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Introduction. Leishmaniasis is still a neglected tropical disease that can endanger more than 350 million people among 98 countries. Leishmania can survive in fibroblasts as latent inactive forms. This study was conducted to evaluate the role of superparamagnetic iron oxide nanoparticles (SPIONs) in cell culture for tracking the labeled Leishmania major in fibroblasts. Methods. Dextran-coated SPIONs were used for labeling L. major in co-culture of fibroblasts with the parasite. To quantify and trace SPION-labeled Leishmania, Prussian blue staining was undertaken. Fibroblast characterization was undertaken by real time polymerase chain reaction. Transmission electron microscope (TEM) was used for confirming the entry of the labeled L. major to the cytoplasm and the nucleus of the fibroblast. Results. Fibroblasts were spindle-shaped and adherent to culture flasks. Promastigotes were with thin elongated lance-like morphology with an anterior kinetoplast and an emergent free flagellum. Prussian blue staining revealed that internalized SPIONs were localized within cytoplasm and nucleus of the fibroblasts after 24 hours of culture. Prussian blue staining successfully showed the presence of iron (stained blue) in labeled L. major within the fibroblasts. This finding was confirmed by TEM, and labeled L. major was detected in the fibroblast cytoplasm and nucleus too. Conclusion. We can conclude that SPIONs are safe, inexpensive, easy to use, and accurate, and a fast method to label Leishmania parasite in cells that the parasite can be latent, such as fibroblasts. These findings can open a new window in diagnosis, pathogenesis, and treatment of cutaneous leishmaniasis and can be added to the literature. [ABSTRACT FROM AUTHOR]

In this study, a new series of spiro indolin-1,2-diazepine were designed, synthesized, and screened for their cholinesterase inhibitory activities. A novel, green, high-yielding approach was constructed to synthesize spiro indolin-1,2-diazepine derivatives through a cascade reaction of different isatins, malononitrile and 1,1-enediamines (EDAMs) via sequential four-component reactions to produce the target compounds with good to excellent yields. Next the inhibitory potencies of all derivatives were determined spectroscopically at 415 nm using the modified Ellman method. The results of the in vitro screening indicated that 5l with spiroindolin-1,2-diazepine core bearing 5-NO2 at R1 and 4-OH at R2 was the most potent and selective AChE inhibitor with an IC50 value of 3.98 ± 1.07 µM with no significant inhibition against BChE while 5j was the most active analog against both AChE and BChE enzymes. The structure–activity relationships suggested the variation in the inhibitory activities of derivatives was affected by different substitutions on the indolinone ring as well as the phenyl moiety. The enzyme kinetic studies of the most potent compound 5l at five different concentrations and acetylthiocholine substrate (0.1–1 mM) by Ellman's method revealed that it inhibited AChE in a mixed mode with a Ki of 0.044 μM. A molecular docking study was performed via induced fit docking protocol to predict the putative binding interaction. It was shown that the moieties used in the initial structure design play a fundamental role in interacting with the enzyme's binding site. Further, molecular dynamics simulations with the Schrödinger package were performed for 5l in a complex with AChE and revealed that compound 5l formed the stable complex with the enzyme. The MTT toxicity assessments against the neuroblastoma cell line were executed, and no toxicity was seen for 5l under the tested concentrations. [ABSTRACT FROM AUTHOR]

To assess in vitro and in vivo tracking of iron oxide labeled stem cells transfected by lipofectamine using magnetic resonance imaging (MRI), rat dental pulp stem cells (DPSCs) were characterized, labeled with iron oxide nanoparticles, and then transfected with lipofectamine to facilitate the internalization of these nanoparticles. Cell proliferation, viability, differentiation, and apoptosis were investigated. Prussian blue staining and MRI were used to trace transfected labeled cells. DPSCs were a morphologically spindle shape, adherent to culture plates, and positive for adipogenic and osteogenic inductions. They expressed CD73 and CD90 markers and lacked CD34 and CD45. Iron oxide labeling and transfection with lipofectamine in DPSCs had no toxic impact on viability, proliferation, and differentiation, and did not induce any apoptosis. In vitro and in vivo internalization of iron oxide nanoparticles within DPSCs were confirmed by Prussian blue staining and MRI tracking. Prussian blue staining and MRI tracking in the absence of any toxic effects on cell viability, proliferation, differentiation, and apoptosis were safe and accurate to track DPSCs labeled with iron oxide and transfected with lipofectamine. MRI can be a useful imaging modality when treatment outcome is targeted. [ABSTRACT FROM AUTHOR]

