인테그린 α₂와 상피성장인자수용체 차단항체의 저해작용을 통한 구강편평상피암 세포의 선택적 제거

최연식,김규천,윤식,황대석,김철훈,전영찬,변준호,신상훈,김욱규,Choi, Yeon-Sik,Kim, Gyoo-Cheon,Yoon, Sik,Hwang, Dae-Seok,Kim, Cheol-Hun,Jeon, Young-Chan,Byun, June-Ho,Shin, Sang-Hun,Kim, Uk-Kyu 대한악안면성형재건외과학회 2013 Maxillofacial Plastic Reconstructive Surgery Vol.35 No.3

Purpose: This study was to find efficacy of integrin alpha2 (${\alpha}_2$) and epidermal growth factor receptor (EGFR) as tumor marker of oral squamous cell carcinoma (SCC) and clarify the selective cell death effect of anti-integrin ${\alpha}_2$ and anti-EGFR on SCC cells, additionally testify conjugated gold nanoparticles (GNP) with air plasma for selective cell death of oral SCC. Methods: Expression of integrin ${\alpha}_2$, EGFR on human SCC cells (SCC25) were examined by western blot. SCC25 cells were treated with anti-integrin ${\alpha}_2$, anti-EGFR and analysed by Hemacolor staining, immunoflorescence staining, FACS flow cytometry. Conjugated GNP with integrin ${\alpha}_2$, EGFR antibody were treated by air plasma on SCC cells. Results: Integrin ${\alpha}_2$ and EGFR were over-expressed on SCC25 cells than normal lung WI-38 cells. The cell viability rate of SCC25 cells treated with anti-integrin ${\alpha}_2$, anti-EGFR was lower than WI-38 cells. The concentration changes of nucleus, releasing cytochrome c and apoptosis inducing factor (AIF) from mitochondria to cytosol were observed. The changes of proteins related with apoptosis were observed. Increase of bax, bcl-xL, activation of caspase-3, -7, -9, and fragmentation of PARP, DFF45 and decrease of lamin A/C in SCC25 cells were observed. In FACS, increase of sub-$G_1$ and S phase was observed. Cell cycle related proteins, Such as cyclin D1, cyclin dependent kinase (CDK) 4, cyclin A, cyclin E, CDK 2, p27 were decreased. After SCC25 cells treated with conjugatged GNP-Integrin ${\alpha}_2$, GNP-EGFR, additionally air plasma, the cell death rate was significantly increased. Conclusion: Integrin ${\alpha}_2$, EGFR were over-expressed in oral SCC cells. Anti-integrin ${\alpha}_2$, anti-EGFR in SCC25 cells induced apoptosis selectively. When GNP-anti integrin ${\alpha}_2$, GNP-anti EGFR were treated with air plasma on SCC25 cells, cancer cells were died more selectively. GNP-anti integrin ${\alpha}_2$, GNP-anti EGFR with air plasma could be treatment choice of oral SCC.

골다공증 유도 백서 경골에 티타늄 임플랜트 매식 시 지방조직유래 줄기세포 주입과 저출력 초음파 적용이 골치유에 미치는 영향

이광호,최연식,신상훈,정인교,김규천,김철훈,김욱규,Lee, Kwang-Ho,Choi, Yeon-Sik,Shin, Sang-Hun,Chung, In-Kyo,Kim, Gyoo-Cheon,Kim, Cheol-Hun,Kim, Uk-Kyu 대한악안면성형재건외과학회 2011 Maxillofacial Plastic Reconstructive Surgery Vol.33 No.3

