The effect of decellularized cartilage matrix scaffolds combined with endometrial stem cell-derived osteocytes on osteochondral tissue engineering in rats.

Since decellularized tissues may offer the instructive niche for cell differentiation and function, their use as cell culture scaffolds is a promising approach for regenerative medicine. To repair osteochondral tissues, developing a scaffold with biomimetic structural, compositional, and functional characteristics is vital. As a result of their heterogeneous structure, decellularized articular cartilage matrix from allogeneic and xenogeneic sources are considered appropriate scaffolds for cartilage regeneration. We developed a scaffold for osteochondral tissue engineering by decellularizing sheep knee cartilage using a chemical technique. DNA content measurements and histological examinations revealed that this protocol completely removed cells from decellularized cartilage. Furthermore, SEM, MTS assay, and H&E staining revealed that human endometrial stem cells could readily adhere to the decellularized cartilage, and the scaffold was biocompatible for their proliferation. Besides, we discovered that decellularized scaffolds could promote EnSC osteogenic differentiation by increasing bone-specific gene expression. Further, it was found that decellularized scaffolds were inductive for chondrogenic differentiation of stem cells, evidenced by an up-regulation in the expression of the cartilage-specific gene. Also, in vivo study showed the high affinity of acellularized scaffolds for cell adhesion and proliferation led to an improved regeneration of articular lesions in rats after 4 weeks. Finally, a perfect scaffold with high fidelity is provided by the developed decellularized cartilage scaffold for the functional reconstruction of osteochondral tissues; these types of scaffolds are helpful in studying how the tissue microenvironment supports osteocytes and chondrocytes differentiation, growth, and function to have a good osteochondral repair effect.

Simultaneous impact of atorvastatin and mesenchymal stem cells for glioblastoma multiform suppression in rat glioblastoma multiform model.

Glioblastoma multiform (GBM) is known as an aggressive glial neoplasm. Recently incorporation of mesenchymal stem cells with anti-tumor drugs have been used due to lack of immunological responses and their easy accessibility. In this study, we have investigated the anti-proliferative and apoptotic activity of atorvastatin (Ator) in combination of mesenchymal stem cells (MSCs) on GBM cells in vitro and in vivo. The MSCs isolated from rats and characterized for their multi-potency features. The anti-proliferative and migration inhibition of Ator and MSCs were evaluated by MTT and scratch migration assays. The annexin/PI percentage and cell cycle arrest of treated C6 cells were evaluated until 72 h incubation. The animal model was established via injection of C6 cells in the brain of rats and subsequent injection of Ator each 3 days and single injection of MSCs until 12 days. The growth rate, migrational phenotype and cell cycle progression of C6 cells decreased and inhibited by the interplay of different factors in the presence of Ator and MSCs. The effect of Ator and MSCs on animal models displayed a significant reduction in tumor size and weight. Furthermore, histopathology evaluation proved low hypercellularity and mitosis index as well as mild invasive tumor cells for perivascular cuffing without pseudopalisading necrosis and small delicate vessels in Ator + MSCs condition. In summary, Ator and MSCs delivery to GBM model provides an effective strategy for targeted therapy of brain tumor.

Electrospun cellulose acetate/gelatin nanofibrous wound dressing containing berberine for diabetic foot ulcer healing: in vitro and in vivo studies.

Functional wound dressing with tailored physicochemical and biological properties is vital for diabetic foot ulcer (DFU) treatment. Our main objective in the current study was to fabricate Cellulose Acetate/Gelatin (CA/Gel) electrospun mat loaded with berberine (Beri) as the DFU-specific wound dressing. The wound healing efficacy of the fabricated dressings was evaluated in streptozotocin-induced diabetic rats. The results demonstrated an average nanofiber diameter of 502 ± 150 nm, and the tensile strength, contact angle, porosity, water vapor permeability and water uptake ratio of CA/Gel nanofibers were around 2.83 ± 0.08 MPa, 58.07 ± 2.35°, 78.17 ± 1.04%, 11.23 ± 1.05 mg/cm2/hr, and 12.78 ± 0.32%, respectively, while these values for CA/Gel/Beri nanofibers were 2.69 ± 0.05 MPa, 56.93 ± 1°, 76.17 ± 0.76%, 10.17 ± 0.21 mg/cm2/hr, and 14.37 ± 0.42%, respectively. The antibacterial evaluations demonstrated that the dressings exhibited potent antibacterial activity. The collagen density of 88.8 ± 6.7% and the angiogenesis score of 19.8 ± 3.8 obtained in the animal studies indicate a proper wound healing. These findings implied that the incorporation of berberine did not compromise the physical properties of dressing, while improving the biological activities. In conclusion, our results indicated that the prepared mat is a proper wound dressing for DFU management and treatment.

