骨形成蛋白二合併使用人工骨塊用於植體周圍區域骨再生之動態螢光分析

Evaluation Of Rhbmp-2 Based Artificial Bone Composite In Peri-Implant Alveolar Bone Regeneration : Dynamic Fluorescence Analysis

黃士哲(Shih-Je Huang)

指導教授 ： 林立德

國立臺灣大學/牙醫專業學院/臨床牙醫學研究所/碩士(2015年)

https://doi.org/10.6342/NTU.2015.10664

摘要

在現今臨床牙科領域裡面，人工牙根的使用已經趨於普及，讓我們臨床上的治療更加順利。但同時也經常遇到病患植牙區域骨質不足，而無法直接置放人工牙根，需要進一步進行手術來增加骨量，特別是垂直方向上的骨量。現今文獻上有許多增加骨量的術式，包括牽引成骨術、引導式骨再生術、自體骨移植和使用生長因子和選擇適合的載體。但是現在對於垂直齒槽骨再生及和植體周圍骨缺損的骨再生，在現代牙科植體學中還是一項困難的挑戰。本研究的目的即是研究加入骨形成蛋白二的人工合成骨塊，修復植體周圍骨缺損的潛力和效益性。
本實驗中使用了健康的米格魯雄性成犬，六個表面經多孔鈦金屬氧化處理的人工植體（TIUNITE, 4.0 X 8.5 MM; NOBEL BIOCARE, SWEDEN）植入到下顎骨的齒槽骨缺損（高度為四毫米、長度為十毫米），人工牙根植入的深度為四毫米，留下四點五毫米曝露在齒槽骨上植體周圍的骨缺損。這些骨缺損為隨機分布到五個組別：負控制組、人工骨塊組、其他三組分別為每毫升五、二十、五十微克骨形成蛋白二於人工骨塊。 為了觀察及量化新生骨的形成過程，實驗動物的螢光骨染色標定分別於手術當天注射四環黴素、三週後注射鈣黃綠素、六週後注射茜草素紅。在大部分骨缺損區的癒合情形都相當良好，除了某些區域有軟組織的併發症，而最後有螺絲帽露出的情形，但並沒有其他的發炎反應。術後八週將動物犧牲後取下標本，並製作成組織玻片於螢光顯微鏡下觀察。螢光顯微鏡下觀察可發現骨質生成順序，由人工骨塊和齒槽骨相接觸開始朝向植體表面生長，此外對於骨形成蛋白二濃度越高的組別，早期骨質生成所產生的螢光範圍越廣，也代表著早期骨生成速度更快。
本研究建立的實驗動物模型是為了評估在人工植體周圍的垂直高度骨再生能力，人工骨塊於本實驗中扮演的角色是當做一個骨引導作用的支架和骨形成蛋白二的載體，且能對骨再生有效的影響，而骨形成蛋白二的濃度和時間點對於骨再生的質量及範圍有顯著的影響。另一方面我們建立硬組織玻片處理方式與骨染劑之螢光顯微鏡觀察方法，提供在骨組織工程後續樣本處理與螢光觀察研究獲得更精準的方法，讓後續實驗研究得以順利進行。

