Mold fabrication and biological assessment of porous DNA-chitosan complexes.

A previous study revealed that DNA-chitosan complex prepared from the reaction between native DNA and chitosan in aqueous solution has suitable porosity for cell seeding, is nontoxic, and causes only a mild soft-tissue response. This simple and easy fabrication method for porous DNA-chitosan complex provides for a wide variety of applications as a scaffold material. The present study evaluated whether rinsing with PBS solution can fabricate DNA-chitosan complex in a mold and the histopathological responses of rat soft tissues to fabricated DNA-chitosan complexes. DNA-chitosan complex paste was prepared by mixing distilled water and freeze-dried water-rinsed DNA-chitosan complex powder. A DNA-chitosan complex disk could be fabricated by rinsing with PBS buffer and subsequently freeze-drying the DNA-chitosan complex paste in the mold. Thus, a wide range of applications of DNA-chitosan complex for tissue engineering can be anticipated using the present easy fabrication method. The porosity of the disk was 85%, and many pores were visible in the DNA-chitosan complex (before fabrication) and in the fabricated DNA-chitosan disk. The values of the complex disks gradually reduced in the tissues although 60% of disks remained in the tissues. In conclusion, an easy fabrication method for making porous DNA-chitosan complex disks was developed. It was found that the fabrication method can delay the biodegradation of the DNA-chitosan complex disk without serious tissue responses in vivo. DNA-chitosan complex is promising as a scaffold material, and a wide range of applications of DNA-chitosan complex for tissue engineering are anticipated.

Fabrication, characterization, and biological assessment of multilayered DNA-coatings for biomaterial purposes.

This study describes the fabrication of two types of multilayered coatings onto titanium by electrostatic self-assembly (ESA), using deoxyribosenucleic acid (DNA) as the anionic polyelectrolyte and poly-d-lysine (PDL) or poly(allylamine hydrochloride) (PAH) as the cationic polyelectrolyte. Both coatings were characterized using UV-vis spectrophotometry, atomic force microscopy (AFM), X-ray photospectroscopy (XPS), contact angle measurements, Fourier transform infrared spectroscopy (FTIR), and for the amount of DNA immobilized. The mutagenicity of the constituents of the coatings was assessed. Titanium substrates with or without multilayered DNA-coatings were used in cell culture experiments to study cell proliferation, viability, and morphology. Results of UV-vis spectrophotometry, AFM, and contact angle measurements clearly indicated the progressive build-up of the multilayered coatings. Furthermore, AFM and XPS data showed a more uniform build-up and morphology of [PDL/DNA]-coatings compared to [PAH/DNA]-coatings. DNA-immobilization into both coatings was linear, and approximated 3microg/cm(2) into each double-layer. The surface morphology of both types of multilayered DNA-coatings showed elevations in the nanoscale range. No mutagenic effects of DNA, PDL, or PAH were detected, and cell viability and morphology were not affected by the presence of either type of multilayered DNA-coating. Still, the results of the proliferation assay revealed an increased proliferation of primary rat dermal fibroblasts on both types of multilayered DNA-coatings compared to non-coated controls. The biocompatibility and functionalization of the coatings produced here, will be assessed in subsequent cell culture and animal-implantation studies.