ABSTRACT
Objective: The purpose of this study was to evaluate in vitro cytotoxicity of gold nanorods [GNRs] on the viability of spermatogonial cells [SSCs] and mouse acute lymphoblastic leukemia cells [EL4s]
Materials and Methods: In this experimental study, SSCs were isolated from the neonate mice, following enzymatic digestion and differential plating. GNRs were synthesized, then modified by silica and finally conjugated with folic acid to form F-Si-GNRs. Different doses of F-Si-GNRs [25, 50, 75, 100, 125 and 140 microM] were used on SSCs and EL4s. MTT [3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide] proliferation assay was performed to examine the GNRs toxicity. Flow cytometry was used to confirm the identity of the EL4s and SSCs. Also, the identity and functionality of SSCs were determined by the expression of specific spermatogonial genes and transplantation into recipient testes. Apoptosis was determined by flow cytometry using an annexin V/propidium iodide [PI] kit
Results: Flow cytometry showed that SSCs and EL4s were positive for Plzf and H-2kb, respectively. The viability percentage of SSCs and EL4s that were treated with 25, 50, 75, 100, 125 and 140 microM of F-Si-GNRs was 65.33 +/- 3.51%, 60 +/- 3.6%, 51.33 +/- 3.51%, 49 +/- 3%, 30.66 +/- 2.08% and 16.33 +/- 2.51% for SSCs and 57.66 +/- 0.57%, 54.66 +/- 1.5%, 39.66 +/- 1.52%, 12.33 +/- 2.51%, 10 +/- 1% and 5.66 +/- 1.15% for EL4s respectively. The results of the MTT assay indicated that 100 microM is the optimal dose to reach the highest and lowest level of cell death in EL4s and in SSCs, respectively
Conclusion: Cell death increased with increasing concentrations of F-Si-GNRs. Following utilization of F-Si-GNRs, there was a significant difference in the extent of apoptosis between cancer cells and SSCs
ABSTRACT
Multiple sclerosis [MS] is an immune-mediated demyelinating disease of the central nervous system [CNS]. Stem cell transplantation is a new therapeutic approach for demyelinating diseases such as MS which may promote remyelination. In this study, we evaluate the remyelinating potential of adipose mesenchymal stem cells [ADSCs] and their effect on neural cell composition in the corpus callosum in an experimental model of MS. This experimental study used adult male C57BL/6 mice. Cultured ADSCs were confirmed to be CD73[+], CD90[+], CD31[-],CD45[-], and labeled by PKH26. Animals were fed with 0.2% w/w cuprizone added to ground breeder chow ad libitum for six weeks. At day 0 after cuprizone removal, mice were randomly divided into two groups: the ADSCs-transplanted group and the control vehicle group [received medium alone]. Some mice of the same age were fed with their normal diet to serve as healthy control group. Homing of ADSCs in demyelinated lesions was examined by fluorescent microscope. At ten days after transplantation, the mice were euthanized and their cells analyzed by luxol fast blue staining [LFB], transmission electron microscopy and flow cytometry. Results were analyzed by one-way analysis of variance [ANOVA]. According to fluorescent cell labeling, transplanted ADSCs appeared to survive and exhibited homing specificity. LFB staining and transmission electron microscope evaluation revealed enhanced remyelination in the transplanted group compared to the control vehicle group. Flow cytometry analysis showed an increase in Olig2 and O4 cells and a decrease in GFAP and Iba-1 cells in the transplanted group. Our results indicate that ADSCs may provide a feasible, practical way for remyelination in diseases such as MS
Subject(s)
Male , Animals, Laboratory , Mesenchymal Stem Cell Transplantation , Adipose Tissue , Cuprizone , MiceABSTRACT
Previous studies have demonstrated the potential of monotherapy with either mesenchymal stem cells [MSCs] or estrogen in autoimmune and cuprizone models of multiple sclerosis [MS]. The aim of this study was to examine the effects of co-administration of 17beta-estradiol [E2] and adipose-derived mesenchymal stem cells [ADSCs] on remyelination of corpus callosum axons in a cuprizone model of MS. Forty eight male C57BL/6 mice were fed cuprizone [0.2%] for 6 weeks. At day 0 after cuprizone removal, animals were randomly divided into four groups. The E2 monotherapy, ADSCs monotherapy, E2/ADSCs combined therapy and vehicle control. Some mice of the same age were fed with their normal diet to serve as healthy control group. E2 pellets, designed to release 5.0 mg E2 over 10 days, were implanted subcutaneously. 10[6] PKH26 labeled ADSCs were transplanted into lateral tail. The extent of demyelination, remyelination, and cell type's composition of host brain were examined at 10 days post-transplantation in the body of the corpus callosum. Transplanted cells migrated to the corpus callosum injury. Histological examination revealed efficacy of intravenous ADSCs transplantation in remyelination of mouse cuprizone model of MS can be significantly enhanced by E2 administration. Flow cytometry showed that the mean percentages of expression of Iba-1, Olig2 and O4 were significantly increased in E2/ADSCs combined therapy in comparison with ADSCs monotherapy. In conclusion, the findings of this study revealed that E2 administration enhanced efficacy of intravenous ADSCs transplantation in remyelination of corpus callosum axons in mouse cuprizone model of MS
ABSTRACT
Recent clinical studies of treating traumatic brain injury [TBI] with autologous adult stem cells led us to compare effect of intravenous injection of bone marrow mesenchymal stem cells [BMSC] and bone marrow hematopoietic stem cell mobilization, induced by granulocyte colony stimulating factor [G-CSF], in rats with a cortical compact device. Forty adult male Wistar rats were injured with controlled cortical impact device and divided randomly into four groups. The treatment groups were injected with 2 x 10[6] intravenous bone marrow stromal stem cell [n = 10] and also with subcutaneous G-CSF [n = 10] and sham-operation group [n = 10] received PBS and [bromodeoxyuridine [Brdu]] alone, i.p. All injections were performed 1 day after injury into the tail veins of rats. All cells were labeled with Brdu before injection into the tail veins of rats. Functional neurological evaluation of animals was performed before and after injury using modified neurological severity scores [mNSS]. Animals were sacrificed 42 days after TBI and brain sections were stained by Brdu immunohistochemistry. Statistically, significant improvement in functional outcome was observed in treatment groups compared with control group [P<0.01]. mNSS showed no significant difference between the BMSC and G-CSF-treated groups during the study period [end of the trial]. Histological analyses showed that Brdu-labeled [MSC] were present in the lesion boundary zone at 42[nd] day in all injected animals. In our study, we found that administration of a bone marrow-stimulating factor [G-CSF] and BMSC in a TBI model provides functional benefits