Human gingival fibroblasts are necessary for oral homeostaslS These cells are fundamental in tissue healing and tissuc remodeling processes under a response to physiological actions such as mastication, Collagen and elastin, that are extracell ular glycoprotein of gingival fibroblast, are found in all animals, '1γpe 1 collagen is most dominant protein found in human gingival fibroblasts , Matrix metalloproteinase-1(MMP-1) has a role play in destruction of metabolism of extracellular matrix(ECM) and MMP-1 can destroy many ECMs as well as non-ECM molecules MMP-1’s local activation is conytolled by tissue inhibitor of metalloproteinase-1(TIMP-1) , Therefore, it is important to have a balance between in both s ituations MMPs and TIMPs of increased 0 1' decreased extracelluar matrix molecules, The purpose of trus study is to find out the effect of physical stimulus to human gingival fibroblast on mRNA, proteins of collagen 1, elastin, MMP-1, and TIMP-1 Healthy human gingival fibroblasts were separated and cultur때 in DMEM(Dulbeco’s Modified Eagle’s Medium) , When the sample reached to confluence state, it was separated with 0,25% t rypsin and 0‘ 53mM ethylendiaminetetraacetic aCld Separated cells were centrifuged in a cell culturing flask at 1000rpm for 30, 60, 120 mmutes Then it was forced by 35g/cm' continuously, The obtained results that expression of mRNA using histological study and Reverse Transcription Polymerase Chain Reaction (RT-PCR) , expression of protein using Enzyme-Linked Immunosorbent Assay(ELISA) for this study, At 30minutes after cen trifuging, there were s pindl e shaped gingival fibroblasts with long processes parallel to other cells in the control group , However the cell density was simil ar to compared group, At 60minutes after centrifuging, spindle shaped human gingival fibroblast with relatively long process, less densely packed, At 120minutes after centrifuging, cell processes were lengthened 2-3 times‘ and cell density was lower, At 30-60 minutes after centrifuging, it was increased by 1,3-1,7 times in expressoin of collagen 1 mRNA as compared with comparison group, However, there was no change in elastin, TIMP-l, and MMP-1, At 120 minutes after centrifuging, The revealed collagen 1 mRNA was increased 3 times as compared with comparison group, It was increased 2 times in elastin , 12 times in TIMP-1 as compared with comparison group, However, there was no change in MMP-l. At 30-60 minutes after centrifuging, it was increased by 1.1 times in revealing of protein revealing in collagen 1, TIMP-1 But there was no cha nge in elastin, MMP-1 At 120 minutes after centrifuging, it was increased by 1,2 times in revealing collagen 1 protein, 11 times in elastin, 12 times in TIMP-l, but there was no change in MMP-l. ln conclu s ion, it increased in revelation of collagen 1 ,elastin and TIMP-1 by continous stimulus in human gi ngival fibroblast, But there was no change in revelation of MMP-l Therefore, th is type of pressu re is one of the components for healing of gingiva l fibroblast

Laser is used to prevent the early dental caries in dental f ield and to apply for treatment of stomatitis and hyper sens it ivity , and laser mass Recently it is reported that laser i1'r adiation affect on soft tissue treatment and bone 1'emodelling after dental implantation. The purpose of this study was to examine laser irradia ti on effect on activity of normal human osteoblast on titanium plate in vitro by various laser wave length, and to observe morphologic change of NHost on LiLa nium plate and to a nalysis concentration of Ca"++ , IP and ALP NHost were cultured in DMEM containing 10% FBS, and observed by in verted microscope 1"or attatchment to the surface of titanium plate. Ca ++, I.P. , and a lkaline phosphat ase(ALP) concent ration in medium was calculated during 4 weeks, which was treated with Wilcoxon rank, Anova test and linear regression. The obtained results were as follows Morphologic changes showed rapid growth rate of NHost ++ at 3 days of laser lrradiatlOn ln spite of laser wave type, Ca" and P concentration was decreased at 2 weeks and was the hig hest at 3 weeks, but decreased at 4 weeks In spite of laser wave type, ALP concentration was decreased at 2 weeks but was increased at 3-4 weeks, From the aboving results, in spite of laser wave type, there were rapid growth rat e of NHost a nd no significant of Ca"++ , IP and P concen tr ation but these concentration showed predominant change than that of control