關鍵字

骨形成蛋白二 ； 人工植體 ； 人工骨塊 ； 垂直高度齒槽骨再生 ； 臨界性骨缺損 ； 動物活體試驗 ； 螢光顯微鏡

並列摘要

INTRODUCTION: IN TODAY'S CLINICAL DENTAL FIELD, ARTIFICIAL TOOTH ROOT DENTAL IMPLANTS USE HAS BECOME POPULAR. BUT ALSO LESS FREQUENTLY ENCOUNTERED IN PATIENTS WITH IMPLANT REGION, THE NEED FOR FURTHER SURGERY TO INCREASE BONE MASS, PARTICULARLY BONE MASS IN THE VERTICAL DIRECTION. BUT NOW FOR THE VERTICAL ALVEOLAR BONE REGENERATION AND REPAIR BONY DEFECTS AROUND THE IMPLANT, IT IS STILL A DIFFICULT CHALLENGE. THE PURPOSE OF THIS STUDY IS BONE MORPHOGENETIC PROTEIN ADDED TO SYNTHETIC BONE BLOCK TO REPAIR BONY DEFECTS AROUND THE IMPLANTS.
MATERIAL AND METHODS : THE EXPERIMENT USED A HEALTHY MALE ADULT DOGS BEAGLE, SIX SURFACE OXIDATION TREATMENT BY POROUS TITANIUM METAL ARTIFICIAL IMPLANTS (TIUNITE, 4.0 X 8.5 MM; NOBEL BIOCARE, SWEDEN) IMPLANTED INTO THE JAW BONE OF THE ALVEOLAR BONE DEFECT (A HEIGHT OF FOUR MILLIMETERS AND A LENGTH OF 10MM), THE ARTIFICIAL TOOTH ROOT IMPLANTATION DEPTH IS FOUR MILLIMETERS, LEAVING 4.5 MM EXPOSED ON THE ALVEOLAR BONE IMPLANTS AROUND THE BONE DEFECT. THESE BONE DEFECTS RANDOMLY DISTRIBUTED TO FIVE GROUPS: NEGATIVE CONTROL GROUP, BONE BLOCK GROUP, THE OTHER GROUPS WERE 5, 20, 50 UG/ML OF BONE MORPHOGENETIC PROTEIN IN BONE TWO BLOCKS. IN ORDER TO OBSERVE AND QUANTIFY NEW BONE FORMATION, BONE FLUORESCENT DYE CALIBRATION LABORATORY ANIMALS WERE INJECTED TETRACYCLINE AT THE DAY OF SURGERY, THREE WEEKS AFTER THE INJECTION OF CALCEIN, SIX WEEKS AFTER INJECTION OF ALIZARIN RED. AFTER EIGHT WEEKS AFTER SURGERY TO REMOVE THE SACRIFICIAL ANIMAL SPECIMENS, AND MADE INTO TISSUE SLIDES OBSERVED UNDER FLUORESCENT MICROSCOPE.
RESULTS : IN THE CASE OF MOST OF THE HEALING BONE DEFECTS ARE QUITE GOOD, EXCEPT FOR SOME AREAS OF SOFT TISSUE COMPLICATIONS. OBSERVE THE TEETH SECTION UNDER THE FLUORESCENT MICROSCOPE, FROM THE OUTER RING OF TOOTH PULP TO THE INSIDE, IS YELLOW, GREEN, RED FLUORESCENT DENTIN DEPOSITION LINES. THE LINES ARE CORRESPOND TO FLUORESCENT DRUG ADMINISTRATION SEQUENCE. WEEK 0 TETRACYCLINE (YELLOW FLUORESCENT); WEEK 3 CALCEIN (GREEN); WEEK 6 ALIZARIN RED (RED). AND IT’S CORRESPOND TO SECONDARY DENTIN DEPOSITION FROM OUTSIDETO INSIDE. THE GROUP WITH BMP-2 APPLICATION SHOWS HIGHLY BONE REMODELING IN BONE GRAFT MATERIAL. AND THE CONTROL GROUP WITHOUT BMP-2 APPLICATION SHOWS LOTS OF BONE GRAFT MATERIAL MAINTAIN THE SPACE AND DID NOT BE RESORBED.
CONCLUSION : THE ARTIFICIAL BONE BLOCK WITH BMP-2 USED IN PERI-IMPLANT BONY DEFECT SHOWS GREAT BONE REMODELING IN BONE BLOCK. AND THE HIGHER BMP-2 DOSE HAS EARLIER AND FASTER BONE REMODELING SYMPTOM IN SIXTH WEEK. IN CONTROL GROUP, THE BONE BLOCK CAN MAINTAIN THE SPACE AND WILL NOT BE WASHED OUT. IN ADDITION THE EXPERIMENT TO CREATE A HARD TISSUE SLIDES PRODUCTION PROCESSES, THE WAY OF FOLLOW-UP OBSERVATION WITH FLUORESCENT BONE DYE.

並列關鍵字

BONE MORPHOGENETIC PROTEIN-2(BMP-2) ； DENTAL IMPLATNS ； ARTIFICIAL BONE BLOCK ； VERTICAL RIDGE AUGMENTATION ； ANIMAL STUDY ； FLUORESCENT MICROSCOPE

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