• Title/Summary/Keyword: 구강외과

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GENE EXPRESSION PATTERNS INDUCED BY $TAXOL^{(R)}$ AND CYCLOSPORIN A IN ORAL SQUAMOUS CELL CARCINOMA CELL LINE USING CDNA MICROARRAY (cDNA Microarray를 이용한 구강편평세포암종 세포주에서 $Taxol^{(R)}$과 Cyclosporin A로 유도된 유전자 발현양상)

  • Kim, Yong-Kwan;Lee, Jae-Hoon;Kim, Chul-Hwan
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.3
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    • pp.202-212
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    • 2006
  • It is well-known that paclitaxel($Taxol^{(R)}$), which is extracted from the pacific and English yew, has been used as a chemotherapeutic agent for ovarian carcinoma and advanced breast carcinoma and Cyclosporin A, which is highly lipophilic cyclic peptide and isolated from a fungus, has been also used as an useful immunosuppressive drug after transplantation and is associated with cellular apoptosis. Since 1953, in which James Watson, Rosalind Franklin and Francis Crick discovered the double helical structure of DNA, a few kinds of techniques for identifying gene expression have been developed. In postgenomic period, many of researchers have used the DNA microarray which is high throughput screening technique to screen large numbers of gene expression simultaneously. In this study, we searched and screened the gene expression in the oral squamous cell carcinoma cell lines treated with $Taxol^{(R)}$, cyclosporin or cyclosporin combined with $Taxol^{(R)}$ using cDNA microarray. The results were as following; 1. It was useful that the appropriate concentration of Cyclosporin A and $Taxol^{(R)}$ used in oral squamous cell carcinoma cell line was under 1${\mu}g/ml$ and 3${\mu}g/ml$. 2. In the experimental group in which $Taxol^{(R)}$ and $Taxol^{(R)}$ + Cyclosporin A were used, the cell growth was extremely decreased. 3. In the group in which Cyclosporin A was used, the MTT assay was rarely decreased which means the activity of succinyl dehydrogenase is remained in mitochondria but in the group in which the mixture of Cyclosporin A and $Taxol^{(R)}$ were used, the MTT assay was extremely decreased. 4. In the each group in which Cyclosporin A(3 ${\mu}g/ml$) and $Taxol^{(R)}$(1 ${\mu}g/ml$) were used, the cell arrest was appeared in $G_2/M$ phase and in the group in which $Taxol^{(R)}$(3 ${\mu}g/ml$) was used, the cell arrest was appeared in both S phase and $G_2/M$ phase. 5. In the oral squamous cell carcinoma cell line treated with $Taxol^{(R)}$, several genes including ANGPTL4, RALBP1 and TXNRD1, associated with apoptosis, SUI1, MAC30, RRAGA and CTGF, related with cell growth, HUS1 and DUSP5, related with cell cycle and proliferation, ATF4 and CEBPG, associated with transcription factor, BTG1 and VEGF, associated with angiogenesis, FDPS, FCER1G, GPA33 and EPHA4 associated with signal transduction and receptor activity and AKR1C2 and UGTA10 related with carcinogenesis were detected in increased levels. The genes that showed increaced expression in the oral squamous cell carcinoma cell line treated with Cyclosporin A were CYR61, SERPINB2, SSR3 and UPA3A which are known as genes associated with cell growth, carcinogenesis, receptor activity and transcription factor. The genes expressed in the HN22 cell line treated with cyclosporin combined with $taxol^{(R)}$ were ALCAM and GTSE1 associated with cancer invasiveness and cell cycle regulation.

A STUDY ON THE MANDIBULAR MOVEMENT CHANGES AFTER ORTHOGNATHIC SURGERY IN SKELETAL CLASS III PATIENTS (악교정 수술 후 골격성 III급 부정교합 환자의 하악운동 변화에 관한 연구)

