• 제목/요약/키워드: 구강악안면외과

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DMBA로 유도된 햄스터 협낭암종에서 p53 유전자 변이와 mdm-2 단백의 발현에 관한 연구 (STUDY ON MUTATION OF P53 AND EXPRESSION OF MDM-2 IN DMBA INDUCED CARCINOMA OF HAMSTER BUCCAL POUCH)

  • 박용선;김경욱;이재훈;김창진
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제27권5호
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    • pp.373-384
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    • 2001
  • Cellular proliferation is an intricately regulated process mediated by the coordinated interactions of critical growth control genes. Two of these factors in mammalian cells are the p53 and mdm-2 genes. A protein product of the mem-2 oncogene has been recently shown to associate with the protein encoded by the tumor suppressor gene p53. The p53 tumor suppressor protein is stabilized in response to DNA damage and other stress signals and causes the cell to undergo growth arrest or apoptosis, thus preventing the establishment of mutations in future cellular generations. Mutation or loss of p53 is a very common event in tumor progression. It occurs in about 50% of all tumors analysed including of colon, lung, breast and liver. The cellular mdm-2 gene, which has potential transforming activity that can be activated by overexpression, is amplified in a significant percentage of human sarcoma and in other mammalian tumors. Proteins encoded by the mdm-2 gene are able to bind to the p53 protein and, when overexpressed, can inhibit p53's transcriptional activation function, thus mdm-2 can act as a negative regulator of p53 function. Experimental study was performed to observe the relationship between p53 gene mutation and mdm-2 protein expression and apply the results to the clinical activity. 36 golden syrian hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek(control side) was treated with mineral oil as the same manner to the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were examined for histology and immunohistochemistry observation, and were completely dissected by microdissection and DNA from both tissue were isolated by proteins K/phenol/chloroform extraction. Segments of the hamster p53 exons 5, 6, 7 and 8 were amplified by PCR using the oligonucleotide primers, and then confirmational change was observed by SSCP respectively. The results were as follows : 1. Dysplasia at 6 weeks, carcinoma in situ at 8 weeks and invasive carcinoma from 10 weeks could be observed in experimental groups. 2. p53 mutations were detected in 10 of the 36(28%) and the exons 6(6 of the 10 : 60%) was the most hot spot area among the highy conserved region(exons 5, 6, 7 & 8). 3. Immunohistochemical study confirmed 22 of the 36(61%) of p53 expression involving 10 of p53 mutations. 4. mdm-2 expression of was showed in 3 of the 36(8%) involving 1 of the 22 of p53 expression and 2 of the 14 of p53 non-expression. From the above results, mutation of p53 gene or expression of p53 protein may have the influence of the DMBA induced carcinoma of hamster buccal pouch but the expression of mdm-2 protein may not have relationship with tumorigenesis.

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방사선 조사후 매식한 임프란트와 관주에 따른 골치유에 대한 비교연구 (INFLUENCE OF TOPICAL IRRIGATION USING THE HA & PURE Ti IMPLANTS ON BONE FORMATION;A STUDY ON THE IRRADIATED RABBIT TIBIA)

