Abstract
The surface molecular chemistry, such as amine functionality, of biomaterials plays a crucial role in the osteogenic activity of relevant cells and tissues during hard tissue regeneration. Here, we examined the possibilities of creating amine functionalities on the surface of titanium by using the nonthermal atmospheric pressure plasma jet (NTAPPJ) method with humidified ammonia, and the effects on human mesenchymal stem cell (hMSC) were investigated. Titanium samples were subjected to NTAPPJ treatments using nitrogen (N-P), air (A-P), or humidified ammonia (NA-P) as the plasma gas, while control (C-P) samples were not subjected to plasma treatment. After plasma exposure, all treatment groups showed increased hydrophilicity and had more attached cells than the C-P. Among the plasma-treated samples, the A-P and NA-P showed surface oxygen functionalities and exhibited greater cell proliferation than the C-P and N-P. The NA-P additionally showed surface amine-related functionalities and exhibited a higher level of alkaline phosphatase activity and osteocalcin expression than the other samples. The results can be explained by increases in fibronectin absorption and focal adhesion kinase gene expression on the NA-P samples. These findings suggest that NTAPPJ technology with humidified ammonia as a gas source has clinical potential for hard tissue generation.
Highlights
Osteoporosis, trauma, and cancer cause damage to and loss of hard tissue, and such conditions are increasing as the baby boomer generation ages [1,2,3,4,5]
Biomaterials are commonly used in hard tissue regeneration, and titanium (Ti) and Ti alloys are commonly used in orthopedics and dentistry [6,7]
The results showed that N-P, A-P, and NA-P showed reversion in the hydrophilic state with respect to time after initial exposure, though the contact angle values were still lower than C-P at 24 h
Summary
Osteoporosis, trauma, and cancer cause damage to and loss of hard tissue, and such conditions are increasing as the baby boomer generation ages [1,2,3,4,5]. The observed chemical changes improved the cell proliferation of the test groups relative to C-P, and the presence of chemical functionality, such as C=O and O-H groups, further enhanced the effects as shown for the A-P and NA-P treatments This finding highlights the advantage of NTAPPJ over other technologies, such as ultraviolet treatments for dental implants, because these treatments reduce only the carbon content but do not form other functional groups [29,31,34]. Because OCN represents the most specific marker in osteoblast maturation, these findings indicate that the NA-P samples were the most potent in terms of osteogenic differentiation of hMSCs. The difference between the NA-P and the C-P and other test groups results from the presence of nitrogen-related functional groups, such as N-O, N-H, and N-Ti, which favor osteogenic differentiation of stem cells [49] because the positive charge of the chemical functional groups increases affinity for fibronectin [50,51]. Future studies are currently planned to determine possible clinical applications of the technology as a bioactive surface treatment for biomaterials, including Ti
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