홍화 추출물이 치주인대세포, 조골세포 활성도에 미치는 영향

류인철,이용무,구영,배기환,정종평,Rhyu, In-Chul,Lee, Yong-Moo,Ku, Young,Bae, Ki-Whan,Chung, Chong-Pyoung 대한치주과학회 1997 Journal of Periodontal & Implant Science Vol.27 No.4

Safflower(Carthamus tinctorius $Linn\acute{e}$ has been traditionally used for the treatment of blood stasis, and Dipsasi Radix has been used as a drug for fracture in Chinese medicine. The purpose of present study was to examine the biologic effects of safflower extract and Disasi radix extracts on the periodontal. ligament cells and osteoblastic cells and on the wound healing of rat calvarial defect. The ethanolic extract of safflower blossom, safflower seed and Dipsasi Radix(125, 250, and 500 ${\mu}g/ml$) were prepared as test group, and PDGF-BB(lOng/ml) and unsafonifiable fraction of Zea Mays L.(125, 250, and 500 ${\mu}g/ml$) were employed as positive control. The effects of each agents on the growth and survival, ALPase activity, expression of PDGF-BB receptor, chemotactic response of PDL cell and ATCC human osteosarcoma MG63 cells in vitro were examined. The tissue regenerative effect of each extracts was evaluated by histomorphometric measuring of newly formed bone on the 8mm defect in rat calvaria after oral administration of 3 different dosages groups : 0.02, 0.1 and 0.35g/kg, per day. It was also employed the same dosages of unsaponifiable fraction of Zea Mays L. as positive controls. Safflower blossom extract, safflower seed extract, and Dipsasi Radix extract stimulate the cellular activity of MG63 cells in concentration range of $125-500{\mu}g/ml$, and safflower bolssom extract and safflower seed extract stimulate also the cellular activity of periodontal ligament cells in concentration range of $250-500{\mu}g/ml$. In activity of ALPase, $250-500{\mu}g/ml$ of safflower blossom extracts showed significant stimulating effects on MG63 cells, and the same concentration range of safflower seed extracts showed significant effect on periodontal ligament cells. In the recovery on PDGF-BB receptor expression which was depressed by $IL-1{\beta}$, $125-250{\mu}g/ml$ of safflower blossom extracts and $250-500{\mu}g/ml$ of safflower seed extracts showed significant increasing effect on MG63 cells, and $500{\mu}g/ml$ of safflower blossom extract and $250-500{\mu}g/ml$ of safflower seed extracts showed significant effect on periodontal ligament cells. In chemotactic response, among all tested group, safflower seed extracts only were chemotactic to MG63 cells and periodontal ligament cells in concentration range of $125-500{\mu}g/ml$. Also in the view of bone regeneration in rat calvarial defect model, the only group that was orally administrated 0.35g/kg, day of safflower seed extract showed significant new bone formation. These results suggested that safflower extracts might have a potential possibilities as an useful drug for adjunct to treatment for regeneration of periodontal defect.

우로수데옥시콜릭산이 치주질환 억제에 미치는 영향

박상현,한승민,최상목,구영,류인철,한수부,이학모,김문무,김상년,정종평,Park, Sang-Hyun,Han, Seoung-Min,Choi, Sang-Mook,Ku, Young,Rhyu, In-Chul,Han, Soo-Boo,Lee, Hak-Mo,Kim, Moon-Moo,Kim, Sang-Nyun,Chung, Chong-Pyoung 대한치주과학회 1999 Journal of Periodontal & Implant Science Vol.29 No.1

