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Analysis of Quorum Sensing-Related Phenotypes of Pseudomonas aeruginosa Clinical Isolates (녹농균 임상균주의 쿼럼 센싱 관련 표현형 분석)

  • Jung, Kyung-Ju;Choi, Yu-Sang;Ha, Chang-Wan;Shin, Jeong-Hwan;Lee, Joon-Hee
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.240-247
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    • 2010
  • Pseudomonas aeruginosa is a Gram (-) opportunistic human pathogen causing a wide variety of infections on lung, urinary tract, eyes, and burn wound sites and quorum sensing (QS), a cell density-sensing mechanism plays an essential role in Pseudomonas pathogenesis. In order to investigate the importance of QS in the Pseudomonas infections of Korean patients, we isolated 189 clinical strains of P. aeruginosa from the patients in Pusan Paik Hospital, Busan, South Korea. The QS signal production of these clinical isolates was measured by signal diffusion assay on solid media using reporter strains. While most clinical strains (79.4%) produced the QS signals as similar level as a wild type strain, PAO1 did, where LasR, the initial QS signal sensor-regulator was fully activated, a minority of them (4.2%) produced much less QS signals at the level to which LasR failed to respond. Similarly, while 72.5% of the clinical isolates produced QS signals enough to activate QscR, an another QS signal sensor-regulator, some few of them (9%) produced the QS signals at much lower level where QscR was not activated. For further analysis, we selected 74 clinical strains that were obtained from the patients under suspicion of Pseudomonas infection and investigated the total protease activity that is considered important for virulence. Interestingly, significant portion of them showed very low protease activity (44.6%) or no detectable protease activity (12.2%). When the biofilm-forming ability that is considered very important in chronic infection was examined, most isolates showed lower biofilm-forming activity than PAO1. Similarly, significant portion of clinical isolates showed reduced motility (reduced swarming activity in 51.4% and reduced twitching activity in 41.9%), or non-detectable motility (swarming-negative in 28.4% and twitching-negative in 28.4%). Our result showed that the clinical isolates that produced QS signals at the similar level to wild type could have significantly reduced activities in the protease production, biofilm formation, and motility, and some clinical isolates had unique patterns of motility, biofilm formation, and protease production that are not correlated to their QS activity.

Effect of Chitosan-Trimer on the Prevention of Postoperative Intraperitoneal Adhesion Formation in Rats (랫트에서 Chitosan-Trimer가 복강유착에 미치는 영향)

  • Kwon, Eun-ju;Jang, Kwang-ho;Jang, In-ho
    • Journal of Veterinary Clinics
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    • v.18 no.3
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    • pp.257-264
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    • 2001
  • This study was performed to investigate the effects of chitosan-trimer (CT) on the prevention of postoperative adhesion formation in the rate model. All animals divided into PBS (control), 1% CT, 3% CT, and chitin treated group. The mean adhesion score in 1% CT group (1.03$\pm$0.63), 3% CT group (0.64$\pm$0.53) and chitin group (1.67$\pm$0.71) was found to be lower than that in control group (2.07$\pm$0.81). More favorable adhesion prevention was achieved in 3% CT group (0.64$\pm$0.53) in comparison with the control group, 1% CT group, and chitin group without any hemorrhagic complications. A statistically significant difference was observed in adhesion formation between control group and 3% CT group (p<0.001). In control group, 44 of 45 sites (97.7%) formed adhesions between the intestine defects. In contrast, 3% CT was effective in reducing the incidence of adhesion formation to 17 to 45 sites (62.2%) (p<0.05). The locations of adhesions were observed in serosa-serosa (60%), serosa-mesentery (13.3%), serosa-connective tissue of testis (10%), omentum-liver (10%), serosa-omentum (3.3%), serosa-cecum (3.3%), and serosa-incision (0%). On the results of histological analysis, grade of inflammation and fibrosis at the sites of postoperative peritoneal adhesion formation were not significantly different in all groups. But, 3% CT showed the lowest score of inflammation and fibrosis. In 3% CT group, the rate of increase of plasma fibrinogen was significantly lower compared with that in control group from pre-operation to 10 days later (p<0.05). There were no appreciable difference in the CBC, leukocyte differential counts and total protein concentrations among four groups. In conclusion, our data suggested that CT should be effective on reducing adhesion formation in experimental rat models. The results also showed that 3% CT does not adversely affect normal wound healing and healthy recovery after operation.