  • Nahm, Dong-Seok;Suhr, Cheong-Hoon;Yang, Won-Sik;Chang, Young-Il
    • The korean journal of orthodontics
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    • v.27 no.2
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    • pp.273-282
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    • 1997
  • The purpose of this study was to evaluate changes in mandibular movement patterns after orthognathic surgery in skeletal Class III patients. The sample consisted of 20 Class III malocclusion patients(9 males, 11 females). Just before and after(2-7months) surgery, maximum opening & closing movement, mandibular border movement on sagittal, frontal and horizontal planes were recorded using Sirognathograph & BioPak EGN. On each record, 21 items were measured and statistically analyzed. The results were as follows 1. Angle of protrusive movement on sagittal plane showed greatest change after surgery. Also, as the incisal guidance was established by surgery, straight path of protrusive movement became curved line. 2. Maximum opening distance and maximum antero-posterior distance on maximum opening & closing movement, maximum opening distance on sagittal plane, angle of left lateral excursion on frontal plane were statistically significant after snrgery(p<0.01). 3. Maximum width of lateral excursion on frontal plane, distane of right lateral excursion and angle of maximum left lateral excursion on horizontal plane were statistiraily significant after surgery(p<0.05). 4. Maximum opening distance and maximum antero-posterior distance on maximum opening & closing movement showed significant differences according to post-surgical time(p<0.05). More recovery of range of movement occured in 5-7month group than in 2-3month group. 5. As the occlusal interferences were removed by orthognathic surgery, irregular opening & closing path became smooth curve.

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FABRICATION OF MYOMUCOSAL FLAP USING CULTURED ORAL EPITHELIUM IN RABBIT MODEL (가토모델에서 배양 구강상피를 이용한 근-점막 피판의 형성에 관한 연구)

  • Shin, Young-Min;Chung, Hun-Jong;Ahn, Kang-Min;Park, Hee-Jung;Sung, Mi-Ae;Kim, Soung-Min;Hwang, Soon-Jung;Kim, Myung-Jin;Jahng, Jeong-Won;Kim, Sung-Po;Yang, Eun-Kyung;Song, Kye-Yong;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.3
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    • pp.226-237
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    • 2005
  • Purpose : Extensive defect of oral and maxillofacial area is usually reconstructed with composite flap including skin paddle. However, if the defects are lined with only skin components, the mucosa's role in mastication and texture are not restored. Furthermore, stiffness and hair-growing prevent denture rehabilitation and good oral hygiene. This study was performed to overcome the disadvantages of composite soft tissue flaps including the skin and to make a model for myo-mucosal flaps. Materials and methods : Buccal mucosa sized $0.5\times1.0\;cm^2$ from New Zealand rabbit (around 1.5kg) was harvested and cultivated by the modification of Rheinwald and Green's keratinocyte culture method. Cultured mucosa was grafted on the fascia of latismus dorsi as form of mucosal sheet. After 7, 10, 14 days, the myomucosal flap was excised and evaluated under light microscope with H & E and immunohistochemical staining. As control group, harvested buccal mucosa from rabbit was transplanted to gracilis muscle(n=6). Results : From 7 days after prelamination, the basal layer of the grafted mucosa resembled that of normal mucosa. As control group, transplanted mucosa had original shape but there's slight inflammatory reaction. Prelaminated mucosa has 19.8$\pm$4.59 cell layers and some samples have more than 20 layers. The expression rate of PCNA was relatively strong (42.9%$\pm$14.1) at the basal layer of grafted mucosa and the laminin was found at the basal layer. On the contrary, prelaminated mucosa at 10 days showed moderate expression rate of PCNA(32.4%$\pm$4.62). We found the mucosal layer was somehow disappeared and there is strong inflammatory reaction. After 14 days prelamination, the grafted oral keratinocytes were almost disappeared and expression of PCNA was not observed. Conclusion : We can make 75 fold large mucosal($3850mm^2$) sheet from small samples of mucosa $(50mm^2)$. Epithelial sheet that grafted on the fascia of muscle underwent differentiation and proliferation. But after 10, 14 days, there was strong inflammatory reaction and the grafted mucosa was destroyed from surface layer. In rabbit model, transfer of fascio-mucosal flap should be done from 7 to 10 days after prelamination.