Purpose: Osteoporosis, is a major health problem for the elderly and post-menopausal women and shown to alter the properties of bone as well as impair bone healing around titanium implants in both human and animals. The objective of this study was to examine the effect of LIPUS with adipose-derived stem cells on the healing process around a titanium implant in rats with osteoporosis. Methods: Sixteen osteoporosis-induced rats were divided into two groups: an adipose-derived stem cell injected with Low Intensity Pulsed Ultrasound (LIPUS) application group and a control group. Titanium screw implants (diameter, 2.0 mm: length, 3.5 mm, Cowell Medi, Korea) were placed into both tibia of 16 rats, on 8 rats as the control group and the other 8 rats as the experimental group. Rats were sacrificed at different intervals from 1, 2, 4 and 8 weeks after implantation for histopathologic and immunohistochemical analyses. Results: Histopathological analysis revealed newly formed bone in experimental group earlier than that in control group. Especially at 1 week after implantation, more amounts of new bone matrix and collagen around the implant in the experimental group were seen compared with the control group. Immunohistochemical analysis showed that the levels of osteoprotegerin (OPG) expression in the experimental group were increased at early stages compared with that of control group until 2 weeks after implantation. But after 2 weeks, the expression level of OPG similar in both groups. The expression levels of receptor activator of nuclear factor kB ligand (RANKL) were stronger in the experimental group than the control group until 2 weeks after implantation. After 4 weeks, expression of RANKL in experimental group was similar to the control group. Conclusion: The results of this study suggest that LIPUS with Adipose-Derived Stem Cells in implantation could promote bone healing around titanium implants in rats with osteoporosis.

설암의 술전 조직표본에서 악성도와 혈관내피세포성장인자 발현과의 상관관계

변준호,박봉욱,정인교,김종렬,김욱규,박봉수,김규천,Byun, June-Ho,Park, Bong-Wook,Chung, In-Kyo,Kim, Jong-Ryoul,Kim, Uk-Kyu,Park, Bong-Soo,Kim, Gyoo-Cheon 대한악안면성형재건외과학회 2005 Maxillofacial Plastic Reconstructive Surgery Vol.27 No.6

Angiogenesis is important for the growth and metastasis of solid tumors. Some growth factors, inflammatory cytokines, and angiogenin are known to promote tumor angiogenesis. Among them, Vascular endothelial growth factor (VEGF) is the most intriguing factor in regard to tumor angiogenesis. Inhibition of VEGF activity by neutralizing antibodies or by the introduction of dominant negative VEGF receptors into endothelial cells of tumor-associated blood vessels resulted in the inhibition of tumor growth and in tumor regression, indicating that VEGF is a major initiator of tumor angiogenesis. VEGF promotes angiogenesis through their receptors, Flt-1 and Flk-1/KDR. on vascular endothelial cells. These two receptors were usually believed to be expressed specifically on vascular endothelial cell. Several reports have now shown that VEGF is not only significantly associated with microvessel density but also has prognostic value in both node-negative and node-positive oral squamous cell carcinoma. For many years several histologic features of the neoplasms are being considered when assessing the influence of malignancy grading on recurrence and prognosis. Among the characteristics investigated, degree of keratinization, nuclear pleomorphism, mode of invasion, microscopic depth of invasion, intravascular invasion, lymphocyte infiltration, and number of mitoses have been considered as important prognostic factors. So, this study was conducted to evaluate the correlation of vascular endothelial growth factor expression with malignancy in paraffin-embedded biopsy specimens from 11 patients with tongue cancers. Our results showed that high immunoreactivity specimens of VEGF expression were significantly lower keratinization degree and more pronounced nuclear pleomorphism than in low immunoreactivity specimens. Thus, VEGF expression could be used as a prognostic marker in tongue cancer.

압축력을 병용한 하악골 신장술

황영섭,허준,김욱규,박성진,황대석,김용덕,정인교,김규천,Hwang, Young-Seob,Heo, June,Kim, Uk-Kyu,Park, Seong-Jin,Hwang, Dae-Seok,Kim, Yong-Deok,Chung, In-Kyo,Kim, Kyu-Cheon 대한악안면성형재건외과학회 2006 Maxillofacial Plastic Reconstructive Surgery Vol.28 No.6

