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백서두개골 결손부에서 키토산/흡수성 콜라겐 전달체의 골재생

Other Titles
 Effect of chitosan/ACS on bone regeneration in rat calvarial defects 
Authors
 김수경 
Issue Date
2003
Description
치의학과/석사
Abstract
[한글] 치주 치료의 최종 목적은 진행되는 치주 질환의 증상을 제거하는 것뿐만 아니라 이미 파괴된 지지조직을 기능적으로 재생시키는데 있다. 현재 파괴된 치주조직의 재생을 위해 다양한 종류의 골이식재를 이용한 골이식술과 차단막을 이용한 치주조직 유도재생술이 행해지고 있으나, 아직까지 각각의 한계점을 가지고 있다. 치주조직의 재생을 위해 사용되는 생약제제는 생체거부반응이나 생분해 시 나타날 수 있는 독성의 위험이 적고 그 효과가 지속적이며 임상에 응용될 경우 경제적으로 사용될 수 있다. 최근 키토산은 이러한 생체 적합성과 항균 작용, 창상 치유 촉진 등의 생물학적 작용, 우수한 기계적 특성으로 관심이 증가되고 있으며, 특정 전구 세포 (예; 조골세포)의 이주와 분화를 증진시키는 기질 역할을 할뿐만 아니라 섬유모세포와 같이 골 형성을 방해하는 세포의 기능을 억제함으로써 직, 간접적으로 골재생을 증진시킨다고 보고 되었다. 본 실험에서는 키토산에서 정제한 순수 키토산 용액을 사용하였다. 그러나 이러한 키토산의 형태는 액상이므로 결손부에서 유지되기가 어렵다. 따라서 결손부에서의 키토산의 송달, 유지, 점진적 유지를 위해서는 운반체의 이용이 필수적이다. 백서의 두개골 천공 모형에서는 비교적 공간 유지가 용이하여 본 실험에서는 키토산 용액을 흡수성 콜라겐 스폰지에 적셔서 결손부에 이식하였다. 실험 모델로는 수컷 백서 두개골 결손부를 선택하였고, 두개골의 임계크기 결손은 지름 8 mm의 원형 결손이다. 두개골에 아무 처치도 하지 않은 군을 음성 대조군으로 하고, 흡수성 콜라겐 스폰지 (Absorbable collagen sponge: ACS)만을 처치한 군을 양성대조군으로 설정하였으며, 순수키토산 용액을 ACS에 적셔 이식한 군을 실험군으로 설정하였다. 백서 두개골에 키토산/ACS를 적용하고, 술후 2주, 8주에 희생하여 치유 결과를 조직학적, 조직 계측학적으로 비교 관찰하여 다음과 같은 결론을 얻었다. 1. 실험군 (키토산/ACS)은 술후 2주부터 혈관증식이 진행되고 조골세포의 침윤으로 신생골 형성이 진행되는 양상을 보였고, 8주에서는 ACS는 거의 흡수되었으며 신생골 형성량이 증가하였고, 많은 골성조직층이 관찰되었다. 2. 술후 2주에 신생골 형성량은 실험군과 양성대조군 (ACS만 이식한군), 음성대조군(술후 무 처치군)이 각각 8.7±0.8, 13.6± 2.3, 4.8±0.7%로, 실험군과 양성대조군에서 음성대조군에 비해 높게 나타났으나 통계학적인 유의차는 없었다. (p<0.01). 3. 술후 8주에 신생골 형성량은 실험군과 양성대조군, 음성 대조군이 각각 62.2±6.1, 17.4±2.5, 8.2±1.4로 실험군이 양성대조군과 음성대조군에 비해 현저하게 높게 나타났으며 통계학적으로 유의한 차이를 보였다. (p<0.01). 이상의 실험 결과에서, 키토산은 ACS를 운반체로 사용했을 때 백서의 두개골 천공 결손부에서 효과적인 골재생을 나타냈다.
[영문] The ultimate objective of periodontal treatment is to get rid of an on-going periodontal disease and further regenerate the supporting tissue, which is already destroyed, functionally. Currently, the bone grafting operation using various kinds of bone grafting materials and the operation for induced regeneration of periodontal tissue using the blocking membrane are performed for regeneration of the destroyed periodontal tissue. However, there are respective limitations Galenical preparations, which are used for regeneration of periodontal tissue, has less risk of rejective reaction or toxicity that may be incidental to degradation and their effect is sustainable. Thus, in case they are applicable to a clinic, they can be used economically. Chitosan has such compatibility, biological actions including antibacterial activity, acceleration of wound treatment, etc., and excellent mechanical characteristics, which has recently aroused more interest in it. Also, it has been reported that it promotes osteogenesis directly or indirectly by functioning as a matrix to promote migration and differentiation of a specific precussor cell (for example, osteoblast) and further inhibiting the function of such a cell as fibroblast to prevent osteogenesis. In this study, the pure chitosan solution, which was obtained by purifying chitosan, was used. However, since this chitosan is of a liquiform, it is difficult to sustain it in a defective region. It is, therefore, essential to use a carrier for delivering chitosan to, and sustaining it gradually in the defective region. In the calvarial defect model of the Sprague-Dawley rat, it is relatively easy to maintain a space. Therefore, in this study, the chitosan solution with which ACS was wetted was grafted onto the defective region. For an experimental model, a calvarial defect of rat was selected, and a critical size of the defective region was a circular defect with a diameter of 8 mm. A group in which no treatment was conducted for the calvarial defect was set as a negative control group. Another group in which treatment was conducted with ACS only was set as a positive control group (ACS group). And another group in which treatment was conducted by grafting the pure chitosan solution onto the defective region through ACS which was wetted with the chitosan solution was set as an experimental group (Chitosan/ACS group). Chitosan was applied to the Sprague-Dawley rat''s calvarial bone by applying ACS which was wetted with the chitosan solution, and each Sprague-Dawley rat was sacrificed respectively 2 weeks and 8 weeks after the operation for such application. Then, the treatment results were compared and observed histologically and histometrically. Thereby, the following conclusions were obtained. 1. In the experimental group, a pattern was shown that from 2 weeks after the operation, vascular proliferation proceeded and osteogenesis proceeded through osteoblast infiltration, and at 8 week after the operation, ACS was almost absorbed, the amount of osteogenesis was increased and many osteoid tissue layers were observed. 2. At 2 weeks after the operation, each amount of osteogenesis appeared to be 8.70.8 %, 13.62.3 % and 4.80.7 % respectively in the experimental group, the positive control group and the negative control group. Accordingly, it appeared to be higher in the Experimental group and the positive control group than in the negative control group, but there was no significant difference statistically (p<0.01). 3. At 8 weeks after the operation, each amount of osteogenesis appeared to be 62.26.1 %, 17.42.5 % and 8.21.4 % respectively in the experimental group, the positive control group and the negative control group. Accordingly, it appeared to be substantially higher in the experimental group than in the positive control group and the negative control group, and there was a significant difference statistically (p<0.01). As a result of conducting the experiment, when ACS was used as a carrier for chitosan, chitosan showed effective osteogenesis in the perforated defective region of the Sprague-Dawley rat''s calvarial bone.
URI
http://ir.ymlib.yonsei.ac.kr/handle/22282913/128401
Appears in Collections:
2. 학위논문 > 2. College of Dentistry (치과대학) > 석사
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