구강암 유전자 치료를 위한 재조합 HSCC-1 아데노바이러스의 개발

김창현,김진우,김명진,표성운,Kim, Chang-Hyen,Kim, Jin-Woo,Kim, Myung-Jin,Pyo, Sung-Woon 대한악안면성형재건외과학회 2005 Maxillofacial Plastic Reconstructive Surgery Vol.27 No.2

In spite of the ongoing advances, standard therapies for oral cancer still has some limitations in efficacy and in ability to prolong survival rate of advanced disease and result in significant functional defect and severe cosmetic deformity. Currently gene therapy using tumor suppressor gene is considered as a potent candidate for new therapeutic approaches that can improve efficacy and reduce complications. The purpose of this research is to identify the role of adenoviral vector to transfer HCCS-1 tumor suppressor gene in oral cancer cells and to find out whether there is a possibility for it to serve in the field of gene therapy. The human SCC-25 cell line was used for transfection. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transduced with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the tranfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1). DNA extracted from Ad5CMV-HCCS-1 revealed HCCS-1 gene is incorporated. The transduction efficiencies were over than 50% of SCC-25 cells with a MOI of 2 and over 95% with a MOI of 50. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transduced SCC-25 cells. There was no or very low transcription HCCS-1 mRNA in wild and Ad5CMV-LacZ transduced SCC-25 cells. Cells transduced with Ad5CMV-HCCS-1 showed significant growth inhibition. By day 6, Ad5CMV-HCCS-1 treated cell count was decreased to 30% of mock-infected cells, while that of Ad5CMV-LacZ treated cells was 90% of mock-infected cells (p<0.05). Finally, these result suggest that the Ad5CMV-HCCS-1 has potential as a gene therapy tool for oral cancer.

Cone Beam Computed Tomography의 두부계측분석을 통한 골격성 제3급 부정교합 환자에서 악교정 수술 후 입술과 주위 연조직의 수직적 변화

이종민,강주완,이종호,김창현,박재억,Lee, Jong-Min,Kang, Joo-Wan,Lee, Jong-Ho,Kim, Chang-Hyen,Park, Je Uk 대한악안면성형재건외과학회 2012 Maxillofacial Plastic Reconstructive Surgery Vol.34 No.5

Purpose: The aim of this study is to evaluate the vertical changes of the lip and perioral soft tissue, following orthognathic surgery in skeletal class III patients by a cephalometric analysis of a cone beam computed tomography (CBCT). Methods: A total of 20 skeletal class III patients, who had bimaxillary surgery with Le Fort 1 osteotomy and bilateral sagittal split ramus osteotomy, were included in this study. The surgical plan for maxilla was posterosuperior impaction with the anterior nasal spine, as the rotation center. Further, the surgical plan for mandible was also posterosuperior movement. The soft tissue changes between lateral cephalogram and CBCT were compared. And the correlations between independent variables and dependent variables were evaluated. Results: There were no significant differences of the soft tissues changes between lateral cephalogram and CBCT. Upper lip philtrum length (SnLs), nasolabial angle increased and upper lip vermilion length (LsStms), lower lip length (StmiB'), lower lip vermilion length (StmiLi), lower lip philtrum length (LiB') and soft tissue lower facial height (SnMe') decreased after surgery. Change of SnLs (${\Delta}$SnLs) was influenced by vertical change of menton (${\Delta}$MeV), and change of LsStms (${\Delta}$LsStms) was influenced by upper lip thickness (ULT). Change of StmiLi' (${\Delta}$StmiLi') were influenced by preoperative overjet. Change of StmiB' (${\Delta}$StmiB') were influenced by preoperative overjet, vertical change of lower incisor (${\Delta}$L1V) and horizontal change of posterior nasal spine (${\Delta}$PNSH). Change of LiB' (${\Delta}$LiB') was influenced by ${\Delta}$L1V and ${\Delta}$PNSH. Change of SnMe' (${\Delta}$SnMe') was influenced by ${\Delta}$MeV, horizontal change of upper incisor (${\Delta}$U1H) and horizontal change of lower incisor (${\Delta}$L1H). ${\Delta}$Nasolabial angle was influenced by change of ULT (${\Delta}$ULT). Conclusion: Both soft tissues and hard tissues can be evaluated by CBCT. Posterosuperior rotation of maxillomandibular complex resulted in increase of upper lip philtrum length and nasolabial angle, while the upper lip vermilion length, lower lip philtrum length, lower lip vermilion length, and soft tissue lower facial height showed a decrease.