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Results of Postoperative Radiation Therapy of Rectal Cancers - with the Emphasis of the Overall Treatment Time - (직장암의 수술 후 방사선치료의 성적 - 예후 인자와 전체 치료기간이 미치는 영향에 관한 고찰 -)

  • Kim Joo-Young;Lee Myung-Hag;Lee Kyu-Chan
    • Radiation Oncology Journal
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    • v.16 no.3
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    • pp.303-310
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    • 1998
  • Purpose : To evaluate the results of the treatment of locally advanced but resectable rectal cancers and to analyze prognostic factors. especially with the emphasis on the treatment time factor. Materials and Methods : There were 71 patients with rectal cancer who had been treated by curative surgical procedure and postoperative radiotherapy from August 1989 to December 1993. The minimum follow up period was 24 months and the median follow-up was 35 months Radiation therapy had been given by 6 MV linear accelerator by parallel opposing or four-box portals. Whole pelvis was treated up to 5040 cGy in most cases. Systemic chemotherapy had been given in 94$\%$ of the patients, mostly with 5-FU/ACNU regimen. Assessment for the overall and disease-free survival rates were done by life-table method and prognostic factors by Log-Rank tests. Results : Five-year overall survival, disease-free survival were 58.8$\%$ and 57$\%$, respectively. Two-year local control rate was 76.6$\%$. Stage according to Modified Astler-Coller (MAC) system, over 4 positive lymph nodes, over 6weeks interval between definitive surgery and adjuvant radiotherapy and over 7 days of interruption during radiotherapy period were statistically significant, or borderline significant prognostic factors. Conclusion : The treatment results of patients with rectal cancers are comparable to those of other large institutes. The treatment results for the patients with bowel wall penetration and/or positive regional lymph nodes were still discouraging for their high local recurrence rate for the patients with MAC 'c' stage diseases and high distant metastases rate even for the patients with node-negative diseases. Maybe more effective regimen of chemotherapy would be needed with proper route and schedule. To maximize postoperative adjuvant treatment. radiotherapy should be started at least within 6 weeks after surgery and preferably as soon as wound healing is completed. Interruption of treatment during radiotherapy course affects disease-free survival badly, especially if exceeds 7 days. So, the total treatment period trout definitive surgery to the completion of radiotherapy should be kept as minimal as possiable.

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Process Development of a Virally-Safe Acellular Bovine Amniotic Membrane for Biological Dressing (바이러스 안전성이 보증된 무세포 소 양막 생물창상피복재 제조 공정 개발)

