사람태아 혀상피의 Cytokeratin 발현에 관한 면역조직화학적 연구

박성식,양연식,배근영,남광일 대한해부학회 1997 Anatomy & Cell Biology Vol.30 No.1

사람 식도-위 이행부 상피의 화생에 관한 면역조직화학적 연구

양대열,남광일,박성식 대한해부학회 1998 Anatomy & Cell Biology Vol.31 No.1

진주종 상피의 전자현미경적 소견

전병훈,백승찬,송계용 인제대학교 1995 仁濟醫學 Vol.16 No.4

다양한 임상상과 합병증을 야기하는 진주종성 중이염에서 관찰되는 진주종 상피는 조직학적으로 중이장내에는 없는 각질형 편평상피세포로 구성되어 있다. 진주종 상피의 광학현미경 관찰 결과에서 진주종 상피는 각질형 중층 편평상피층외에 비각질형 중층 편평상피층 및 그들간의 이행부위가 관찰되어 이 부위 각각의 미세구조를 전자현미경으로 관찰하는 것이 진주종 병인연구에 도움이 될 것으로 생각되어 본 연구를 시행하였다. 그 결과 각질형 편평상피에서는 각질유리질 과립과 당김세사와 교소체 등 피부상피와 같은 구조를 가지고 있었으며 비각질형 편평상피에서는 각질유리질 과립과 당김세사가 거의 관찰되지 않았으나 교소체는 뚜렸하였다. 반면에 이행부위라고 생각되는 상피에서는 당김세사와 각질유리 질 과립이 소수 관찰되었고 일부에서는 거대한 과립도 관찰되었다. 기저막은 염증이 심하고 증식성이 강한 곳에서는 형성이 불완전하였으나 각질형 상피로 갈수록 점차적인 기저막의 완전한 형성을 관찰하였다. 또한 기저층세포와 결체조직내의 염증세포 및 섬유모세포와의 직접적인 접촉은 발견되지 않고 피부의 상피와 미세구조상 별다른 유의한 차이가 없는 것으로 보아 성숙한 진주종상피는 표피와 구조적으로 동일한 것으로 보였다. 이와같은 소견에서 비각질형 진주종상피가 활발한 증식과 분화를 통해 성숙한 각질형 상피로 이행할 수 있는 것으로 생각되었다. The cholesteatoma was known to composed of be keratinizing striated squamous cells on the histology. On the previous study, we had found the cholesteatoma consisted of nonkeratinizing striated squamous epithelium, keratinizing epithelium, and transitioning zone on light microscope. We tried the electromictoscpoic exam to elucidate the pathogenesis of the cholesteatoma. As a result, the keratinizing epithelium had many keratohy-alinene granules, tonofilaments, obvious desmosomes like as the skin, the nonkeratininzing epithelium had few keratohyailne granulse and tonofilaments, and many obvious desmosomes. The transitional zone was composed of a few of keratohyaline granules and tonofilaments, large granule in the certain region. The incomplete basement membrane formation was found at the severe inflammatory and hyperproliferating zone, but completely organized basement membrane was toward the keratinizing epithelium. Fully matured keratinzing epithelium of cholesteatoma was similar to the epithelium of the skin, because the basement membrane was intact, and the basal cells did not contact the inflammatory cells and fibroblasts, and the epithelium of the cholesteatoma was not different from the epithelium of the skin, ultramicroscopically. Based upon the observations, we thought that the nonkeratinizing epithelium of the cholesteatoma was continually proliferating and differentiating toward the matured keratinizing epithelium.

Involvement of laminin and integrins in adhesion and migration of junctional epithelium cells

Kinumatsu, T.,Hashimoto, S.,Muramatsu, T.,Sasaki, H.,Jung, H-S,Yamada, S.,Shimono, M. Blackwell Publishing Ltd 2009 Journal of periodontal research Vol.44 No.1