The purpose of this study was to investigate the biomechanical, histologic findings of distracted regenerate and TMJ response in modified distraction osteogenesis (DO) technique combined with compression force as biomechanical stimulation method which has been suggested in 2002, and developed thereafter by authors. This study was performed with two experiments. First experiment was designed to explore the optimal ratio of compression force versus distraction force for the new DO technique. Second experiment was planned to evaluate the reaction of TMJ tissue, especially condyle, disc after application of the DO technique with compression force. Total 52 New Zealand adult male-rabbits with 3.0kg body weight were used for the study. For the first study, 30 adult male-rabbits underwent osteotomy at one side of mandibular body and a external distraction device was applied on each rabbit with same manner. In the control group of 10 rabbits, final 8 mm of distraction with 1 mm rate per day was done with conventional DO technique after 5 latency days. For the experimental group of 20 rabbits, a compression force with 1 mm rate per day was added to the distracted mandible on 3-latency day after over-distraction (over-lengthening). As the amount of the rate of compression versus distraction, experimental subgroup I (10 rabbits) was set up as 2 mm compression versus 10 mm distraction (1/5) and experimental subgroup II (10 rabbits) was set up as 3 mm compression versus 11 mm distraction (about 1/3). All 30 rabbits were set up to obtain final 8 mm distraction and sacrificed on postoperative 55 day to analysis on biomechanical, and histologic findings of the bone regenerates. For second study, 22 adult male-rabbits were used to evaluate TMJ response after the DO method application with compression force. In the control group, 10 rabbits was used to be performed with conventional DO method, on the other hand, in a experimental group of 10 rabbits, 10 mm distraction with 2 mm compression (1/5 ratio) was done. The remaining 2 rabbits served as the normal control group. Histomorphologic examinations on both condyle, histological studies on condyle, disc were done at 1, 2, 3, 4, 7 weeks after distraction force application. The results were as follows: 1. On the bone density findings, the experimental group II (force ratio - 1/3) showed higher bone density than the other experimental group (force ratio - 1/5) and control group (control group - $0,2906\;g/cm^2$, experimental group I - $0.2961\;g/cm^2$, experimental group II - $0.3328\;g/cm^2$). 2. In the histologic findings, more rapid bone maturation like as wide lamellar bone site, more trabeculae formation was observed in two experimental groups compared to the conventional DO control group. 3. In morphologic findings of condyle, there were no differences of size and architecture in the condyle in the control and experimental groups. 4. In histologic findings of condyles, there were thicker fiberous and proliferative layers in experimental group than those of control group until 2 weeks after distraction with compression force. But, no differences were seen between two groups on 3, 4, 7 weeks after compression. 5. In histologic findings of disc, more collagen contents in extracellular matrix, more regular fiber bundles, and less elastin fibers were seen in experimental group than control group until 2 weeks after distraction with compression. But, no differences were seen between two groups on 3, 4, 7 weeks after distraction with compression. From this study, we could identify that the new distraction osteogenesis technique with compression stimulation might improve the quality of bone regeneration. The no remarkable differences on TMJ response between control and experimental groups were seen and TMJ tissues were recovered similarly to normal TMJ condition after 3 weeks.

사이즈를 조절한 홍삼분말의 첨가가 어묵의 품질 특성에 미치는 영향

심도완(Do-Wan Shim),강군(Jun Jiang),김진효(Jin-Hyo Kim),김원우(Won-Wu Kim),강위수(Wie-Soo Kang),최원석(Won-Seok Choi),허선진(Sun-Jin Hur),김동영(Dong-Young Kim),김규천(Kyu-Cheon Kim),이광호(Kwang-Ho Lee) 한국식품영양과학회 2012 한국식품영양과학회지 Vol.41 No.10

