Yuda Heru Fibrianto

Human umbilical mesenchymal stem cells conditioned medium promote primary wound healing regeneration.

Aim: This research was conducted to clarify the capability of human umbilical mesenchymal stem cells conditioned medium (HU-MSCM) to promote regenerations of primary wound healing on the incision skin injury. Materials and Methods: In this study, two approaches in vitro and in vivo already done. On in vitro analysis, tube formation was performed using HU vein endothelial cells in the presence of HU-MSCM, in some experiments cells line was incubated prior the presence of lipopolysaccharide and HU-MSCM then apoptosis assay was performed. Furthermore, in vivo experiments 12 female rats (Rattus norvegicus) were used after rats anesthetized, 7 mm wound was made by incision on the left side of the body. The wound was treated with HU-MSCM containing cream, povidone iodine was run as a control. Wound healing regenerations on the skin samples were visualized by hematoxylin-eosin staining. Results: In vitro models elucidate HU-MSCM may decreasing inflammation at the beginning of wound healing, promote cell migration and angiogenesis. In addition in vivo models show that the incision length on the skin is decreasing and more smaller, HE staining describe decreasing of inflammation phase, increasing of angiogenesis, accelerate fibroplasia, and maturation phase. Conclusions: Taken together our observation indicates that HU-MSCM could promote the acceleration of skin tissue regenerations in primary wound healing process.

The structural and functional recovery of pancreatic β-cells in type 1 diabetes mellitus induced mesenchymal stem cell-conditioned medium

Aim: Various studies have shown that secreted factors alone in culture medium without stem cell are capable of repairing tissues by itself in various conditions involving damaged tissue/organ. Therefore, this study was aimed to investigate the role of human umbilical cord mesenchymal stem cell-derived conditioned medium (CM) on the recovery of pancreatic β-cells in Wistar rats (Rattus norvegicus) with type 1 diabetes mellitus. Materials and Methods: The 0.05 ml CM induction was applied to the diabetic group of rats in weeks 1, 2, 3, and 4. 1 week after each CM induction, insulin concentration was analyzed using ELISA. The pancreas was divided into 3 regions, processed by paraffin method, stained with hematoxylin-eosin, and immunohistochemical method for insulin. Results: This study indicated the decrease in the total number of islets and insulin concentration after the injection of single dose of alloxan. The exocrine acini were also damaged. Microscopic observation detected the presence of small islets in the diabetic group 1 week after the first 0.05 ml CM induction. The number and size of the islets increased in line with the CM doses and time of inductions. Immunohistochemically, the presence of low intensity of insulin-positive cells could be recognized at the splenic and duodenal regions of the pancreas, but not gastric region, 1 week after the first and second 0.05 ml CM induction. The intensity of staining and the number of insulin-positive cells increased dramatically in 1 week after the third and fourth 0.05 ml of CM induction in all regions of the pancreas. The data of insulin blood concentration showed clear differences between the second and the fourth induction of 0.05 ml CM induction. Conclusions: This study showed very strong evidence on the role of human umbilical cord mesenchymal stem cell-derived CM in recovering the pancreatic β-cells damage in Wistar rats (R. norvegicus) with type 1 diabetes mellitus, structurally and functionally.

Effects of secretome on cisplatin-induced testicular dysfunction in rats

Background: Testicular dysfunction is a degenerative disorder characterized by failure in the synthesis of reproductive hormones and spermatogenesis. Secretome derived from the human umbilical mesenchymal stem cell (MSC) has been reported to repair some degenerative disorders. Aim: This study aimed to investigate the effect of secretome derived from the human umbilical MSCs on cisplatin-induced testicular dysfunction in rats. Materials and Methods: Thirty-six male Wistar rats were divided into the control and secretome-treated groups. In the secretome-treated group, testicular dysfunction was induced by 3 mg/kg BW of cisplatin intraperitoneally 3 times with 3-day intervals. The secretome-treated group was divided according to dose: Low-dose (0.2 mL/kg BW) and high-dose (0.5 mL/kg BW) groups. Secretomes were injected intraperitoneally once a week for 3 weeks. 1 week after the injection of secretome, the cauda epididymis of the rats was removed for spermatozoa evaluation and histological examination. Result: After the injection of secretome, the sperm motility of the high-dose group showed thin wave-like, rare, and slow movements. No abnormal sperm morphology was observed in all the treated groups. The number of spermatozoa increased gradually in the high-dose group after the injection of secretome. The developmental stages of the spermatogenic cells were complete in both spermatozoa groups after the injection of secretome. However, the spermatozoa in the seminiferous tubules of the high-dose group were denser. Vimentin and cytokeratin immunoreactivities were very strong in the high-dose group 1 week after the second secretome injection. Conclusion: High-dose secretome derived from the human fetal umbilical cord could increase the number and motility of sperms in rats with cisplatin-induced testicular dysfunction. The administration of high-dose secretome was effective 1 week after the second dose, as indicated by very strong immunoreactivity for vimentin and cytokeratin. Moreover, secretome could promote the regeneration of the seminiferous tubules of both the groups.

