We aimed to evaluate in vivo bone response and in vitro apatite formation to titanium (Ti) implants, coated with double-stranded DNA (DNA-d) or single-stranded DNA (DNA-s), and to compare the influence in different structure of DNA, double strand and single strand on bone response and apatite formation. The bone responses to multilayered DNA-d/protamine or DNA-s/protamine coating implants were evaluated after implantation into the extracted sockets of rat maxillary molars. Apatite formation on either coating surface after immersion in simulated body fluid (SBF) was evaluated using the quartz crystal microbalance (QCM) method. DNA-d/protamine and DNA-s/protamine coatings produced more roughened and hydrophilic surfaces than untreated Ti. Animal experiments showed that higher bone-to-implant ratios were achieved 3 and 6 weeks after implantation using DNA-d/protamine and DNA-s/protamine coatings compared with Ti. QCM measurements revealed that each coating contributed to significant earlier apatite formation in SBF. We conclude that both DNA-d/protamine and DNA-s/protamine coatings enhanced early bone formation. We suggest that a DNA-multilayer coating is useful for the surface modification of a Ti implant.
DNA/protamine complex has been reported to enhance bone formation in animal experiments. Another notable property of the complex is that it becomes a paste after being knead with water. Conventionally, DNA/protamine paste is prepared by kneading the DNA/protamine complex with water; however, conventional DNA paste has problems with regards to the preparation method, handling, moldability, etc. In the present study, we developed a new method for preparing DNA pastes, called the on-the-spot preparation method. DNA/protamine paste was prepared by mixing DNA into viscous liquid protamine or mixing gelatin into viscous liquid DNA. The viscosity of the DNA/protamine and DNA/gelatin paste was better than that of the conventional DNA/protamine-complex paste. The DNA/protamine and DNA/gelatin pastes were injected into the extracted sockets of rat maxillary molars. Microfocus X-ray computed tomography (micro-CT) and histological observation of nondecalcified sections after 2 weeks of injection revealed that the DNA/protamine and DNA/gelatin paste enhanced the formation of new bone in the extracted sockets. Quantitative analysis for calcification degree and newly formed bone mass showed there was a significant difference between DNA/protamine paste-and DNA/gelatin paste-injected specimens. The bone formation ability of DNA/gelatin paste is better than that of the DNA/protamine paste in the extracted sockets of rat maxillary molars. It is suggested that DNA paste prepared by on-the-spot preparation method will be useful candidate materials for bone regeneration.
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