<P>Background and Objective: </P><P>The junctional epithelium attaches to the enamel surface with hemidesmosomes (of which laminin-5 and integrin-&agr;<SUB>6</SUB>&bgr;<SUB>4</SUB> are the main components) in the internal basal lamina. Laminin-5 is also involved in cell motility with integrin-&agr;<SUB>3</SUB>&bgr;<SUB>1</SUB>, although their functions have not yet been clarified. The purpose of this study was to determine the functions of those adhesive components between the tooth and the junctional epithelium during cell migration. Because an idea has been proposed that directly attached to tooth cells (DAT cells) may not contribute to cell migration, 5-bromo-2-deoxyuridine staining was performed to confirm cell migration.</P><P>Material and Methods: </P><P>We investigated laminin-&ggr;<SUB>2</SUB> (contained only in laminin-5), integrin-&bgr;<SUB>4</SUB> (involved in cell–extracellular matrix contact) and integrin-&agr;<SUB>3</SUB> (inducing cell migration) in the junctional epithelium, oral gingival epithelium and gingival sulcus epithelium of 6-wk-old ICR mice using laser microdissection, quantitative real-time reverse transcription-polymerase chain reaction, immunofluorescence and 5-bromo-2-deoxyuridine staining.</P><P>Results: </P><P>Laminin and integrins were clearly immunolocalized in the basal lamina of all epithelium. Quantitative analysis of laminin and integrin mRNAs by laser microdissection showed that they were more highly expressed in DAT cells than in basal cells in the oral gingival epithelium. In particular, a 12-fold higher expression of laminin-5 was observed in the junctional epithelium compared with the oral gingival epithelium. 5-Bromo-2-deoxyuridine staining showed rapid coronal migration of DAT cells.</P><P>Conclusion: </P><P>These results suggest that the abundant expression of laminin-5 and integrin-&agr;<SUB>6</SUB>&bgr;<SUB>4</SUB> is involved in the attachment of DAT cells to teeth by hemidesmosomes. Abundant expression of laminin-5 and integrin-&agr;<SUB>3</SUB>&bgr;<SUB>1</SUB> might assist in DAT cell migration, confirmed by 5-bromo-2-deoxyuridine staining during the turnover of junctional epithelium.</P>

In Situ Hybridization and Immunohistochemical Study of Epidermal Growth Factor Receptor in the Human Inflamed Gingival Epithelium

Bae, Chang,Kim, Young Ho THE CATHOLIC UNIVERSITY OF KOREA 1997 Bulletin of The Catholic Research Institutes of Me Vol.25 No.-

The authors investigated the expression of EGFR in the normal and inflamed human gingival epithelium to Study the role of EGFR in the inflammation of the gingival epithelium by using in situ mRNA Hybridization and immunohistochemistry. The results showed: (1) The expression of EGFR mRNA in the normal gingival epithelium on in situ mRNA hybridization was mainly localized on the basal cell layer, and the spinous layer and the granular layer were weakly positive. The cornified layer wag negative. (2) The expression of EGFR mRNA in the inflamed gingival epithelium on in situ mRNA hybridization was evenly and homogeneously distributed with the high staining intensity in the whole layers of the gingival epithelium except the cornified layer. (3) The expression of EGFR protein in the normal gingival epithelium on immunohistochemistry wag mainly localized on the cornified layer and the granular layer, and the spinous layer was weakly positive, The basal cell layer was negative, (4) The expression of EGFR protein in the inflamed gingival epithelium on immunohistochemistry was evenly and homogeneously distributed with the high staining intensity in the whole layers of the gingival epithelium. In conclusion, the up-regulation of EGFR in the inflamed gingival epithelium can be regarded as responses to the possible damages to the oral environment to maintain the homeostatic conditions.

Reconstruction of Rabbit Corneal Epithelium on Lyophilized Amniotic Membrane Using the Tilting Dynamic Culture Method

Ahn, Jae-Il,Lee, Doo-Hoon,Ryu, Yang-Hwan,Jang, In-Keun,Yoon, Mun-Young,Shin, Youn Ho,Seo, Young-Kwon,Yoon, Hee-Hoon,Kim, Jae-Chan,Song, Kye-Yong,Yang, Eun-Kyung,Kim, Ki-Ho,Park, Jung-Keug Blackwell Publishing Inc 2007 Artificial Organs Vol.31 No.9

<P>Abstract: </P><P>Rabbit corneal epithelium was reconstructed using tilting dynamic culture with a self-manufactured, amniotic membrane (AM) supporter and a lyophilized amniotic membrane (LAM). Rabbit corneal epithelial (RCE) cells were cultured and cryopreserved after isolation from the limbus. The second- and third-passage RCE cells were plated onto the epithelial side of the LAM of Ahn's AM supporter. Two days later, the air–liquid interface culture was maintained with third-passage RCE cells for 6 days and second-passage corneal epithelial cells for 9 days. The average viability of thawed RCE cells, assessed using trypan blue dye exclusion, was 77.42%. The reconstructed corneal epithelium was characterized by histological (hematoxylin and eosin) and immunohistochemical staining (proliferating cell nuclear antigen) for light microscopy, and by reverse transcriptase-polymerase chain reaction, glucose assay, and transmission electron microscopy. The basal layer of the reconstructed corneal epithelium was well formed, and the epithelium was tightly constructed due to the increase in cell proliferation and differentiation caused by the tilting dynamic culture, as opposed to static culture. Tilting dynamic culture was useful for the reconstruction of the epithelium using easily damaged epithelial cells and resulted in more stratum cell layers. Moreover, cytokeratin (CK3) mRNA expression in tilting dynamic cultured third-passage RCE cells seeded onto AM was greater than in static cultured third-passage RCE cells. The morphology of the reconstructed corneal epithelium on LAM by tilting dynamic culture for 9 days resembled that of the skin epidermis. This was thought to be because the tilting dynamic culture not only accelerated the proliferation and differentiation of cells by physical or mechanical stimulation, but also ensured that the supply of medium was delivered to the basal cells more efficiently. Thus, the reconstruction of the corneal epithelium using LAM and tilting dynamic culture was considered to be a good in vitro model for autologous or allogeneic transplantation of corneal epithelium and skin epidermis in patients with damaged epithelia. </P>