  • 홍성팔;이건주;차용두;오세종;현정민;최동주;박영주;박준우
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제26권1호
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    • pp.59-72
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    • 2000
  • In this study, the rate of bone formation and the pattern of bone to implant contact surface around HA coated implant and pure Ti implant inserted into the irradiated tibia of rabbit were compared. Sixteen mongrel mature male rabbits were used as experimental animal. Each rabbit received 15 Gy of irradiation. Four weeks after irradiation, two holes were prepared on the tibia of each rabbit for placement of HA coated type and pure Ti type implants. Prior to implant placement, one group received steroid irrigation and the control group was similarly irrigated with normal saline. This was immediately followed by placement of the two different types of implants. Postoperatively, tetracycline was injected intramuscularly for 3 days. For fluorescent labelling, 3 days of intramuscular alizarine red injection was given. 2 weeks before sacrifice, followed by intramuscular calcein green on the last 3 days before specimen collection. Each rabbit was sacrificed on the second, fourth, sixth and eighth week after the implantation. The specimens were observed by the light microscope and the fluorescent microscope. The results were as follows; 1. All implants inserted into the irradiated tibia of rabbit were free from clinical mobility and no signs of bony resorption were noted around the site of implant placement. 2. Under the light microscope, new bone formation proceeded faster around implants that received steroid irrigation compared to the control group irrigated with saline. Bone to implant contact surface was greater in the steroid irrigated group than the saline irrigated group. Therefore, better initial stabilization was observed in the group pretreated with steroid irrigation. 3. Under the light microscope. HA coated implants showed broader bone to implant contact surface than pure Ti implants, and HA coated implants had better bone healing pattern than pure Ti implants. 4. In the steroid pretreated group, acceleration of bone formation was demonstrated by fluorescent microscopy around the 2, 4 weeks group and the 6 weeks HA coated implant group. The difference in the rate of bone formation proved to be statistically significant(P<0.05). Faster bone formation was noted in the saline irrigated group in the 6 weeks pure Ti implants and 8 weeks group. The difference was not statistically significant(P<0.05). 5. For the rabbits that were sacrificed on the second and fourth week after the implant placements, the rates of bone formation around HA coated implants proceeded faster than those around pure Ti implants under the fluorescent microscopy. For the rabbits that were sacrificed on the sixth week after the implant placements, the rates of bone formation around pure Ti implants proceeded faster than those around HA coated implants under the fluorescent microscopy. But this result did not show statistical significance (P<0.05) For the rabbits that were sacrificed on the eighth week after the implant placements, the rates of bone formation around HA coated implants proceeded faster than those around pure Ti implants under the fluorescent microscopy. This result was statistically significant (P<0.05).

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성견 하악골 절단 후 기계적 골 견인에 의해 형성된 골 신장부에 대한 시기별 조직학적 변화 (HISTOLOGICAL CHANGES IN THE ELONGATED BONE AFFECTED BY OSTEODISTRACTION OF THE MANDIBLE IN THE DOG)

  • 백선호;안병근;박영주;박희건;박준우;이건주;이용찬;조병욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제27권5호
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    • pp.404-416
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    • 2001
  • Purpose : Traditionally, the treatement of choice has been a bone grafting procedure to increase the length of bone in case of actual length discrepancy. But, bone grafting procedure has many disadvantages, for example, graft resorption, donor site morbidity, and so on. So, many trials have been performed to avert the use of autogenous bone graft via introducing new materials or methods. And, one of those trials has been realized by the development of a technique inducing bone lengthening by osteotomy (or corticotomy) and slow gradual distraction of the osteotomized segments. This new technique of bone lengthening dates back to the early 20th century. But, the majority of information concerning the biology of new bone formation during bone lengthening and technical details of the procedure were produced by extensive clinical and experimental studies performed by Ilizarov, a Russian surgeon. According to Ilizarov, with adequate blood supply, preservation of periosteum, rigid fixation of the osteotomized segments, and proper rate and rhythm of distraction, intramembranous bone rapidly develops within the distraction gap in the limb lengthening procedure. In the limb lengthening, many orthopedic surgeons try to observe the biologic and clinical principles recommended by Ilizarov. In the oral and maxillofacial region, however, not a few studies must be performed to apply this surgical technique in the clinical cases. Besides, the mechanism of bone formation in the distraction gap is not clear, yet. The purpose of this experiment was to scrutinize serially the histological changes in the elongated bone affected by osteodistraction of the mandibular body in an adult canine model. In addition, it was performed to confirm the presence of specific region(s) which was important in the bone formation in the gap through the observation of the expression pattern of osteocalcin and osteonectin with the immunohistochemical examination. Materials and Methods : The experimental and control specimens were obtained from seven adult male mongrel dogs weighing over 20kg. The distractors were custom-made linear extraoral devices and bicortical fixation screws were 2.3mm in diameter, 50mm in total length, 15mm in screw length. The distractors were devised to produce a linear gap of 0.75mm between two bony segments every $360^{\circ}$ turn of the rotation rod of the device. The mandibular body of the right side of each animal was corticotomized perpendicular to the occlusal plane and then two bony segments were separated completely by careful manipulation of the segments with bone forceps. The left side of each animal was left intact. This side was served as control. At sixth day after osteotomy and fixation of the segments were performed, distraction of the segments was commenced with a rate of 1.1mm/day and a rhythm of two/day for ensuing 7 days. The animals were euthanized at the 16th. 29th, and 44th day after the osteotomy. The bony specimens were decalcified, embedded in paraffin, sectioned $5{\mu}m$ thick and stained with H&E. The prepared specimens were examined under the light microscope. And, immunohistochemical examinations using anti-osteocalcin antibody (OC1, Biodesign, USA) and anti-osteonectin antibody (Haematologic Technologies Inc., Essex, VT) to locate the expressions of osteocalcin and osteonectin, respectively, were performed. Results : 1. New bone was observed already at the 16th. day after osteotomy. This suggests that new bone formation in osteodistraction was commenced at an early stage of the regenerative process. But, radiologically and microscopically, bony union was not completed in the distraction gap at the 44th. day after osteotomy. Therefore, rigid fixation must be maintained between the bony fragments till the complete bony union is confirmed clinically rather than one month or so after the completion of distraction.