The aim of this study was to track dental pulp stem cells (DPSCs) labeled with dextran-coated superparamagnetic iron oxide nanoparticles (SPIONs) using magnetic resonance imaging (MRI). Dental pulp was isolated from male Sprague Dawley rats and cultured in Dulbecco's modified Eagle's medium F12 (DMEM-F12) and 10% fetal bovine serum. Effects of SPIONs on morphology, viability, apoptosis, stemness, and osteogenic and adipogenic differentiation of DPSCs were assessed. Prussian blue staining and MRI were conducted to determine in vitro efficiency of SPIONs uptake by the cells. Both non-labeled and labeled DPSCs were adherent to culture plates and showed spindle-shape morphologies, respectively. They were positive for osteogenic and adipogenic induction and expression of cluster of differentiation (CD) 73 and CD90 biomarkers, but negative for expression of CD34 and CD45 biomarkers. The SPIONs were non-toxic and did not induce apoptosis in doses less than 25 mg/mL. Internalization of the SPIONs within the DPSCs was confirmed by Prussian blue staining and MRI. Our findings revealed that the MRI-based method could successfully monitor DPSCs labeled with dextran-coated SPIONs without any significant effect on osteogenic and adipogenic differentiation, viability, and stemness of DPSCs. We provided the in vitro evidence supporting the feasibility of an MRI-based method to monitor DPSCs labeled with SPIONs without any significant reduction in viability, proliferation, and differentiation properties of labeled cells, showing that internalization of SPIONs within DPSCs were not toxic at doses less than 25 mg/mL. In general, the SPION labeling does not seem to impair cell survival or differentiation. SPIONs are biocompatible, easily available, and cost effective, opening a new avenue in stem cell labeling in regenerative medicine. [ABSTRACT FROM AUTHOR]

Keratin was extracted from chicken feather to produce scaffolds with poly(3-hydroxybutyrate) (P(3HB)) for tissue engineering applications. The scaffolds with different concentrations (10%, 20%, and 30%) of keratin were produced by electrospinning. The keratin presence in the produced scaffold was confirmed by FTIR analysis. The surface wettability of the P(3HB)/keratin scaffold was much higher compared to neat P(3HB) scaffold because of hydrophilic nature of the keratin. All the scaffolds showed over 80% porosity with an average fiber diameter of less than 750 nm. Mechanical properties of the P(3HB) scaffolds with up to 20% keratin were superior compared to neat P(3HB) scaffold. However, the mechanical properties of the P(3HB)/keratin scaffolds were deteriorated when the keratin concentration was 30% in the scaffold. The observed mechanical properties reduction was mainly attributed to the poor interaction between the keratin and P(3HB) matrix. Due to better biocompatibility of keratin, cell culture studies suggested that the P(3HB)/keratin scaffolds can have better cell attachment and proliferation compared to the neat P(3HB) scaffold. Owing to good biological response and mechanical properties, the P(3HB)/keratin scaffolds with an optimal concentration of keratin (20%) could be used as potential material for tissue engineering application. [ABSTRACT FROM AUTHOR]

In the present study, poly(3-hydroxybutyrate)-based composite scaffolds were prepared with multi-walled carbon nanotubes and hydroxyapatite nanoparticles for hard tissue engineering applications by electrospinning. All the prepared scaffolds showed connective porous structure, which were suitable for cell proliferation and migration. The mechanical properties of the poly(3-hydroxybutyrate) scaffold were improved by 0.5% of carbon nanotube addition, whereas the addition of hydroxyapatite nanoparticles up to 10% had an insignificant effect in tensile strength. However, scanning electron microscopy and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay results suggested that the mesenchymal stem cells attachment and their metabolic activities on the surface of the poly(3-hydroxybutyrate) scaffolds with hydroxyapatite were enhanced compared to poly(3-hydroxybutyrate) scaffolds. In addition, after 6 weeks of in vivo biocompatibility results in a model of rat indicated better tissue reactions for the scaffolds that contained hydroxyapatite. Overall, poly(3-hydroxybutyrate) composite scaffolds with 10% hydroxyapatite and 0.5% carbon nanotube showed optimal performances for the potential scaffold for hard tissue engineering application. [ABSTRACT FROM AUTHOR]

Covering surgical wounds with biomaterials, biologic scaffolds, and mesenchymal stem cells (MSCs) improves the healing process and reduces postoperative complications. This study was designed to evaluate and compare the effect of MSC-free/MSC-seeded new collagen/poly(3-hydroxybutyrate) (COL/P3HB) composite scaffold and human amniotic membrane (HAM) on the colon anastomosis healing process. COL/P3HB scaffold was prepared using freeze-drying method. MSCs were isolated and characterized from rat adipose tissue. After biocompatibility evaluation by MTT assay, MSCs were seeded on the scaffold and HAM by micro-mass seeding technique. In total, 35 male rats were randomly divided into five groups. After the surgical procedure, cecum incisions were covered by the MSC-free/MSC-seeded scaffold or HAM. Incisions in the control group were only sutured. One month later, the healing process was determined by stereological analysis. The Kruskal-Wallis followed by Dunn's tests were utilized for statistical outcome analysis (SPSS software version 21). COL/10% P3HB scaffold showed the best mechanical and structural properties (7.86 MPa strength, porosity more than 75%). MTT assay indicated that scaffold and especially HAM have suitable biocompatibility. Collagenization and neovascularization were significantly higher, and necrosis was considerably lower in all treated groups in comparison with the controls. MSC-seeded scaffold and HAM significantly decrease inflammation and increase gland volume compared with other groups. The MSC-seeded HAM was significantly successful in decreasing edema compared with other groups. Newly synthesized COL/P3HB scaffold improves the colon anastomosis healing; however, the major positive effect belonged to HAM. MSCs remarkably increase their healing process. Further investigations may contribute to confirming these results in other wound healing. • The novel COL/P3HB composite scaffold has a suitable biocompatibility. • COL/P3HB scaffold and HAM significantly improve the healing of colon anastomosis. • MSCs significantly improve the healing effect of both COL/P3HB scaffold and HAM. • The maintenance condition of COL/P3HB scaffold is simpler (4 °C) than HAM (−80 °C). [ABSTRACT FROM AUTHOR]