HISTOMORPHOLOGIC CHANGE AND REGENERATION OF THE ADVENTITIA-STRIPPED AND FROZEN RAT FEMORAL ARTERY:HISTOLOGIC, HISTOMORPHOMETRIC, IMMUNOHISTOCHEMICAL AND SCANNING ELECTRON MICROSCOPIC STUDY (백서 대퇴동맥 외막층 제거후 혈관동결시 조직형태학적 변화 및 재생에 관한 연구)

  • Kim, Tae-Deug;Kim, Kyung-Wook;Lee, Jae-Hoon;Kim, Chang-Jin
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.25 no.4
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    • pp.281-294
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    • 1999
  • Vasospasm causes microvascular surgery to fail as a main factor in the loss of transferred flap dye to the diminution of blood flow in reconstruction surgery. Although there has been extensive research to resolve the vasospasm problem, no one has reached an ideal solution to date. However, cryotherapy, which is often used for destruction of tumor lesions, is being presented as a new way of releasing vasospasm. After making a histomorphometric measurement on vasodialation during the course of 1, 3 and 7 days, 2 and 4 weeks, and 5 months periods and observing the change of blood vessel in a histologic, immunohistochemical, and scanning electronic microscopic approach, the results were as follows : 1. Vascular inner diameters of the experimental 1 and 3 days groups were measured $476.3{\pm}28.20{\mu}m$, $497.15{\pm}48.79{\mu}m$ respectively showing statistically meaningful vasodilation(P<0.05), which continued by the experiment 4 weeks group. However, in the experimental 5 months group, the vascular inner diameter appeared similar to the control groups. Even though the thickness of smooth muscular layers come out to be thinner in all the experimental groups compared to the control group, it was difficult to find any statistical meaningfulness. In addition, the vascular external diameters of every experimental groups were shown to be longer than the control group. 2. In light microscopic view, severe injury was evident on the smooth muscular layer cell from the experimental 1 day group, started recovering partially from the experimental 7 days group, and was mostly restored in the experimental 4 weeks group and layer of adventitial stripping were nearly recoverd 2 weeks group. 3. The PCNA positive cells of smooth muscular layer were observed from the experimental 7 days group and had a tendency to increase by the experimental 2 weeks group. In the experimental 4 weeks and 5 months group, the number of PCNA possitive cells observed was comparable to the control group. 4. ${\alpha}$-SMA level of smooth muscular layer cells, having been significantly lower than the control group in the severly damaged experimental 1 day group. It was seen to be increased in the experimental 7 days group and turned out to show similar ${\alpha}$-SMA level in 4 weeks to the control group. 5. In the view of SEM, the endothelial cells were destructed and falling off, and also present the appearance of flattening in the experiment 1 day group. The endothelial layer cells started partially recovering from the 7 days group after the freezing injury. On 4 weeks and 5 months, the endothelial cells were fully coverd the damaged area, also it's appearance is similar to control group. In conclusion, the vascular freezing after the removal of adventitia caused damages to smooth muscular layer cells, and brought about vasodilation, which continued by the 4th week. The smooth muscular layer cells started partially reviving from the 7rd day after the damage by vascular freezing, and recovered their similar figure to the control group's 4 weeks later. This was considered the result of cells which surround the damaged blood vessel being influxed into the smooth muscular layers. Therefore, this local freezing injury on the blood vessel was thought to be applied clinically to relieve severe vasospasm which cannot be treated by vasodilation drug, a microvascular surgery.

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STABILITY OF OBWEGESER II METHOD IN MANDIBULAR PROGNATHIC OR ANTERIOR OPEN BITE PATIENTS (하악전돌증 및 개교합 환자에 있어 Obwegeser Ⅱ method의 안정성)