Ursodeoxycholic acid(UDCA) is a hydrophilic gall bladder acid and has been used as a effective drug for liver disease related to in1munity. This drug inhibits secretions of IL-2, IL-4, and $IFN-{\gamma}$ from T-cells and production of immunoglobulin from B-cells. Also it has been reported that UDCA inhibits production of IL-1 related to the progression of periodontal disease and activation of collagenases. The purpose of the present study was to elucidate the effects of UDCA on inhibition of periodontal disease progression using clinical, microbiological and histometrical parameters. Twelve pure bred, 16 month-old-beagle dogs were used in the study. After ligature-induced periodontal diseases were formed, experimental drugs were applied twice a day and then the results of clinical, microbiological, and histometrical parameters were measured at baselie(initiation of experiment) , 4weeks and 8weeks. The gel with UDCA(concentration 0.5%, 5% 3 dogs in each) was applied to experimental group, chlorhexidine to positive control group(3dogs) and the gel without UDCA(base) to negative control group. After induction of general anesthesia, the maxillary 2nd, 3rd premolars and 1st molar and the mandibular 2nd, 3rd, 4th premolars and 1st molar were ligated in one side selected randomly and were not ligated in the opposite side. The plaque index(PI), gingival index(GI), pocket depth(PD) and gingival crevicular fluid(GCF) volum were measured clinically. The PI and GI were measured at 3 buccal points of all experimental teeth and the GCF was measured only at the 3rd premolar in the maxilla and the 4th premolar in the mandible. In the microbiological study, the samples extracted from the 3rd premolar of the maxilla and the 4th premolar of the mandible at the center of buccal surface were analyzed aerobics, anaerobics and Streptococcus colony forming units, After clinical and microbiological examination at 8weeks, the dogs were sacrificed by carotid artery perfusion. The samples were fixed and sectioned including interproximal area, and the distance from cementoenamel junction(CEJ) to alveolar crest was measured. The results were that PI, GI and PD increased until 4 weeks and decreased at 8 weeks in three groups but the differences between all the groups were not significant. The 0.5% UDCA in non-ligated group showed remarkable decrease of GCF. The experimental group applied 5% UDCA decreased the number of aerobics and anaerobics. The distance from CEJ to alveolar crest was greater in the negative control group on both ligated and non-ligated sides, but the differences were not significant stastically.

Poly(alpha-hydroxy acids) 제제 생분해성 차폐막의 치주조직 재생유도능력에 관한 조직학적 장기관찰

류인철,구영,정종평,한수부,최상묵,Rhyu, In-Chul,Ku, Young,Chung, Chong-Pyoung,Han, Soo-Boo,Choi, Sang-Mook 대한치주과학회 1997 Journal of Periodontal & Implant Science Vol.27 No.3

The recent trend of research and development on guided tissue regeneration focuses on the biodegradable membranes, which eliminate the need for subsequent surgical removal. They have demonstrated significant and equivalent clinical improvements to the ePTFE membranes. This study evaluate guided tissue regeneration wound healing in surgically induced intrabony periodontal defects following surgical treatment with a synthetic biodegradable membranes, made from a copolymer of glycolide and lactide, in 8 beagle dogs. After full thickeness flap reflection, exposed buccal bone of maxillary and mandibular canine and premolar was removed surgically mesiodistally and occlusoapically at $6mm{\times}6mm$ in size for preparation of periodontal defects. In experimental sites a customized barrier was formed and fitted to cover the defect. Flap was replaced slightly coronal to CEJ and sutured. Plaque control program was initiated and maintained until completion of the study. In 4, 8, 16 and 24 weeks after surgery, the animals were sacrificed and then undecalcified specimens were prepared for histologic evaluation. Histologic examination indicated significant periodontal regeneration characterized by new connective tissue attachment, cementum formation and bone formation. These membranes showed good biocompatibility throughout experiodontal period. The barriers had been completely resorbed with no apparent adverse effect on periodontal wound healing at 24 weeks. These results implicated that present synthetic biodegradable membrane facilitated guided tissue regeneration in periodontal defect.

수종의 상용 세치제들의 향균 및 향염효과 비교연구

류인철,Rhyu, In-Chul 대한치주과학회 1996 Journal of Periodontal & Implant Science Vol.26 No.2