재조합 HCCS-1 아데노바이러스를 이용한 구강암 세포주의 세포사멸 유발

김창현(Chang-Hyen Kim),이동주(Dong-Ju Lee),이일규(ll-Kyu Lee),김명진(Myung-Jin Kim),김진우(Jin-Woo Kim),표성운(Sung-Woon Pyo) 대한구강악안면외과학회 2005 대한구강악안면외과학회지 Vol.31 No.4

Despite advances in surgery, radiotherapy, and chemotherapy, the survival of patients with oral squamous cell carcinoma has not significantly improved over the past several decades. Gene therapy is currently under investigation and shows us new possibility of cancer curing method. This experiment was undergone to find out the cell growth inhibition effect and evidence of apoptosis by HCCS-1(human cervical cancer suppressor-1), one of the candidates of tumor suppressor gene, transducted to human oral cancer cell line. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transducted with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the transfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay using cell count kit(CCK). To show the evidence of apoptosis, DNA fragmentation assay and flow cytometry(FACS) were performed. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1), and importation efficiency was 20% at 2 MOI(multiplicity of infection), 80% at 20 MOI. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transducted cell lines. As a result of CCK, when comparing to control subjects, transducted group showed 50% growth inhibition. In DNA fragmentation assay, according to increasing of MOI, DNA volume was diminished. In FACS analysis, DNA distribution showed fragmentation. This results imply that HCCS-1gene has growth inhibition effect in human oral cancer cell lines through apoptosis induction.

인간협지방대유래 줄기세포의 골분화에 대한 심바스타틴의 효과

김창현 ( Chang Hyen Kim ),최기수 ( Ki Soo Choi ),이일규 ( Il Kyu Lee ),박재억 ( Je Uk Park ),표성운 ( Sung Woon Pyo ) 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.1

Simvastatin, a cholesterol-lowering drug, has been widely used for hyperlipidemic patients. Recently, it has been reported that they promote bone formation. Also, Human buccal fat pad-derived stem cells(hBFDSCs) have been shown to differentiate into osteoblast. Here, we investigated the effect of simvastatin on osteogenic differentiation in primary cultured hBFDSCs. hBFDSCs were isolated from the buccal fat pad and cultured. After cell expansion and three passages, the cell sorting for STRO-1, CD105 and CD34 were performed to characterize the HBFSCs. Osteogenic differentiation was induced on a monolayer culture with osteogenic media(OM), media including 100 ng/mL of BMP-2 and media including 1 ?M of simvastatin, respectively. After 7 to 28 days, alkaline phosphatase(ALP), Alizarin Red S, von Kossa staining and RT-PCR analysis for osteocalcin(OC) expression were performed for confirmation of osteogenesis. Flow cytometric analysis and biochemical assays demonstrated that hBFDSCs might be a distinguished stem cells, and mineralization was accompanied in all groups. However, with simvastatin, the hBFDSCs presented most efficient staining pattern of ALP and Alizarin red and were not inferior for the OC expression to other experimental groups. These results suggest that simvastatin has the potential stimulatory effect on bone formation through osteogenic differentiation and may be effective for the bone tissue regeneration.

