BackgroundEpididymal sperm maturation occurs via interactions between sperm and proteins secreted by the epididymal epithelium. Although this is an important process, the genes that encode the involved proteins remain largely uncharacterized. Previous studies have demonstrated that the genes involved in sperm maturation are regulated by androgen. Spag11a is an epididymal gene that is influenced by androgen. However, little is known about the putative role of this gene in the sperm maturation process. The objective of this study was to characterize Spag11a in the mouse epididymis.MethodsIn silico analyses were performed to predict signal peptides and functional domains. Spag11a expression was measured by quantitative real-time RT-PCR. Western blots and immunocytochemistry were performed to determine protein expression.ResultsSPAG11A is a member of the beta defensin protein family and constitutes a secretory protein. Spag11a was expressed exclusively in the epididymis. Moreover, it exhibited region-specific expression in the caput, which is typical for genes that are involved in creating a suitable microenvironment for sperm maturation. Mouse Spag11a was regulated by androgen. A significant decrease of Spag11a expression was observed at third day following a gonadectomy (P < 0.001). Interestingly, testosterone replacement therapy was able to maintain the expression almost at the normal level, indicating a dependency on androgen. Besides androgen, testicular factors influenced Spag11a expression in a different way. This was revealed by efferent duct ligation in which Spag11a was transiently up-regulated at the third day following the ligation before returning to the normal level at day 5. Spag11a regional expression was also observed at protein level detected by western immunoblotting which revealed a clear band in the caput but not in other regions. The prediction that SPAG11A is a secretory protein was confirmed by immunocytochemical analyses indicating cell-specific expression mainly in the caput principal cells and detection of the protein in epididymal luminal fluid and spermatozoa.ConclusionsBased on the characteristics of Spag11a, it is likely that this gene has a specific role in epididymal sperm maturation. Further studies using functional assays are necessary to confirm this finding.

Objectives: The objectives of this research was to measure the relative expression levels of vascular endothelial growth Factor-A (VEGF-A) in the deep dermal burns treated with human adipose-derived stem cell (hADSC) in collagen gel in each observational day (days 7, 14, 21, and 28).Methods: This study used 20 male Sprague Dawley rats, divided into four groups of observation days. Each rat received three burn injuries and then given different treatments (hADSC in collagen gel, collagen gel, and control). Deep dermal burn injury on the dorsal was made by placing a metal plate with 250°C for 15 s. Relative expression level of VEGF-A measurement with quantitative reverse transcription polymerase chain reaction.Results: On the 7th day, the relative expression level of VEGF-A in the wound treated with hADSC in collagen gel was significantly different from the scaffold collagen and control group (p<0.05), whereas the control and scaffold collagen group was not significantly different. The relative expression level of VEFG-A in wound treated with hADSC in collagen gel, collagen gel only, and control was (mean±standard error of the mean) 17.93±4.37, 7.54±2.63, and 5.44±1.59, respectively. On the next observation days, the result showed that the relative expression level of VEGF-A was not significantly different between the three treatments. The relative expression level of VEGF-A has decreases from day 7 to 28 days. The decrease of VEGF-A relative expression level hADSC in collagen gel group was significantly different on the 7th day to the 21st and 28th days (p<0.05).Conclusion: The provision of hADSC in scaffold collagen increases the relative expression of VEGF-A early in the wound healing process compared to the without a hADSC group.

Angiogenesis in the burn wound healing process is important. The application stem cell therapy in burn wound problem is expected to increase formation of new blood vessel so that the distribution of oxygen and nutrients working well. The purpose of this study was to known effect of human Adipose Derived Stem Cell (hADSC) in collagen gel on the average number of blood vessels based on time dependent. This study used 25 male rats Sprague Dawley who had received three internal wounds with different treatments. The treatments used hADSC in collagen gel, collagen gel only and without treatment. The rats were divided into 5 groups based on the duration of observation day 0, 7, 14, 21 and 28. Rat without burn wound were used as control. Observation blood vessels number using hematoxylin-eosin staining and calculated of average number blood vessels from five filed in five mice. Statistical analysis used ANOVA one way with significance value p<0.05. The average number blood vessels increase significantly until day 21 and decreased on day 28, both in untreated burn wound, collagen gel and hADSC in collagen gel. The treatment with hADSC in collagen gel was not significantly different from the collagen gel group or without treatment at day 7, 14 and 21. Even though the average of blood vessels of treatment with hADSC in collagen gel was not significantly different but our research had showed that treatment with hADSC in collagen gel was higher number of blood vessels than another group. Decrease of blood vessel number at 28 days indicated start of near homeostasis state and treatment hADSC in collagen gel has significantly decreased than another group. Our research showed that angiogenesis was time dependent and hADSC in collagen gel have not a role in increase of number blood vessels but we have indicated that hADSC might be have a role in qualitative improvement of angiogenesis.

Human adipose derived stem cells (hADSC) is expected to be an efficient solution for healing deep-dermal burns that help in wound closure. This study used 20 male Sprague dawley rats. Each rat received three wounds with different treatments (control, hADSC in collagen gel and collagen gel). Wound closure was observed at day 7th, 14th, 21th and 28th (sacrificed day) macroscopically, wound areas were measured and collagen densities were evaluated. Reduction of wound area showed significant difference between hADSC in collagen gel group and control group (p 0.001) and between hADSC in collagen gel group and collagen gel group. The hADSC in collagen gel group had a higher collagen density compared to control and collagen gel group. Application of hADSC in bovine collagen gel in rat deep-dermal burn models provided faster and better wound healing compared with control and bovine collagen gel group.

Pulsed Electromagnetic Fields (PEMF) is reported to encourage the healing of nonunion fractures. However, the mechanism by which this occurs is still not known. Wnt signaling pathways are believed to be important signaling pathways in bone formation. This study will evaluate the healing of delayed union femur fracture, given PEMF exposure. 48 Spraque Dawley rats were fracturized at the left femoral shaft. These rats were randomized into two groups: A control group (24 rats) and the PEMF group (24 rats). The PEMF group was given PEMF exposure of 1.6 mT, with a frequency of 50 Hz for 4 h every day for 5, 10, 18 and 28 days, while the control group was not given PEMF exposure. Consequently, on days 5, 10, 18 and 28 days after fracture, 6 rats from each group were sacrificed. Callus bone was used for histological and RT-PCR examination on the expression of Wnt10b, Wnt5a and β-catenin. Blood samples were taken to examine Alkaline Phosphatase (ALP) activity using the ELISA method. Hematoxylin Eosin (HE) staining results showed that in the initial phase of healing, fibrous tissue in the fracture gap of the PEMF group was less compared to the control group. In the PEMF group, ALP activity increased significantly on day 10. This is thought to be related to an increase in osteoblast activity in a bone matrix formation. Furthermore, RT-PCR examination results showed that Wnt10b, Wnt5a and β-catenin gene expression was higher in the PEMF group compared to the control group. It can be concluded that PEMF exposure is thought to accelerate delayed union fracture healing through the Wnt signal pathway.