ABSTRACT
To investigate of the effects of a poly L-lactic acid [PLLA] nanofiber scaffold on proliferation of frozen-thawed neonate mouse spermatogonial stem cells. Spermatogonial cells were isolated from neonatal 3-6-day-old NMRI mice testes by two steps enzymatic digestion and differential plating. The isolated spermatogonial cells were divided into four culture groups: 1] fresh spermatogonial cells, 2] fresh spermatogonial cells seeded onto PLLA 3] frozen-thawed spermatogonial cells, 4] frozen-thawed spermatogonial cells seeded onto PLLA. Cells in all groups were cultured in DMEM supplemented with 5% FCS and 10 ng/ml GDNF for 3 weeks. Diameter and number of clusters which were determined during the culture and semi-quantitative RT-PCR were carried out at the end of 3rd week for all culture groups. Presence of spermatogonia at the culture was determined by reverse transcription polymerase chain reaction [RT-PCR] for several important spermatogonial markers [PLZF, Oct4, GFRalpha-1, VASA, ITGA6 and ITGB1]. The significancy of the data was analyzed using Repeated Measures and ANOVA tests. The findings indicated that the viability rate of the fresh cell [control 1 and exprimental 1] and the frozen cells after thawing [control 2 and exprimental 2] were 89.25 +/- 2.2 and 63 +/- 3.56, respectively and the differences were significant [p<0.001]. In vitro culturing of spermatogonial cells on PLLA significantly increased the formation of cell clusters in comparison with those of the control groups [p
ABSTRACT
Background and Purpose: Eyes are among the most sensitive organs to chemical agents especially Sulfur mustard or Hun Distilled. Therapeutic effects of anti-inflammatory drugs have been shown on the decrease of epithelium of corneal injuries after being exposed to sulfur mustard. The aim of the present study was to evaluate the protective effects of topically applied Bethametazone - Diclofenac Na in rabbits
Materials and Methods: In this experimental research, thirty six rabbits were used. Animals were randomly divided into six equal groups [6 rabbits in each group] including control, solution and mustard groups and prophylaxis groups included Betamethasone, Diclofenac Na; Betamethasone-Diclofenac Na were applied before being exposed to the sulfur mustard solution. Animals were kept for 2 weeks and the drugs were used 3 times a day for 2 weeks. Slit-lamp examinations were performed under anesthesia before exposure and subsequently at days 1, 2, 5, 7, and 14 after sulfur mustard exposure by ophthalmologists. At the end of the 14th day, specimens of cornea were obtained for ultrastructural evaluation of corneal epithelium. Statistical analysis was performed by one way analysis of variance [ANOVA] with Tukey's test using SPSS 13 software
Results: No significant differences were found between the control and solvent groups as far as the variables were concerned. Corneal epithelial defect and severe changes in ultrastructure of corneal epithelial surface was found in the sulfur mustard group. Clinically, corneal epithelial defect in prophylactic Diclofenac Na group [4.2 +/- 1.32] decreased significantly when compared to the mustard group [68.7 +/- 8.42] [P=0.034]. Betamethasone-Diclofenac Na group [4.2 +/- 1.17] also decreased significantly when compared to the mustard group [68.7 +/- 8.42] [P=0.031]. The corneal epithelial defect was not seen in prophylactic Betamethasone group. Ultrastructural damage of corneal epithelial surface and their microvillus in Betamethasone group was similar to the control group
Conclusion: Betamethazone is capable of protecting corneal epithelial defect in the eyes of rabbits exposed to sulfur mustard