후각 소실된 마우스에서 스테로이드가 후각상피의 재생에 미치는 영향

박희태,김용민,나기상 대한비과학회 2009 Journal of rhinology Vol.16 No.2

Background and Objectives:Various chemicals can affect the function of olfaction and steroids have been used for the treatment of olfactory dysfunction. In this study, we investigated the effect of chronic dexamethasone treatment on olfactory epithelium injured by 3-methylindole (3-MI). Subjects and Methods:0.75 mg/kg of dexamethasone and 0.15 mL of normal saline were administered to each of the 12 mice belonging to the experimental and control group respectively every other day from 1 week, before a single intraperitoneal administration of 175 mg/kg 3-MI, to 4 weeks after 3-MI injection. Three mice from each group were sacrificed every week, and olfactory epithelium was examined after H & E and immunohistochemical staining. Results:On H & E staining, the height of the olfactory epithelium and the polarity of the cells showed no difference between the two groups. On olfactory marker protein (OMP) staining, the number of OMP-immunoreactive (IR) olfactory receptor cells was significantly increased in the experimental groups from 2 and 4 weeks after 3-MI injection compared with the control group. On proliferating cell nuclear antigen (PCNA) staining, PCNA-IR basal cells were significantly reduced in groups that received dexamethasone from 2 weeks to 3 weeks after injection compared with the control group. Conclusion:Dexamethasone shows no protective effect in early necrosis of olfactory epithelium by 3-MI, but showed positive effect on the regeneration of the olfactory receptor cells of olfactory epithelium. Background and Objectives:Various chemicals can affect the function of olfaction and steroids have been used for the treatment of olfactory dysfunction. In this study, we investigated the effect of chronic dexamethasone treatment on olfactory epithelium injured by 3-methylindole (3-MI). Subjects and Methods:0.75 mg/kg of dexamethasone and 0.15 mL of normal saline were administered to each of the 12 mice belonging to the experimental and control group respectively every other day from 1 week, before a single intraperitoneal administration of 175 mg/kg 3-MI, to 4 weeks after 3-MI injection. Three mice from each group were sacrificed every week, and olfactory epithelium was examined after H & E and immunohistochemical staining. Results:On H & E staining, the height of the olfactory epithelium and the polarity of the cells showed no difference between the two groups. On olfactory marker protein (OMP) staining, the number of OMP-immunoreactive (IR) olfactory receptor cells was significantly increased in the experimental groups from 2 and 4 weeks after 3-MI injection compared with the control group. On proliferating cell nuclear antigen (PCNA) staining, PCNA-IR basal cells were significantly reduced in groups that received dexamethasone from 2 weeks to 3 weeks after injection compared with the control group. Conclusion:Dexamethasone shows no protective effect in early necrosis of olfactory epithelium by 3-MI, but showed positive effect on the regeneration of the olfactory receptor cells of olfactory epithelium.

Immunohistochemical Analysis of Interleukin-8 and Intercellular Adhesion Molecule-1 in Human Gingival Epithelium

Huang, George T.-J.,Zhang, Xinli Korean Academy of Oral Biology and the UCLA Dental 1999 International Journal of Oral Biology Vol.24 No.1

The neutrophil chemoattractant interleukin-8 (IL-8) and the intercellular adhesion molecule-1 (ICAM-1) may play important roles in recruiting and retaining neutrophils in gingival epithelium during infection and inflammation. To locate the in situ expression of these two molecules in gingival epithelium, immunohistochemical staining with specific antibodies was performed on human gingival samples. The results revealed a distinct distribution and level of immunoreactivities with either anti-IL-8 or anti-ICAM-1 antibodies in gingival epithelium. IL-8 expression was constitutive in gingival epithelium, including jumctional and sulcular epithelia, with higher levels in the basal and prickle cell layers. ICAM-1 was only observed in the sulcular and junctional epithelia, with stronger staining in the outer layers (facing the tooth surface). These data suggest that constitutive expression of IL-8 in gingival epithelium may perform some as yet unidentified function other than to attract neutrohpils, and that the concentration gradient formed by ICAM-1 with higher levels towards the outer layers, may facilitate the transmigration of leukocytes through junctional and sulcular epithelia into the gingival sulcus.