  • Bae, Jung-Eun;Kim, Chang-Kyong;Kim, Sung-Po;Yang, Eun-Kyung;Kim, In-Seop
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.420-427
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    • 2010
  • A process for manufacturing virally-safe bovine amniotic membrane(BAM) has been developed for biological dressing. BAM was harvested from a healthy bovine placenta, and then the epithelium was removed. The remaining stromal layer was consecutively disinfected with 70% ethanol and 0.05% sodium hypochlorite. The stromal layer was incubated in a decellularization solution containing 0.25%(w/v) trypsin to remove the cellular components. The resulting acelluar BAM was lyophilized to preserve its biochemical and structural integrity. The BAM was packed and exposed to 25 kGy of gamma irradiation for sterilization purpose. Histological, electron microscopical, and biochemical observations showed that the acellualr BAM had intact structural integrity of three dimensional collagen fibers and contained several growth factors, accelerating wound healing, such as EGF (Epidermal growth factor), KGF (Keratinocyte growth factor), and FGF (Fibroblast growth factor). Bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), bovine parainfluenza virus type 3 (BPIV-3), and bovine parvovirus (BPV) were chosen as the biological indicators for validation of viral safety of the acellular BAM. Samples from relevant stages of the production process were spiked with each virus and subjected to viral inactivation processes. Viruses were recovered from the samples and then titrated immediately. All the viruses tested were completely inactivated to undetectable levels within 1 h of 70% ethanol treatment. Enveloped viruses such as BHV, BVDV, and BPIV-3 were more effectively inactivated than BPV by 0.05% sodium hypochlorite treatment. BHV, BVDV, and BPIV-3 were completely inactivated to undetectable levels by 25 kGy of gamma irradiation. Also BPV was effectively inactivated by 25 kGy of gamma irradiation. The cumulative log reduction factors of BHV, BVDV, BPIV-3, and BPV were ${\geq}$13.30, ${\geq}$14.32, ${\geq}$15.22, and ${\geq}$7.57, respectively. These results indicate that the production process for acelluar BAM has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.

The Effect of Safflower Seed Extract on the Bone Formation of Calvarial Bone Model in Sprague Dawley rat (백서 두개골 결손부에서 홍화씨 추출물의 골조직 재생 유도 효과)

  • Kim, Sung-Tae;Jhon, Gil-Ja;Lim, So-Hyoung;Cho, Kyoo-Sung;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.835-852
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    • 2000
  • The ultimate goal of periodontal therapy is the regeneration of periodontal tissue and repair of function. For more than a decade there have been many efforts to develop materials and methods of treatment to promote periodontal wound healing. Recently many efforts are concentrated on the regeneration potential of material used in oriental medicine. In some in vitro and in vivo experiments, there have been many evidences that these materials have an effect on bone regeneration. The purpose of this study was to evaluate histologically and radiologically in Sprague-Dawley rats the effects of safflower seed extracts on the regeneration of the calvarial defects surgically produced. So in this study, the critical size defects were surgically produced in the calvarial bone of 30 Sprague-Dawley rats using the 8mm trephine bur. The safflower seed extract was applied into the defect of each rat in experimental group, whereas nothing was applied into the defect of each rat in control group. Rats were sacrificed at 2, 4, 8 weeks following operation and histomorphometric and radiodensitometric analysis were performed. 1. The newly formed bone length was $102.91{\pm}22.05$, $178.29{\pm}24.40$ at 2 week in the each control, experimental group, $130.95{\pm}39.24$, $242.62{\pm}50.33$ at 4 week and $181.53{\pm}76.35$, $240.36{\pm}22.00$ at 8 week($unit,{\mu}m$). In the 2, 4 week, there were statistically significant difference between control and experimental group(P<0.05). 2. The newly formed bone area was $2962.06{\pm}1284.48$, $10648.35{\pm}1284.48$ at 2 week, $5103.25{\pm}1375.88$, $9706.78{\pm}1481.81$ at 4 week, $8046.02{\pm}818.99$, $12057.06{\pm}740.47$ at 8 week($unit,{\mu}m^2$). In every week, there were statistically significant difference between control and experimental group(P<0.05). 3. The radiopacity was $14.26{\pm}.33$, $25.47{\pm}4.33$ at 2 week, $20.06{\pm}9.07$, $26.61{\pm}2.78$ at 4 week, $22.99{\pm}3.76$, $27.29{\pm}1.54$ at 8 week(unit, %). In the 2 week, there was statistically significant difference between control and experimental group(P<0.05). In conclusion, the results of the present study suggest that safflower seed extract initially has an effect on the newly formed bone area, length and radiopacity when it is applied to the calvarial defect of Sprague - Dawley rat. Then. the material has an effect on newly formed bone area and length.