고품질 어묵 개발을 위하여 홍삼(red ginseng)을 크기가 다른 미세 분말화하여 튀김 어묵을 제조한 다음 색조 변화, 보존성, 가열감량, 지방산화, 총균수와 같은 물리, 화학, 생물학적 특성을 조사하였다. 홍삼분말이 첨가된 어묵의 관능검사를 실시한 결과, 1%와 850 μm 크기의 홍삼분말첨가가 최적으로 나타났다. 홍삼분말이 들어간 어묵의 경우 색도의 변화에서 L값은 감소하는 경향을 나타냈으며, a값과 b값은 증가하는 경향을 보였다. 이는 대체적으로 첨가물이 들어간 다른 어묵들과 비슷한 경향을 보였다. 그리고 물성검사에서 홍삼분말 90 μm 처리군에서만 경도와 씹힘성이 유의적으로 증가하는 것을 확인할 수 있었다. 그리고 가열감량에서도 홍삼분말이 들어간 어묵이 시중 어묵에 비해서 가열했을 때 어묵 자체의 무게 변화가 더 적은 것을 확인하였다. 지방산패도 측정결과 홍삼분말이 들어간 어묵이 일반 어묵에 비해 지질산화 억제효능이 높은 것을 확인하였다. 특히 사이즈가 가장 작은 10 μm의 홍삼분말을 첨가했을 때 지질산화 억제 효능이 높은 것으로 나타났다. 본 연구결과 1% 수준의 홍삼 분말을 어묵에 첨가하는 것이 풍미 증가와 이화학적인 안정성뿐만 아니라 저장성 또한 증가시킬 수 있는 것으로 나타났다. 또한 홍삼의 사이즈를 조절하여 첨가함으로써 그 기능성을 더욱 향상시킬 뿐만 아니라 생리활성 효능을 가진 홍삼이 함유된 고품질의 어묵을 생산할 수 있는 것으로 나타났다. Fried fish pastes containing different size and amounts of red ginseng powder (RGP) were manufactured, and their physico-chemical and biological properties, including color changes, preservation, weight loss after heating, lipid oxidation, and total colony count were analyzed to improve fish paste quality. Sensory evaluation of fish pastes containing RGP was carried out, and the results showed that 1% and 850 μm sized RGP additives were most preferred. In the color change test, decreased L<SUP>*</SUP> (lightness), increased a<SUP>*</SUP> (redness), and increased b<SUP>*</SUP> (yellowness) values were observed, which was similar to other additive-containing commercial fish pastes. In the physical properties test, hardness and chewiness significantly increased in the 90 μm sized RGP group. Weight loss of fish pastes containing RGP after heating was attenuated compared to commercial fish pastes. Fish paste containing RGP showed an inhibitory effect on lipid oxidation. Especially, the 10 μm sized RGP group showed the most significant inhibitory effect on lipid oxidation and reduced total microbes during storage. Therefore, 1% addition of RGP can give rise to high quality fish pastes through improvement of sensory evaluation and physico-chemical properties. Moreover, functionally and physiologically improved fish pastes can be produced by adding different amounts of RGP.

골다공증 유도 흰쥐에서 칼슘과 비타민 D 섭취가 티타늄 임플랜트 주위의 골 형성에 미치는 영향

이재열(Jae-Yeol Lee),정석영(Seok-Young Jeong),신상훈(Sang-Hun Shin),김규천(Gyoo-Cheon Kim),김용덕(Yong-Deok Kim),정인교(In-Kyo Chung),김욱규(Uk-Kyu Kim) 대한구강악안면외과학회 2008 대한구강악안면외과학회지 Vol.34 No.3

Purpose: The purpose of this study was to observe the effect of calcium and vitamin D to the titanium implant osseointegration in the osteoporosisinduced animal model. Material and method: Thirty-two rats, 10 weeks of age, were divided into two groups: experimental group was ingested additional calcium and vitamin D , and a control group was not. Titanium screw implant(diameter, 2.0 mm; length, 3.5 mm; pitch-height 0.4 mm) were placed into tibia of 32 rats, 16 in the control group and 16 in the experimental group. The rats were sacrificed at 1, 2, 4 and 8 weeks after implantation for histopathologic examination, histomorphometric analysis and immunohistochemistry with fibronectin and collagen type I antibody. Result: In histopathological findings, newly formed bone was seen at 2 weeks and became lamellar bone at 4 weeks, and mature trabecullar bone was seen at 8 weeks in experimental group. In control group, thickness of regenerated bone increased till 4 weeks gradually and trabecullar bone was seen at 8 weeks. In histomorphometric analysis, marrow bone density increased significantly in experimental group compared to control group. Fibronectin immunoreactivity was strong at 2 weeks in experimental group and reduced after 4 weeks gradually. But it was maintained continuously from 2 to 8 weeks in control group. Collagen type I immunoreactivity was very strong from 2 to 4 week in experimental group. And the amount of Collagen type I expression was more abundant in experimental group. Conclusion: The results of this study suggest that calcium and vitamin D supplementation promote bone healing around titanium implants in osteoporosis induced animals.

