| Abstract: | 在矯正的方面的研究當中，如何加速牙齒移動一直是矯正醫師想要克服的問題之一。為了加速牙齒移動，有許多學者提出不同的方法，包括比較侵入性或是非侵入性的方法。其中有醫師研究認為，在局部注射藥物像是前列腺素，應該有助於骨組織的吸收代謝，可以加速牙齒移動的速度。而矯正的患者最常會被注射的局部藥物，就是在拔牙的時候所注射的局部麻醉劑。因此本次研究想要了解，局部麻醉葯物會不會對牙周組織細胞的成骨作用造成影響。
本次實驗旨在進行局部麻醉劑之細胞模式研究，實驗所使用的細胞是牙周韌帶細胞以及人類骨母細胞 （MG63）兩種細胞株進行研究；藥物的部分則是選用兩種牙科常見的局部麻醉劑：Septanenest with adrenaline成分是Articaine hydrochloride, Septodont Co.，簡稱Ｓ（＋）；Scandonest 3%成分Mepivacaine hydrochloride, Septodont Co.，簡稱Ｓ（—）。分別將含有不同濃度的局部麻醉劑溶液加入細胞培養皿中與細胞一起培養24小時，觀察PDL細胞以及MG63的細胞外觀、局部麻醉劑的細胞毒性（MTT）以及發炎因子COX-2、IL-1、IL-6、TNF-α的表現量，並汲取MG63細胞培養皿中的溶液（Extraction Protein A）。另外實驗將不同濃度的局部麻醉劑加入PDL細胞培養皿中培養24hr後：汲取PDL細胞培養皿中的溶液加入MG63細胞培養皿中培養24小時，之後汲取培養皿中的溶液（Extraction protein B）；同時加入細胞培養溶液至原PDL培養皿中繼續培養24小時，之後汲取PDL細胞培養皿中的溶液加入MG63細胞培養皿中培養24小時，再汲取培養皿中的溶液（Extraction protein C）。利用細胞墨點法分析細胞溶液（Extraction Protein A/ B/ C）中的成骨細胞分化蛋白RANKL、ALP、OPG的表現。
研究的結果顯示，在細胞外觀以及細胞毒性的分析中，局部麻醉劑的細胞毒性會隨濃度增加而升高；細胞的活性降低，細胞外觀也都有呈現逐漸皺縮的趨勢。在發炎因子的檢測方面，局部麻醉劑皆會增加PDL細胞和MG63發炎因子的表現。在成骨細胞分化因子的表現上，MG63細胞在含有局部麻醉劑的環境下會增加RANKL的表現量。PDL細胞在含有局部麻醉劑的環境下所釋出的影響因子幾乎不會影響MG63細胞分化因子的表現。但隨後繼續培養的PDL細胞則會會使得MG63細胞的RANKL表現增加、OPG的表現降低。綜合以上之結果，局部麻醉劑會影響牙周組織細胞，促進骨吸收的作用。
In the study of orthodontics, how to accelerate tooth movement has always been one of the problems that orthodontists want to overcome. In order to accelerate tooth movement, many scholars have proposed different methods, including more invasive or non-invasive methods. Among them, doctors believe that local injection of drugs like prostaglandins should contribute to the absorption and metabolism of bone tissue and accelerate the movement of teeth. The localized drug that is most often injected by the corrected patient is the local anesthetic injected at the time of tooth extraction. Therefore, this study wants to understand whether local anesthetics will affect the osteogenesis of periodontal tissue cells.
The purpose of this experiment was to conduct a cell model study of local anesthetics. The cells used in the experiment were periodontal ligament cells and human osteoblasts (MG63). The drug part was selected from two types of dental common. Local anesthetic: Septanenest with adrenaline ingredients is Articaine hydrochloride, Septodont Co., S(+); Scandonest 3% ingredient Mepivacaine hydrochloride, Septodont Co., S(-). The local anesthetic solution containing different concentrations was added to the cell culture dish and cultured with the cells for 24 hours to observe the cell appearance of PDL cells and MG63, the cytotoxicity (MTT) of local anesthetics, and the inflammatory factors COX-2, IL-1, IL. -6, the amount of TNF-α expression, and draw the solution in the MG63 cell culture dish (Extraction Protein A). In addition, different concentrations of local anesthetic were added to the PDL cell culture dish for 24 hrs: the solution in the PDL cell culture dish was added to the MG63 cell culture dish for 24 hours, and then the solution in the culture dish (Extraction protein B) was taken; The cell culture solution was added to the original PDL culture dish for further 24 hours, and then the solution in the PDL cell culture dish was added to the MG63 cell culture dish for 24 hours, and the solution in the culture dish (Extraction protein C) was extracted. The expression of osteoblast differentiation proteins RANKL, ALP, and OPG in cell extract (Extraction Protein A/B/C) was analyzed by cell dot method.
The results of our study showed that in the analysis of cell appearance and cytotoxicity, the cytotoxicity of local anesthetics increased with increasing concentration; the activity of cells decreased, and the appearance of cells also showed a tendency to shrink. In the detection of inflammatory factors, local anesthetics increase the performance of PDL cells and MG63 inflammatory factors. In the expression of osteoblast differentiation factors, MG63 cells increase the expression of RANKL in the presence of a local anesthetic. The effect factors released by PDL cells in a local anesthetic environment hardly affect the expression of MG63 cell differentiation factors. However, the subsequent culture of PDL cells will increase the RANKL expression of MG63 cells and decrease the performance of OPG. Based on the above results, local anesthetics affect the periodontal tissue cells and promote bone resorption. |
