| Abstract: | 將牙根管通道系統及其表面可能的通道封閉是逆向充填或是牙根修復材料最
為主要的目的。因此良好的生物相容性也是必備的要素，除了不會造成細胞毒性之
外，最好能具有引導新骨再生、牙週組織附著、牙骨質再生的能力。
因此本研究目的：
1. 於體外試驗，探討自行合成之MTA之對細胞發炎免疫方面反應。
2. 於體外試驗，探討自行合成之MTA於細胞內對於骨生成因子之訊息變化。
3. 於體內試驗，探討自行合成之MTA於動物體內植入後之生物體反應。
研究材料與方法：
第一與第二年為體外試驗。依過去的製程MTA燒結出，配製成所需的溶液，細胞
採用U2OS 細胞株。於細胞發炎ELISA分析，以材料作用於細胞後，分析有關於負責活化
骨細胞之細胞素如； IL 1α,1β和IL6之變化。同時也分析osteocalcin，alkalaine phosphatase
之活性。以鹼性磷酸脢(alkaline phosphatase [ALP] activity)分析材料與細胞作用後之活性。
以RTPCR方法，觀測細胞COX-2的表現。以西方墨點分析探討細胞內訊息蛋白COX-2
protein、ERKinase、 JNK kinase、caspase 3 、alkaline phosphatase, osteonidogen, osteonectin,
和 osteopontin之的表現，以了解自行研發材料的生物活性機轉。第三年為作用於老鼠體內
的試驗，以60隻Sprague Dawley rat作為植入之對象，以合成之MTA為實驗組，另外以
Hydroxy apatite 作為控制組，作為比較組織反應之差異性。植入部位為parietal bone和
subcutaneous位置，觀察記錄其組織之發炎反應，並紀錄之。所有結果居已統計軟體分析
比較結果。
研究結果顯示
CS材料對於U2OS細胞具有良好之生物相容性，隨著濃度與作用時間改變，其細胞
生長現象良好。在發炎因子COX-2表現上，CS與MTA一起比較顯示於第三天有明顯COX-
2表現，第七天則無差異。 Interleukin 的表現方面，IL1α，IL1β和IL6的表現，CS和MTA的
表現無差異，但比控制組的表現高。CS材料對於MG63細胞具有良好之生物相容性(Fgure
6.)，隨著濃度與作用時間改變，其細胞生長現象良好。MTA與CS材料之ERKinase 表現較
控制組佳，代表有良好之細胞增殖反應；對於骨誘導生成因子ALP, Type I collegenase,
osteocalcin, Bone sialoprotein 和 osteopontin之表現也都呈現明顯反應。不論是MTA或是CS
材料於動物體內植入第六週均有呈現發炎反應，到第十二周，發炎效應就降低。組織埋入
與切片結果顯示材料對於生物體據有良好生物相容性。
結語：
本計劃顯示本研究室自行合成之鈣矽類材料據有良好之生物相容性，日後可以應用
於臨床上
The root end filling material is used on the root apex or pulp perforation repair. The material will
contact with tooth and surround tissue. It is needed material have good biocompatibility. The
ideal material should have good physical properties, biocompatibility and may have
osteoconduction or tissue regeneration ability. The purpose of present project was to 1.
Investigate the calcium silicate (CS) and MTA inflammation or immune reaction. 2. The CS and
MTA osteoconduction ability. 3. The material implantation reaction on rate. Material and
methods: Follow the previous study, CS were fabricated from our laboratory. The CS material
after preparation were compared with MTA. The control group in present were blank group. The
MTT assay were to study the cell survival rate. The western blot assay and RTPCR assay were
used to identify following marker expression. They were included: COX-2 protein、ERKinase、
JNK kinase、caspase 3 、alkaline phosphatase, osteonidogen, osteonectin, osteopontin proteins.
The material after preparation were implanted into rat submucosa and parietal bone. The HE stain
were to evaluate the inflammation reaction. The data were all statistically analysed to compare
the difference. The result showed as follows: 1. The CS and MTA were all biocompatible with
U2OS cells. It can promote the cell growth. The ERK Kinase showed increasing expression . 2.
The inflammatory marker COX2 were appeared when initial contact with cells. And were
decreasing as culture time increased. It represent no inflammation reaction happened. 3.The
expression of IL1α，IL1β, IL6 showed MTA and CS were have different degrees changes. The
outcome showed materials were no immune reaction to cell. 4. The MG63 bone cell were also
biocompatible with CS and MTA. The bone marker expression were obviously appeared on ALP,
Type I collegenase, osteocalcin, Bone sialoprotein ,osteopontin proteins. 5. The implantation test
on rat, the subcutaneous tissue showed mild inflammation reaction, not in parietal bone
implantation. No dominant bone formation were found in bone tissue.
Conclusion: The CS material were biocompatible with cell in vitro study and in animal in vivo
study. This material can be used in clinical after further clinical study test. |