Natural biomacromolecule based composite scaffolds from silk fibroin, gelatin and chitosan toward tissue engineering applications.

Natupolymer-based scaffolds can increase cell affinity to biomaterials and improve cell responses. Silk fibroin, chitosan and gelatin that mimic the properties of natural extra-cellular matrix (ECM) were chosen due to their biocompatibility, biodegradability and less immunogenic reactions. We prepared composite scaffolds with different blending ratios of silk fibroin-chitosan-gelatin by freeze-drying technique. Silk fibroin was extracted from the Bombyx mori silkworm. The scaffolds were characterized by scanning electron microscopy (SEM), surface wettability, swelling measurements, In Vitro enzymatic degradation measurements and tensile test. The composite scaffolds showed pore sizes from 125 µm to 175 µm, good interconnectivity between pores and suitable porosity which are desirable for cell growth. The addition of chitosan-gelatin to silk fibroin increased water uptake and degradation rate and reduced mechanical strength but silk fibroin affect reversely on the degradation and mechanical strength of composite scaffolds. Biocompatibility of scaffolds was demonstrated by MTT-assay and hematoxylin-eosin (H&E) staining which lead to the growth and adhesion of endothelial cells. In this study, the fabricated composite scaffolds have the potential for tissue engineering applications.

A facile two step heat treatment strategy for development of bioceramic scaffolds for hard tissue engineering applications.

In the present study, a two-step sintering (TSS) method has been used to improve the mechanical properties, biocompatibility, drug release, and osteogenesis abilities of hardystonite (HT) ceramic scaffolds for tissue engineering and drug delivery applications. The average particle size of HT scaffold is kept lower than 80 nm and is reached higher than 130 nm by using two-step and conventional sintering methods, respectively. The compressive strengths of the prepared nanocrystalline HT scaffolds were found to be significantly higher than those of the micro-structure HT and currently available hydroxyapatite scaffolds. A comparative analysis of cell viability and live/dead staining of human mesenchymal stem cells (hMSCs) in nano- and micro-structured HT scaffolds and their drug release potentiation was carried out. The results showed that the nano-structured HT scaffolds have higher cell viability, biocompatibility and longer-term doxorubicin (DOX) release potential than the micro-structured ones. The results of quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) analyses showed that the expression of adhesion and differentiation supporting genes were significantly higher in nano-structured HT scaffolds as compared to the micro-structured ones. The results of qRT-PCR also showed that the mRNA expression level of ERK1/2 and P38 MAPK from hMSCs were significantly higher in nano-structured HT scaffolds than the micro-structured ones. These results potentially open new aspects for using nano-structured scaffolds in bone tissue engineering applications.

The effect of purmorphamine on differentiation of endometrial stem cells into osteoblast-like cells on collagen/hydroxyapatite scaffolds.

We assessed the effect of purmorphamine along with collagen/hydroxyapatite scaffold in inducing osteogenesis of human endometrial stem cells (hEnSCs). The adhesion, viability, proliferation, and differentiation of cells on scaffold were assayed with SEM, MTT, real time-PCR, and ALP assay, respectively. The results were shown good integration of cells with scaffold. Also, qRT-PCR of differentiated cells shows that osteoblast cell markers are expressed after 21d in 2D and scaffold groups while in the scaffold group the expression of these markers were higher than the 2D group. Based on our findings, collagen/hydroxyapatite scaffold with PMA has the potential role in osteogenic differentiation of hEnSCs.