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전치부 개방교합을 동반한 골격성 제3급 부정교합 환자에 대한 양측 하악지 시상분할 골절단술후 안정성에 관한 임상적 분석 (CLINCAL ANALYSIS OF SKELETAL STABILITY AFTER BSSRO FOR CORRECTION OF SKELETAL CLASS III MALOCCLUSION PATIENTS WITH ANTERIR OPEN BITE)

  • 김현수;권대근;이상한;김진수;강동화;장현중
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제33권2호
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    • pp.152-161
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    • 2007
  • This study was conducted to patients visited oral maxillo-facial surgery, KNUH and the purpose of the study was to assess skeletal and dento-alveolar stability after surgical-orthodontic correction treated by skeletal Class III malocclusion patients with open bite versus non-open bite. This retrospective study was based on the examination of 40 patient, 19 males and 21 females, with a mean age 22.3 years. The patients were divided into two groups based on open bite and non-open bite skeletal Class III malocclusion patients. The cephalometric records of 40 skeletal Class III malocclusion patients (open bite: n = 18, non-open bite: n = 22) were examined at different time point, i.e. before surgery(T1), immediately after surgery(T2), one year after surgery(T3). Bilateral sagittal split ramus osteotomy was performed in 40 patients. Rigid internal fixation was standard method used in all patient. Through analysis and evaluation of the cephalometric records, we were able to achieve following results of post-surgical stability and relapse. 1. There was no significant statistical differences between open bite and non-open bite with skeletal Class III malocclusion patients in maxillary occlusal plane angle of pre-operative stage(p>0.05). 2. Mean vertical relapses of skeletal Class III malocclusion patients with open bite were $0.02{\pm}1.43mm$ at B point and $0.42{\pm}1.56mm$ at Pogonion point. In skeletal Class III malocclusion patients with non-open bite, $0.12{\pm}1.55mm$ at B point and $0.08{\pm}1.57mm$ at Pogonion point. There was no significant statistical differences between open bite and non-open bite with skeletal Class III malocclusion patients in vertical relapse(p>0.05). 3. Mean horizontal relapses of skeletal Class III malocclusion patients with open bite were $1.22{\pm}2.21mm$ at B point and $0.74{\pm}2.25mm$ at Pogonion point. In skeletal Class III malocclusion patients with non-open bite, $0.92{\pm}1.81mm$ at B point and $0.83{\pm}2.11mm$ at Pogonion point. There was no significant statistical differences between open bite and non-open bite with skeletal Class III malocclusion patients in horizontal relapse(p>0.05). 4. There were no significant statistical differences between open bite and non-open bite with skeletal Class III malocclusion patients in post-surgical mandibular stability(p>0.05). and we believe this is due to minimized mandibular condylar positional change using mandibular condylar positioning system and also rigid fixation using miniplate 5. Although there was no significant relapse tendency observed at chin points, according to the Pearson correlation analysis, the mandibular relapse was influenced by the amount of vertical and horizontal movement of mandibular set-back(p=0.05, r>0.304).