It is known that some natural extracts from plants have a various range of antimicrobial and antiinflammatory activity. There are lots of clinical trials to develop toothpastes containing natural extracts for prevention of dental caries and gingival inflammation. The purpose of this study was to evaluate antimicrobial and antiinflammatory activity of magnolol containing toothpastes and other commercial toothpastes. Eleven kinds oftoothpastes were used. They include magnolol, sanguinarine, Myrrha, Mori radicis cortex,Cimicifugae rhizoma, sodium fluoride, aminocaprolactic acid etc. Six strains of bacteria were used for this test, ego Porphylomonas gingivalis, Prevotellain-termedia, Actinobacillus actinomy cetemcomitans, Streptococcus mutans, Stretococcus sanguis, and Actinomyces species. Antimicrobial activity was determined by an agar dillution method and a broth microdillution method. Antiinflammatory activity was assessed by the inhibition of $PGE_2$ production from gingival fibroblast with the addition of rHIL-1 and centrifuged solution of toothpastes. Control group was only rHIL-1 additive sample. $PGE_2$ enzyme immunoassay systemfAmersham, In. Buckinghamshire, U.K). $PGE_2$ level was measured by ELISA reader with 450 nm, The results from the study revealed that toothpastes containing natural extracts generally had high antimicrobial and antiinflammatory activity. Especially magnolol containing toothpaste showed higher antimicrobial activity than other toothpastes, and sanguinarine containing toothpaste showed particularly high antimicrobial activity in A. actinomicetemcomitans and A. viscosus. In some degree all toothpastes inhibited $PGE_2$ production, but magnolol containing toothpaste was potent inhibitor of $PGE_2$. Sodium chloride containing toothpaste had also effective result. The results suggested that toothpastes containing natural extracts were promising in plaque control and prevention of dental caries and gingivitis.

흡수성 차폐막의 치주조직 재생에 혈소판유래 성장인자가 미치는 영향

류인철,배규현,설양조,구영,이승진,한수부,최상묵,정종평,Rhyu, In-Chul,Bae, Kyoo-Hyun,Seol, Yang-Jo,Ku, Young,Lee, Seung-Jin,Han, Soo-Boo,Choi, Sang-Mook,Chung, Chong-Pyoung 대한치주과학회 1999 Journal of Periodontal & Implant Science Vol.29 No.3

The ultimate objective of periodontal treatment is to stop disease progression and to regenerate destroyed periodontal tissues and thereby regain normal function. Growth factors are naturally found polypetides which stimulate many cellular activities pertaining to wound healing by acting as signal molecule in controlling cell movement, proliferation, and matrix production. Platelet derived growth factor (PDGF) is 28,000-35,000 Da molecular weight dimeric protein with 2 long positively charged polypeptide chains connected by sulfide bonds. The purpose of this study is to evaluate histologically the initial guided tissue regeneration in a periodontal defect f a beagle dog treated with a biodegradable membrane formed with polylactic acid (poly-L-lactic acid) and polyglycolic acid loaded with 200ng/$cm^2$ platelet derived growth factor. 2 beagle dogs were used in he experiment. $5mm{\times}6mm$ alveolar bone defect was formed in upper and lower canines and third premolars and a reference notch was placed. PDGF-BB non-containing membrane was used as control. Each defect was randomly assigned to the test roup or the control group. The dogs were sacrificed 3 weeks after membrane placement. Toluidine blue and multiple staining was done for histological analysis. In the 3 week specimen in the control group, no new one formation could be seen. Small amount f bone resorption below the notch could be seen. In the notch, loose connective tissue with infiltration of inflammatory cells could be seen. Also thin discontinuous new cementum could be seen and the membrane still retained its structure. Where PDGF-BB containing membrane was used, new bone formation could be seen in the notch at weeks and also continuous thin cementum could be seen. PDL cells were observed between new bone and new cementum and some were attached to bone and cementum. These results suggest that new bone and cementum formation seen when PDGF-BB loaded membrane was used was due to inhibition of downgrowth of epithelial cells and also due to continuous release of the growth factor. Further study on the resorption characteristics of the membrane nd the release characteristics of the PDGF-BB is necessary. Also, development of a membrane easier to use clinically is necessary.

Polyethyleneimine-응축 BMP-2 발현 유전자를 이용한 골형성 효과

정희선,김경화,박윤정,김태일,이용무,구영,류인철,이동수,이승진,정종평,한수부,설양조,Cheong, Hee-Sun,Kim, Kyoung-Hwa,Park, Yoon-Jeong,Kim, Tae-Il,Lee, Yong-Moo,Ku, Young,Rhyu, In-Chul,Lee, Dong-Soo,Lee, Seung-Jin,Chung, Chong-Pyoung,Han, 대한치주과학회 2007 Journal of Periodontal & Implant Science Vol.37 No.4

