新式膠原蛋白骨填料於齒槽骨脊保存術之功效性評估：動物試驗

Efficacy of novel β-TCP/Collagen composites in ridge preservation: Animal Study

楊倚弦(Yi-Hsien Yang)

指導教授 ： 郭彥彬

國立臺灣大學/醫學院/臨床牙醫學研究所/碩士(2017年)

https://doi.org/10.6342/NTU201702775

摘要

臨床牙科處置上，常常面臨拔牙後齒槽骨發生寬度跟高度減少的問題。若病人想回復缺牙區的功能，會面臨重建已喪失的齒槽骨脊高度跟寬度的問題。因此如何在拔牙當時適當保留齒槽骨高度跟寬度，其應用術式與材料是需要研究的課題。目前雖有膠原蛋白骨填料應用於臨床拔牙窩或骨缺損的修補，但其臨床之降解期往往有過長的問題，往往在組織學上會發現較多非成熟性的骨小樑及剩餘的骨填料；本研究應用新式膠原蛋白骨填料，材料萃取自豬體的膠原蛋白，經由升溫脫水、冷凍乾燥的處理方式，使結構產生交聯作用；β型三鈣磷酸鹽因其化學計量上類似骨質之非晶狀生物前驅物，為生物可降解，其降解產物能被人體重組以形成新骨礦物質，利於骨沈積之發生。此外，此類混合β型三鈣磷酸鹽及膠原蛋白的產品，在實際手術操作上，也具有不錯之生物相容性及良好操作性質。本研究之目的，希望能開發具備生物相容性良好及操作便利特質的膠原蛋白骨填料，以動物實驗確認其保存拔牙後齒槽脊的功效。實驗動物為六隻成犬，於不同時間點拔牙，同時使用所開發之膠原蛋白骨填料及市售產品進行齒槽骨脊保存的相關比較。實驗方法測試於米格魯犬之下顎第三、四小臼齒之拔牙窩進行齒槽骨保存術後，觀察於第四、八及十二周之空間維持與骨再生之效果。每隻動物先拔除下顎第三、四小臼齒，並於頰側骨板各製造一個高四公釐、寬八公釐單壁之骨缺損，以隨機分配決定各個缺損所放置之實驗材料組別。第一組實驗組放置「含(10% collagen， 90% β-TCP)的海昌膠原蛋白填骨材料（“Horien” Collagen Bone Graft Matrix）」，第二組實驗組放置「含(20% collagen， 80% β-TCP)的海昌膠原蛋白填骨材料（“Horien” Collagen Bone Graft Matrix）」，而陽性對照組放置「“吉士德”百歐士膠原質 (“Geistlich ” Bio-Oss® Collagen)」，陰性對照組不放置任何材料；藉以評估不同的膠原蛋白填骨材料於齒槽骨保存之效果。
實驗結果呈現四個組別使用之材料在術後癒合皆無不良反應。組織形態測量的結果顯示，新生骨面積百分比（New bone area ％）的數值在四組排序由多至少分別為實驗組T2（38.18±6.85％）、陽性對照組實驗組（32.37±10.74％）、實驗組T1（25.02±19.50％）、陰性對照組（24.97±8.91％），各組間並無統計顯著差異。微斷層掃瞄之結果顯示，新生骨體積百分比（new bone volume %）在實驗組和陽性對照組非常接近（分別為71.68±12.63% 與82.40±14.14％），並皆高於陰性對照組（31.18±12.57％），有統計上的顯著差異。於螢光顯微鏡的觀察，骨生長的方向大致從缺損外圍往中心生長，而各組中骨生長的方式略有不同，實驗組似乎在術後癒合的前期前期還是後期?有加速骨生長的傾向，螢光顯微鏡中在四組中新生骨面積百分比（New bone area ％）的估算排序由多至少分別為陽性對照組實驗組（15.24±8.85％）、實驗組T1（13.88±3.15％）、實驗組T2（12.42±8.37％）、陰性對照組（6.33±3.73％），彼此間並無統計顯著差異。
整體而言，此β-TCP/Collagen composites之使用，具有保存拔牙後齒槽骨的潛力，因樣本數較小，統計上不易達到顯著差異，後續仍需更多研究證實其功效。另外透過骨螢光標定技術，除了能呼應本實驗其他結果，也使我們進一步觀察到不同組別中骨缺損的新骨生成模式。