구순구개열 환아에서의 치조골이식 (TOOTH MOVEMENTS TO THE SITE OF ALVEOLAR BONE GRAFT)

  • 조해성;박재홍;김광철;최성철;이긍호;최영철
    • 대한소아치과학회지
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    • 제34권1호
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    • pp.140-149
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    • 2007
  • 치조파열 및 구개열과 같은 선천성 기형은 이환된 환자에게 기능적, 심미적으로 많은 문제점을 야기하므로 정상으로 회복시켜 주는 것은 중요한 문제이다. 장기간 방치될 경우 영양장애, 구강위생 불량, 호흡기 감염, 언어 장애, 악안면 변형, 그리고 정신적인 문제 등이 복합적으로 발생할 수 있다. 따라서 구순구개열 환자의 치료는 여러 전문 치료 분야의 복합적 인 접근이 필요하다. 골이식은 구순구개열 환자에 있어 중요한 치과치료 단계이다. 치아의 맹출과 치열의 안정화를 위해 치열궁은 골결손이 없이 완전해져야 한다. 또한 치열궁의 파열이 있는 부위로는 정상적인 교정적 치아이동이 곤란하다. 따라서 구순구개열 환자에 있어 골이식은 광범위하게 적용되고 있는 외과적 술식이다. 치조골을 이식함으로써 치조열은 안정화되고, 견치 또는 절치가 이식부위로 이동할 수 있게 된다. 그리고 골 이식 후, 교정을 통해 치조열 부위의 공간을 폐쇄함으로써 보철치료 없이 치열을 재형성 할 수 있다. 골이식술에는 다양한 이식재료가 사용되었다. 자가골을 이식할 경우 장골이 가장 선호되며, 그밖에 경골, 늑골, 두개골, 하악골을 이용하기도 한다. 그리고 골이식은 골이식 시기에 따라 일차골이식, 조기 이차골이식, 이차골이식, 만기 이차골이식으로 구분할 수 있다. 이차골이식은 혼합치열기 말경에 시행되는 것을 말하며 가장 좋은 것으로 받아들여지고 있다. 영구 견치가 맹출되기 전에 이차골이식을 시행하면 인접치아의 맹출과 보존에 도움이 된다. 본 증례에서는 치조구개파열 환자의 장골에서 골을 채취하여 골 이식을 시행한 후, 골이 채워진 이식 부위에 인접 치아가 성공적으로 이동하거나 치축이 개선되고 교정치료를 통해 치열궁 배열의 만족스러운 결과를 얻었기에 이를 보고하는 바이다.

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인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구 (SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT)

  • 최원재;안강민;황순정;정필훈;김명진;김남열;유상배;장정원;김현만;김중수;김윤희;김성민;이종호
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제31권3호
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

당뇨유도 백서 경골에 티타늄 임플란트 매식 시 지방조직 유래 줄기세포 주입 후 저출력 초음파 적용이 골치유에 미치는 영향 (Effect on bone healing by the application of low intensity pulsed ultrasound after injection of adipose tissue-derived stem cells at the implantation of titanium implant in the tibia of diabetes-induced rat)

