유전자재조합 인간 골형성단백2 및 생흡수성고분자를 이용한 골형성유도체의 개발

이장희,김각균,안강민,김종원,이종호 大韓顎顔面成形再建外科學會 1999 Maxillofacial Plastic Reconstructive Surgery Vol.21 No.4

We tested the bone regenerating capacity and histologic response of bioresorbable matrix-type implant, which was made with Poly(lactide-co-glycolide)(PLGA) and bone apatite for the carrier of bone morphogenetic protein(BMP). The critical size defect of 8mm in diameter was created at the calvaria of SD rats(n=18), and repaired with polymer implant with 15㎍ of rhBMP-2(n=9) or without it(n=9). At 2 weeks, 1 months after implantation, the animals were sacrified(3 animals at every interval and group) and histologically evaluted. The calvarial defect which was repaired with polymer with BMP healed with newly formed bone about 70% of total defect. But that without BMP showed only 0 to under 30% bony healing. Inflammtory response was absent in both group through the experimental period, but there's marked foreign body giant response though it was a little less significant in polymer with BMP group. As the polymer was resorbed, the space was infiltrated and replaced by fibrovascular tissue, not by bone. In conclusion, our formulation of bioresorbable matrix implant loaded with bone morphogenetic protein works good as a bone regenerating material. However, it is mandatory to devise our system to have better osteoinductive and osteoconductive property, and less mutinucleated giant cell response.

Whole-genomic DNA probe를 이용한 구강 Streptococcus 균종의 식별

이영호,이장희,김각균,최선진 대한구강생물학회 1992 International Journal of Oral Biology Vol.16 No.1

DNA hybridization with whole-genomic DNA probe and restriction pattern analysis were done in search for a useful method for identification of the species of mutans group streptocci. Dot blots of genomic DNA of five strains (serotypes a, b, c and g) of mutans streptococci and on strain of Streptococcus sanguis were hybridized with ^32P-labeled whole-genomic DNA of Streptococcus mutans strain 10449 (serotype c) as a probe. Also, genomic DNAs of each strain were digested with two restriction enzymes (EcoRI and HindIII) and the restriction patterns were compared on 0.7% agarose gel after electrophoresis. Whole-genomic DNA probe of Strep. mutans strain 10449 was able to detect 3ng of homologous DNA per dot and was also able to detect the same amount of genomic DNA of a different strain of the same serotype with about the same intensity as shown on the autoradiogram. Genomic DNAs of differnet serotypes of mutans streptococci and of Strep. sanguis strain MPC1 were hybridized weakly with the probe at 50-100ng per dot. Genomic DNAs of six bacteria of two differnet genera (Bacteroides gingivalis and intermedius) were not hybridized with the probe at any concentration used in experiment. Restriction-digested patterns of genomic DNAs were all different from strain to strain, indicating genomic structures between each serotypes of mutans streptococci.

Hydroxyapatite 비드에 부착한 [^3H] - 표지된 세균의 씬틸레이션 측정에 관한 연구

이시영,이장희,최선진 대한구강생물학회 1989 International Journal of Oral Biology Vol.13 No.2

This study investigated the possible involvement of sample self absorption when [^3H]-labeled Streptococcus mutans cells that adhered to spheroidal hydroxyapatite beads were counted by scintillation spectrophotometer. To determine whether or not the beads cause self absorption of the energy of β particles, the beads-adhered bacterial cells were removed and counted alone. Two procedures were employed to remove bacterial cells from the beads. First, bacteria-adhered beads were dissolved with 0.33 N HCl, the dissolved solution was filtered to collect bacteria, and the filter was counted. Second, the bacteria adhered to beads were desorbed with potassium phosphate buffer, bacteria were collected on filter, and the filter was counted. Whatman glass microfibre filters GF/A and GF/F were used and filtration was carried out either with suction or without suction (i.e., by blotting). When the radioactivity was counted by blotting on GF/F filter, the count rate was increased to 280% in HA and 240% in SHA as compared with CPM of the control. Count rate of the bacteria was lower when filtered with suction than without suction. However, there was no difference in count rate when intact cell suspensions filtered either with suction of without were counted. Though HCl-treated cells were observed as intact by Gram staining, the above results seem to indicate that cells were damaged when treated with HCl, thus some of the DNA leaked out of the cells when filtered with suction. When the buffer-desorbed bacterial cells were filtered by blotting and counted, the count rate was increased to 290% in HA and 350% in SHA as compared with CPM of the control. The results of the present study mean that a high count rate is obtainable when the sample self absorption is removed. Therefore, the sample self absorption should be considered in order to measure the correct number of bacteria adhered to HA beads.