The Significant Effect of Conditioned Medium of Umbilical Cord Mesenchymal Stem Cells in Histological Improvement of Cartilage Defect in Wistar Rats

Materials and Methods: Twenty-four of 3-month old male rats that were divided into 6 groups (3 control groups and 3 treatment groups) with different evaluation time (month 2, month 3, and month 4). Treatment in each group was repeated 5 times. The cartilage defect was induced manually and mechanically at the medial condyle of rats’ right femur using a Kirschner Wire (D=1.0 mm; h=1.0 mm). The umbilical cord stem cell cultures were obtained from pregnant rats (19 days). Rats in treatment groups were injected with mesenchymal stem cell-conditioned medium (1 mL/kg BW) 5 times with interval a week after cartilage defect. Histopathological examination of chondrocytes formation and fibrosis tissues was done through Haematoxylin Eosin staining. Data were assessed with O’driscoll scoring and analyzed statistically using Kruskal-Wallis test with SPSS 16.0 software statistically using Kruskal-Wallis Test with SPSS 16.0 software.

The types of endocrine cells in the pancreas of Sunda porcupine (Hystrix javanica).

Aim: To identify the types of endocrine cells in the pancreas of the Sunda porcupine (Hystrix javanica) and its immunolocalization. Materials and Methods: Five adult H. javanica were used without sexual distinction. The presences of endocrine cells (glucagon, insulin, somatostatin, and pancreatic polypeptide [PP]) in pancreatic tissues were detected using the avidin-biotin-peroxidase complex method. Results: The fusiform, round, and oval form endocrine cells were detected in the islets of Langerhans and exocrine parts. Most of the insulin cells were found in the central area, glucagon cells were identified in the central and peripheral areas, and somatostatin and PP cells were detected in the mantle area of the islets of Langerhans. Glucagon and somatostatin cells were also detected in smaller numbers of peripheral parts of the islet. In all of the islet parts, glucagon endocrine cells were most prevalent cell type and then, somatostatin, insulin, and PP. In the exocrine parts, PP, somatostatin, glucagon, and insulin endocrine cells were found in the inter-acinus part with moderate, moderate, a few and rare numbers, in that order. In the pancreatic duct, glucagon and somatostatin cells were found between epithelial cells in rare numbers. Conclusion: The pancreas of Sunda porcupine (H. javanica) contains four types of major pancreatic endocrine cells with approximately similar distribution patterns to the other rodents, except for abundant glucagon cells in the peripheral area of the islets of Langerhans.

WITHDRAWN: Effects of various glycerol concentrations and thawing temperatures on CASA parameters and acrosomal integrity of frozen–thawed canine spermatozoa

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The Influence of the Captivity in Bottlenose Dolphin Erythrocytes Profile from The Waters of The Java Sea

In the Java Sea, there are a lot of dolphins, that often snagged fishing nets. While In Indonesia, bottlenose dolphins are mammals protected water. The aims of this research was to study the erythrocytes profile in bottlenose dolphins before and after the captivity. Seven dolphins were used in this research. Blood samples were taken out from the superficial vein of tail for examination of the number of erythrocytes, hemoglobin levels and value of hematokrit. Sampling was done when the dolphins appointed from the waters of the Java Sea and after experiencing dolphin captivity in PT. WSI, Kendal, Central Java. The data before and after captivity were compared and tested with the statistical analysis on the level of significance of the paired t-test 95%. The results showed the significant increase in the number of erythrocytes, hemoglobin levels and the values of hematokrit significantly. It was concluded that captivity done by WSI Corporation, Kendal, Central Java did increase the profile of erythrocytes in bottlenose dolphins. Keyword : Bottlenose dolphins, captivity, profile erythrocytes, superficial vein of tail, paired t-test