Purpose: Osteoarthritis (OA) as a progressive destructive disease of articular cartilage is the most common joint disease characterized by reduction of joint cartilage thickness, demolition of cartilage surface and new bone formation. To overcome these problems, the purpose of the current research was to evaluate and compare the in vivo effects of synovial membrane-derived mesenchymal stem cell (SMMSCs), platelet-rich plasma (PRP) and conditioned medium (secretome) on collagenase II-induced rat knee osteoarthritis (KOA) remedy. Methods: For the first step, SMMSCs were isolated and characterized. Also, secretome was collected from SMMSCs culture. Furthermore, PRP was collect from the rat heart venous blood. Second, two injection of collagenase II with an interval of 3 days was performed in the knee intra-articular space to induce osteoarthritis. Two weeks later, animals were randomly divided into 6 groups. Control group without treatment, positive group: taken an intra-articular sodium hyaluronate injection (0.1 ml), treatment groups taken an intra-articular injection of; treatment 1: SMMSCs (5 × 106), treatment 2: SMMSCs (5 × 106)/secretome (50 µl), treatment 3: SMMSCs (5 × 106)/PRP (50 µl), and treatment 4: SMMSCs (5 × 106)/ secretome (50 µl)/ PRP (50 µl). Three months later, rats were killed and the following assessments were executed: radiography, histopathology, and immunohistochemistry. Results: Our findings represented that a combination of the SMMSCs/secretome/PRP had a considerable effect on glycosaminoglycans (GAGs) and collagen II contents, articular cartilage preservation, compared with other groups. In addition, combination of the SMMSCs with PRP and secretome showed the lowest expression of mmp3, while SOX9 had the highest expression in comparison with other groups. Also, SMMSCs-injected groups demonstrated better results compared with positive and control groups. Conclusions: Injecting a combination of the SMMSCs/secretome/PRP resulted in better efficacy in terms of joint space width, articular cartilage surface continuity and integrity, sub-chondral bone and ECM constituents such as collagen II. Indeed, transplantation of this combination could be considered as a preliminary therapy for clinical trial study in the future. [ABSTRACT FROM AUTHOR]

Uterine fibrosis is an acquired disorder leading to menstrual irregularities, implantation impairment, and abortion. Mesenchymal stromal cells (MSCs) have antifibrotic properties through chemokine secretion. MSC-conditioned media (MSC-CM) contain paracrine components—exosomes—with a great potential for repairing damaged tissue or preventing fibrosis. The main goal of this study was to evaluate the preventive effects of bone marrow-derived MSC-CM (BM-MSC-CM) on uterine fibrosis after uterine curettage in rabbits. This study included 12 female rabbits (24 uterine horns in total). Excised uteri of each of the 12 female rabbits were randomly divided into four groups of intact negative control, curettage positive control, BM-MSC injection, and BM-MSC-CM injection in the way that two corresponding uteri from a rabbit were allocated to different groups. The MSC-CM were collected from cultivated BM-MSCs 48 hours after having been washed three times and replaced in serum-free media. Through a surgical approach, the caudal parts of the uteri were submitted to traumatic endometrial curettage, except for the intact negative uteri. After suturing the uterine walls, BM-MSCs or BM-MSC-CM were injected in the curettage site. Endometrial regeneration was histologically evaluated 30 days after treatment. Based on the evaluation of histomorphometric indices, curettage with or without preventive injections increased the growth of endometrial layers. However, the amount of fibrotic tissue in the CM and the BM-MSC injection groups was the same as the normal control groups, and all were less than the curettage group. A single injection of CM of MSCs after 30 days prevented the fibrotic tissue formation induced by curettage in endometrial layers of rabbits. Injecting BM-MSC-CM immediately after curettage prevented and reduced the uterine fibrosis similar to BM-MSCs in a rabbit model. [ABSTRACT FROM AUTHOR]

Background: Animals can play an important role in preparing tissues for human through the development of xenotransplantation protocols. The most common problem with liver transplantation like any other organ transplantation is organ supply shortage. Objective: To evaluate the in utero xenotransplantation of mouse bone marrow-derived stromal/stem cells (BMSCs) to the liver of rat fetus to produce mouse liver tissue. Materials and Methods: BMSCs were isolated and confirmed from enhanced green fluorescent protein (eGFP)-genetic labeled mice. Using a microinjection protocol, mice BMSCs were injected into the liver of rat fetuses in utero on day 14 of pregnancy. After birth, livers were collected and the presence of mice eGFP-positive cells in rat livers was evaluated through polymerase chain reaction. Results: The eGFP mRNA was detected in the liver of injected infant rats. BMSCs of adult mice were capable to remain functional probably as hepatocyte-like cells in liver of infant rats after in utero xenotransplantation. Conclusion: BMSCs have the potential for intrauterine xenotransplantation for the treatment of liver dysfunction before birth. This method can also be used for xenoproduction of liver tissue for transplantation. [ABSTRACT FROM AUTHOR]

