CAJ | 학술논문

目的：建立Beagle犬的种植体周围骨缺损模型，构建含有质粒pcDNA3．1 flag-Osx的基因活化基质，并与自体骨髓基质干细胞混合，观察Osx基因活化技术对种植体周围骨的形成和矿化的作用。方法：术前抽取Bea-gle犬骨髓，培养骨髓基质干细胞，扩增质粒pcDNA3．1 flag-Osx。雄性Beagle犬15只，随机分为3个组，每组5只，第1组为4周组，第2组为8周组，第3组为12周组。每只动物拔除左侧下颌第2、4前磨牙及右侧第2前磨牙，于拔牙创颊侧制备骨缺损，植入奥齿泰GSII种植体3枚，分别填入A组：倍骼生； B组：倍骼生+BMSCs+空载体pcDNA3.1flag； C组：倍骼生+BMSCs+pcDNA3.1flag-Osx。应用GBR技术，在材料表面覆盖Bio-gide胶原膜。分别于4、8、12周后处死动物，取出标本，应用影像学、组织学、组织形态计量学等手段与对照组比较。各指标用均数±标准差（X±s）来表示。应用统计软件SPSS17.0进行单因素的方差分析。 P<0.05为差异有统计学意义。结果：1、硬组织观察：骨缺损处均有新骨形成。 C组最多， B组其次， A组最少。2、 Micro-CT数据结果显示：第1组：除骨体积分数（BVF）外，其余指标B组与A组均有显著差异（P<0.05）。骨小梁间隙（Tb.Sp.）、骨小梁数目(Tb.N)、 BVF、骨小梁的体积(BV) C组与B组间差异有显著性（P<0.05）。各指标C组与A组间差异均有显著性（P<0.05）。第2组：除BVF外，其余指标B组与A组均有显著差异（P<0.05）。 Tb.Sp.、骨小梁厚度(Tb.Th)、 Tb.N.、 BV C组与B组间差异有显著性。各指标C组与A组间差异均有显著性（P<0.05）。第3组：各指标B组与A组均有显著差异（P<0.05）。除BVF外其余指标C组与B组间差异有显著性（P<0.05）。各指标C组与A组间差异均有显著性（P<0.05）。结论：骨髓基质干细胞与Osx质粒构成的基因活化材料，有明显的骨诱导作用，与不含Osx基质的对照组相比成骨速度更快，新生骨量更多，明显促进了骨组织再生。目的：建立Beagle犬的種植體週圍骨缺損模型，構建含有質粒pcDNA3．1 flag-Osx的基因活化基質，併與自體骨髓基質榦細胞混閤，觀察Osx基因活化技術對種植體週圍骨的形成和礦化的作用。方法：術前抽取Bea-gle犬骨髓，培養骨髓基質榦細胞，擴增質粒pcDNA3．1 flag-Osx。雄性Beagle犬15隻，隨機分為3箇組，每組5隻，第1組為4週組，第2組為8週組，第3組為12週組。每隻動物拔除左側下頜第2、4前磨牙及右側第2前磨牙，于拔牙創頰側製備骨缺損，植入奧齒泰GSII種植體3枚，分彆填入A組：倍骼生； B組：倍骼生+BMSCs+空載體pcDNA3.1flag； C組：倍骼生+BMSCs+pcDNA3.1flag-Osx。應用GBR技術，在材料錶麵覆蓋Bio-gide膠原膜。分彆于4、8、12週後處死動物，取齣標本，應用影像學、組織學、組織形態計量學等手段與對照組比較。各指標用均數±標準差（X±s）來錶示。應用統計軟件SPSS17.0進行單因素的方差分析。 P<0.05為差異有統計學意義。結果：1、硬組織觀察：骨缺損處均有新骨形成。 C組最多， B組其次， A組最少。2、 Micro-CT數據結果顯示：第1組：除骨體積分數（BVF）外，其餘指標B組與A組均有顯著差異（P<0.05）。骨小樑間隙（Tb.Sp.）、骨小樑數目(Tb.N)、 BVF、骨小樑的體積(BV) C組與B組間差異有顯著性（P<0.05）。各指標C組與A組間差異均有顯著性（P<0.05）。第2組：除BVF外，其餘指標B組與A組均有顯著差異（P<0.05）。 Tb.Sp.、骨小樑厚度(Tb.Th)、 Tb.N.、 BV C組與B組間差異有顯著性。各指標C組與A組間差異均有顯著性（P<0.05）。第3組：各指標B組與A組均有顯著差異（P<0.05）。除BVF外其餘指標C組與B組間差異有顯著性（P<0.05）。各指標C組與A組間差異均有顯著性（P<0.05）。結論：骨髓基質榦細胞與Osx質粒構成的基因活化材料，有明顯的骨誘導作用，與不含Osx基質的對照組相比成骨速度更快，新生骨量更多，明顯促進瞭骨組織再生。
목적：건립Beagle견적충식체주위골결손모형，구건함유질립pcDNA3．1 flag-Osx적기인활화기질，병여자체골수기질간세포혼합，관찰Osx기인활화기술대충식체주위골적형성화광화적작용。방법：술전추취Bea-gle견골수，배양골수기질간세포，확증질립pcDNA3．1 flag-Osx。웅성Beagle견15지，수궤분위3개조，매조5지，제1조위4주조，제2조위8주조，제3조위12주조。매지동물발제좌측하합제2、4전마아급우측제2전마아，우발아창협측제비골결손，식입오치태GSII충식체3매，분별전입A조：배격생； B조：배격생+BMSCs+공재체pcDNA3.1flag； C조：배격생+BMSCs+pcDNA3.1flag-Osx。응용GBR기술，재재료표면복개Bio-gide효원막。분별우4、8、12주후처사동물，취출표본，응용영상학、조직학、조직형태계량학등수단여대조조비교。각지표용균수±표준차（X±s）래표시。응용통계연건SPSS17.0진행단인소적방차분석。 P<0.05위차이유통계학의의。결과：1、경조직관찰：골결손처균유신골형성。 C조최다， B조기차， A조최소。2、 Micro-CT수거결과현시：제1조：제골체적분수（BVF）외，기여지표B조여A조균유현저차이（P<0.05）。골소량간극（Tb.Sp.）、골소량수목(Tb.N)、 BVF、골소량적체적(BV) C조여B조간차이유현저성（P<0.05）。각지표C조여A조간차이균유현저성（P<0.05）。제2조：제BVF외，기여지표B조여A조균유현저차이（P<0.05）。 Tb.Sp.、골소량후도(Tb.Th)、 Tb.N.、 BV C조여B조간차이유현저성。각지표C조여A조간차이균유현저성（P<0.05）。제3조：각지표B조여A조균유현저차이（P<0.05）。제BVF외기여지표C조여B조간차이유현저성（P<0.05）。각지표C조여A조간차이균유현저성（P<0.05）。결론：골수기질간세포여Osx질립구성적기인활화재료，유명현적골유도작용，여불함Osx기질적대조조상비성골속도경쾌，신생골량경다，명현촉진료골조직재생。