Naked DNA and standard vectors have been previously used for gene delivery. Among these, PEI can efficiently condense DNA and has high intrinsic endosomal activities. The aim of this study is to investigate whether the cationic polycation PEI could increase the transfection efficiency of BMP expressing DNA using a vector-loaded collagen sponge model. BMP-2/pcDNA3.1 plasmid was constructed by subcloning human BMP-2 cDNA into the pcDNA3.1 plasmid vector. PEI/DNA complexes were prepared by mixing PEI and BMP-2/pcDNA3.1 and the constructed complexes were loaded into the collagen sponges. In vitro studies, BMSCs were transfected with the PEI/BMP-2/pcDNA3.1 complexes from collgen sponge. The level of secreted BMP-2 and alkaline phosphatase activities of transfected BMSCs were significantly higher in PEI/BMP-2/pcDNA3.1 group than in BMP-2/pcDNA3.1 group (p<0.05). Transfected BMSCs were cultured and mineralization was observed only in cells treated with PEI/BMP-2/pcDNA3.1 complexes. In vivo studies, PEI/BMP-2/pcDNA3.1/collagen, BMP-2/pcDNA3.1/collagen and blank collagen were grafted in skeletal muscle of nude mice. Ectopic bone formation was shown in PEI/BMP-2/pcDNA3.1/collagen grafted mouse 4 weeks postimplantation, while not in BMP-2/pcDNA3.1 grafted tissue. This study suggests that PEI-condensed DNA encoding for BMP-2 is capable of inducing bone formation in ectopic site and might increase the transfection rate of BMP-2/pcDNA3.1. As a non-viral vector, PEI offers the potential in gene therapy for bone engineering.

임플랜트 주위 연조직세포의 세포-기질 접착

이석원,류인철,한종현,이재봉,Lee Suk-Won,Rhyu In-Chul,Han Chong-Hyun,Lee Jai-Bong 대한치과보철학회 2006 대한치과보철학회지 Vol.44 No.1

The importance of soft tissue response to implant abutments has become one of the major issues in current implant dentistry. To date, numerous studies have emphasized on maintaining connective tissue barriers in quantity, as well as in quality fir the long term success of dental implants. The cells mainly consisting the soft tissue around dental implants are fibroblasts and epithelial cells. The mechanism of the fibroblasts adhesions to certain substrata can be explained by the 'focal adhesion' theory. On the other hand, epithelial cells adhere tn the substratum via hemidesmosomes. The typical integrin-mediated adhesions of cells to certain matrix are called 'cell-matrix adhsions'. The focal adhesion complex of fibroblasts, in relation to the cell-matrix adhsions, consists of the extracellular matrix(ECM) such as fibronectin, the transmembrane proteins such as integrins, the intracellular cytoplasmic proteins such as vinculin, talin, and more, and the cytoskeletal structures such as filamentous actin and microtubules. The mechanosensory function of integrins and focal adhesion complexes are considered to play a major role in the cells adhesion, migration, proliferation, differentiation, division, and even apoptosis. The '3-D matrix adhesions' defined by Cukierman et al. makes a promising future for the verification of the actual process of the cell-matrix adhesions in vivo and can be applied to the field of implant dentistry in relation to obtaining strong soft tissue attachment to the implant abutments.

임산부의 치주 질환 활성도와 조산과의 상관관계에 관한 연구

최은정,구영,류인철,함병도,윤보현,한수부,정종평,최상묵,Choi, Eun-Cheong,Ku, Young,Rhyu, In-Chul,Hahm, Byung-Do,Yoon, Bo-Hyun,Han, Soo-Boo,Chung, Chong-Pyoung,Choi, Sang-Mook 대한치주과학회 2000 Journal of Periodontal & Implant Science Vol.30 No.1