Many researchers are interested in wound healing in the t reatment of burns, prevention of post surgical adhesions and cosmetic s urgery by excess collagen production and scar formatlOn Synthetic epidermal substi tutes with cultured epi thelial cells seem to be an attractive strategy since keratinocytes have been demonstrated to modulate fibroblast growth and collagen synthesis. Bioa bsorbable and biocompatible chitosan structurally mimics hyaluronic acid. Recently, a bio compatible synthesi zecl ch itosa n-PVP(polyvinyl pyrrolidone) hydrogels demonstrated in vitro biocompat ibi li ty for bio medical applications . However. there is no re port on this hydrogeJ"s ability to modulate human gingival fibroblast growth. The purpose of this study were to investigate different growth modulation between human gingival fibroblast and normal human oral keratinocyte by chitosan- PVP hydrogel, and to apply this biocompatible synthetic polymer to oral and maxillofacial wound healing. We have synthesized a hydrogel from chitosan-PVP and examined its effect on human gingival fibroblast growth modulation in vitro. Non-toxic and biocompatible hydrogel with human gingival fi broblasts and epithelial cells was tested by MTT assay. HGF showed a higher growth proliferation than that of NHOK after cell seeding. In MTT assay, 30% hydrogel leach out products showed a higher cellular viability in NHOK than that of any other products. In MTT assay, 30% hyclrogel leach out products showed relatively lower cellular viability of HGF ln growth profile, NHOK showed about 7 fo lcls higher than HGF after 1 day, while about 2 fo lds higher after 5 days. And also NHOK showed above about 70% cell ular via bility from 1 to 7 days. It suggested that Chitosan-PVP hydrogel would inhibit relatively the growth of HGF and s timulate the growth of NHOK_ This phenomenon may prove to be of use in wound management 0 1' oral and maxillofacial area as epitheli al substitutes.

본 논문의 목적은 인간작업모델(The Model of Human Occupation: MOHO)을 이론적 틀로 하여 개발된 근로 자역할인식 면담도구(Worker Role Interview: WRI)를 소개하고, 향후 국내에서의 적극적인 사용을 제안하기 위한 것이다. WRI를 이용한 면담조사를 준비하기 위하여 2006년 9월부터 12월까지 WRI Version 9.0의 매뉴얼을 한국어로 1차 번역을 실시하였고, 감수와 수정 과정을 통하여 면담조사 권장질문지와 점수표, 판정기준을 마련하였다. 면담조사를 위하여 작업치료실이 있는 3곳의 산재전문병원을 방문하여 연구의 취지를 설명하였고, 그 중 한 곳에서 직업복귀 가능성이 있고 연구의 취지에 동의하는 산재 환자를 소개받아 2007년 2월 27일 면담조사를 실시하였다. WRI의 평가결과는 최소 17점에서 최대 68점으로 점수화 할 수 있는 데, 클라이언트의 평가 결과 총점 48점으로 전체적으로 직업복귀에 부정적인 측면 보다는 긍정적인 측면이 더 큰 것으로 나타났다. 특히 개인적 사유나 자신의 직업에 대한 가치, 흥미와 관련되어 있는 의지 영역에서 직업복귀를 강하게 지지하는 반응을 보였는데, 이는 클라이언트의 직업복귀에 대한 동기가 강하므로 직업복귀에 성공할 가능성이 높다는 전제 하에 재활서비스가 제공되어야 함을 의미한다. 인간작업모델이라는 새로운 작업치료의 패러다임을 임상에서 적용할 수 있는 간편한 면담평가도구로써 WRI는 심리적, 사회적, 환경적 변인을 통한 직업복귀의 가능성을 예측한다는 점에서 의미가 있으며, 작업치료사가 환자의 기능적 손상이나 장해에만 관심을 두는 것이 아니라 보다 폭넓은 관점에서 환자를 이해하고 접근하여 성공적인 재활프로그램을 계획하는데 도움이 되리라 기대된 다.