Objective: To build the beagle dog peri-implant bone defect model, and construct Gene Activated Matrix (GAM)containing plasmid pcDNA3.1 flag-Osx and mixed with autologous Bone Marrow Stromal Cells (BMSC);to evaluate the effect of Osx gene activation technology on bone formation and minerali-zation. Methods: With the marrow extracted prior to operation, bone marrow stromal cells were culti-vated and plasmid pcDNA3.1 flag-Osx amplified. Fifteen male beagle dogs were randomly whilst evenly divided into three groups, with group 1 being a four-week group, group 2 as an eight-week group and group 3 a twelve-week group. The left mandibular 2nd and 4th premolar and the rihgt mandibular 2nd premolar were extracted, based on which the operations of trapezoidal incisions and bone defects were made. Each of the three bone defects was implanted with an OSSTEM GSII, and filled respectively with:Category A, Perioglas;Category B, Perioglas+BMSCs+empty vector pcDNA3.1flag;and Category C, Perioglas+BMSCs+pcDNA3.1flag-Osx. Each material was covered with Bio-Gide collagen membrane via Guided Bone Regeneration (GBR) technique. Finally in 4, 8, and 12 weeks after the operation, ani-mals in group 1, 2 and 3 were killed respectively to obtain the specimens. The specimens were analyzed by comparison with the control group utilizing histological, imageological, and histomorphometric mea-surements. Each index is weighted by Mean ±Standard deviation (X ±s). The index differences between groups at each time sample are analyzed using the statistical package SPSS 17.0 by one way analysis of variance (ANOVA). P<0.05 were considered statistically significant. Results:1. Sclerous tissue exami-nation. Newly formed bone appeared at every bone defect under tests, among which Group C has the richest new bones, Group B has less and Group A has the least. 2.Results from Micro-CT analysis:Group 1: Except BVF, all the other indexes were significant different between category B and A (P<0.05). In terms of Trabecular Spacing (Tb.Sp), Trabecular Number (Tb.N), Bone Volume Fraction (BVF) and Bone Volume (BV), significant difference existed between category C and category B (P<0.05). For each index, there was significant difference between category C and A (P<0.05). Group 2: Except BVF, all the other indexes were significant different between category B and A (P<0.05). In terms of Tb.Sp, Tb.N, BV and Trabecular Thickness(Tb.Th), significant difference existed between category C and cate-gory B (P<0.05). For each index, there was significant difference between category C and A (P<0.05). Group 3: For each index, there was significant difference between category B and A (P<0.05). Except BVF, all the other indexes were significant different between category C and B ( P<0.05) . For each in-dex, there was significant difference between category C and A ( P<0.05) . Conclusions: Superior to the solutions without applying Osx plasmid, gene activated matrix combined with bone marrow stromal cells and Osx plasmid offer a faster bone issue promoting speed. These materials have significant bone induction effect which leads to more neoformative bone mass regenerated.