사람태아 이행상피의 Cytokeratin 발현에 관한 면역조직화학적 연구

남광일,김성근,박성식 대한해부학회 1996 Anatomy & Cell Biology Vol.29 No.3

염증성 치은 상피와 치낭의 표피성장인자 수용체의 발현 및 실험적 치아이동에 미치는 영향에 관한 연구

김영호,배창 대한치과교정학회 1997 대한치과교정학회지 Vol.27 No.2

동소 mRNA 보합결합법과 면역조직 화학적 염색법을 이용하여 정상 치은 상피와 염증성 치은 상피의 표피성장인자 수용체의 발현을 관찰하여 치은 상피의 염증에 있어서 표피성장인자 수용체의 역할을 연구하고, 타액에 노출되지 않는 치낭 조직에 있어서 표피성장인자 수용체의 발현을 관찰하여 다음과 같은 결과를 얻었다. 1. 동소 mRNA 보합결합법상 정상 치은 상피에서 EGFR mRNA는 거의 기저세포층에 국한되어 나타났으며, 극세포층은 약양성을 보였고 과립층과 각화층에서는 발현되지 않았다. 2. 면역조직 화학적 염색법상 정상 치은 상피에서 EGFR 단백은 거의 각화층과 과립층에국한되어 나타났으며, 극세포층은 약양성을 보였고 기저세포층에서는 발현되지 않았다. 3. 동소 mRNA 보합결합법상 염증성 치은 상피에서 EGFR mRNA는 각화층을 제외한 전층에 걸쳐서 균일하게 분포하였다. 4. 면역조직 화학적 염색법상 염증성 치은 상피에서 EGFR 단백은 기저세포층에서 각화층에 걸쳐서 균일하게 분포하였다. 5. 치낭 조직에서는 동소 mRNA 보합결합법과 면역조직 화학적 염색법 모두에서 Malassez 상피세포 잔존물에 강하게 염색이 되었고, 그 외의 주위 조직에서는 발현되지 않았다. 이상의 결과에서 볼 때, 염증성 치은 상피에서 EGFR의 과발현과 치낭 조직의 Malassez 상피세포 잔존물에서 다량의 EGFR이 존재하는 것으로 미루어, 이는 구강 환경에 가해질 수 있는 손상에 대하여 생체의 항상성을 유지하기 위한 반응으로 여겨진다. Epidermal growth factor(EGF), a single chain polypeptide of 53 amino acids with a molecular weight of 6,045 Da, was first isolated from the male mouse submandibular glands. EGF stimulates cellular proliferation and differentiation in several tissues and accelerates the rate of wound healing. EGF is bound to the specific receptor(EGFR) on the cell membrane of its target cell. EGFR is a transmembrane glycoprotein with a molecular weight of 170,000 Da and is detectable on a large variety of cell types. The authors investigated the expression of EGFR in the normal and inflamed human gingival epithelium to study the role of FGFR in the inflammation of the gingival epithelium, and the expression of EGFR in the dental follicle by using in situ mRNA hybridization and immunohistochemistry. The results were as follows; 1. The expression of EGFR mRNA in the normal gingival epithelium on in situ mRNA hybridization was mainly localized on the basal cell layer, and the spinous layer was weakly positive. The granular and cornified layers ere negative. 2. The expression of EGFR protein in the normal gingival epithelium on immunohistochemistry was localized on the cornified and granular layers, and the spinous layer was weakly positive. The basal cell layer was completely negative. 3. The expression of EGFR mRNA in the inflamed gingival epithelium on in situ mRNA hybridization was evenly and homogeneously distributed in the whole layers of the gingival epithelium except the cornified layer. The staining intensity appeared to increase progressively from the basal cell layer to the cornified layer. 4. The expression of EGFR protein in the inflamed gingival epithelium on immunohistochemistry was evenly and homogeneously distributed in the whole layers of the gingival epithelium. The staining intensity appeared to increase progressively from the cornified layer to the basal cell layer. 5. Strong positive reaction was seen in the epithelial cell rests of Malassez, whereas only background staining was seen in other cells of the dental follicle. In conclusion, the up-regulation of EGFR in the inflamed gingival epithelium and the high amounts of EGFR in the epithelial cell rests of Malassez in the dental follicle can be regarded as responses to the possible damages to the oral environment to maintain the homeostatic conditions.