cDNA array를 이용한 피골세포 분화관련 유전자의 탐색

조영준,이장희,이창섭,이상호 大韓小兒齒科學會 2002 大韓小兒齒科學會誌 Vol.29 No.2

RAW 264.7과 이것이 분화한 파골세포양 세포에서 파골세포 분화관련유전자의 전체적인 유전자 발현을 조사하기 위해 cDNA array 방법을 사용하였다. 1176 cDNA spot grid가 있는 Mouse Atlas cDNA array 결과를 확인하기 위하여 역전사 효소 중합반응 검사를 시행하였다. cDNA array 결과 6개의 유전자가 2.5% 이상 발현이 증가하였으며(PKC beta Ⅱ, POMC, PTEN 등), 16개의 유전자가 2.5%이상 발현이 감소하였다(Osteopontin, Cyclin D1, Cathepsin C, PTMA 등). PKC beta Ⅱ유전자의 역전사-효소 중합 반응 검사 결과 이 유전자를 확인할 수 있었다. 파골세포 분화 결과 RAW 264.7 세포주에 비해 파골세포양 세포에서 PKC beta Ⅱ 유전자가 발현이 많았다. 파골세포 분화 관련 유전자는 RAW 264.7 세포주의 파골세포 분화와 연관을 보였다. To examine the global gene expression of osteoclastogenesis-related genes in RAW 264.7 and its differentiated OCLs through the use of Atlas Mouse cDNA Array 2.1 membranes printed with 1176 well-characterized mouse genes involved in biology. Both samples were screened in parallel using cDNA expression arrays. The array re-sults were additionally validated using RT-PCR. The results of cDNA arrays showed that 6 genes were up-regu-lated >2.5-fold (PKC beta Ⅱ, POMC, PTEN, etc) and 16 genes were down-regulated >2.5-fold (Osteopontin, Cyclin D1, Cathepsin C, PTMA, etc) in both samples at the mRNA level. RT-PCR analysis of PKC beta Ⅱ of these differentially expressed genes gave result consistent with cDNA array findings. The result of osteoclastoge-nesis showed that the PKC beta Ⅱ gene was overexpressed in OCLs compared with RAW264.7 cell line. Osteoclastogenesis-related genes are differentially expressed in RAW264.7 cell line and its differentiated OCLs. its gene overexpression correlates with osteoclast differentiation in RAW264.7 cell line.

고정성 교정장치 장착 환자의 타액내 구강미생물군의 수준에 관한 연구

김동훈,이장희,김광원 대한치과교정학회 1993 대한치과교정학회지 Vol.23 No.3

This study was done to evaluate the effect of fixed orthodontic patients on the level of oral streptococci, Streptococcus mutans, lactobacilli, yeasts in saliva. 35 patients wearing bands were compared with age-matched 35 non-banded control group by conlony counting method on the specially designed culture medium. The following results were obtained ; 1. The colony forming unit(CFU) of total streptocci per militer of saliva in subjects with or without orthodontic treatment showed no significant statistical difference between them(p>0.05). 2. The colony forming unit(CFU) of total Streptococcus mutans per mililiter of saliva in subjects with orthodontic treatment showed significantly higher than those without orthodontic treatment(p<0.05) 3. The colony forming unit(CFU) of total lactobacilli per mililiter of saliva in sujects with or without orthodontic treatment showed no significant statistical difference between them but higher tendency in those with orthodontic treatment(p=0.052). 4. The colony forming unit(CFU) of total yeasts per mililiter of saliva in subjects with or without orthodontic treatment showed no significant statistical difference between them(p>0.05).