  • Jung, Chang-Wook;Nam, Jeong-Hun;Lee, Sang-Han;Kwon, Tae-Geon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.1
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    • pp.25-33
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    • 2004
  • The purpose of this study was to evaluate the postoperative stability of the severe open bite or mandibular prognathic patients after mandibular set back surgery by Obwegeser II method. There were 19 patients who had been undergone Obwegeser II method. The horizontal and vertical position of the cephalometric points were measured preoperation and immediate postoperation, postoperative 1 month, postoperative 6 months ; were analyzed by linear measurement to evaluate changes in skeletal landmark and the relapse was compared between open bite group and prognathism group. By the operation, horizontal change of B was $6.84{\pm}4.35mm$ and vertical change of B was $6.28{\pm}3.25mm$ in open bite group and horizontal change of B was $14.20{\pm}4.81mm$ and vertical change of B was $1.99{\pm}2.66mm$ in prognathism group, horizontal change of Pog was $3.82{\pm}5.71mm$ and vertical change of Pog was $5.38{\pm}2.11mm$ in open bite group and horizontal change of Pog was $13.24{\pm}5.99mm$ and vertical change of Pog was $1.91{\pm}0.94mm$ in prognathism group. Between immediate postoperation and postoperative 1 month, all skeletal landmarks change was no statistical difference (p>0.05) and there were no statistical difference between open bite group and prognathism group except x-Me landmark (p>0.05). Between postoperative 1 month and 6 months, horizontal change of B was $0.12{\pm}1.35mm$ and vertical change of B was $1.47{\pm}1.48mm$ in open bite group and horizontal change of B was $1.43{\pm}1.35mm$ and vertical change of B was $0.82{\pm}1.99mm$ in prognathism group, horizontal change of Pog was $0.13{\pm}1.40mm$ and vertical change of Pog was $0.88{\pm}1.71mm$ in open bite group and horizontal change of Pog was $1.08{\pm}1.74mm$ and vertical change of Pog was $0.47{\pm}1.57mm$ in prognathism group (p>0.05) and there were no statistical difference between open bite group and prognathism group (p>0.05). Between immediate postoperation and postoperative 6months, horizontal change of B was $0.24{\pm}1.17mm$ and vertical change of B was $1.87{\pm}1.63mm$ in open bite group and horizontal change of B was $1.54{\pm}1.55mm$ and vertical change of B was $1.04{\pm}1.96mm$ in prognathism group, horizontal change of Pog was $0.91{\pm}1.46mm$ and vertical change of Pog was $1.18{\pm}2.05mm$ in open bite group and horizontal change of Pog was $0.96{\pm}1.62mm$ and vertical change of Pog was $1.23{\pm}2.35mm$ in prognathism group (p>0.05) and there were statistical difference between open bite group and prognathism group in x-B, x-Pog, x-Gn, x-Me (p<0.05). Obwegeser II method is considered as one of the best operation when surgical correction of severe open bite or severe mandibular prognathism is needed.

The study of bone density assessment on dental implant sites (임플란트 식립 부위의 골밀도 평가에 관한 연구)

  • Park, Su-Won;Jang, Soo-Mi;Choi, Byoung-Hwan;Son, Han-Na;Park, Bong-Chan;Kim, Chang-Hwan;Son, Jang-Ho;Sung, Iel-Yong;Lee, Ji-Ho; Cho, Yeong-Cheol
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.417-422
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    • 2010
  • Introduction: Bone density is one of the important factors for the long term success of endosseous implants. The bone density varies from site to site and from patient to patient. A preoperative evaluation of the bone density is quite useful to oral surgeons for planning dental implantation. More accurate information on the bone density will help surgeons identify suitable implant sites, thereby increase the success rate of dental implantation. This study examined the correlation between the bone density measured preoperatively by computed tomography (CT) and the implant primary stability measured by resonance frequency analysis. Furthermore, the effects of the implant sites, gender, age and generalized systemic disorder patients on the bone density and primary implant stability were examined. Materials and Methods: One hundred and fourteen patients were selected. None of the patients had undergone a tooth extraction or bone graft history in the previous year. Preoperatively, the patients underwent CT scanning to evaluate the Hounsfield unit (HU), and resonance frequency analysis (RFA) was used to evaluate the implant primary stability at the time of implant installation. All implants were 4.0 mm diameter and 11.5 mm length US II. All patients were recorded and the HU and implant stability quotient (ISQ) value were evaluated according to the sites, gender and age. Results: The highest HU values were found in the mandibular anterior site ($827.6{\pm}151.4$), followed by the mandibular molar site ($797{\pm}135.1$), mandibular premolar site ($753.8{\pm}171.2$), maxillary anterior site ($726.3{\pm}154.4$), maxillary premolar site ($656.7{\pm}173.8$) and maxillary molar site ($621.5{\pm}164.9$). The ISQ value was the highest in the mandibular premolar site ($81.5{\pm}2.4$) followed by the mandibular molar site ($80.0{\pm}5.7$), maxillary anterior site ($77.4{\pm}4.1$), mandibular anterior site ($76.4{\pm}11.9$), maxillary premolar site ($74.2{\pm}14.3$) and maxillary molar site ($73.7{\pm}7.4$). The mean HU and ISQ value were similar in females and males. (HU: P=0.331, ISQ: P=0.595) No significant difference was also found in the age group respectively. However, the correlation coefficients between the variables showed a closed correlation between the HU and ISQ value. Conclusion: These results showed close correlation between the bone density (HU) and primary stability value (ISQ) at the time of implant installation (Correlation coefficients=0.497, P<0.01). These results strengthen the hypothesis that it might be possible to predict and quantify the initial implant stability and bone density from a presurgical CT diagnosis.

