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배양된 사람 치주인대세포와 골수유래간엽줄기세포의 분화에 미치는 법랑기질유도체 (Enamel Matrix Derivative, EMD)의 영향
박상규,주성숙,권용대,최병준,김영란,이백수,Park, Sang-Gyu,Jue, Seong-Suk,Kwon, Yong-Dae,Choi, Byung-Joon,Kim, Young-Ran,Lee, Baek-Soo 대한악안면성형재건외과학회 2009 Maxillofacial Plastic Reconstructive Surgery Vol.31 No.4
Introduction: Enamel matrix derivative (EMD) is a protein which is secreted by Hertwig root sheath and plays a major role in the formation of cementum and attachment of peridontium. Several studies have shown that EMD promoted the proliferation and differentiation of preosteoblasts, osteoblasts and periodontal ligament cells in vitro: however, reports showing the inhibition of osteogenic differentiation by EMD also existed. This study was designed to simultaneously evaluate the effect of EMD on the two cell lines (human mesenchymal stem cells: hMSC, human periodontal ligament derived fibroblasts: hPDLCs) by means of quantitative analysis of some bone related matrices (Alkaline phosphatase : ALP, osteopontin ; OPN, osteocalcin ; OC). Materials and Methods: hMSCs and hPDLCs were expanded and cells in the 4${\sim}$6 passages were adopted to use. hMSc and hPDLCs were cultured during 1,2,7, and 14 days with 0, 50 and 100 ${\mu}g/ml$ of EMD, respectively. ALP activity was assessed by SensoLyte ALP kit and expressed as values of the relative optical density. Among the matrix proteins of the bony tissue, OC and OPN were assessed and quantification of these proteins was evaluated by means of human OC immunoassay kit and human OPN assay kit, respectively. Results: ALP activity maintained without EMD at $1,2^{nd}$ day. The activity increased at $7^{th}$ day but decreased at $14^{th}$ day. EMD increased the activity at $14^{th}$ day in the hPDLCs culture. In the hMSCs, rapid decrease was noted in $7^{th}$ and $14^{th}$ days without regard to EMD concentrations. Regarding the OPN synthesis in hPDLCs, marked decrease of OPN was noted after EMD application. Gradual decrease tendency of OPN was shown over time. In hMSCs, marked decrease of OPN was also noted after EMD application. Overall concentration of OPN was relatively consistent over time than that in hPDLCs. Regarding the OC synthesis, in both of hPDLCs and hMSCs, inhibition of OC formation was noted after EMD application in the early stages but EMD exerted minimal effect at the later stages. Conclusion: In this experimental condition, EMD seemed to play an inhibitory role during the differentiation of hMSCs and hPDLCs in the context of OC and OPN formation. In the periodontium, there are many kinds of cells contributing to the regeneration of oral tissue. EMD enhanced ALP activity in hPDLCs rather than in hMSCs and this may imply that EMD has a positive effect on the differentiation of cementoblasts compared with the effect on hMSCs. The result of our research was consistent with recent studies in which the authors showed the inhibitory effect of EMD in terms of the differentiation of mineral colony forming cells in vitro. This in vitro study may not stand for all the charateristics of EMD; thus, further studies involving many other bone matrices and cellular attachment will be necessary.
PCR 과 Southern hybridization 을 이용한 구지뽕나무와 무궁화의 클론감별
박상규,류장발 ( Sang Gyu Park,Jang Bal Ryu ) 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.41 No.1
Polymerase chain reaction (PCR) and Southern hybridization analyses were carried out to identify clones of silk worm thorn (Cudraria tricuspidata) and Rose of sharon (Hibiscus syriacus) which look like one tree with two ar three, branches or two or three different trees. For PCR five different PCR primers (17∼24 nucleotides) are derived from CaMV 35S promoter, nopaline synthase terminator and coding region of thylakoid membrane protein gene. In the case of silk worm thorn, about 500 bp of PCR product was produced from DNAs of one tree or branch in the presence of 35S primer alone. Southern hybridization analysis of genomic DNAs hybridized with ^(32)P labeled PCR product showed that the same size of DNA fragments were hybridized with different intensities. In addition, PCR analyses using 20 different primers of OPERON 10-mer kits showed that only OPA01 primer produced PCR products of different size. These results indicate that two different trees of silk worm thorn combined to one tree. In the case of the Rose of Sharon, the same size of PCR products were produced from three different samples but Southern hybridization with the above PCR product as a probe did not show any hybridized bands. PCR analyses in the presence of OPERON 10-mers showed OPA04 and OPA13 produced different products including same sizes of products. These, results indicate that three different trees of the Rose of Sharon seem to be derived from the tree.
유기금속 화학증착법을 이용한 탄탈륨 산화 박막의 층덮힘 특성 연구
박상규,윤종호,남갑진,Park, Sang-Gyu,Yun, Jong-Ho,Nam, Gap-Jin 한국재료학회 1996 한국재료학회지 Vol.6 No.1
본 연구에서는 PET(PentaEthoxy Tanatalum:Ta(OC2H5)5) 유기금속 화합물 전구체를 사용하여 차세대 초고집적회로 제조시 고유전체 물질로 유망한 Ta2O5 박막을 열화학증착 방법에 의하여 증착하였다. 본 증착실험을 통하여 여러 가지 운속기체, 기판온도, 반응압력 등의 공정변수가 층덮힘에 미치는 영향을 고찰하였으며 Monte Carlo 전산모사 결과와 기판온도 변화에 따른 층덮힘 패턴의 변화에 대한 실험결과를 비교하여 부착계수를 산출하였다. 운송기체로는 N2, Ar, He을 바꿔가며 실험하였으며 He>N2>Artns으로 층덮힘이 양호한 것으로 나타났다. 이는 운송기체의 종류에 따라 운동량 확산도, 열 확산도, 물질 확산도 등의 이동현상 특성값들이 다르기 때문이라 생각된다. 기판온도의 증가는 운송기체의 종류에 관계없이 층덮힘을 악화시켰으며 도랑내부에서의 Knudsen 확산과 표면반응물의 탈착에 비해 표면반응이 보다 지배적인 역할을 담당함을 알 수 있었다. 또한 질소를 운송기체로 사용한 경우에 부착계수의 겉보기 활성화 에너지는 15.9Kcal/mol로 나타났다. 그리고 3Torr 이하에서 반응압력이 증가하는 반응압력이 증가하는 경우에는 물질 확산도의 감소 효과 때문에 층덮힘이 악화되었다. 본 연구결과 3Torr, 35$0^{\circ}C$에서 He 운송기체를 이용한 경우가 가장 우수한 층덮힘을 얻을 수 있는 최적 공정 조건임을 알 수 있었다.