Antiosteoporotic Constituents from Salvia miltiorrhiza

Lee Hyang-Hee,Han Song-Hee,Lee Zang Hee,Kim Hong-Hee,Woo Eun-Rhan 대한약학회 2004 大韓藥學會 總會 및 學術大會 Vol.2004 No.1

Caveolin-1 Regulates Osteoclastogenesis and Bone Metabolism in a Sex-dependent Manner

Lee, Yong Deok,Yoon, Soo-Hyun,Park, Cheol Kyu,Lee, Jiyeon,Lee, Zang Hee,Kim, Hong-Hee American Society for Biochemistry and Molecular Bi 2015 The Journal of biological chemistry Vol.290 No.10

<P>Lipid raft microdomains have important roles in various cellular responses. Caveolae are a specialized type of lipid rafts that are stabilized by oligomers of caveolin proteins. Here, we show that caveolin-1 (Cav-1) plays a crucial role in the regulation of osteoclastogenesis. We found that caveolin-1 was dramatically up-regulated by receptor activator of nuclear factor κB ligand (RANKL), the osteoclast differentiation factor. Knockdown of <I>Cav-1</I> reduced osteoclastogenesis and induction of NFATc1, the master transcription factor for osteoclastogenesis, by RANKL. Consistent with the <I>in vitro</I> results, injection of caveolin-1 siRNA onto mice calvariae showed reduction in RANKL-induced bone resorption and osteoclast formation. Moreover, <I>Cav-1</I><SUP>−/−</SUP> female mice had higher bone volume and lower osteoclast number compared with wild type mice. However, <I>Cav-1</I><SUP>−/−</SUP> male mice had both osteoclast and osteoblast numbers higher than wild type mice with no difference in bone volume. The sex dependence in the effect of <I>Cav-1</I> deficiency was partly attributed to decreased receptor activator of nuclear factor κB and increased cFms expression in osteoclast precursors of female and male mice, respectively. Taken together, these data demonstrate that Cav-1 has a complicated but critical role for osteoclastogenesis.</P>

POSTERS / PM-001 : ROLE OF THE TRAF PROTEIN IN THE ACTIVATION OF THE JNK PATHWAY BY RANK

(Zang Hee Lee) 한국생화학분자생물학회 (구 한국생화학회) 1999 6th IUBMB SEOUL CONFERENCE Vol.- No.-

5-Lipoxygenase mediates RANKL-induced osteoclast formation via the cysteinyl leukotriene receptor 1.

Lee, Jung-Min,Park, Hyojung,Noh, A Long Sae Mi,Kang, Ju-Hee,Chen, Ling,Zheng, Ting,Lee, Juhyun,Ji, Sun-Young,Jang, Chang-Young,Shin, Chan Soo,Ha, Hyunil,Lee, Zang Hee,Park, Hea-Young,Lee, Dong-Seok,Yi American Association of Immunologists 2012 Journal of Immunology Vol. No.

<P>5-Lipoxygenase (5-LO) catalyzes the formation of two major groups of leukotrienes, leukotriene B4 and cysteinyl leukotrienes (CysLTs), and it has been implicated as a promising drug target to treat various inflammatory diseases. However, its role in osteoclastogenesis has not been investigated. In this study, we used mouse bone marrow-derived macrophages (BMMs) to show that 5-LO inhibitor suppresses RANKL-induced osteoclast formation. Inhibition of 5-LO was associated with impaired activation of multiple signaling events downstream of RANK, including ERK and p38 phosphorylation, and IκB degradation, followed by a decrease in NFATc1 expression. Ectopic overexpression of a constitutively active form of NFATc1 partly rescued the antiosteoclastogenic effect of 5-LO inhibitor. The knockdown of 5-LO in BMMs also resulted in a significant reduction in RANKL-induced osteoclast formation, accompanied by decreased expression of NFATc1. Similar effects were shown with CysLT receptor (CysLTR)1/2 antagonist and small RNA for CysLTR1 in BMMs, indicating the involvement of CysLT and CysLTR1 in 5-LO-mediated osteoclastogenesis. Finally, 5-LO inhibitor suppressed LPS-induced osteoclast formation and bone loss in the in vivo mouse experiments, suggesting a potential therapeutic strategy for treating diseases involving bone destruction. Taken together, the results of this study demonstrate that 5-LO is a key mediator of RANKL-induced osteoclast formation and possibly a novel therapeutic target for bone-resorption diseases.</P>

Methaol extract of Theaceae supresses nitrite synthesis and expression of inflammatory cytokines

Lee Tae-Hoon,Chung Dae-Kyun,Kim Hong-Hee,Lee Zang-Hee,Kim Ji-Young 한국작물학회 2006 한국작물학회 학술발표대회 논문집 Vol.2006 No.-