  • 정태영;박상준;황대석;김용덕;이수운;김욱규
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제37권4호
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    • pp.301-311
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    • 2011
  • Introduction: This study examined the effect of the application of low intensity pulsed ultrasound on bone healing after an injection of adipose tissue-derived stem cells (ADSCs) during the implantation of a titanium implant in the tibia of diabetes-induced rats. Materials and Methods: Twelve Sprague-Dawely rats were used. After inducing diabetes, the ADSCs were injected into the hole for the implant. Customized screw type implants, 2.0 mm in diameter and 3.5 mm in length, were implanted in both the tibia of the diabetes-induced rats. After implantation, LIPUS was applied with parameters of 3 MHz, 40 mW/$cm^2$, and 10 minutes for 7 days to the left tibiae (experimental group) of the diabetesinduced rats. The right tibiae in each rat were used in the control group. At 1, 2 and 4 week rats were sacrificed, and the bone tissues of both tibia were harvested. The bone tissues of the three rats in each week were used for bone-to-implant contact (BIC) and bone area (BA) analyses and the bone tissues of one rat were used to make sagittal serial sections. Results: In histomorphometric analyses, the BIC in the experimental and control group were respectively, $39.00{\pm}18.17%$ and $42.87{\pm}9.27%$ at 1 week, $43.74{\pm}6.83%$ and $32.27{\pm}6.00%$ at 2 weeks, and $32.62{\pm}11.02%$ and $47.10{\pm}9.77%$ at 4 weeks. The BA in experimental and control group were respectively, $37.28{\pm}3.68%$ and $31.90{\pm}2.84%$ at 1 week, $20.62{\pm}2.47%$ and $15.64{\pm}2.69%$ at 2 weeks, and $11.37{\pm}4.54%$ and $17.69{\pm}8.77%$ at 4 weeks. In immunohistochemistry analyses, Osteoprotegerin expression was strong at 1 and 2 weeks in the experimental group than the control group. Receptor activator of nuclear factor kB ligand expression showed similar staining at each week in the experimental and control group. Conclusion: These results suggest that the application of low intensity pulsed ultrasound after an injection of adipose tissue-derived stem cells during the implantation of titanium implants in the tibia of diabetes-induced rats provided some positive effect on bone regeneration at the early stage after implantation. On the other hand, this method is unable to increase the level of osseointegration and bone regeneration of the implant in an uncontrolled diabetic patient.

백서 편측 설신경 손상 후 미각 및 설유두의 변화에 대한 연구 (CHANGE OF TASTE PREFERENCE AND TASTE BUD AFTER UNILATERAL LINGUAL NERVE TRANSECTION IN RAT)

  • 김윤태;전승호;염학렬;강진한;안강민;김성민;장정원;박경표;이종호
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제31권6호
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    • pp.515-525
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    • 2005
  • Purpose of study: Lingual nerve damage can be caused by surgery or trauma such as physical irriatation, radiation, chemotherapy, infection and viral infection. Once nerve damage occurred, patients sometimes complain taste change and loss of taste along with serious disturbance of tongue. The purpose of this study was to evaluate the effects of unilateral lingual nerve transection on taste as well as on the maintenance of taste buds. Materials & Methods: Male Sprague-Dawley rats weighing 220-250g received unilateral transection of lingual nerve, subjected to the preference test for various taste solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) with two bottle test paradigm at 2, 4, 6, or 8 weeks after the operation. Tongue was fixed with 8% paraformaldehyde. After fixation, they were observed with scanning electron microscope(JSM-$840A^{(R)}$, JEOL, JAPAN) and counted the number of the dorsal surface of the fungiform papilla for changes of fungiform papilla. And, Fungiform papilla were obtained from coronal sections of the anterior tongue(cryosection). After cryosection, immunostaining with $G{\alpha}gust$(I-20)(Santa Cruz Biotechnology, USA), $PLC{\beta}2$(Q-15)(Santa Cruz Biotechnology, USA), and $T_1R_1$(Alpha Diagnostic International, USA) were done. Immunofluorescence of labeled taste bud cells was examined by confocal microscopy(F92-$300^{(R)}$, Olympus, JAPAN). Results: The preference score for salty and sweet tended to be higher in the operated rats with statistical significance, compared to the sham rats. Fungiform papilla counting were decreased after lingual nerve transaction. In 2 weeks, maximum differences occurred. Gustducin and $T_1R_1$ expressions of taste receptor in 2 and 4 weeks were decreased. $PLC{\beta}2$ were not expressed in both experimental and control group. Conclusion: This study demonstrated that the taste recognition for sweet and salty taste changed by week 2 and 4 after unilateral lingual nerve transection. However, regeneration related taste was occurred in the presence of preserving mesoneurial tissue and the time was 6 weeks. Our results demonstrated that unilateral lingual nerve damage caused morphological and numerical change of fungiform papilla. It should be noted in our study that lingual nerve transection resulted in not only morphological and numerical change but also functional change of fungiform papillae.

미성숙 매복지치의 치낭, 치수, 치근유두 조직에서 다능성 줄기세포의 분리와 특성화에 대한 연구 (Isolation and characterization of human dental tissue-derived stem cells in the impacted wisdom teeth: comparison of dental follicle, dental pulp, and root apical papilla-derived cells)

  • 송정호;박봉욱;변준호;강은주;노규진;신상훈;김욱규;김종렬
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제36권3호
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    • pp.186-196
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    • 2010
  • Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.