Introduction: Cell‐based therapy is considered as promising strategy to cure stroke. However, employing appropriate type of stem cell to fulfill many therapeutic needs of cerebral ischemia is still challenging. In this regard, the current study was designed to elucidate therapeutic potential of epidermal neural crest stem cells (EPI‐NCSCs) compared to bone marrow mesenchymal stem cells (BM‐MSCs) in rat model of ischemic stroke. Methods: Ischemic stroke was induced by middle cerebral artery occlusion (MCAO) for 45 minutes. Immediately after reperfusion, EPI‐NCSCs or BM‐MSCs were transplanted via intra‐arterial or intravenous route. A test for neurological function was performed before ischemia and 1, 3, and 7 days after MCAO. Also, infarct volume ratio and relative expression of 15 selected target genes were evaluated 7 days after transplantation. Results: EPI‐NCSCs transplantation (both intra‐arterial and intravenous) and BM‐MSCs transplantation (only intra‐arterial) tended to result in a better functional outcome, compared to the MCAO group; however, this difference was not statistically significant. The infarct volume ratio significantly decreased in NCSC‐intra‐arterial, NCSC‐intravenous and MSC‐intra‐arterial groups compared to the control. EPI‐NCSCs interventions led to higher expression levels of Bdnf, nestin, Sox10, doublecortin, β‐III tubulin, Gfap, and interleukin‐6, whereas neurotrophin‐3 and interleukin‐10 were decreased. On the other hand, BM‐MSCs therapy resulted in upregulation of Gdnf, β‐III tubulin, and Gfap and down‐regulation of neurotrophin‐3, interleukin‐1, and interleukin‐10. Conclusion: These findings highlight the therapeutic effects of EPI‐NCSCs transplantation, probably through simultaneous induction of neuronal and glial formation, as well as Bdnf over‐expression in a rat model of ischemic stroke. [ABSTRACT FROM AUTHOR]

Background: Various methods were introduced to overcome the autograft shortage in burn wound care, including cell transplantation and tissue engineering. Aims: To evaluate the healing effect of allogenic human Wharton's jelly stem cells (hWJSCs) seeded onto acellular dermal matrix (ADM) in rat burn injuries. Patients and Methods: Human Wharton's jelly stem cells provided from umbilical cord tissue were characterized before transplantation, and the growth kinetic was determined. Skin samples from cosmetic surgeries were used for preparation of ADM. Forty male Sprague Dawley rats were randomly divided into 4 equal groups. Third‐degree burn was induced for all animals by exposing to hot water using a 2 cm ring for 10 seconds. Group 1 was burned rats that did not receive any treatment. After burn injury, the second group received silver sulfadiazine (SSD), the third group was treated just by using ADM, and the fourth group received 2 × 106 hWJSCs seeded onto ADM. The animals were euthanized for histologic evaluation after 7, 14, and 21 days. Results: Human Wharton's jelly stem cells were characterized to be spindle shape and positive for osteogenic and adipogenic induction and for mesenchymal markers but lacked hematopoietic markers. Population doubling time (PDT) was 40.1 hours with an increasing growth trend until day 6th. Macro‐ and microscopically, the healing was mild in ADM group and moderate in ADM + hWJSCs group after 21 days. Conclusion: Allogenic hWJSCs seeded onto ADM improved the healing process in burn wounds denoting to their therapeutic and anti‐inflammatory effects in burn wounds that can be added to the literature. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD) is a neurodegenerative disorder that leads to cognitive decline and memory loss. Unfortunately, there is no effective treatment for this condition, so there is a growing interest in developing new anti-AD agents. In this research project, a series of phenyl-quinoline derivatives were designed as potential anti-AD agents. These derivatives were substituted at two different positions on benzyl and phenyl rings. The structures of the derivatives were characterized using techniques such as IR spectroscopy, 1H NMR, 13C NMR, and elemental analysis. During the in vitro screening, the derivatives were tested against both acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). It was observed that most of the derivatives showed higher selectivity against BChE compared to AChE. Among the derivatives, analog 7n (with a methoxy group at R1 and a 4-bromine substituent at R2 exhibited the highest potency, with a 75-fold improvement in the activity compared to the positive control. Importantly, this potent analog demonstrated no toxicity at the tested concentration on SH-SY5Y cells, indicating its potential as a safe anti-AD agent. The level of GSK-3β was also reduced after treatments with 7n at 50 μM. Overall, this study highlights the design and evaluation of phenyl-quinoline derivatives as promising candidates for developing novel anti-AD agents. [ABSTRACT FROM AUTHOR]