설암에서 신부가화학요법후 미세혈관밀도에 대한 종양관련 대식세포의 역할

박봉욱(Bong-Wook Park),정인교(In-Kyo Chung),김종렬(Jong-Ryoul Kim),김욱규(Uk-Kyu Kim),박봉수(Bong-Soo Park),김규천(Gyoo-Cheon Kim),변준호(June-Ho Byun) 대한구강악안면외과학회 2006 대한구강악안면외과학회지 Vol.32 No.3

Preoperative neoadjuvant chemotherapy using cisplatin and 5-FU is generally given in oral and maxillofacial cancer. At tissue level both inflammation and fibrosis occur after chemotherapy. The cellular changes mimic those of a granulating wound, with activated macrophages and fibroblasts replacing the malignant cells as they are erradicated. Stromal cells, together with extracellular matrix components, provide the microenvironment that is pivotal for tumor cell growth, invasion, and metastatic progression. Vascular endothelial growth factor(VEGF), an important regulator of angiogenesis in cancer, induces mitogenesis of vascular endothelial cells, and vascular permeabilization and microvessel formation in a tumor are associated with tumor nutrition and oxygenation. Also, they are associated with chemotherapeutic drug delivery. Oxygen delivery to tumor appears to rely on a network of microvessels, On the other hand, the tumor microvessel is clearly an important factor in chemotherapeutic drug delivery to cancer cells, and the efficacy of drug delivery can be high in richly vascularized tumors. So, this study was conducted to evaluate the effect of neoadjuvant chemotherapy on microvessel density from 11 patients with tongue cancers. Our results showed that neoadjuvant chemotherapy was seemed to decrease VEGF expression in tumor cells, however, it did not significantly alter VEGF expression in tumor-associated macrophages. Also, Neoadjuvant chemotherapy had little effect on the microvessel density using CD34, and tumor-associated macrophage level using CD68. Thus, tumorassociated macrophages seem to be the key factor associated with the maintenance of microvessel density after neoadjuvant chemotherapy in tongue cancer.

구강 편평세포 암종에서의 CD44 발현

박상준(Sang Jun Park),박혜련(Hae Ryoun Park),김규천(Gyoo Cheon Kim),박봉수(Bong Soo Park),김태규(Tae Kyu Kim) 대한구강악안면외과학회 2000 대한구강악안면외과학회지 Vol.26 No.2

The cell surface glycoprotein CD44 is a kind of adhesion molecule, which binds hyaluronic acid, type I collagen and fibronectin. Although there have been numerous reports on the expression and the function of CD44 in lymphocytes and macrophages, very little is known about its distribution and definite role in epithelial tissue, especially in oral epithelial one. The present study was performed to investigate the distribution and expression of the CD44 in human gingiva and squamous cell carcinoma(SCC) arising in human gingiva. And the authors compared CD44 expression with histopathologic grade of SCC. The results were as follows: 1. The CD44 was strongly expressed in granular, spinous and basal layers of normal marginal and attached gingiva, in spinous and basal layers of normal sulcular gingiva, and in all epithelial layers of normal junctional gingiva. 2. In SCC of gingiva, the CD44 was expressed in all but one case. In most of the cases the CD44 was expressed at cell membrane and the degree of expression was relatively strong. 3. In low-grade SCC of gingiva, the CD44 was strongly expressed, especially at the basal and spinous layers of abundantly keratinized cancer nests. In high-grade SCC of gingiva, the CD44 expression tended to be weak but was strong at cells showing individual keratinization. This study suggest that the CD44 expression of normal and cancerous gingival epithelium is associated with the degree of proliferation and differentiation of epithelial cells.