Purpose We designed this study for the purpose of determining the relationship between periodontal disease activity and PLBW, using the evaluation of probing pocket depth, loss of attachment, gingival index, gingival crevicular fluid amount and subgingival microflora. Methods A total of 100 volunteer mothers(mean age 30.44) at the Department of Obstetrics and Gynecology Seoul National University Hospital were selected for this study.Pregnancy outcomes were categorized into cases and controls in two ways. our definition was based on the following; Group 1 : Any PLBW cases Vs. All NBW controls Group 2 : PLBW cases Vs. NBW controls A periodontal exam was performed on the Ramfjord( #16, 21, 24, 36, 41, 44) teeth and Clinical evaluation consisted of probing pocket depth, loss of attachment, gingival index and gingival crevicular fluid amount. Subgingival plaque samples were collected by three sterile #35 paper points. The total number of anaerobic colonies and aerobic bacteria were enumerated after incubation. Antisera to P. gingivalis, P. intermedia, A. actinomycetemcomitans were produced in white rabbits with live whole cells suspensions. The specific fluorescent bacteria obtained by immunofluorescence and total cell counts obtained by dark-field microscopy were counted on four fields. The percent of each specific microorganism in the total cell count was determined. Results Any PLBW and PLBW cases showed significantly greater probing depth and attachment loss than all NBW and NBW controls. Cases group had significantly increased anaerobic bacterial counts compared with control group and no differences in the other microbes. This study confirmed that periodontal disease is a statistically significant risk factor for PLBW by investigating clinical parameters and subgingival plaque analysis.

불화나트륨을 함유한 저작성 정제의 치태제거 및 치은염 완화 효과에 관한 임상시험

배규현,설양조,류인철,한수부,최상묵,정종평,Bae, Kyoo-Hyun,Seol, Yang-Jo,Rhyu, In-Chul,Han, Soo-Boo,Choi, Sang-Mook,Chung, Chong-Pyoung 대한치주과학회 1999 Journal of Periodontal & Implant Science Vol.29 No.2

The objective of this study was to evaluate the effect a chewable tablet containing sodium flouride and lauroyl sodium sulfate on removing plaque and inhibiting gingival inflammation. A randomized parallel study was designed. 100 voluteers participated in the study. There were two test groups each with 30 subject. Test group A was instructed to brush once in the morning, and to use the tablet once in the afternoon and once in the evening. Test group B was instructed to use the tablet three times a day without brushing. There were two control groups each with 20 subjects. Control group A was instructed to brush once in the morning only. Control group B was instructed not to brush all. Two weeks before the test period, the subjects received through tooth cleaning and polishing. At baseline, GI, PI, BOP, and GCF of the Ramfjord teeth were measured in all groups. Bacteria culture was done with the plaque sampled from tooth with the deepest pocket. After 5 days, clinical indices were measured and the bacterial culture was repeated. Control group B was dropped from the study after this period. All the other groups remained and the indices and the culture was repeated after 2 weeks, and 3weeks. Also whether the oral mucosa showed signs of irritation was monitored throughout the test period. Test group A showed less PI, GI, BOP, probing depth, GCF than test group B or control group A. Especially, PI and the BOP was significantly less than that of the group that was instructed to brush once a day. This implies that the added use of this tablet aids in plaque removal in people who brush just once a day. Test group A showed increase of cocci, decrease of motile rods, and decrease of spirochetes after 14-21 days. And this was significantly different from the control group A. At no time of the test period did any of the subjects show signs of irritation of the oral mucosa or adverse reactions. Following conclusions could be obtained from this study. This chewable tablet for enhanced oral hygiene could be used as an adjunct to oral hygiene in people who do not brush adequately. The use of this tablet decreased the number of subgingival bacteria, and this could be effective in plaque removal and for prevention of gingival inflammation.

교원섬유 기질을 이용한 3차원 세포 배양 모델에 기계적인 힘을 가하는 경우 기질을 통하여 세포에 가해지는 힘의 특성에 대한 비교 분석 연구

김현종(Hyeon-Jong Kim),류인철(In-Chul Rhyu),박준우(Jun-Woo Park),김성곤(Seong-Gon Kim) 대한구강악안면외과학회 2009 대한구강악안면외과학회지 Vol.34 No.2

Different kinds of forces can be applied to the biological tissue. The analysis of the applied force is highly important to explain the mechanism of cellular response. In this study, the applied force to the collagen gel was analyzed by the finite elements analysis. The model received two different kinds of static force (compression and tension). The force range was 50g to 400g. In results, von Mises stress was concentrated in the peripheral region in the compression model. It was concentrated in the central area in the tension model. However, the compressive force was high in the peripheral area of the compression model and the tensional force was also high in the same area of the tension model. In conclusion, the applied force could be different to the region and it should be considered in the experiment to analyze the effects of the mechanical force on the cells.