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The Bone Regenerative Effects of Chitosan on the Calvarial Critical Size Defectin Sprague Dawley Rats (백서 두개골 결손부에서 키토산의 골조직 재생 유도 효과)

  • Jung, Ui-Won;Suh, Jong-Jin;Choi, Seong-Ho;Choi, Kyoo-Sung;Chai, Jung-Kiu;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.851-870
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    • 2000
  • The major goals of periodontal therapy is the functional regeneration of periodontal supporting structures already destructed by periodontal disease as well as the reduction of signs and symptoms of progressive periodontal disease. There have been many efforts to develop materials and therapeutic methods to promote periodontal wound healing. There have been increasing interest on the chitosan made by chitin. Chitin is second only to cellulose as the most abundant natural biopolymer. It is a structural component of the exoskeleton of invertebrates(e.g., shrimp, crabs, lobsters), of the cell wall of fungi, and of the cuticle of insects. Chitosan is a derivative of chitin made by deacetylation of side chains. Many experiments using chitosan in various animal models have proven its beneficial effects. The aim of this study is to evaluate the osteogenesis of chitosan on the calvarial critical size defect in Sprague Dawley rats. An 8 mm surgical defect was produced with a trephine bur in the area of the midsagittal suture. The rats were divided into two groups: Untreated control group versus experimental group with 50mg of soluble chitosan gel. The animals were sacrificed at 2, 4 and 8 weeks after surgical procedure. The specimens were examined by histologic, histomorphometric and radiodensitometric analyses. The results are as follows: 1. The length of newly formed bone in the defects was $102.91{\pm}25.46{\mu}m$, $219.46{\pm}97.81{\mu}m$ at the 2 weeks, $130.95{\pm}39.24{\mu}m$, $212.39{\pm}89.22{\mu}m$ at the 4 weeks, $181.53{\pm}76.35{\mu}m$ and $257.12{\pm}51.22{\mu}m$ at the 8 weeks in the control group and experimental group respectively. At all periods, the means of experimental group was greater than those of control group. But, there was no statistically significant difference between the two groups. 2. The area of newly formed bone in the defects was $2962.06{\pm}1284.48{\mu}m^2$, $5194.88{\pm}1247.88{\mu}m^2$ at the 2 weeks, $5103.25{\pm}1375.88{\mu}m^2$, $7751.43{\pm}2228.20{\mu}m^2$ at the 4 weeks and $8046.20{\pm}818.99{\mu}m^2$, $15578.57{\pm}5606.55{\mu}m^2$ at the 8 weeks in the control group and experimental group respectively. At all periods, the means of experimental group was greater than those of control group. The experimental group showed statistically significant difference to the control group at the 2 and 8 weeks. 3. The density of newly formed bone in the defects was $14.26{\pm}6.33%$, $27.91{\pm}6.65%$ at the 2 weeks, $20.06{\pm}9.07%$, $27.86{\pm}8.20%$ at the 4 weeks and $22.99{\pm}3.76%$, $32.17{\pm}6.38%$ at the 8 weeks in the control group and experimental group respectively. At all periods, the means of experimental group was greater than those of control group. The experimental group showed statistically significant difference to the control group at the 2 and 8 weeks. These results suggest that the use of chitosan on the calvarial defects in rats has significant effect on the regeneration of bone tissue in itself

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The Bone regenerative effects of tetracycline blended chitosan membranes on the calvarial critical size defect in Sprague dawley rats (백서 두개골 결손부에서 항생제를 함유한 키토산 차단막의 골재생 유도 효과)

