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        배양된 사람치은각화상피세포의 미세구조

        권용대,이백수,주성숙,Kwon, Yong-Dae,Lee, Baek-Soo,Jue, Sung-Sook 대한악안면성형재건외과학회 1999 Maxillofacial Plastic Reconstructive Surgery Vol.21 No.3

        In oral and maxillofacial surgery, there are many cases requiring the graft of epidermal tissues such as maxillectomy, and vestibuloplasty. There have been so many challenges for the culture of the epidermal tissue. Observing the ultrastructure of the cultured human oral kertinocytes, we could compare this findings with that of in vivo ones. With that, we could find the differencies and similarities between cultured cells and in vivo ones, and evaluate the clinical applications of cultured tissue. Human gingiva was obtained and the specimen was explanted on 24-well plate. Two types of culture media were used in this culture system. One was for the growth of the keratinocytes (Media I), and the other was for the stratification (Media II). Media I had special ingredients for the epidermal growth. Those were 0.5% dimethyl sulfoxide (DMSO), 30ng/ml of epidermal growth factor (EGF), 30ng/ml of cholera toxin, and $5{\mu}g/ml$ of transferrin. We cultured the oral keratinocytes for 3 weeks, and at that time the cultured keratinocytes were processed to prepare the specimen for the TEM study. The results were as follows.; 1. In the phase contrast micrograph, epidermal outgrowth firstly appeared on the 3rd day after explantation, and the growing keratinocytes were activley mitotic, and had polygonal shape and increased N/C ratio. 2. In the phase contrast micrograph, the outer most cells exhibited areas where broad cytoplasmic processes extended out onto the culture subtratum(fan-like appaearance). 3. In the TEM micrographs, the cultured keratinocytes showed stratification. The cells were in elongated form, and there were no morphologic differencies among the layers usually found in the in vivo gingiva. 4. Most of cellular organelles underwent lysis, and keratohyaline granules were seen. Tonofibrils were dispersed in the cytoplasm. 5. The cells were interconnected by desmosomes, and their frequency of distribution was considered to be lower than that of in vivo keratinocytes. 6. We could conclude the cultured oral keratinocytes exhibited signs of terminal differentiation.

      • KCI등재

        인간 치주인대 유래 섬유모세포에 대한 흡수성 교원질 차폐막의 생체적합성

        권용대(Yong-Dae Kwon),이백수(Baeksoo Lee),주성숙(Sung-Sook Jue) 대한구강악안면외과학회 2006 대한구강악안면외과학회지 Vol.32 No.3

        Purpose: This study is designed to evaluate biocompatibility of three types of absorbable collagen GBR membrane in vitro. Material and Method: The human PDL fibroblasts culture was obtained through typical way and the cells used in the experiment was forth passage. The membranes examined were Experimental group A, B, C. All the 3-experimental groups were made of bovine pericardium and the membranes were excised into 5×5mm respectively. The samples of the membranes were fixed on the 24-well plate with the double-sided adhesive tape. Then, 2ml of cell suspension which included 2×104cells was inoculated into the 24-well plate, and the cells were cultured for 1 week. Cellular viability and the alkaline phosphatase activity were measured with ELISA. The membranes in the culture were processed to examine with SEM. Results: The survival rate was highest in control and Experimental group A is the next, group B and group C in order of the value. The values are analyzed for statistical difference using Wilcoxon test. All the values of experimental groups are significantly lower than those of control, and the vaules among the experimental groups significantly differ from each other. Alkaline phosphatase level was identical order with the viable cell rate. SEM examination revealed that the PDL fibroblasts adherent on culture dish (control) and group A were spindle-shaped, but on group B and C, the cells were round-shaped without processes.

      • KCI등재

        고농도 포도당 환경에서 EMD(Enamel Matrix Derivatives)가 인간 치주인대 세포와 뼈모세포양 세포(MC3T3-E1)에 미치는 영향

        이백수(Baek-soo Lee),김선욱(Sun-Wook Kim),주성숙(Sung-Sook Jue),권용대(Yong-Dae Kwon) 대한구강악안면외과학회 2008 대한구강악안면외과학회지 Vol.34 No.5

        Purpose: This study was designed to evaluate effect of EMD on proliferation of HPDLCs and MC3T3-E1 cells in high glucose condition in vitro. Material and method: The Human PDL fibroblasts(HPDLCs) were obtained through typical way and the cells used in this experiment were divided in 4 groups. 1x104/ml HPDLCs suspension was cultured in typical DMEM and assigned to group 1. The cells cultured in DMEM which included 400mg/dl glucose are allocated to group 3. Group 2 and 4 are established by adding EMD to group 1 and 3 respectively. These control and experimental groups had been cultured for 24 and 48 hours, and MTT assay was conducted. The differences of each group in cellular proliferation was evaluated. The same experiment was conducted for preosteoblast (MC3T3-E1) with adding 25㎍/ml EMD. Results: EMD had the same effect on both PDL cells and MCT3T3-E1 cells. The experimental group had more meaningful differences and active cellular proliferation than the control group did. The EMD accelerated cellular proliferation not only in normal glucose condition but also in high glucose condition. The same results were observed via MTT assay; EMD-added experimental group had more meaningful differences and showed higher cellular activity than control group did. Each experimental and control group was inspected for statistical significance through Kruskal-Wallis Test. Statistical significances were observed among these groups. (SPSS 12.0 Chicago, IL, USA, p=0.008, p=0.011) Conclusion: EMD is considered to accelerate proliferation of PDL cells and MC3T3-E1 cells in high glucose condition as well as normal glucose condition.

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