일반총설 : 치과에서 사용되는 이식재의 최신 연구 동향

김창현 ( Chang Hyen Kim ),박민규 ( Min Kyu Park ),이일규 ( Il Kyu Lee ),박재억 ( Je Uk Park ) 한국조직공학과 재생의학회 2008 조직공학과 재생의학 Vol.5 No.1

BMP-2에 의한 협부 지방 성체 줄기세포의 골형성

김창현(Chang-Hyen Kim),박철헌(Cheol-Hun Park),이일규(Il-Kyu Lee),표성운(Sung-Woon Pyo) 대한구강악안면외과학회 2008 대한구강악안면외과학회지 Vol.34 No.4

Bone morphogenetic proteins (BMPs) in combination with stem cells gain more significance for their use in bone tissue engineering. The mesenchymal stem cell can be differentiated into osteoblast by the treatment of BMP. The aim of this study is to characterize the osteogenic differentiation process of adult stem cells derived from buccal fat pad according to BMP-2 within culture media and decide the appropriate concentration of BMP-2 to facilitate osteogenesis. The authors procured the stem cell from buccal fat pad and analyzed for presence of stem cell by flow cytomety against CD-34, CD-105 and STRO-1. The buccal fat derived stem cells (BFDC) were treated by application of the different concentration with BMP-2 of 0, 10, 50, 100 and 200ng/ml, respectively. And their ability to differentiate into osteogenic pathway were checked by alkaline phosphatase(ALP) staining, Alizarin red staining and RT-PCR for osteocalcin(OC) gene expression at 7, 14 and 21day of culture. Flow cytometric analysis and biochemical assays demonstrated that BFDC might be a distinguished stem cells, and mineralization was accompanied in proportion to BMP-2 concentration. However, with 100ng/ml concentration of BMP-2, the BFDC demonstrated most efficent staining pattern of ALP and Alizarin red. The feasibilty of the osteogenic differentiation in the group of both 50ng/ml and 200ng/ml of BMP-2 showed similar activity and relatively weaker than that of 100ng/ml. These results suggest that the BMP-2 stimulate osteogenesis by BFDC effectively and that bone induction might be controlled through negative regulatory feedback in higher concentration.

전신질환자의 치과 치료 중/후 발생 가능한 합병증과 대처법

김창현(Change-Hyen Kim),신희진(Hee-Jin Shin),권영욱(Young-Wook Kwon),박재억(Je-Uk Park) 대한치과의사협회 2010 대한치과의사협회지 Vol.48 No.1

Nowadays, medically compromised patients who could not receive dental treatments in the past are able to go through minor oral surgeries with adequate preoperative measures. Thorough understanding of the systemic disease and its complications is needed as well as the management them. Frequent complications of surgical procedures are bleeding, infection, delayed healing, systemic reactions by stress and they can be aggravated due to the patients’ systemic conditions. Therefore, understanding of the systemic disease of patient visiting dental office and treatment modification according to the systemic status is needed. Also consultation to the medical doctor is imperative, through which perioperative risk and complications can be reduced. Among the high frequency complications of dental treatment of medically compromised patients, bleeding, infection, delayed healing, systemic reactions by stress will be discussed with the management of each one.

상이한 골질과 제원에 따른 짧은 임프란트의 응력 분포: 3차원 유한 요소 분석

김한구,김창현,표성운,Kim, Han-Koo,Kim, Chang-Hyen,Pyo, Sung-Woon 대한악안면성형재건외과학회 2009 Maxillofacial Plastic Reconstructive Surgery Vol.31 No.2

The use of short implants has been accepted risky from biomechanical point of view. However, short implants appear to be a long term viable solution according to recent clinical reports. The purpose of this study was to investigate the effect of different diameter and length of implant size to the different type of bone on the load distribution pattern. Stress analysis was performed using 3-dimensional finite element analysis(3D-FEA). A three-dimensional linear elastic model was generated. All implants modeled were of the various diameter(${\phi}4.0$, 4.5, 5.0 and 6.0 mm) and varied in length, at 7.0, 8.5 and 10.0 mm. Each implant was modeled with a titanium abutment screw and abutment. The implants were seated in a supporting D2 and D4 bone structure consisting of cortical and cancellous bone. An amount of 100 N occlusal load of vertical and $30^{\circ}$ angle to axis of implant and to buccolingual plane were applied. As a result, the maximum equivalent stress of D2 and D4 bones has been concentrated upper region of cortical bone. As the width of implant is increased, the equivalent stress is decreased in cancellous bone and stress was more homogeneously distributed along the implants in all types of bone. The short implant of diameter 5.0mm, 6.0mm showed effective stress distribution in D2 and D4 bone. The oblique force of 100N generated more concentrated stress on the D2 cortical bone. Within the limitations of this study, the use of short implant may offer a predictable treatment method in the vertically restricted sites.