Cell behavior of the transformed cells is known to affect by interaction with extracellular matrix (ECM) proteins and integrin. To investigate the alterations of both integrin expression and cell-matrix interaction during neoplastic conversion of human oral kerationcytes, we studied expression levels of integrin subunits by flow cytometry and cellular responses to the ECM proteins in normal human oral keratinocytes (NHOKs), HPV-immortalized HOK-16B line, and three oral cancer cell lines established from HOK-16B line, CTHOK-16B-BaP, CTHOK-16B-DMBA, and CTHOK-16B-Dexa lines. The expression levels of α and β integrin subunits were shown decreased tendency in human oral keratinocytes undergoing immortalization and tumorigenic transformation except CTHOK-16B-DMBA line tested. Although αuβ6 integrin is known to be highly expressed in squamous cell carcinomas, and the altered integrin expression is suspected to be associated with cellular carcinogenesis, αu integrin subunit and αuβ6 egrin did not express in oral cancer cell lines tested. Cell behavior to the ECM proteins in HOK-16B line was generally similar to that of exponentially proliferating NHOKs. The adhesion activity profiles of type I collagen were very similar to that of its laminin counterparts, but fibronectin showed minimal adhesion activity under our conditions compared to the BSA control. The ability of the CTHOK-16B-BaP line to spread upon type I collagen and laminin markedly decreased, but migration was notably increased on type I collagen. In contrast, CTHOK-16B-DMBA and CTHOK-16B-Dexa lines spread less but migrated more upon type I collagen than immortalized HOK-16B line. These data indicate that downregulation of integrin subunits causes the changes of cellular responses to the ECM proteins during neoplastic conversion of human oral keratinocytes, and that cellular responses to the ECM proteins in oral cancer cell lines established by exposing different carcinogens are variable according to chemical carcinogens treatment.

본 연구는 사람 H-transferase가 과발현하는 돼지 체세포주를 개발하는데 있다. 돼지 세포에 사람 H-transferase 유전자를 발현시키는 것은 이종간 장기 이식에 있어서 초급성 거부 반응을 방지하기 위한 한 가지 방법이다. 본 연구에서는 과발현 벡터를 구축하기 위하여 사람 H-transferase을 HepG2 세포로부터 동정하였으며, 이 유전자를 CMV promoter를 이용하여 발현할 수 있도록 포유동물 발현 벡터인 pRc/CMV 벡터에 삽입하였다. 또한, 돼지 산자의 귀 세포를 이용하여 체세포를 수립한 후 jetPEI DNA transfection reagent를 이용하여 벡터를 도입하였고, 300 μg/ml의 G418로 12일간 선별하였다. PCR을 이용하여 선별된 colony들을 분석한 결과, 벡터가 도입되었음을 확인하였고, RT-PCR을 이용하여 사람 H-transferase mRNA가 발현하는 것을 확인하였다. 본 연구에서 확립된 세포주는 사람 H-transferase가 과발현하는 형질 전환 돼지의 생산에 이용될 수 있을 것으로 생각된다.

본 연구에서는 hG-CSF의 발현을 유도적으로 조절하기 위한 FIV-Tet-On lentivirus vector system을 구축하고자 하였다. hG-CSF는 호중성구 계열 세포의 증식과 분화, 생존에 영향을 미치는 물질로서, 이 유전자의 발현을 증가시키기 위하여 FIV-Tet-On vector 상의 hG-CSF나 rtTA2SM2 서열의 3' 위치에 WPRE 서열을 도입하였다. 구축된 각각의 vector는 293FT 세포에 일시적으로 transfection하여 virus를 생산하였으며, 이 virus를 일차 배양 세포인 CEF와 PFF에 감염시켰다. 각 세포에 전이된 hG-CSF의 발현 양상을 관찰하기 위하여 doxycycline을 첨가하거나 첨가하지 않은 배지에서 배양한 후 quantitative real-time PCR, Western blot과 ELISA를 이용하여 hG-CSF 유전자의 발현 정도를 비교 측정한 결과, CEF에서는 WPRE가 hG-CSF의 3' 위치에 도입된 경우에 발현량과 유도율이 가장 높은 것으로 나타났고, PFF에서는 rtTA 서열의 3'위치에 도입된 경우에 발현량과 유도율이 가장 큰 것으로 확인되었다. 이 FIV-Tet-On vector system은 형질 전환 동물의 생산이나 유전자 치료에서 문제시되는 외래 유전자의 지속적인 과다 발현에 의한 개체의 생리적인 부작용을 최소화하기 위한 해결 방법으로 제시될 수 있을 것이다.