Objective(s): Adipose tissue-derived mesenchymal stem cells (AT-MSCs) with more potent immunomodulatory effects, greater proliferative potential and secretion of growth factors and cytokines in comparison with bone marrow derived MSCs are more appropriate for cell therapy. The aims of the present study were to evaluate the histomorphometric effect of AT-MSCs allotransplantation on regeneration of germinal layer cells of seminiferous tubules in busulfan-induced azoospermic hamsters. Materials and Methods: In the present experimental case-control study, AT-MSCs were isolated from adipose tissue of two female and six male donor albino hamsters, and testes of the males were simultaneously used as negative control group. Six mature male recipient hamsters received two doses of busulfan with three weeks interval to stop endogenous spermatogenesis. Right testis of hamsters was intratubular injected with AT-MSCs via efferent duct 35 days after induction of azoospermia and was used as cell therapy group. The left testis without cell therapy was served as azoospermia group. Results: After 35 days, testes and epididymis in all groups were removed for histological evaluation. Histomorphometric analyses of AT-MSCs-treated testes and epididymis showed that the epithelial tissue of seminiferous tubules was normally repaired in most cell-treated seminiferous tubules, and spermatozoa were present in epididymis tubes in comparison with intact testes. The untreated seminiferous tubules and epididymis tubes of azoospermia group were empty. Conclusion: Allotransplanted AT-MSCs could successfully induce spermatogenesis in azoospermic seminiferous tubules of hamster. Therefore, AT-MSCs can be suggested as an attractive candidate in cell transplantation of azoospermia. [ABSTRACT FROM AUTHOR]

Background: Endometrial mesenchymal stem stromal cells (EnMSCs) are critical for uterine function, repair, and regeneration. Objective: This study introduced isolation technique of EnMSCs and compared the characteristics of EnMSCs in mature and immature ewes. Materials and Methods: Endometrial tissue samples from the uterus of 10 ewes were collected from the slaughterhouse. Endometrial cells were isolated from tissue using cold incubation and then chopping and treating was performed with collagenase type I. Isolated cells were cultured in cell culture medium and then attached cells to flasks were harvested as EnMSCs and subcultured. To enumerate the cells, the population doubling time (PDT) was determined and 2.2×104 cells in passage 4 were seeded into 24-well culture plates to compare the growth curves of isolated cells. Reverse transcription polymerase chain reaction (RT-PCR) was performed for detection of CD34 and CD73 markers. The osteogenic and adipogenic potential of isolated cells were determined using differentiation tests. Results: EnMSCs adhered to the flasks and displayed spindle-shape. Based on findings of the cell count and the growth curves, the EnMSCs growth was significantly more prominent in immature ewes in comparison to mature sheep. The PDT of EnMSCs in immature ewes was about 21 hr whereas this time period was two times higher (45 hr) in mature sheep. RT-PCR analyses of EnMSCs were positive for CD73 and negative for CD34. EnMSCs were differentiated into osteoblasts and adipocytes. Conclusion: Based on mesenchymal stem cells characters confirmed in EnMSCs, they can be a candidate for cell therapy and regenerative medicine. [ABSTRACT FROM AUTHOR]

The endometrial layer of the uterus contains a population of cells with similar characteristics of mesenchymal stem cells (MSCs). In the present study, caprine endometrial mesenchymal stromal stem cells (En-MSCs) characters and differentiation potential to chondrogenic, osteogenic, and adipogenic cell lines as well as their growth kinetics in breeding and anestrous stages were evaluated. En-MSCs were enzymatically isolated from endometrial layer of the uterus of adult goats and were cultured and subcultured until passage 4. The growth kinetics and population doubling time (PDT) of caprine En-MSCs in breeding and anestrous stages were determined. En-MSCs in passage 4 were used for the karyotyping and differentiation into chondrocytes, osteocytes, and adipocytes. The PDT in anestrus phase was 40.6 h and in cyclic goats was 53 h. En-MSCs were fibroblast-like in all passages. The number of chromosomes was normal (2n=60) with no chromosomal instability. Chondrogenic, osteogenic, and adipogenic differentiation of En-MSCs was confirmed by staining with Alcian blue, Alizarin red, and Oil Red O, respectively. Caprine En-MSCs demonstrated to be an alternative source of MSCs for cell therapy purposes in regenerative medicine. [ABSTRACT FROM AUTHOR]

Objective(s): Nowadays much effort is being invested in order to diagnose the mechanisms involved in neural differentiation. By clarifying this, making desired neural cells in vitro and applying them into diverse neurological disorders suffered from neural cell malfunctions could be a feasible choice. Thus, the present study assessed the capability of fetal brain extract (FBE) to induce rat bone marrow-derived mesenchymal stem cells (BM-MSCs) toward neural cells. Materials and Methods: For this purpose, BM-MSCs were collected from rats and cultured and their mesenchymal properties were confirmed. After exposure of the BM-MSCs to fetal brain extract, the cells were evaluated and harvested at days 3 and 7 after treatment. Results: The BM-MSCs that were exposed to FBE changed their appearance dramatically from spindle shape to cells with dendrite-like processes. Those neural like processes were absent in the control group. In addition, a neural specific marker, vimentin, was expressed significantly in the treatment group but not in the negative control group. Conclusion: This study presented the FBE as a natural neural differentiation agent, which probably has required factors for making neurons. In addition, vimentin overexpression was observed in the treated group which confirms neuron-like cell differentiation of BM-MSCs after induction. [ABSTRACT FROM AUTHOR]

