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골다공증 유발 쥐에서 혈소판 농축 혈장이 골 재생에 미치는 영향
조종문,강정경,서규원,류재준,Cho, Jong-Moon,Kang, Jeong-Kyung,Suh, Kyu-Won,Ryu, Jae-Jun 대한치과보철학회 2010 대한치과보철학회지 Vol.48 No.1
연구 목적: 본 연구의 목적은 실험용 쥐의 초기 골 재생 과정에서 혈소판 농축 혈장 (platelet-rich plasma; PRP)이 난소호르몬 분비 유무에 따라 각각 어떤 효과를 나타내는지 확인 비교해 보는 것이다. 연구 재료 및 방법: 실험용 쥐 40마리 중 20마리에는 난소절제술 (ovariectomy; OVX)을 시행하여 골다공증을 유발시킨 상태에서 골 이식을 하였고, 나머지 20마리에는 난소절제술 없이 골 이식을 하였다. 또, 난소절제술을 시행한 쥐 중 10마리와 난소절제술을 시행하지 않은 쥐 중 10마리에는 골 이식 시 골전도성 합성골 이식재인 $MBCP^{TM}$ (Micro-& macro-porous biphasic calcium phosphate)에 혈소판 농축 혈장을 첨가하여 적용하고, 아래와 같은 실험군으로 구분하였다. A군; 10마리의 non-OVX 쥐/골 이식재 ($MBCP^{TM}$). B군; 10마리의 non-OVX 쥐/골 이식재($MBCP^{TM}$)+PRP. C군; 10마리의OVX 쥐/골 이식재($MBCP^{TM}$). D군; 10마리의OVX 쥐/골 이식재($MBCP^{TM}$)+PRP. 모든 실험동물의 두 개관 정중부에 직경 8 mm 원형의 임계 크기 결함을 한 개씩 인위적으로 형성한 후, 임계 크기 결함 내부에 골 이식재 및 혈소판 농축 혈장을 적용하여 골 이식술을 실시하였다. 골이식 시행 4주 후에 실험 동물을 희생시켜 표본을 제작한 후, 광학현미경상을 관찰하고 기존에 형성한 임계 크기 결함 내부에 새롭게 침착된 신생골의 면적을 측정하여 그 측정값을 통계 분석하였다. 결과: 신생골 면적 측정값을 각 군 간 비교하여 다음과 같은 결과를 얻었다. 1. 난소절제술이 시행되지 않은 정상 쥐에서는 혈소판 농축 혈장의 사용이 골 재생에 유의한 효과를 나타내지는 않았다 (p>.05). 2. 난소절제술이 시행된 골다공증 유발 쥐에서는 혈소판 농축 혈장의 사용이 골 재생에 유의한 효과를 나타내었다 (P<.05). 3. 혈소판 농축 혈장이 사용되지 않은 경우, 난소절제술이 시행된 골다공증 유발 쥐는 정상 쥐보다 골 재생 능력이 유의할 만큼 감소하였다 (P<.05). 4. 혈소판 농축 혈장이 사용된 경우, 난소절제술이 시행된 골다공증 유발 쥐라고 하더라도 정상 쥐보다 골 재생 능력이 유의할 만큼 감소하지는 않았다 (P>.05). 결론: 이상의 결과를 토대로, 골전도성 합성골 이식재와 함께 쓰인 혈소판 농축 혈장은 정상 쥐에서보다 난소절제술이 시행된 골다공증 유발 쥐에서 골 재생 및 치유에 더 큰 효과가 있음을 알 수 있었다. Purpose: The aim of this experimental study is to observe the effect of platelet-rich plasma (PRP) on early bone regeneration of rats both in normal condition and in osteoporosis induced by ovariectomy. Material and methods: Total 40 Sprague-Dawley female rats were divided into 4 groups. A 8-mm-diameter calvarial critical-sized defect (CSD) was made by drilling with trephine at the center of calvaria in cranium of every rat. Every CSD was augmented with an osteoconductive synthetic alloplastic substitute ($MBCP^{TM}$) and PRP as follows. Group A; 10 non-ovariectomized rats grafted with only $MBCP^{TM}$. Group B; 10 non-ovariectomized rats grafted with $MBCP^{TM}$ and PRP. Group C; 10 ovariectomized rats grafted with only $MBCP^{TM}$. Group D; 10 ovariectomized rats grafted with $MBCP^{TM}$ and PRP. At 4 weeks after graft, every rat was sacrificed. And histomorphometric analysis was performed by measuring calcified area of new bone formation within the CSD. Results: Comparing four groups, results were obtained as follows. 1. In non-ovariectomized groups, PRP showed a positive effect somewhat but not significant (P > .05). 2. In ovariectomized groups, PRP showed a positive effect significantly (P < .05). 3. In PRP untreated groups, ovariectomy diminished bone regeneration significantly (P < .05). 4. In PRP treated groups, ovariectomy diminished bone regeneration somewhat but not significant (P > .05). Conclusion: Within the limitation of this study, it can be concluded that PRP in combination with an osteoconductive synthetic alloplastic substitute has an effect on bone regeneration more significantly in ovariectomized osteoporotic rats than in normal rats.
조종한,윤근철,문 경,이재담 大韓成形外科學會 1996 Archives of Plastic Surgery Vol.23 No.3
A cartilage is the most commonly used tissue in reconstructive and esthetic surgery as an implant and it is closely related with ossification process. There were few reports of primary culture of chondrocytes in Korea because of complicated and time consuming procedures, and frequent fibroblast contamination. These previous reports were limited to monolayer cell culture, so no report exists about high density three dimensional primary culture of rat chondrocytes. We performed primary culture of rat chondrocytes using growth cartilage at the costochondral junction in young male Sprague-Dalwey rat of 80-120 gram body weight and culturing in 6.7×10⁴/360μl cell density with stainless steel Peni-cylinder in the center of the 35mm culture dish. Characterization with inverted microscopy, toluidine blue staining, and measuring alkaline -phosphatase activity was performed. To evaluate the effects of bovine pituitary extract (BPE) and fibroblast growth factor (FGF) on chondrocyte growth, the BPE or the FGF was added to the culture media. BPE is known to promote cell growth, and the FGF is the known active component of BPE. [³H]- thymidine incorporation was used as an index of DNA synthesis during cell proliferation. With adding the BPE or the FGF to the culture media, increased DNA synthesis was defected in comparison with the control. The effect of the BPE may be partially contributed by the FGF because the FGF is one of the active components of the BPE. To evaluate effect of the FGF,[³H]-proline incorporation was used as an index of collagen synthesis. At the level of 0.01ng/ml of the FGF concentration, the amount of collagen synthesis was almost same with the control, but it decreased with increasing the FGF concentration above 0.1ng/ml. From these data, we found that the FGF stimulates growth of chondrocytes and inhibits differentiation of chondrocytes. We consider that it can modulate ossification process with these actions.