phase of estrous cycle and pre-implantation.

In this experimental study 42 NMRI female mice were divided in seven different groups. Ovulation in NMRI female mice was stimulated by injecting hMG and hCG. Estrus phase was considered as stimulated and un-stimulated groups. The other groups of mice were mated, and the day of vaginal plug formation was considered as the day 1 of pregnancy. The mice of all groups were sacrificed on different days of pre-implantation period and their uterine horns were fixed and avidin- biotin complex method of immunohistochemistry (IHC) was applied.

In estrus group, Gal-3 immunoreactivity in luminal epithelium was strong, in stromal cells very strong, in glandular epithelium very weak and endothelial cells very strong. No identifiable difference was observed in un-stimulated and stimulated estrus phase. In test groups, days 1-2, insignificant difference of Gal-3 expression was observed. On day 3, luminal epithelium and stromal cells showed significant decrease in comparison to estrus and day 1 (p=0.001). On the 4th and 5th days, luminal epithelium and stromal cells showed significant decrease in comparison to estrus phase and days 1-3 (p=0.0001).

The data suggested that successful implantation is probably associated with the downregulation of Gal-3 in the mouse endometrium at the beginning of pregnancy.