Aspirin effects on MG-63 cell proliferation on different modified titanium surfaces under fluid shear stress

doi:10.3969/j.issn.2095-4344.2017.22.019

Abstract

Abstract:

BACKGROUND: Recent research has shown that the proper concentration of aspirin can increase the proliferation and osteogenic ability of MG-63 cells.

OBJECTIVE: To explore the effects of different concentrations of aspirin on osteoblast proliferation on the implant-cell interface under fluid shear stress.

METHODS: (1) MG-63 cells were cultured in low-glucose DMEM containing 10% fetal bovine serum and different concentrations of aspirin (0, 0.023, 0.046, 0.0625, 0.125, 0.25, 0.5, 1.0, 2.0, 4.0 mmol/L) for 1-7 days. Then cell proliferation was detected using MTS method. (2) MG-63 cells were cultured on three different surfaces: glass slide, PT titanium surface and SLA titanium surface. After 3 days of culture with aspirin at a concentration of 0 or 0.5 mmol/L, the cells were subjected to fluid shear stress. MTS test was applied to estimate the proliferation of MG-63 cells at 0, 0.5, 1, 2, 4 hours after stress application.

RESULTS AND CONCLUSION: (1) After 1-3 days of culture, 0.023, 0.046, 0.5 mmol/L aspirin promoted the proliferation of MG-63 cells, while after 1-7 days of culture, 1, 2, 4 mmol/L aspirin inhibited the proliferation of MG-63 cells. (2) Under the fluid shear stress, aspirin showed significant effects on the cell proliferation as confirmed by one-way analysis of variance (F=8.349, P=0.004), and 0.5 mmol/L aspirin inhibited the cellular proliferation of MG-63 cells. However, surface modification and stress loading time showed no significant effects on the cell proliferation (F=2.826, P=0.064; F=0.893, P=0.406). (3) Under the fluid shear stress, surface modification showed no significant effect on the cell proliferation of MG-63 cells cultured with 0.5 mmol/L aspirin (F=1.803, P=0.171). Under the fluid shear stress, 0.5 mmol/L aspirin significantly inhibited the proliferation of MG-63 cells on the glass slide (P=0.003), while PT and SLA titanium surfaces showed on inhibitory effect on the cell proliferation (P=0.891, P=0.051). The present results demonstrate that the cell proliferation of MG-63 is related with aspirin in a concentration-dependent manner. In addition, different titanium surfaces may decrease the sensitivity of MG-63 cell proliferation to aspirin.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Aspirin, Biomechanics, Titanium, Osteoblasts, Cell Proliferation, Tissue Engineering

CLC Number:

R318

Liang Zhong-lang, Liu Chang-hong, Li Shi-yi, Yang Xiao-yu.

Aspirin effects on MG-63 cell proliferation on different modified titanium surfaces under fluid shear stress

[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(22): 3555-3560.

Figures/Tables 4

从不同浓度阿司匹林药物因素分析，所有适宜浓度组(0.023，0.046，0.062 5，0.125，0.25，0.5 mmol/L)与 0 mmol/L浓度组比较差异均无显著性意义；中高浓度、高于治疗剂量浓度组(1，2，4 mmol/L)与0 mmol/L浓度组比较差异均有显著性意义(P < 0.004)。其中细胞A值的均数由高到低排列为：0.023，0.046，0，0.062 5，0.125，0.25，0.5，1，2，4 mmol/L。在培养第1天，0.023，0.046，0.5 mmol/L浓度组细胞A值高于0 mmol/L浓度组(P=0.006，P=0.025，P=0.016)；在培养第2天，0.023，0.046，0.0625，0.5 mmol/L阿司匹林细胞A值高于0 mmol/L浓度组(P=0.002，P=0.006，P=0.019，P=0.002)；培养第3天后，适宜浓度阿司匹林A值与0 mmol/L浓度组比较差异无显著性意义，提示在作用早期，0.023，0.046，0.062 5，0.5 mmol/L的阿司匹林可促进MG-63人成骨样细胞的增殖。根据药物梯度筛查的原则，实验选取了适宜浓度的最大值0.5 mmol/L阿司匹林来进行流体力学实验。 2.3 流体剪切力作用下阿司匹林对细胞增殖能力影响的检测见图4。析因设计的方差分析显示，阿司匹林药物处理因素与材料表面处理间不存在交互效应(F=2.193，P=0.115)。所以阿司匹林药物处理因素和材料表面处理因素对细胞增殖的影响各自改用单因素方差分析。阿司匹林药物处理因素对细胞增殖影响的差异有统计学意义(F=8.349，P=0.004)，0 mmol/L浓度组A值大于0.5 mmol/L浓度组(2.260±0.244，2.148±0.276)，提示在流体剪切力作用下，0.5 mmol/L阿司匹林可降低细胞的增殖活性。从材料表面处理因素分析，不同材料表面处理对细胞增殖影响的差异无统计学意义(2.252±0.273，2.219± 0.273，2.140±0.244，F=2.826，P=0.064)。从流体剪切力加载时间因素分析，加载时间对细胞增殖影响的差异无统计学意义(2.198±0.246，2.220±0.377，2.254±0.245，2.139±0.245，2.208±0.179，F=0.893，P=0.406)。 2.4 流体剪切力作用下0.5 mmol/L阿司匹林对不同材料表面细胞增殖的影响在0.5 mmol/L阿司匹林干预下，不同材料对细胞增殖影响的差异无统计学意义(2.150±0.222，2.214±0.321，2.080±0.270，F=1.803，P=0.171)。在光滑载玻片上，0 mmol/L阿司匹林组的细胞A值高于0.5 mmol/L阿司匹林组(2.355±0.284，2.150±0.222，F=9.712，P=0.003)。在高度抛光和喷砂酸碱处理钛板上，0，0.5 mmol/L阿司匹林组的细胞A值比较差异无显著性意义(2.224±0.220，2.214±0.321，F=0.019，P=0.891；2.202±0.201，2.080±0.270，F=3.962，P=0.051)。 "

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