不同參數之二氧化碳雷射對牙周韌帶細胞的影響

The Different Irradiation Parameters of Carbon Dioxide Laser Effects on Periodontal Ligament Cells

何俊德(Chun-Te Ho)

指導教授 ： 高嘉澤

中山醫學大學/口腔醫學院/牙醫學系/博士(2022年)

https://doi.org/10.6834/csmu202200156

摘要

研究目的
本研究的目的是使用負壓培養來模擬矯正移動牙齒張力側的環境，受機械力刺激後的牙周韌帶（Periodontal Ligment , PDL）細胞以二氧化碳（CO2 ）雷射照射做對照比較，評估不同 PBS（Photobiostimulation）參數對培養的 PDL 細胞之影響，進一步分析被照射的PDL細胞，其骨形成物和發炎物質的差異。
材料和方法
將加有一般培養基和分化培養基上負壓培養的PDL細胞，以能量密度為5J/cm 2 或10J/cm 2 的CO 2 雷射連續照射。在第 1、5 和 7 天照射的 PDL 細胞，以Presto Blue 測定存活率，以反轉錄聚合酶鏈反應（RT-PCR）測量細胞的生物標記物，鹼性磷酸酶 (ALP)、骨唾液酸生成素 (BSP)、骨橋蛋白 (OPN)、骨鈣素 (OC)、基質金屬蛋白酶3（MMP-3）和環氧合酶2（COX-2）的表現。
結果
PDL細胞存活率顯示分化培養基組高於一般培養基組。細胞形態呈現為梭型。雷射照射組的發炎標記物在第一天有較明顯表現，在第七天表現下降（p<0.05）。成骨標記物在不同時間呈現高度表現（p<0.05）。 Col I和OPN基因在第一天表現較高，Col I基因的高度表現一直持續到第七天。 OC基因表現在第七天較高表現。

結論
本研究結果顯示使用負壓培養來模擬矯正移動牙齒張力側的環境
，發現PDL細胞可被誘導進入成骨途徑，減少細胞發炎。

關鍵字

光生物刺激 ； PDL 細胞 ； CO2 雷射 ； 存活率 ； 成骨 ； 發炎

並列摘要

Background/ Purpose
Photobiostimulation (PBS) can affect cellular functions. The objective of the present study was to evaluate the cellular changes in periodontal ligament (PDL) cells that received different carbon dioxide (CO 2 ) laser irradiation parameters under negative pressure culture.

Materials and Methods
The negative pressure-cultured PDL cells on normal medium and differentiation medium were subjected to continuous irradiation with a CO 2 laser at an energy density of 5 J/cm 2 or 10 J/cm 2 . The irradiated PDL cells were harvested at Days 1, 5 and 7, and their viability was analyzed by the Presto Blue assay and the biologic markers alkaline phosphatase (ALP), bone sialopoietin (BSP), osteopontin (OPN), osteocalcin (OC), matrix metalloproteinase-3 (MMP-3) and cyclooxygenase-2 (COX-2) expression by reverse transcription-polymerase chain reaction (RT–PCR).

Results:
The PDL cell viability showed that the differentiation medium groups were higher than the normal culture groups. The cell morphologies were all expressed as spindle type. The inflammatory markers in the laser-irradiated groups were higher on the first day and decreased on the seventh day (p<0.05). Osteogenesis markers were highly expressed at different time periods (p<0.05). The Col I and OPN genes were highly expressed on the first day, and the Col I high expression lasted until the seventh day. The OC gene was highly expressed on the seventh day.

Conclusions:
A low-dose CO 2 laser continuously irradiating cultured PDL cells can induce osteogenesis and reduce cell inflammatory expression.

並列關鍵字

Photobiostimulation ； PDL cell ； CO2 laser ； viability ； osteogenesis ； inflammation

參考文獻

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