비스포스포네이트 연관 악골괴사의 치료 방식과 투약 휴지기가 치료 예후에 미치는 영향

이승현(Seung-Hyun Rhee),박군효(Kun-Hyo Park),이재열(Jae-Yeol Lee),김용덕(Yong-Deok Kim),신상훈(Sang-Hun Shin),정인교(In-Kyo Chung),김규천(Gyoo-Cheon Kim),김욱규(Uk-Kyu Kim) 대한치과의사협회 2015 대한치과의사협회지 Vol.53 No.2

Objective: The purpose of this study is to compare the surgical treatment with conservative treatment and to evaluate the effectiveness of drug holiday in bisphosphonate related osteonecrosis of the jaw (BRONJ) patients who were diagnosed as stage 2. Patients and Method: From January 2012 to October 2014, seventy two patients who visit to Pusan National University of Dental Hospital were diagnosed as stage 2 of BRONJ. All the patients had taken computed tomography(CT) and panoramic radiography. The surgical treatment including sequestrectomy of necrotic bone and curettage of soft tissue around the sequestrum were performed to fifty patients. Twenty two patients underwent conservative treatment such as antibiotics medications, mouth rinsing and follow up checking for every two weeks. Prognosis of treatment was classified into 3 groups ? response, unresponse, and worsens according to clinical, radiographic symptoms. P value less than 0.05 were regarded as significant. Results: In surgery group, forty five patients (90%) were healed without recurrence or any complication and five patients (10%) showed the wound dehiscence or infection. In conservative treatment group, fifteen patients (68.2%) were healed without any complication, four patients (18.2%) did not show improvement and three(4.2%) patients°Ø symptoms increased. P value among groups was 0.014. Evaluation of preoperative drug holiday in surgery group did not show the effectiveness(p=0.478). Conclusion: Statistically, the prognosis of patients with stage 2 BRONJ treated with surgery was significantly better than conservative treatment. There was no statistical difference for the preoperative drug holiday.

칼슘과 비타민 D 섭취가 티타늄 임플랜트 주위의 골 형성에 미치는 영향

박광일(Kwang-Il Park),이재열(Jae-Yeol Lee),황대석(Dae-Seok Hwang),김용덕(Yong-Deok Kim),김규천(Gyoo-Cheon Kim),신상훈(Sang-Hun Shin),김욱규(Uk-Kyu Kim),정인교(In-Kyo Chung) 대한구강악안면외과학회 2007 대한구강악안면외과학회지 Vol.33 No.2

The purpose of this study was to observe the effect of calcium and vitamin D to the titanium implant osseointegration in animal model. 32 rats, 10 weeks of age, were divided into two group: additional calcium and vitamin D supplementation group and a control group. Titanium screw implant(diameter, 2.0mm; length, 3.5mm; pitch-height 0.4 ㎜) were placed into tibia of 32 rats, 16 in the control group and 16 in the experimental group. The rats were sacrificed at different time interval(1, 2, 4, and 8 weeks after implantation) for histopathologic observation, histomorphometric analysis and immunohistochemistry with osteocalcin and osteopontin antibody. Histopathologically findings, newly formed bone was seen at 1 weeks and became lamellar bone at 2 weeks, and mature trabecullar bone was seen at 4 weeks experimental group. In control group, thickness of regenerated bone increased till 4 weeks gradually and trabecullar bone was seen at 8 weeks. By histomorphometric analysis, bone marrow density was increased significantly at 1 and 2 weeks in experimental group compared to control group. Osteocalcin immunoreactivity was strong at 1 week experimental group and reduced after 4 weeks gradually. But it was continuously weakly from 1 to 4 weeks in control group. Osteopontin immunoreactivity was very strong in newly formed bone from 2 to 8 weeks experimental group. And the amount of osteopontin expression was more abundant in experimental group. The results of this study suggest that calcium and vitamin D supplementation promotes bone healing around dental implants