  • Chae, Gyung-Joon;Kim, Tae-Gyun;Jung, Ui-Won;Lee, Soo-Bok;Jung, Yong-Sik;Lee, Yong-Keun;Kim, Chang-Sung;Chae, Jung-Kiu;Cho, Kyoo-Sung;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.35 no.4
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    • pp.1019-1037
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    • 2005
  • The major goals of periodontal therapy are the functional regeneration of periodontal supporting structures already destructed by periodontal disease as well as the reduction of signs and symptoms of progressive periodontal disease. There have been many efforts to develop materials and therapeutic methods to promote periodontal wound healing. There have been increasing interest on the chitosan made by chtin. Chitosan is a derivative of chitin made by deacetylation of side chains. Chitosan has been widely studied as bone substitution and membrane material in periodontology. Many experiments using chitosan in various animal models have proven its beneficial effects. Tetracycline has been considered for use in the treatment of chronic periodontal disease and gingivitis. The aim of this study is to evlauate the osteogenesis of tetracycline blended chitosan membranes on the calvarial critical size defect in Sprague Dawley rats. An 8mm surgical defect was produced with a trephine bur in the area of the midsagittal suture. The rats were divided into five groups: Untreated control group versus four experimental group. Four types of membranes were made and comparative study was been done. Two types of non-woven membranes were made by immersing non-woven chitosan into either the tetracycline solution or chitosan-tetracycline solution. Other two types of sponge membranes were fabricated by immersing chitosan sponge into the tetracycline solution, and subsequent freeze-drying. The animals were sacrificed at 2 and 8 weeks after surgical procedure. The specimens were examined by histologic analyses. The results are as follows: 1. Clinically the use of tetracycline blended chitosan membrane showed great healing capacity. 2. The new bone formations of all the experimental group, non-woven and sponge type membranes were greater than those of control group. But, there was no significant difference between the experimental groups. 3. Resorption of chitosan membranes were not shown in any groups at 2 weeks and 8 weeks. These results suggest that the use of tetracycline blended chitosan membrane on the calvarial defects in rats has significant effect on the regeneration of bone tissue in itself. And it implicate that tetracycline blended chitosan membrane might be useful for guided tissue regeneration.

A Study on the Change of the Palatal Length after Palatoplasty

  • Ryu Sun-Youl;Kim Sun-Kook;Kim Tae-Hee;Hwang Ung;Kook Min-Suk;Han Chang-Hun
    • Korean Journal of Cleft Lip And Palate
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    • v.7 no.1
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    • pp.25-34
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    • 2004
  • The present study was carried out to investigate the change of the palatal length after palatoplasty in congenital cleft palate. With the data from one hundred and twelve patients with cleft palate who had been treated at the Department of Oral and Maxillofacial Surgery of Chonnam University Hospital over a period of 10 years(April 1995 to April 2004). The epidemiological characteristics, the method of palate repair, the postoperative complications and the extent of palatal lengthening were investigated and analyzed statistically. Incomplete cleft palate occurs more frequently than complete cleft palate. Male were affected 2.1 times more than female in complete cleft palate, and female were affected 1.2 times more than male in incomplete cleft palate. Dorrance method and Wardill V-Y method were frequently used in repair of incomplete cleft palate. Wardill V-Y method, Furlow double opposing Z-plasty, two flap method, and Perko method were widely used in repair of complete cleft palate. The extent of palatal lengthening was greater in the incomplete cleft palate group(5.84 mm) than in the complete cleft palate group(4.71 mm), and in the Furlow double opposing Z-plasty group(5.70 mm) than in the push back palatoplasty group(5.33 mm). But no significant difference was noted. Palatal fistula and wound dehiscence were popular postoperative complications in cleft palate. These results indicate that the extent of palatal lengthening, which contributing to speech function, is a range of 3.5% to 24.0%(average 10.8%) after palatoplasty in cleft palate patients.