Multilineage properties of mesenchymal stem cells (MSCs) cause their contribution in repair of different tissue. Adipose tissue is considered a good source of MSCs because of its abundance and easy access. The aims of this study were to characterize and determine the differentiation properties and growth kinetics of adipose tissue-derived stem cells (AT-SCs) from Syrian hamster. Fat tissue was isolated from the abdominal and pelvic regions of the hamster and by using type I collagenase, and cells were isolated and cultured. AT-SCs were characterized by morphology, cell surface markers, and adipogenic and osteogenic differentiations. AT-SCs were transferred to 24 well plates, and the growth kinetic was assessed by evaluation of the population doubling time (PDT). To show the chromosome stability of the AT-SCs, karyotyping was done. Hamsters' AT-SCs showed fibroblast like morphology and expressed CD73 and CD29 but not CD45. AT-SCs could be differentiated to adipocytes and osteoblasts verified by Alizarin Red and Oil Red O staining methods. The PDT of the passages 2 and 3 of the AT-SCs were 51.6 and 80.6 h, respectively. The karyotyping was normal until passage 4. Hamsters' AT-SCs showed mesenchymal properties denoting to these cells as a good source for the cell transplantation purposes in experimental and preclinical studies in hamster as an animal model. [ABSTRACT FROM AUTHOR]

Objectives: To determine the effect of bone marrow-derived mesenchymal stem cells (BMSCs) on regeneration of bone marrow and intestinal tissue and survival rate in experimental mice with acute radiation syndrome (ARS). Methods: Forty mice were randomly divided into two equal groups of A receiving no BMSC transplantation and B receiving BMSCs. BMSCs were isolated from the bone marrow and cultured in DMEM media. Both groups were irradiated with 10 Gy (dose rate 0.28 Gy/ min) 60CO during 35 minutes with a field size of 35?35 for all the body area. Twenty-four hours after ? irradiation, 150?10³ cells of passage 5 in 150 µl medium were injected intravenously into the tail. Animals were euthanized one and two weeks after cell transplantation. They were evaluated histologically for any changes in bone marrow and intestinal tissues. The survival rate in mice were also determined. Results: A significant increase for bone marrow cell count and survival rate were observed in group B in comparison to group A. Histological findings denoted to a healing in sample tissues. Conclusion: BMSCs could significantly reduce the side effects of ARS and increase the survival rate and healing in injured tissue. As such their transplantation may open a window in treatment of patients with ARS. [ABSTRACT FROM AUTHOR]

One of the readily available sources of mesenchymal stem cells (MSCs) is menstrual blood-derived stem cells (Men-SCs), which exhibit characteristics similar to other types of MSCs. This study was performed to determine the growth kinetics, plasticity, and characterization of Men-SCs in women. During spring 2014 in the southern Iranian city of Shiraz, menstrual blood (5 mL) was obtained from 10 women on their third day of menstruation in 2 age groups of 30 to 40 and 40 to 50 years old. Ficoll was used to separate the mononuclear cell fraction. After the Men-SCs were cultured, they were subcultured up to passage 4. Growth behavior and population doubling time were evaluated by seeding 5xl04cells into 12-and 24-well culture plates, and the colonies were enumerated. The expression of CD44, CD90, and CD34 was evaluated. The osteogenic potential was assessed by alizarin red staining. The Men-SCs were shown to be plastic adherent and spindle-shaped. Regarding the growth curves in the 12- and 24-well culture plates, it was demonstrated that in the women aged between 30 and 40 years, population doubling time was 55.5 and 62 hours, respectively, while these values in the women aged between 40 and 50 years were 70.4 and 72.4 hours, correspondingly. Positive expression of CD44 and CD90 and negative expression of CD34 were noted. In the osteogenic differentiation medium, the cells differentiated toward osteoblasts. As human Men-SCs are easily collectable without any invasive procedure and are a safe and rapid source of MSCs, they can be a good candidate for stem cell banking and cell transplantation in women. [ABSTRACT FROM AUTHOR]

Nowadays, regenerative medicine is going to find its true position among different kinds of treatment measures. In this regard, cell therapy brings hopes for the treatment of various diseases. As it is obvious, the role of nervous system for the appropriate function of other organs is not negligible. Unfortunately, damage to nerve cells is irreversible. Substitution of malfunctioned neural cells with the normal one is a noteworthy choice. The aim of the present study was to determine the effect of fetal rat brain extract on morphology of bone marrow-derived mesenchymal stem cells (BM-MSCs). Hence, pooled BM-MSCs were obtained from femurs and tibias of rats and cultured until reaching passage 3. BM-MSCs were characterized by the presence of mesenchymal specific stem cell markers and the absence of hematopoietic specific stem cell markers. Also, to confirm the BM-MSC potency, osteogenic and adipogenic differentiation capability was done. Fetal brain extract were prepared from rat in the late second week of gestational period. BM-MSCs were treated with basal culture medium containing 20 % brain extract. Control group was only treated with basal medium. After 3 days, neural-like cells were noticed with round body and significant projections. In some cases, the cells were bipolar and in the remained were multipolar. Cells with such structures were not seen in the control group. As the brain extract is readily available, inexpensive, do not cause any toxicity, and induce neural-like cells, it can be a candidate for neural lineage differentiation purposes. [ABSTRACT FROM AUTHOR]