치성낭에서 c-erbB-2 종양 단백의 발현에 관한 면역조직화학적 연구

표성운,김창현,이원,김영실,Pyo, Sung-Woon,Kim, Chang-Hyen,Lee, Won,Kim, Young-Sil 대한구강악안면외과학회 2001 대한구강악안면외과학회지 Vol.27 No.1

To investigate epithelial cell proliferation reactivity in the odontogenic cysts, the expression of c-erbB-2 oncoprotein by epithelial lining was studied in odontogenic keratocyst(OKC, n=10), dentigerous cyst(DC, n=12), radicular cyst(RC, n=12) and normal dental follicle(n=7). The c-erbB-2 immunoreactivity was studied using a streptavidine- biotin- peroxidase method with polyclonal rabbit antihuman antibody to c-erbB-2 oncoprotein which is known to react with formalin fixed, paraffin-embedded sections and the intensity of staining was determined by manually. In all of 10(100%) OKCs, showed positive expression for c-erbB-2 oncoprotein compared with 10/12(83.3%) in DCs, 11/12(91.7%) in RCs and 5/7(71.4%) in normal dental follicles. The expression within OKC was higher than that of DC, RC and dental follicle but statistically not significant(p>0.05) and but may reflects underlying genetic defect. These results demonstrate differences in c-erbB-2 expression between the epithelial linings of the three major odontogenic cyst types, indicating differences in proliferation activity and differentiation processes within these lesions. And, in particular, these results are able to explain the peculiar aggressive growth pattern of OKC.

30% 과산화수소를 함유한 전문가 미백제의 안전성 평가

김신영,박재억,김창현,양성은,Kim, Sin-Young,Park, Je-Uk,Kim, Chang-Hyen,Yang, Sung-Eun 대한치과보존학회 2010 Restorative Dentistry & Endodontics Vol.35 No.3

30% 과산화수소를 함유한 전문가 미백제인 레메화이트(Remewhite, Remedent Inc., Deurle, Belgium)를 시험군 피험자 37명, 시험용 미백제에서 과산화수소를 제외한 동일한 제재를 대조약으로 대조군 피험자 34명에게 1일 3회씩, 2차례에 걸쳐서 시행하였다. 미백의 임상적 안전성은 전기치수검사, 치은염증지수, 지각과민증, 이상반응, 활력징후 등을 통하여 평가하였다. 미백시행 직후, 14주 후, 26주 후에 평가하여 다음과 같은 결론을 얻었다. 30% 과산화수소를 함유한 전문가 미백제를 임상적 적용시에 전기치수검사 결과에서는 미백 전후에 차이가 없었다. 치은염증지수, 지각과민증 평가 결과에서는 시험군에서 다소 경미한 통증 반응이 나타났으나 시간이 지나면서 사라지는 것을 확인하였다. 따라서 30% 과산화수소를 함유한 전문가 미백제를 사용하여 미백시에 임상적 안전성을 확인할 수 있었다. This study evaluated the safety of an office bleaching gel (RemeWhite, Remedent Inc., Deurle, Belgium) containing 30% hydrogen peroxide. 37 volunteers were received office bleaching with the RemeWhite for 3 times at one visit, total 2 visits. As control group, the same gel in which hydrogen peroxide was not included was applied to 34 volunteers with the same protocol. There was no difference between experimental group and control group using electric pulp test. In the result of gingival inflammation index and tooth sensitivity test, there was mild pain response in experimental group but it disappeared as time went by. Therefore, safety of the office bleaching gel containing 30% hydrogen peroxide was confirmed.