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The Clinical Efficacy of Uvulopalatopharyngoplasty in the Treatment of Obstructive Sleep Apnea Syndrome (폐쇄성 수면 무호흡 증후군 치료에서 구개수구개인두성형술의 임상적 유용성)

  • Moon, Hwa-Sik;Choi, Young-Mee;Park, Young-Hak;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Song, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.6
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    • pp.1366-1381
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    • 1997
  • Background : Uvulopalatopharyngoplasty(UPPP) has become the most common surgical treatment for obstructive sleep apnea syndrome(OSAS). However, the results of this therapeutic modality have been quite variable with successful results by several authors and poor results by others. Until recently, in Korea, there is only a few reports about the clinical efficacy of UPPP. A prospective study was undertaken to evaluate the effectiveness and complications of UPPP. Method : Twenty-six OSAS patients who had undergone UPPP with preoperative and postoperative polysomnographic studies were included in this study. Two definitions of surgical success were used. The responder was defined, using a conventional criteria, as a 50% or more reduction in apnea index(AI) or apneahypopnea index(AHI) after UPPP, or a postoperative AI of <10 or AHI of <20. The initial cure was defined, using our own criteria, as a postoperative AI of <5 or AHI of <10. Complications were categorized in two groups : early(disorders during the first 10 postoperative days) and late. Results : Eighteen patients(69.2%) were responders, and ten patients(38.5%) were considered as initial cure. On the other hand, in five patients (19.2%), postoperative polysomnographic data demonstrated deterioration compared with preoperative data. Reduction rate of AI or AHI following UPPP was not significantly related to the preoperative body mass index, AI or AHI. There was no significant change of sleep architecture before and after UPPP in responder and initial cure groups. Early complications such as pain, dyspnea, bleeding, nasal reflux, dysphagia or wound disruption were observed in all patients. Late complications such as nasal reflux, voice change, dysphagia, loss of taste, pharyngeal dryness or foreign body sensation were discovered in 22 patients (84.6%). However, all early and late complications were of minor importance. Conclusion : The response to UPPP was favorable in approximately 70% of OSAS patient. However, the initial Cure rate of UPPP was relatively low. We suggest that selection of more appropriate surgical candidates and adequate surgical protocol is necessary to obtain a more successful result with UPPP.

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A Comparative Study of Initial Healing Process in White Rats after Gingivectomy using $CO_2$ Laser of different watts (($CO_2$)레이저를 이용한 백서의 치은절제술시 출력에 따른 초기 치유과정의 비교)

  • Cho, Kyoo-Sung;Hong, Sung-Jae;Choi, Seong-Ho;Chai, Jung-Kiu;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
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    • v.27 no.3
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    • pp.603-619
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    • 1997
  • The use of laser in the treatment of soft tissue minimizes hemorrhage, provides better view of the operating field, and thereby minimizes operating time. Also, there will be far less post-operative swelling, pain and scar formation, and sterilizing effect are shown in some portions of the wound site. All these advantages of laser therapy contribute to its widespread use in the field of medicine and dentistry. Regarding such facts, we used CO2 laser of different watts in gingivectomy for white rats to compare initial healing process. For the control group, the least amount of output in performing gingivectomy(4watts) was offered, and for the experimental group, 6watts was given. Animals were sacrificed on the second, third days, 1 weeks, 2 weeks, and 3 weeks after operation, and their specimens were histologically analyzed. The following results were obtained: 1. Blood clot of small size was observed in both the control and experimental groups after two days, and no more thereafter. 2. In both the control and experimental groups, the inflammation zone size was the greatest after two days, and it decreased gradually to become almost invisble by the second week. The experimental group showed larger size of inflammation zone during second and third days: however, there was no difference after one week. 3. Granulation tissue in both the control and experimental groups showed gradual maturation with time, and by the second week, it was almost replaced by normal connective tissue. By the third week, complete healing pattern was observed. The experimental group showed larger granulation tissue than the control group until the third day, but there was no significant difference after one week. 4. In both the control and experimental groups, gingival epithelialization began on the second day. After one week, regeneration of rete peg and partial formation of junctional epithelium were observed: by the second week, keratinization of oral sulcular epithelium began, and it was completed by the third week.

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