Bone marrow is the most important source of mesenchymal stem cells (MSCs). Significant development in the comprehension of MSCs biology has opened the way to their clinical use. The object of this study was to establish a method for karyotyping of canine MSCs. We examined the effect of the increase in passage number on the chromosome and appearance of MSCs. A large 3-year-old mixed breed dog was used. Bone marrow (20 ml) was collected from the proximal humerus under general anesthesia and cultured and passaged in alpha minimal essential medium (αMEM) and fetal bovine serum (FBS). The canine bone marrow MSCs (BM-MSCs) were characterized by reverse transcription polymerase chain reaction (RT-PCR) and flask adherent fibroblast-like morphology. Karyotyping of cells of passages 1 and 8 was performed. The chromosome numbers were counted and evaluated to obtain the percentage of diploid cells. The best time for exposing BM-MSCs with media containing colcemid was 10 h. These cells were positive for the BM-MSCs markers CD90 and negative for hematopoietic markers CD34 and CD45 by RT-PCR. Karyotyping of these cells was normal, and also fibroblast-like cells were observed in every passage. [ABSTRACT FROM AUTHOR]

Mesenchymal stem cells (MSCs) have been isolated from various tissues of different species. The mammalian endometrium has morphological and functional modifications throughout the estrous cycle undergoing periodic proliferation and degeneration. The aim of this study was to isolate, culture, and characterize and to determine adipogenic differentiation of endometrial mesenchymal stem cells (En-MSCs) in heifers. Uteri of healthy heifers were collected from Shiraz Slaughterhouse, Iran and transferred to Stem Cell Laboratory of Stem Cell and Transgenic Technology Research Center to isolate En-MSCs from endometrial tissue samples. The tissue samples were exposed to collagenase type IA to reach the primary culture of En-MSCs. Isolated En-MSCs were sub-cultured up to passage 4. A specified number of En-MSCs were seeded into 12- and 24-well culture plates, and the number of cells was counted to evaluate the growth behavior of isolated cells and the population doubling time (PDT). RT-PCR for CD45 marker (hematopoietic stem cells) and CD73 marker (MSCs) and differentiation to adipocytes were performed for MSCs confirmation of En-MSCs of heifer. After cell culture, spindle-shape En-MSCs were visible adherent to the culture flasks. The cell count and the growth curves using 12- and 24-well culture plates showed that the PDT of En-MSCs was 37, 159.5, 52.9, and 136.3 h after seeding 2.2 × 10 and 20 × 10 (12-well) and 5 × 10 and 10 × 10 (24-well) En-MSCs, respectively. Using RT-PCR, heifer En-MSCs were negative for CD45 marker and positive for CD73 marker. Moreover, after culture of En-MSCs in differentiation medium, the cells differentiated toward adipocytes as verified by positive staining with Oil Red O staining. The morphology, growth kinetic, and differentiation of bovine En-MSCs demonstrated that these cells may be a good choice in bovine cell therapy and if necessary, they can be easily and safely administered in bovine tissue regeneration. [ABSTRACT FROM AUTHOR]

CONTEXT: Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are less invasive than bone marrow mesenchymal stem cells to obtain for cell therapy. AIMS: The aims of this study were to evaluate the germinal cells characteristics and repairs in seminiferous tubules of busulfan-induced azoospermic rats after AT-MSCs transplantation. SETTINGS AND DESIGN: Experimental case-control study. MATERIALS AND METHODS: In the present experimental study, donors AT-MSCs were isolated from subcutaneous adipose tissue of two Sprague-Dawley rats. The recipients (n = 5) were received two doses of 10 mg/kg of busulfan with 21 days interval to stop endogenous spermatogenesis. After induction of azoospermia by busulfan, rats were injected with the AT-MSCs into the efferent duct of right testes. After 60 days, the right testes were injected AT-MSCs were compared to left azoospermic testes. Five untreated male rats served as negative control. STATISTICAL ANALYSIS USED: Stereological indices were analyzed by one-way ANOVA and LSD post-hoc test. The spermatogenesis index was compared using Mann-Whitney U test. RESULTS: After stereological analyses, the seminiferous tubules treated with AT-MSCs had normal morphology. The untreated seminiferous tubules were empty. Spermatogenesis was observed in most cell-treated seminiferous tubules. CONCLUSIONS: The testis of busulfan-induced azoospermic rats accepted transplanted AT-MSCs. The transplanted AT-MSCs could induce spermatogenesis in seminiferous tubules of the rat. [ABSTRACT FROM AUTHOR]