The role of climate on prevalence or eradication of vesical schistosomiasis in Khuzestan Province of Iran.

Climate is defined as the combination of climate and air elements of a given region which is usually measured for a period of decades. De-marton climate classification has been established based on many factors, including elements such as temperature and rainfall. Vesicle schistosomiasis is a parasitic disease caused by Schistosoma haematobium. This parasite lives in the blood vessels of the bladder. The parasite can cause hematuria in human and if not treated properly can lead to vesicale carcinoma. The parasite is distributed only in certain parts of the province and it is highly dispersed along the rivers of Dez, Karkheh and Karun with high emissions. In 1970, the prevalence of infection in infected foci was 23.8 %. Campaign against the parasite began in 1958 but it did not encompass all centers of infection. Preventive measures include diagnosis and treatment of patients, public health promotion, health education, drying swamps and ponds, improving the environment, cementing the irrigation canals, and the use of moluscocide eventually leads to changing the ecological and conditions of parasite and snail inhabits. Application of preventive measures resulted in the reduction of infection level to 0.7 % in 1979. By continuing struggle and intensifying preventive measures and changing ecological and climatic environment, in 2008, the examination of 3400 urine samples of students in Andimeshk district revealed no cases of the vesical schistosomiasis. It is concluded that S. haematobium and vesical schistosomiasis is eliminated from Khuzestan province southwest Iran, but the disease is still prevalent in neighboring Iran's western border country (Iraq) and due to the special conditions of its facilities and the traffic between the two countries, it is necessary to control and eradicate the disease in Iraq by using the experiences of Iran in eliminating the disease.

Control of Cutaneous Leishmaniasis Using Geographic Information Systems from 2010 to 2014 in Khuzestan Province, Iran.

BACKGROUND: Cutaneous leishmanisis (CL) is found worldwide and is considered to be endemic in 88 countries such as Iran. Geographic information system (GIS) is a method that can create, archive, analyze traditional map and place data of the disease distribution. The aim of this study was to produce distributional maps of CL over five years and evaluate the role of GIS in control of CL in Khuzestan province where an endemic area of CL in Iran is. METHODS: CL epidemiological data on the District and village levels for the period 2010-2013 were provided as census by health surveillance system in all counties and in control diseases center (CDC) of Khuzestan province. After collection of CL data, the collected data of CL from 2010 to 2013 were analyzed using GIS. The collected data of CL from 2010 to 2013 was analyzed using GIS. The endemic areas of CL during 2010-2013 were recognized using GIS maps and the control programs of CL were done in these regions based on epidemiological situation and the stratification of risk areas. RESULTS: During the study period, there were 4672 recorded cases of clinical cases of CL by Khuzestan Health Center. Data of GIS referring to CL patients showed that center and eastern districts of Khuzestan had a significant number of cases. In 2014 that control program was done, ten distinct of Khuzestan Province didn't show any cases of the disease. CONCLUSION: In conclusion, analyses of data distributed in the geographic spaces are increasingly appreciated in leishmaniasis control management. GIS tools promoted greater efficiency in making decisions and planning activities in the control of vector born disease such as leishmaniasis.

Evaluation of Motor Neuron-Like Cell Differentiation of hEnSCs on Biodegradable PLGA Nanofiber Scaffolds.