본 연구에서는 SV40 Tag을 마우스 albumin 유전자의 promoter/enhancer 조절 하에 발현하도록 설계된 재조합 유전자를 마우스 1세포기 수정란에 미세 주입하여 형질 전환 마우스를 제작하고 이들의 인체 간암 모델로써의 적합성을 조사하였다. 형질 전환이 확인된 총 11개체의 founder 생쥐들 중 4개체가 간암을 일으켰고, 두 개체는 신장암을, 한 개체는 피부 및 뇌에서 종양을 각각 일으켰다. 이들로부터 외래 유전자를 계대 유전하는 3가계를 얻었다(#1-2, #1-6, #1-11). 이들 가계의 자손들에서 8주령(#1-2, #1-6) 혹은 10주령(#1-11) 시부터 간암이 반복적으로 발생되었으며, #1-11 founder 개체에서 폐로 암세포가 전이된 것 외에는 다른 조직에서의 형태학적 변이가 발견되지 않았다. 간암 발생은 조직학적 변화에 따라 3단계로 나눌 수 있었다. 즉, 출생에서 3주령까지는 간세포의 과량증식을 보이나 세포핵의 이상은 관찰되지 않았으며, 4주령부터 7주령(#1-2, #1-6) 혹은 9주령(#1-11)까지는 diffuse liver cell dysplasia를 나타내지만 tumor nodule은 발견되지 않았고, 그 이후에는 liver dysplasia를 배경으로 간암이 발생하였다. 본 연구에서 작출한 간암 모델 마우스는 인체 간암과 일부 유사한 소견을 보였는 바 인체 간암 기전 연구를 위한 유용한 동물 모델로 이용할 수 있을 것으로 생각된다.

The purpose of this study was to examine the effects of vi tamin D3 and 1'etinoic acid(RA) on the human mesenchymal stem ce!ls(MSC) g1'owth and osteogenic differentiations. Cell proliferation, mineralization, cell cycle, expression of cell cycle regu l atOJγ proteins and markers fo1' osteogenic differenatiaiton were determined by MTI assay, mineralization assay, flow cytomet1'Y‘ and Western blot analysis, respectively. Cell viability was dec1'ease by each vitamin D3 and RA added to MSC. it was more decrease by vitamin D3 and RA. Mineralized nodule formation revealed similar expression pattern with positive cont rol group at vitamin D3 and RA mixed add to MSC. At vitamin D3 and RA mixed add to MSC after 7 days of incubation was increase G1 s tage. after 21 days of incubation was inhibit cell cycle prog1'ess by inc1'ease of sub-G1 Treatment vitamin D3 to MSC inhibits p53 and p21, but inc1'ease pRb. RA inhibit p53, but increase p21 and pRb, vitamin D3 plus RA group was same as added RA group. so two vitamin was effect to inhibited cell growth each different mechanism. Expression of BMP-2 protein was prominent in osteogonic supplement treated g1'oup of MSC at 2 weeks cultivation days, but vi tamin D3 treatment decreased BMP-2 expression rather than in (+) control group. BSP protein was notably increased in the OS compa red to positive controls at 2 weeks cultivation, but similar to that of vitamin D3 group t1'eatment group and was least expressed in plus RA mixed group, at 3 weeks, BSP expression was similar to 1'esult of 2 weeks Collectively, these results shows that vitamin D3 and RA have diffe1'ential effects on the MSCs g1'owth and differ entia tion 211