The effect of silk fibroin and rhBMP-2 on bone regeneration in rat calvarial defect model (백서 두개골결손모델에서 실크단백과 골형성단백 이식체가 골재생에 미치는 영향)

  • Nam, Jeong-Hun;Noh, Kyung-Lok;Pang, Eun-O;Yu, Woo-Geun;Kang, Eung-Sun;Kweon, Hae-Yong;Kim, Seong-Gon;Park, Young-Ju
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.366-374
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    • 2010
  • Introduction: This study evaluated the capability of silk fibroin (SF) and recombinant human bone morphogenetic protein-2 loaded SF (SF-BMP) as a bone defect replacement matrix when grafted in a calvarial bone defect of rats in vivo. Materials and Methods: A total 70 calvarial critical size defects (5.0 mm in diameter) made on 35 adult female Sprague-Dawley rats were used in this study. The defects were transplanted with (1) rhBMP-2 loaded silk fibroin graft (SF-BMP: 0.8+$10\;{\mu}g$), (2) Silk fibroin (SF: $10\;{\mu}g$), and (3) no graft material (Raw). The samples were evaluated with soft x-rays, alkaline phosphatase activity, calcium/phosphate quantification, histological and histomorphometric analysis at postoperative 4 and 8 weeks. Results: The SF-BMP group ($48.86{\pm}14.92%$) had a significantly higher mean percentage bone area than the SF group ($24.96{\pm}11.01%$) at postoperative 4 weeks.(P<0.05) In addition, the SF-BMP group ($40.01{\pm}12.43%$) had a higher % bone area at postoperative 8 weeks than the SF group ($33.26{\pm}5.15%$). The mean ratio of gray scale levels to the host bone showed that the SF-BMP group ($0.67{\pm}0.08$) had a higher mean ratio level than the SF group ($0.61{\pm}0.09$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.168 and P=0.243, respectively) The ratio of the calcium and phosphate contents of the SF-BMP ($0.93{\pm}0.22$) group was lower than that of the SF ($1.90{\pm}1.42$) group at postoperative 4 weeks. However, the SF-BMP group ($0.75{\pm}0.31$) had a higher Ca/$PO_4$ ratio than the SF ($0.68{\pm}0.04$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.126 and P=0.627, respectively) For the bone-specific alkaline phosphatase (ALP) activity, which is recognized as a reliable indicator of the osteoblast function, the SF-BMP ($23.71{\pm}8.60\;U/L$) groups had a significantly higher value than the SF group ($12.65{\pm}6.47\;U/L$) at postoperative 4 weeks.(P<0.05) At postoperative 8 weeks, the SF-BMP ($21.65{\pm}10.02\;U/L$) group had a lower bone-specific ALP activity than the SF group ($16.72{\pm}7.35\;U/L$). This difference was not statistically significant.(P=0.263) For the histological evaluation, the SF-BMP group revealed less inflammation, lower foreign body reactions and higher bone healing than the SF group at postoperative 4 and 8 weeks. The SF group revealed more foreign body reactions at postoperative 4 weeks. However, this immunogenic reaction decreased and the remnant of grafted material was observed at postoperative 8 weeks. For histomorphometric analysis, the SF-BMP group had a significantly longer bone length to total length ratio than those of the SF group at postoperative 4 and 8 weeks.(P<0.05) Conclusion: The rhBMP-2 loaded silk fibroin graft revealed fewer immunoreactions and inflammation as well as more new bone formation than the pure silk fibroin graft. Therefore, silk fibroin may be a candidate scaffold for tissue engineered bone regeneration.