Human endometrium is a high-dynamic tissue that contains human endometrial stem cells (hEnSCs) which can be differentiated into a number of cell lineages. The differentiation of hEnSCs into many cell lineages such as osteoblast, adipocyte, and neural cells has been investigated previously. However, the differentiation of these stem cells into motor neuron-like cells has not been investigated yet. Different biochemical and topographical cues can affect the differentiation of stem cells into a specific cell. The aim of this study was to investigate the capability of hEnSCs to be differentiated into motor neuron-like cells under biochemical and topographical cues. The biocompatible and biodegradable poly(lactic-co-glycolic acid) (PLGA) electrospun nanofibrous scaffold was used as a topographical cue. Human EnSCs were cultured on the PLGA scaffold and tissue culture polystyrene (TCP), then differentiation of hEnSCs into motor neuron-like cells under induction media including retinoic acid (RA) and sonic hedgehog (Shh) were evaluated for 15 days. The proliferation rate of cells was assayed by using MTT assay. The morphology of cells was studied by scanning electron microscopy imaging, and the expression of motor neuron-specific markers by real-time PCR and immunocytochemistry. Results showed that survival and differentiation of hEnSCs into motor neuron-like cells on the PLGA scaffold were better than those on the TCP group. Taken together, the results suggest that differentiated hEnSCs on PLGA can provide a suitable, three-dimensional situation for neuronal survival and outgrowth for regeneration of the central nervous system, and these cells may be a potential candidate in cellular therapy for motor neuron diseases.

Enhancing neuronal growth from human endometrial stem cells derived neuron-like cells in three-dimensional fibrin gel for nerve tissue engineering.

Nerve tissue engineering (NTE) is one of the most promising methods to restore central nerve systems in human health care. Three-dimensional (3D) distribution and growth of cells within the porous scaffold composed of nanofibers are of clinical significance for NTE. In this study, an attempt was made to develop and characterize the use of fibrin gel and human endometrial stem cells (hEnSCs)-derived neuron-like cells simultaneously to support cell behavior especially neuron outgrowth. The structural and mechanical characteristics of fibrin gel scaffold were examined with SEM and rheometer. Also, hEnSCs-derived neuron-like cells were cultured in fibrin gel and were subsequently analyzed with immunofluorescent staining against neuronal markers. In parallel, the survival and growth rates of the cells were determined by MTT assay and neurite extension. At the end, cell-matrix interactions were investigated with SEM and TEM micrographs. Mechanical properties of fabricated scaffold were studied and results indicated appropriate choice of material, SEM and TEM showed excellent integration of cells with nanofibers regarding the relation between cells and fibrin gel. Immunofluorescent staining of fibrin gel after 6 days of cell seeding and culture demonstrated well expanded and incorporated network of neurons. In addition, viability, proliferation, and neuronal growth of seeded cells were analyzed at days 1, 3, and 6. Comparing those results with 2D culture of seeded cells showed positive effect of 3D culture. Taken together, the results suggest that fibrin can provide a suitable, three-dimensional scaffold for neuronal survival and outgrowth for regeneration of the central nervous system.

Three-dimensional culture of differentiated endometrial stromal cells to oligodendrocyte progenitor cells (OPCs) in fibrin hydrogel.

Neural tissue engineering is one of the most promising strategies for treatment of nerve tissue injuries. Three-dimensional (3D) environment mimics in vivo conditions for cells. 3D distribution and growth of the cells within the scaffold are both important for neural tissue engineering. In this study, endometrial stromal cell-derived oligodendrocyte progenitor cells (EnSC-derived OPCs) were cultured in fibrin gel and cell differentiation and viability were evaluated after 8 days of post-culture. The structural and mechanical characteristics of fibrin gel-like scaffold were examined with rheological analysis. EnSCs were isolated from donor tissue and were induced to OPCs with growth factors (FGF2/EGF/PDGF-AA) for 12 days, then were cultured in fibrin gel with Triiodothyronine (T3) medium for another 8 days. The viability of cells was analyzed using MTT assay for a period of 8 days culturing in a fibrin matrix. Structure of fibrin matrix and cell morphology was analyzed with SEM. TEM, immunostaining and quantitative RT-PCR was performed for OPCs markers after cell culturing in fibrin matrix. Cell viability is enhanced in fibrin matrix after 8 days. SEM and TEM show that cells are in good integration with nano-fibers. Moreover, immunohistochemistry and quantitative RT-PCR of OPCs differentiation markers showed that Olig2, Sox10, PDGFRa, CNP, and A2B5 are expressed after 8 days culturing within fibrin matrix. Fibrin can provide a suitable 3-D scaffold for EnSCs differentiated cells for the regeneration of CNS.