關鍵字

β型三鈣磷酸鹽 ； 骨替代物 ； 齒槽骨保存術

並列摘要

Background: Alveolar ridge remodeling is observed after tooth extraction. It would become the problem if future rehabilitation involved implant placement. Therefore , ridge preservation technique was introduced to solve this kind of problem. Although there are many available products of β-TCP(β-tricalcium phosphate)/Collagen composites for clinical usage, such as for periodontal regeneration and repair of bony defect. There is still lack of ideal products fulfilled the requirement of easily handling and predicable clinical outcome. Some popular product such as Bio-Oss® Collagen may retained too long during its degradation. It may degrade relatively slowly, more residual bone filler after bone modeling. β -TCP also has substantial physical strength; it provides a three-dimensional scaffold for bone regeneration against the pressure of tissue shrinkage . Moreover, β-TCP has the potential to function as a reservoir of calcium and phosphate ions for the local tissue during the degradation process, which possibly results in stimulation of osteoblastic function and promotion of bone formation. Above as though better than traditional β-TCP/Collagen composites. In this project, we will perform animal test to identify its clinical efficacy. Different β-TCP/Collagen composites will be tested in their physical property. Afterward proceeding for further animal experiment in observation the dimensional change of the extraction socket. Six dogs will be used in this project, symmetric extraction sockets will be created and covered with commercial or new developed β-TCP/Collagen composites in this project at different time points to identify its efficacy in ridge augmentation. We hope the β-TCP/Collagen composites will be developed and its superiority in guide bone regeneration will also been identified following this project proceeding.
Materials and Methods:
The mandibular third premolars and fourth premolars of six dogs were extracted bilaterally, and buccal dehiscence defects were prepared bilaterally on the buccal side of the mandibular third and fourth premolars. Sixteen defects were assigned to four treatment groups: T1 group (β-TCP/10%Collagen composites );T2 group (β-TCP/20%Collagen composites ) ; positive control (Bio-Oss® Collagen); negative control (no grafting material ). Fluorescence bone labeling was administrated subcutaneously three weeks, five weeks and seven weeks post-operatively. The animals were sacrificed 8 weeks after surgery. For histomorphometric analysis, new bone area were measured. New bone volume was measured using microcomputed tomography(micro-CT). Fluorescence microscopic observation was performed to figure out the possible sequence of new bone formation in each group.
Result: No adverse effect was found in all four groups after ridge preservation surgery. For new bone area (%) of Histomorphometry, the four groups was 25.02±19.50% for the T1 group, 38.18±6.85% for the T2 group , 32.37±10.74% for the positive control group, and 24.97±8.91% for the negative control group. The differences between each group were not statistically significant. For new bone volume (%)analyzed from micro-CT, the result of the T1 group come close to the positive control group (respectively 71.68±12.63 % and 82.40±14.14 %), which is higher than T2 group (62.00±14.39 %) and the negative control group (31.18±12.57 %).Nevertheless, there is statistical significant between positive control、T1 group and negative control group. Under fluorescence microscopy, the pattern of new bone formation usually starts from the border of the defect toward the central part of the defect. However, the timing and the specific area of new bone formation are different in positive control、T1、T2 and negative control groups. It seems that the test group may delay the time of bone formation (compare with negative control group) ,but more bone gain may find after 5 weeks in the test group. For new bone area (%)of fluorescence microscopy calculated separately in the four groups was 13.88±3.15 % for the T1 group, 12.42±8.37 % for the T2 group , 15.24±8.85 % for the positive control group, and 6.33±3.73% for the negative control group. But the differences between each group were not statistically significant.
Conclusion: The β-TCP/Collagen composites seems to have the potential to preserve ridge volume. However, due to limited sample size in this study, there is less possibility that the result between groups achieving statistical significance. More research are needed on investigating the potential of this new β-TCP/Collagen composites for ridge preservation. In addition, the fluorescence bone labeling techniques not only supporting the results of histology and micro-CT , but also assisting us to understand the mechanism of new bone formation in different treatment

並列關鍵字

β-TCP ； bone substitute ； Ridge preservation

參考文獻

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