Although both medical and surgical approaches have been investigated, recurrent symblepharon due to chronic ocular surface inflammation is a difficult disease to manage.This condition can also result in many complications such as cosmetic and visual deteriorations.In this condition, we can use the amniotic membrane transplantation that has a biological contact inhibition effect for the prevention of recurrent symblepharon. We treated successfully a case of a progressive symblepharon involving central cornea from 2-year old. The 14 year-old male patient had suffered from serious cosmetic problem and undergone multiple dissection, argon laser photocoagulation, using cyclosporin eyedrops. He was treated using allograft amniotic membrane, autograft limbal-conjunctiva, and buccal mucosa transplantation. we think that amniotic membrane transplantation was very effective method for the prevention of recurrent symblepharon.
Adolescent ; Allografts ; Amnion* ; Argon ; Autografts* ; Child, Preschool ; Contact Inhibition ; Cornea ; Cyclosporine ; Humans ; Inflammation ; Light Coagulation ; Male ; Mouth Mucosa ; Ophthalmic Solutions

PURPOSE: The cell cycle control system is necessary for the normal growth and differentiation of cells. The purposes of this study were to compare CD99 expression with a known intracellular marker of a specific cell cycle and to evaluate the potential of CD99 as surface marker for this cell cycle. METHODS: We induced arrest of the cell cycle in fetal lung fibroblast by contact inhibition or serum deprivation from culture media. We activated peripheral blood lymphocytes with the treatment of phytohemagglutinin (PHA) and interleukin-2 (IL-2). Next, we synchronized the cell cycle of peripheral blood lymphocytes to the late G1 phase with rapamycin. According to their CD99 expression, the peripheral blood lymphocytes were separated by magnetic bead and analyzed by Western blotting. RESULTS: CD99 expression in fetal lung fibroblast rapidly decreased in cell cycle arrest and recovered soon after G1 activation of the cell cycle. By analyzing chronologic changes of CD99 expression and PI-histogram, we found CD99 expression decreased after passing the G1 checkpoint. G1/S transition was interrupted by potent immunosuppresant, rapamycin. IL-2 receptor remained high after rapamycin treatment in the activated lymphocytes, whereas CD99 expression and propium iodide decreased as compared with the same condition without rapamycin. This suggested that CD99 expression was decreased in the late G1 phase. Retinoblastoma gene (Rb) and CDK-2 are necessary for G1/S transition. We found both of these in CD99+ lymphocyte through Western blotting only. Cyclin B, which has an important role in S/G2/M transition, was only found in CD99-activated lymphocytes. CONCLUSION: CD99 may be a G1 phase specific surface marker.
Blotting, Western ; Cell Cycle Checkpoints ; Cell Cycle* ; Contact Inhibition ; Culture Media ; Cyclin B ; Fibroblasts ; G1 Phase ; Genes, Retinoblastoma ; Interleukin-2 ; Lung ; Lymphocytes ; Receptors, Interleukin-2 ; Sirolimus

CDH-13(T-cadherin), which is one of a kind among the 20 cadherins, can be found mainly in wall of aorta, neuron, spleen, blood vessel etc. It is also called H-cadherin. This structural difference can explain that CDH-13 is thought to play a key role in maintaining mutual relation between extra and intra-cellular environment rather than in cell adhesion. The main function of CDH-13 is to participate in blood vessel function. Additionally, it is known to regulate cell growth and cell contact inhibition. When cells are proliferating, cell surface perceives other cells so that substance such as CDH-13 can inhibit their growth or proliferation resulting in homeostasis without endless proliferation or invasion of connective tissue boundaries. However, tumor cell itself appears to be different from normal cells' growth, invasion or transmission. Therefore, it can be diagnosed that these characteristics are closely related to expression of CDH-13 in tumor cells. This study is to investigate expression of CDH-13 in SCC and its correlation with promoter methylation. 20 of tissue species for the study are excised and gathered from 20 patients who are diagnosed as SCC in department of OMS, dental hospital, dankook university. To find development of CDH-13 in each tissue samples, immunohistochemical staining, RT-PCR gene analysis and methylation specific PCR are processed. The results are as follows. 1.Immunohistochemical staining: In normal oral squamous epithelial tissue, strong expression of CDH-13 was found in cell plasma membrane of basal cell layer. On the other hand, in case of low-differentiated oral SCC, development of CDH-13 was hardly seen. 2.The development of CDH-13 gene: In 9 of samples, expression of CDH-13 gene could be seen and 2 of them showed low expression compared to the others. And rest of the 11 samples showed no expression of CDH-13 gene. 3.Methylation of CDH-13 gene: Among 9 samples which expressed CDH-13 gene, 7 of them showed unmethylation. In addition, among 11 samples without CDH-13 gene expression, 10 showed methylation. According to the results stated above, promoter methylation were found in 13 samples(65%) among 20 of oral SCC samples. In low-differentiated SCC, suppression of gene expression could be seen accompanying promoter methylation. These phenomenon of gene expression was proved by immunohistochemical investigation. Finally, for development of oral SCC, conclusions can be made that suppression of CDH-13 played a main role and suppression of gene expression was originated from promoter methylation. Considering this, it is expected that suppression of CDH-13 from promoter methylation to be utilized as a good diagnostic marker of oral SCC.
Aorta ; Blood Vessels ; Cadherins ; Carcinoma, Squamous Cell ; Cell Adhesion ; Cell Membrane ; Connective Tissue ; Contact Inhibition ; Gene Expression ; Glycosaminoglycans ; Hand ; Homeostasis ; Humans ; Methylation ; Neurons ; Polymerase Chain Reaction ; Spleen