The effect of heat shock protein 70 on inducible nitric oxide synthase during sepsis in rats (백서 패혈증 모델에서 HSP70의 과도 발현이 iNOS의 발현에 미치는 효과에 관한 연구)

  • Lee, Yong-Keun;Ahn, Yung;Leem, Dae-Ho;Baek, Jin-A;Ko, Seung-O;Shin, Hyo-Keun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.5
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    • pp.346-352
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    • 2010
  • Introduction: Heat shock protein70 (HSP70) is a highly conserved family of proteins produced after a variety of stresses. Many studies reported that the overexpression of HSP70 can improve the prognosis of the patients with sepsis through a reduction of the nitric oxide concentration. However, these results only revealed the effect of HSP70 and nitric oxide. No studies have examined the relationship between HSP70 and nitric oxide. The aim of this study was to evaluate the effect of the overexpression of HSP70 on the expression of inducible nitric oxide synthase and the nitric oxide concentration. In addition, the mechanism of the relationship of HSP70 and inducible nitric oxide synthase (iNOS) in sepsis was examined. Materials and Methods: The experiments were performed on male sprague-dawley rats. Sepsis was induced by a cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 hour after the initiation of sepsis. Serum and lung tissues were acquired from the rats 12 hours or 24 hours after the initiation of sepsis. The nitric oxide concentration, the expression of HSP70 in lung, and the gene expression of iNOS in lung were analyzed. The three groups, sham operation, CLP and CLP+GLN, were compared. Results: Compared to the other groups, in CLP+GLN, GLN administered after the initiation of sepsis enhanced the expression of HSP70 in the lung at 12 hours ($47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, P=0.025) and 24 hours ($47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, P=0.004). In CLP+GLN, GLN attenuated the expression of iNOS messenger RNA (mRNA) in the lung at 12 hours ($5,513.73{\pm}1,051.60$ vs. $4,167.17{\pm}951.59$, P=0.025) and 24 hours ($18,740.27{\pm}8,241.20$ vs. $9,437.65{\pm}2,521.07$, P=0.016), and reduced the concentration of nitric oxide in the serum at 12 hours ($0.86{\pm}0.48$ vs. $3.82{\pm}2.53$, P=0.016) and 24 hours ($0.39{\pm}0.25$ vs. $1.85{\pm}1.70$, P=0.025). Conclusion: The overexpression of HSP70 induced by the administration of GLN in sepsis attenuates the expression of the iNOS gene but reduces the nitric oxide concentration.

PERIPHERAL NERVE REGENERATION USING POLYGLYCOLIC ACID CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE TRANSFECTED SCHWANN CELLS IN RAT SCIATIC NERVE (BDNF 유전자 이입 슈반세포와 PGA 도관을 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Gao, En-Feng;Shin, Young-Min;Kim, Yoon-Tae;Hwang, Soon-Jeong;Kim, Nam-Yeol;Kim, Myung-Jin;Jo, Seung-Woo;Kim, Byung-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.6
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    • pp.465-473
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    • 2004
  • Purpose : The essential triad for nerve regeneration is nerve conduit, supporting cell and neurotrophic factor. In order to improve the peripheral nerve regeneration, we used polyglycolic acid(PGA) tube and brain-derived neurotrophic factor(BDNF) gene transfected Schwann cells in sciatic nerve defects of SD rat. Materials and methods : Nerve conduits were made with PGA sheet and outer surface was coated with poly(lactic-co-glycolic acid) for mechanical strength and control the resorption rate. The diameter of conduit was 1.8mm and the length was 17mm Schwann cells were harvested from dorsal root ganglion(DRG) of SD rat aged 1 day. Schwann cells were cultured on the PGA sheet to test the biocompatibility adhesion of Schwann cell. Human BDNF gene was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into E1 deleted region of adenovirus shuttle vector, pAACCMVpARS. BDNF-adenovirus was multiplied in 293 cells and purified. The BDNF-Adenovirus was then infected to the cultured Schwann cells. Left sciatic nerve of SD rat (250g weighing) was exposed and 14mm defects were made. After bridging the defect with PGA conduit, culture medium(MEM), Schwann cells or BDNF-Adenovirus infected Schwann cells were injected into the lumen of conduit, respectively. 12 weeks after operation, gait analysis for sciatic function index, electrophysiology and histomorphometry was performed. Results : Cultured Schwann cells were well adhered to PGA sheet. Sciatic index of BDNF transfected group was $-53.66{\pm}13.43$ which was the best among three groups. The threshold of compound action potential was between 800 to $1000{\mu}A$ in experimental groups which is about 10 times higher than normal sciatic nerve. Conduction velocity and peak voltage of action potential of BDNF group was the highest among experimental groups. The myelin thickness and axonal density of BDNF group was significantly greater than the other groups. Conclusion : BDNF gene transfected Schwann cells could regenerate the sciatic nerve gap(14mm) of rat successfully.

