마이크로그루브 및 열산화 복합 티타늄 표면의 골아세포분화 증진효과
Effect of titanium surface microgrooves and thermal oxidation on in vitro osteoblast responses
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목적: 다양한 크기의 마이크로그루브가 형성된 티타늄 표면에 열산화 처리를 한 복합 표면의 표면특성을 규명하고, 인간치주인대세포 배양 시 표면에 따른 다양한 세포행동들간 차이와 상관관계를 분석하고자 하였다. 재료 및 방법: Grade II 티타늄 디스크를 시편으로 제작하였다. 포토리소그라피를 이용하여 티타늄 시편의 마이크로그루브 크기를 폭/깊이 0/0 ㎛, 15/3.5 ㎛, 30/10 ㎛, 60/10 ㎛로 각각 형성하였다. 평활한 티타늄 표면인 대조군(ST)을 제외한 모든 실험군(ST/TO, Gr15-TO, Gr30-TO, Gr60-TO)에 700℃에서 3시간동안 열산화 처리하고, 주사현미경 사진을 사용하여 표면특성을 평가하였다. 인간치주인대세포를 배양한 후 BrdU (Bromdeoxyuridine) 실험, 알칼리성 인산가수분해효소 활성 실험, 세포외 칼슘 침착 실험을 통해 세포접착, 세포분화 및 골광화를 평가하였다. 통계분석으로는 일요인분산분석과 피어슨상관관계분석(SPSS version 17.0)을 사용하였다. 결과: 열산화를 동반한 마이크로그루브가 형성된 실험군(Gr15-TO, Gr30-TO, Gr60-TO)들은 평활한 대조군(ST)과 단순 열산화 처리 실험군(ST-TO)에 비하여 BrdU 실험, 알칼리성 인산가수분해효소 활성 실험, 세포 외 칼슘 침착 실험 모두에서 유의하게 증가된 활성도를 나타내었다. 특히, Gr60-TO군은 대조군 및 Gr15-TO, Gr30-TO, Gr60-TO 군 등에 비해 가장 증진된 세포접착 및 골아세포분화/골광화를 나타냈다. 결론: 본 연구의 한계 내에서, 열산화 처리 및 마이크로그루브 복합 티타늄 표면은 골아세포분화에 효과적 방법임이 확인되었다. 본 연구에서 규명된 적정한 마이크로그루브 크기와 열산화 처리 조건은 마이크로그루브-열산화 복합 표면 티타늄 임플란트 개발의 기초 확립에 기여할 수 있을 것이다.
Abstract
Purpose: We aimed to investigate the effect of combined various microgrooves and thermal oxidation on the titanium (Ti) and to evaluate various in vitro responses of human periodontal ligament cells (PLCs). Materials and methods: Grade II titanium disks were fabricated. Microgrooves were applied on titanium discs to have 0/0 ㎛, 15/3.5 ㎛, 30/10 ㎛/, and 60/10 ㎛of respective width/depth by photolithography. Thermal oxidation was performed on the microgrooves of Ti substrata for 3 h at 700℃ in air. The experiments were divided into 3 groups: control group (ST), thermal oxidation group (ST/TO), and combined microgrooves and thermal oxidation group (Gr15-TO, Gr30-TO, Gr60-TO). Surface characterization was performed by field-emission scanning microscopy. Cell adhesion, osteoblastic differentiation, and mineralization were analyzed using the bromodeoxyurdine (BrdU), Alkaline phosphatase (ALP) activity, and extracellular calcium deposition assays, respectively. Statistical analysis was performed using the oneway analysis of variance and Pearson’s bivariate correlation analysis (SPSS Version 17.0). Results: In general, the combined microgrooves and thermal oxidation group (Gr15-TO, Gr30-TO, Gr60-TO) showed significantly higher levels compared with the control (ST) or thermal oxidation (ST-TO) groups in the BrdU expression, ALP activity, and extracellular calcium deposition. Gr60-TO group induced highest levels of cell adhesion and osteoblastic differentiation. Conclusion: Within the limitation of this study, we conclude that the Ti surface treatment using combined microgrooves and thermal oxidation is highly effective in inducing the cell adhesion andosteoblastic differentiation. The propose surface is also expected to be effective in inducing rapid and strong osseointegration of Ti oral implants.
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