effect of mesenchymal stem cells and aqueous extract of Elaeagnus Angustifolia on the mechanical properties of articular cartilage in an experimental model of rat osteoarthritis

Introduction: Although, the effect of direct intra-articular injection of bone marrow stem cells [BMSCs] on the repair of articular cartilage and the effect of Elaeagnus angustifolia extract on pain relief in patients with osteoarthritis have been investigated, no studies has been conducted to compare the effects of these two therapeutic methods on the mechanical properties of articular cartilage. In the present stuy, the effect of these two methods on the mechanical strength of knee articular cartilage in a model of rat osteoarthritis has been studied

Methods: In the present research, 48 mature, male Wistar rats were used. Animals were randomly divided into 6 groups of 8 as follows: control group [healthy animals], saline with mono-iodoacetate [MIA], MIA with Elaeagnus angustifolia extract, MIA with BMSCs, and MIA with a combination of Elaeagnus angustifolia extract and BMSCs. Osteoarthritis was induced by injection of 50 and muL solution of MIA in rats of groups 3 to 6. About 500 mg/kg Elaeagnus angustifolia extract was injected intraperitoneally daily for 4 weeks and nonautologous mesenchymal stem cells were injected into the knee joint on the 14th day. Stress-relaxation test was conducted applying 0.1 mm displacement at the rate of 5 mm/min for 1000 seconds. Then, the maximum initial force, instantaneous stiffness,equilibrium force, and equilibrium stiffness were calculated

Results: Induction of osteoarthritis model decreased instantaneous stiffness, maximum initial force, and equilibrium stiffness as compared to the healthy group [P=0.05]. Using Elaeagnus angustifolia extract and bone marrow stem cells increased instantaneous stiffness and equilibrium stiffness compared to MIA group, although this increase was statistically significant only in the BMSCs group [P=0.04 and P=0.026, respectively]. In the BMSCs group, maximum initial force also significantly increased compared to MIA group [P=0.04]

Conclusion: Apparently direct injection of BMSCs into the knee joint with osteoarthritis is more effective in increasing mechanical strength of the cartilage and improving the performance of the weight-bearing joint compared to using Elaeagnus angustifolia extract

Derivation of ES-like cell from neonatal mouse testis cells in autologous sertoli cells co-culture system

Spermatogonial stem cell [SSC] is a self-renewing population of male adult stem cell. SSCs have a differentiation potential which are similar to embryonic stem cells. These Embryonic stem like [ES-like] cells can be a potential source for pluripotent cells for stem cell-based therapy. This study presents an economical and simple co-culture system for pluripotent stem cells generation from neonatal mouse testis. Isolated testicular cells were cultured in DMEM/F12. Characteristics of the isolated cells and obtained ES-like cell were immune-cytochemically confirmed by examining the presence of PLZF, vimentin, Oct4 and Nanog protein. Expression of the pluripotency and germ-cell specific genes was analyzed by qPCR in derived ES-like colony and SSCs respectively. The experiment results indicated that our method of obtaining pluripotent ES-like cells from spermatogonial cells [SCs] is simpler than the described methods. ES-like cells were immunopositive for pluripotency markers. ES-like cell qPCR results indicated significant increase in pluripotency genes expression and significant decrease in germ cell-specific genes expression. The results indicated that ES-like cell with pluripotency characteristic were generated from freshly isolated spermatogonial cells. The pluripotent stem cells provide a cellular reservoir usable for regenerative medicine instead of embryonic stem cells.

Isolation, expansion and purification of mouse spermatogonial stem cells in an autologous sertoli cell co-culture system

This study presents a simple method for isolation, expansion and purification of neonatal mouse Spermatogonial stem cells. We used enzymatic digestion to isolate a cell suspension of spermatogonia and Sertoli cells from neonatal 2-day-old mice. The cells were cultured in DMEM/F12 that contained 10% serum for two weeks. Sertoli and spermatogonia cell characteristics were confirmed by examining for the presence of vimentin and PLZF proteins, respectively. To assess the rate of spermatogonia stem cell expansion, the area and number of colonies were measured during the two weeks of culture. At the end of the second week, we detected spermatogonia cell-specific expressions of the Stra8, Piwill2, DAZL, and Mvh genes. Current results indicated that isolated Sertoli and spermatogonia cells were immunopositive for specific markers. During the culture period, a significant difference was seen in the number and area of Spermatogonial stem cell colonies [P<0.05] at four time points. In addition, Spermatogonial specific gene expression demonstrated that these cells were undifferentiated after two weeks of culture. Our study showed that co-culture of spermatogonia and Sertoli cells from same source provides a convenient and efficient environment. This co-culture, without the addition of external growth factors and chemical manipulations, can be used for proliferation of spermatogonia stem cells

Effect of trolox addition to cryopreservation media on human sperm motility

Sperm parameters and motion kinetics are affected by cryopreservation. The main purpose of the current study was to determine the effect of different concentrations of Trolox as an antioxidant to freezing-thawing procedure on human sperm kinematic parameter. Semen was collected from 20 normal donors and divided into five aliquots prior to cryopreservation. The first aliquot was analyzed by computer-assisted sperm analysis [CASA]. Other aliquots were mixed with cryo-protective agent containing 0, 20, 40, and 80 micro mol Trolox and treated samples were cryopreserved in liquid nitrogen. After two weeks samples were thawed and sperm motion kinematics was measured by CASA. Percent motility [Mot], curvilinear velocity [VCL], straight-line velocity [VSL], average path velocity [VAP], linearity [LIN], and amplitude of lateral head displacement [ALH] were compared before and after freeze. Addition of 40 micro mol Trolox resulted in significantly higher [p<0.05] post thaw VCL, VSL and VAP compared to other groups. Therefore the percentage of post thaw motile spermatozoa were significantly higher [p<0.01]. The supplementation of Trolox significantly improved the post-thawed human semen quality, especially progressive motility and average path velocity

[Study of the prophylactic effects of amifostine on decrease of lung tissue injuries in the acute phase in rats exposed to sulfur mustard]

Inhalation of Sulfur mustard [HD] will cause lung epithelial inflammation and injury. There are different results from the prophylactic effects of amifostine [AM] on protection of lung epithelial tissue against HD. The aim of this study was to evaluate the prophylactic effects of AM on protection of rat lung tissue exposed to HD. In this study twenty Albino Wistar adult male rats weighting 200 +/- 20 grams were used. Rats were divided randomly into 4 groups [5 rats in each group] as below: Normal saline group [NS], AM group, HD group [0.25% HD] and HD+AM group. Normal saline and HD solution were injected by intra tracheal catheter. Animals in AM and HD+AM groups received AM by intra peritoneal injection for 14 days daily. All rats were killed after 14 days; parts of the base of right lungs were removed, fixed and processed for histological evaluation by Toluidine blue, H and E staining and apoptotic cell death study by the TUNEL Apoptosis Detection Kit. In addition, glutathione level was measured in all specimens. No significant differences were revealed between Saline and AM groups in any of the aforementioned tests. Significant reduction of mast cells in lung tissue of the HD+AM group was shown when compared to the HD group. Lung tissue inflammation in the HD group was significantly more severe as compared to HD+AM group. In addition, amifostine in HD+AM group could prevent excess reduction of GSH level. The number of apoptotic cells in the HD group was significantly higher than the HD+AM group. Administration of amifostine before exposure to HD in rats prevents collection of mast cells, and excess reduction of GSH level in lung tissue; in addition it can partially reduce pulmonary edema and alveolar cell death apoptosis