DEVELOPMENT OF BIOCOMPATIBLE DRESSING MATERIAL MADE OF COLLAGEN AND AMNIOTIC MEMBRANE AND WOUND HEALING EXPERIMENT IN RAT (양막과 콜라겐을 이용한 생체 적합 드레싱 소재 개발 및 백서 창상치유 실험)

  • Ahn, Kang-Min;Lee, Ji-Ho;Lee, Ui-Lyong;Lee, Jong-Ho;Lee, Jong-Won;Kim, Sung-Po;Yang, Eun-Kyung;Kim, Ki-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.3
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    • pp.189-199
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    • 2006
  • Purpose of study: Partial thickness skin graft is the golden standard regimen for full-thickness skin defect caused by burn or trauma. However, in case of extensive burns of more than 50% of total body surface area, the donor site is not sufficient to cover all defects. As a second choice, allograft, xenograft and synthetic materials have been used to treat skin defect. Among them the amniotic membrane(AM) was used as a biological dressing for centuries because of its potential for wound healing. In this study, quantification of EGF in AM and effect of AM-collagen complex on full thickness skin defects was examined. Materials & Methods: The concentration of EGF in fresh, deep frozen and freeze-dried AM was evaluated by ELISA. EGF-R immunostaining was performed in freeze-dried AM. SD rats weighing 250${\sim}$300g was used for wound healing experiment. Three full thickness skin defects(28mm diameter) were made on dorsal surface of SD rat. The control group was covered by Vaselin gauze and AM-collagen complex and $Terudermis^{(R)}$. was grafted in two other defects. Healing area, Cinamon's score were evaluated before biopsy. Grafted sites were retrieved at 3 days, 1 week, 2 weeks and 4 weeks after operation. H & E and Factor VIII immunohistochemical stain was performed to evaluate the microscopic adhesion and structural integrity and microvessel formation. Results: 1. EGF concentration of fresh, deep frozen and freeze-dried AM showed similar level and EGF-R was stained in epithelial layer of freeze-dried AM. 2. At 4 weeks after grafting, the healing area of AM-collagen and Terudermis group was 99.29${\pm}$0.71% and 99.19${\pm}$0.77 of original size. However, that of control group was 24.88${\pm}$2.90. 3. The Cinamon's score of AM-Collagen and $Terudermis^{(R)}$. group at 4 weeks was 15.6${\pm}$1.26 and 14.6${\pm}$3.13 and that of control group was 3.7${\pm}$0.95. Significant difference was observed among control and experimental groups(p<0.05). 4. Histologic examination revealed that AM protected leukocyte infiltration and epithelial migration was nearly completed at 4 weeks. $Terudermis^{(R)}$. group showed mild neutrophil infiltration until 2 weeks and completion of epithelization at 4 weeks. Control group showed massive leukocyte infiltration until 4 weeks. 5. Microvessels were increased sharply at 1 week and control group at 1 and 4 week showed significant differences with $Terudermis^{(R)}$. group of same interval(p<0.05) but no differences were found with AM group(p<0.05). Conclusion: EGF and EGF-R were well preserved in freeze-dried AM. AM attached to collagen acted as excellent biologic dressing which had similar effect with $Terudermis^{(R)}$. AM showed anti-inflammatory action and healing was completed at 4 weeks after full-thickness skin defect.