Establishment of Guinea pig fetal fibroblast cells and their biological evaluation before and after cryopreservation were the main purposes of this study. After determination of the proper age of pregnancy by ultrasonography, 30 days old fetuses of Guinea pigs were recovered. Their skins were cut into small pieces (1 mm2) and were cultured. When reaching 80-90% confluence, the cells were passaged. Cells of the second and eighth passages were cultured in 24-well plates (4×104 cells/well) for 6 days and three wells per day were counted. The average cell counts at each time point were then plotted against time and the population doubling time (PDT) was determined. Then, vials of cells (2 × 106 cells/mL) were cryopreserved for 1 month and after thawing, the cell viability was evaluated. The PDT of the second passage was about 23 h and for the eighth passage was about 30 h. The viability of the cultures was 95% in the second passage and 74.5% in the eighth passage. It was shown that the Guinea pig fetal fibroblast cell culture can be established using the adherent culture method while, after freezing, the viability indices of these cells were favorable. [ABSTRACT FROM AUTHOR]

• Secretome of dental pulp stem cells contains mixture of growth factors. • Hair follicle stem cells express variety of trophic factors. • Bone marrow and adipose mesenchymal stem cells are widely used in the cell therapy. • Secretome preconditioning enhanced neurotrophic expression of mesenchymal stem cells. • Secretome of HFSCs promoted the differentiation of BM-MSCs into the glial lineage. During the last 20 years, stem cell therapy has been considered as an effective approach for regenerative medicine. Due to poor ability of stem cells to survive following transplantation, it has been proposed that beneficial effects of stem cells mainly depend on paracrine function. Therefore, the present study was designed to reinforce mesenchymal stem cells (MSCs) to express higher levels of trophic factors especially the ones with the neurotrophic properties. Here, bone marrow (BM)-MSCs and adipose-MSCs were treated with conditioned medium (CM) of dental pulp stem cells (DPSCs) or hair follicle stem cells (HFSCs) for up to three days. The relative expression of five key trophic factors that have critical effects on the central nervous system regeneration were evaluated using qRT-PCR technique. Furthermore, to assess the impacts of conditioned mediums on the fate of MSCs, expression of seven neuronal/glial markers were evaluated 3 days after the treatments. The obtained data revealed priming of BM-MSCs with HFSC-CM or DPSC-CM increases the BDNF expression over time. Such effect was also observed in adipose-MSCs following DPSC-CM treatment. Secretome preconditioning remarkably increased NGF expression in the adipose-MSCs. In addition, although priming of adipose-MSCs with HFSC-CM increased GDNF expression one day after the treatment, DPSC-CM enhanced GDNF mRNA in BM-MSCs at a later time point. It seemed priming of BM-MSCs with HFSC-CM, promoted differentiation into the glial lineage. Our findings showed that MSCs preconditioning with secretome of neural crest-derived stem cells could be a promising approach to enhance the neurotrophic potential of these stem cells. [ABSTRACT FROM AUTHOR]

Transplantation of bone marrow stem cells (BMSCs) has shown to have a vital role in promoting nerve regeneration after SCI. The aim of this study was to investigate the effect of BMSCs transplantation in healing of spinal cord injury (SCI) in mice based on morphologic parameters. Forty two male mice were randomly divided into 3 groups of control with no intervention, experimental SCI without treatment, and experimental SCI transplanted with 2 × 105 BMSCs intravenously. To induce SCI bilaterally, T10 was compressed for 2 min. The animals were sacrificed 3 and 5 weeks after SCI and T7-T11 segments of spinal cord were removed and stained by Giemsa and H&E methods. Stereological assessment estimated the gray and white matter volume, the number of neurons and neuroglia and diameter of central canal. The average amount of gray matter in SCI injury group was significantly lower than control group. An increase in the number of neurons was noted after cell transplantation. The number of neurons in SCI injury group significantly decreased in comparison to the control group. In cell transplantation group, a significant increase in the number of neurons was visible when compared to SCI injury group. The increase in the number of neurons after cell transplantation denotes to the regenerative potential of BMSCs in SCI. These findings can be added to the literature and open a new window when targeting treatment of SCI. [ABSTRACT FROM AUTHOR]

Appropriate selection of suitable materials and methods is essential for scaffolds fabrication in tissue engineering. The major challenge is to mimic the structure and functions of the extracellular matrix (ECM) of the native tissues. In this study, an optimized 3D structure containing poly(3-hydroxybutyrate) (P3HB), multiwalled carbon nanotubes (MCNTs) and curcumin (CUR) was created by electrospinning a novel biomimetic scaffold. CUR, a natural anti-inflammatory compound, has been selected as a bioactive component to increase the biocompatibility and reduce the potential inflammatory reaction of electrospun scaffolds. The presence of CUR in electrospun scaffolds was confirmed by 1H NMR and Fourier-transform infrared spectroscopy (FTIR). Scanning electron microscopy (SEM) revealed highly interconnected porosity of the obtained 3D structures. Addition of up to 20 wt% CUR has enhanced mechanical properties of the scaffolds. CUR has also promoted in vitro bioactivity and hydrolytic degradation of the electrospun nanofibers. The developed P3HB-MCNT composite scaffolds containing 20 wt% of CUR revealed excellent in vitro cytocompatibility using mesenchymal stem cells and in vivo biocompatibility in rat animal model study. Importantly, the reduced inflammatory reaction in the rat model after 8 weeks of implantation has also been observed for scaffolds modified with CUR. Overall, newly developed P3HB-MCNTs-CUR electrospun scaffolds have demonstrated their high potential for tissue engineering applications. [ABSTRACT FROM AUTHOR]