Human epithelial cell line immortalized with Ad12-SV40 hybrid virus was transfected with plasmid containing HPV-16 gene. Among these clones, clone-3 and clone-6 showed neoplastic transformation properties of contact inhibition, anchorage independence and cellular adhesion after 7 subcultures. The results suggest that SV40 gene in the immortalized human cell system be in concert with HPV-16 in the process of neoplastic transformation of human cells. While TGF-Beta1(5ng/ml) inhibited growth of contml cells and clone-1 cells which did not show transformation, there was no significant change on the growth of clone-3 cells with transformation properties. When transcriptional level of fibronectin on control cells and clone-3 cells were analyzed with northern blot technique, transcription of fibronectin an clone-3 cells were higher, as compared with control cells. RNA hybridization techniques were performed to compare trasnscriptional levels of TGF-Beta1 between control cells and clone-3 cells. RNA level on clone-3 cells with transformation properties was higher than on control cells. These studies indicate that TGF-Beta1 is associated with increases of fibronectin, which may lend to changes of TGF-Beta receptor and loss of its inhibitory action on the transformed cells. Thus, it seems that loss of inhibitory action of TGF-Beta which is mediated by changes of fibronectin may account for a possible mechanism of action in the HPV-16 induced transformation of human cells.
Blotting, Northern ; Clone Cells ; Contact Inhibition ; Epithelial Cells ; Fibronectins ; Human papillomavirus 16 ; Humans* ; Plasmids ; Receptors, Transforming Growth Factor beta ; RNA ; Transforming Growth Factor beta* ; Transforming Growth Factor beta1

Primary and secondary corneal endothelial decompensation leads to stromal edema, corneal opacity and loss of visual acuity. The pathogenesis of corneal endothelial decompensation is that adult corneal endothelium in vivo lacks of a robust proliferative response to injury, does not divide sufficiently to replace the lost cells. Previous studies indicate that cell-cell contact inhibition and transforming growth factor-beta2 (TGF-β2) in aqueous humor may be responsible for maintaining human endothelial cells in a non-replicative state in vivo. The results of the experimental investigation by using immunofluorescent staining of the cell cycle-associated proteins and cell proliferation marker Ki67 in corneal endothelium indicate that human corneal endothelial cells in vivo are arrested in the G1-phase and have not exited from the cell cycle. Successful outgrowth in culture of human corneal endothelial cells in vitro and the establishment of the immortalized human endothelial cell line, provide strong evidence that corneal endothelial cells retain proliferative capacity. Experiments with cell culture ex vivo demonstrate that corneal endothelial cells cultured from young donors grow more robustly than those from older donors, and cells cultured from peripheral area of corneas show greater cell density than central regions. Studies have demonstrated that in vitro human corneal endothelia undergo mitotic changes in response to stimulation of growth promoting agents, such as growth factors, EDTA and extracellular matrix. Identification of corneal endothelial stem cells and isolation and culture of human endothelial precursor cells in vitro will be beneficial for further investigation regarding the mechanism of corneal endothelial regeneration as well as corneal endothelial cells in vitro culture.
Aqueous Humor ; metabolism ; Cell Count ; Cell Culture Techniques ; Cell Cycle ; Cell Proliferation ; Cells, Cultured ; Contact Inhibition ; Endothelium, Corneal ; cytology ; Humans ; Stem Cells ; Transforming Growth Factor beta2 ; metabolism

Since NNK is one of the most abundant tobacco-specific alkaloids and a strong carcinogenic nitrosamine, it has been used for evaluating a potential of carcinogenicity in the animal models. The present study has attempted to examine the potential of carcinogenicity of NNK in human epithelial cells, from which the cell type the most of cancers including oral cancer and nasal cavity cancer are originated. The cellular model used for the study is a human keratinocyte cell system immortalized by Ad12-SV40 hybrid virus. The cellular system has successfully been used for the carcinogenicity studies because of its limitless life span, epithelial morphology and non-tumorigenicity. When cells were treated with a variety of NNK concentrations, levels of saturation density and soft agar colony formation were increased in a dose-dependent fashion. Colonies of large cell aggregates were above 5 at the higher doses. The results indicate that exposure of human cells with NNK induced loss of contact inhibition and increases of anchorage independence and cellular adhesion, which are typical characteristics of the neoplatically transformed cells. When cells were exposed with 100uM NNK for 2hr, mRNA levels of IL-1 and PAI-2 were increased in a dose-dependent manner, but expression of TGF- 1 was not affected. While expression of growth regulatory factors were altered with a short-term exposure, there was no alteration of these factors in the NNK-transformed cells. However, mRNA levels of fibronectin were increased both in the short-term treatment and in the transformation. The results suggest that altered expression of extracellular matrix such as fibronectin following short-term exposure might be fixed in the genome and these altered properties be continuously transfered throughout the cell division. Western blot analysis showed a translocation of PKC- from cytosolic fraction to the particulate fraction, indicating a possible role of NNK in the signal transduction pathway. The present study provided an evidence that NNK in the smoking may be associated with epithelial origin cancer such as oral and nasal cavity cancers. In addition, this study suggested that altered expression of extracellular matrix and PKC may play an important role in the carcinogenic mechanism of NNK.
Agar ; Alkaloids ; Blotting, Western ; Cell Division ; Contact Inhibition ; Cytosol ; Epithelial Cells* ; Extracellular Matrix ; Fibronectins ; Genome ; Humans* ; Interleukin-1 ; Keratinocytes ; Models, Animal ; Mouth Neoplasms ; Nasal Cavity ; Plasminogen Activator Inhibitor 2 ; RNA, Messenger ; Signal Transduction ; Smoke ; Smoking