종양수술전 화학요법이 미세수술시 피판생존율에 미치는 영향 (EFFECT OF INDUCTION CHEMOTHERAPY ON FLAP SURVIVAL RATE IN MICROSURGERY)

  • 김욱규;김용덕;변준호;신상훈;정인교
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제29권6호
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    • pp.421-429
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    • 2003
  • 총 24 마리의 백서를 선택하여 종양화학요법제의 하나인 5-FU를 복강내 투여한후 24 시간이 지난후 8마리에서는 하복부 동맥 및 정맥을 근간으로 한 하복부 피부 도상피판($2{\times}3cm^2$)울 좌, 우 2 개씩 형성하고 피판을 허혈상태를 유도하여 유리피판상태를 만들었다. 허혈상태를 1 시간정도유지한후 다시 혈류를 재개시켜 피판으로의 혈류를 재개시키고 상처부를 층별봉합 하였다. 그 후 3 일, 5 일, 7 일째 각각 2마리씩 희생하여 H-E staining하였다. 또한 술후 1 일, 3 일, 5 일, 7 일째 피판생존을 육안으로서 확인하고 피판의 survival rate를 비교하기 위하여 사진촬영하여 digital image를 얻은후 adobe software program을 이용하여 Image, Histogram기능을 써서 60,000 pixels(가로${\times}200$, 세로${\times}300$)상태에서 피판의 Luminosity(광도)정도를 수치화하였다. 마찬가지의 술식으로 5-FU를 투여후 3일이 경과된후 8마리를 동일한 부위에 동일한 피판을 써서 3 일, 5 일째 각각 2 마리에 대해 피판을 채취하여 H-E staining 하여 조직현미경상의 치유과정을 관찰하였다. 대조군으로서 복강내 5-FU를 투여받지않은 백서 8마리에서 동일한 도상 피부피판을 형성하고 3 일, 5 일, 7 일째되는 2마리씩 피판부위를 채취하여 H-E염색하여 조직치유반응을 광학현미경으로 관찰하였고 상기와 동일한 방식으로 피판의 생존율을 계산하여 다음의 결과를 얻었다. 1. 육안적 소견으로 치유의 순은 대조군, 항암 24 시간째군, 항암 3 일군순으로 빨리 치유되었다. 2. 피판생존율을 비교하기 위한 광도(Luminosity)의 차는 3군끼리 모두 유의한 차를 보였으며 평균치 비교에서는 대조군, 항암 24 시간째군, 항암 3 일군순으로 높았다. 3. 광학현미경상 조직치유의 정도는 5개 항목으로 나누어 관찰하였다. 전체적으로 대조군과 항암 24 시간군은 시일이 지날수록 치유가 되는 양상을 보였으나 항암 3 일군은 조직의 염증반응이 더 심해지는 양상을 보였다. epidermis necrosis관찰항목에서는 대조군에 비해 항암제투여군이 경도의 반응을 보였고 항암3 일군에서 가장 심하였다. Inflammation state는 3일째는 대조군에서 가장 높게 나타났으나 시일이 지남에 따라 감소한 반면 항암 3 일군은 시일이 지날수록 심해졌다. dermis fibrosis면에서는 항암 24 시간군에서 가장 크게 나타났다. vessel change는 대조군과 항암 24군에서는 거의 관찰되지 않았고 항암 3 일군에서는 중등도로 관찰되었다.fatty tissue layer thinning은 3 군모두에서 관찰되었고 항암 3 일군이 가장 심하게 나타났다. 이상의 실험결과를 보면 술전 항암제투여가 초기에 시행한 경우에는 조직의 치유에 초기 5 일정도까지는 영향을 미치나 7 일이 지나면 정상범주로 회복함을 알수 있었고 실험결과 항암제 투여후 3 일째 피판 형성한 군에서 피판치유가 늦어진 것으로 관찰되어 인체에서 항암 투여후 수술시기는 인체면역계가 회복하는 시기를 3주이상 경과후 적어도 4주째 수술시기를 정하는 것이 유리하리라 생각되었다.