Sputtered crystalline TiO2 film drives improved surface properties of titanium-based biomedical implants.

Different crystalline phases in sputtered TiO2 films were tailored to determine their surface and electrochemical properties, protein adsorption and apatite layer formation on titanium-based implant material. Deposition conditions of two TiO2 crystalline phases (anatase and rutile) were established and then grown on commercially pure titanium (cpTi) by magnetron sputtering to obtain the following groups: A-TiO2 (anatase), M-TiO2 (anatase and rutile mixture), R-TiO2 (rutile). Non-treated commercially pure titanium (cpTi) was used as a control. Surfaces characterization included: chemical composition, topography, crystalline phase and surface free energy (SFE). Electrochemical tests were conducted using simulated body fluid (SBF). Albumin adsorption was measured by bicinchoninic acid method. Hydroxyapatite (HA) precipitation was evaluated after 28 days of immersion in SBF. MC3T3-E1 cell adhesion, morphology and spreading onto the experimental surfaces were evaluated by scanning electron microscopy. Sputtering treatment modified cpTi topography by increasing its surface roughness. CpTi and M-TiO2 groups presented the greatest SFE. In general, TiO2 films displayed improved electrochemical behavior compared to cpTi, with M-TiO2 featuring the highest polarization resistance. Rutile phase exhibited a greater influence on decreasing the current density and corrosion rate, while the presence of a bi-phasic polycrystalline condition displayed a more stable passive behavior. M-TiO2 featured increased albumin adsorption. HA morphology was dependent on the crystalline phase, being more evident in the bi-phasic group. Furthermore, M-TiO2 displayed normal cell adhesion and morphology. The combination of anatase and rutile structures to generate TiO2 films is a promising strategy to improve biomedical implants properties including greater corrosion protection, higher protein adsorption, bioactivity and non-cytotoxicity effect.

Synthesis of multifunctional chlorhexidine-doped thin films for titanium-based implant materials.

Our goal was to create bio-functional chlorhexidine (CHX)-doped thin films on commercially pure titanium (cpTi) discs using the glow discharge plasma approach. Different plasma deposition times (50, 35 and 20 min) were used to create bio-functional surfaces based on silicon films with CHX that were compared to the control groups [no CHX and bulk cpTi surface (machined)]. Physico-chemical and biological characterizations included: 1. Morphology, roughness, elemental chemical composition, film thickness, contact angle and surface free energy; 2. CHX-release rate; 3. Antibacterial effect on Streptococcus sanguinis biofilms at 24, 48 and 72 h; 4. Cytotoxicity and metabolic activity using fibroblasts cell culture (NIH-F3T3 cells) at 1, 2, 3 and 4 days; 5. Protein expression by NIH-F3T3 cells at 1, 2, 3 and 4 days; and 6. Co-culture assay of fibroblasts cells and S. sanguinis to assess live and dead cells on the confocal laser scanning microscopy, mitochondrial activity (XTT), membrane leakage (LDH release), and metabolic activity (WST-1 assay) at 1, 2 and 3 days of co-incubation. Data analysis showed that silicon films, with or without CHX coated cpTi discs, increased surface wettability and free energy (p < 0.05) without affecting surface roughness. CHX release was maintained over a 22-day period and resulted in a significant inhibition of biofilm growth (p < 0.05) at 48 and 72 h of biofilm formation for 50 min and 20 min of plasma deposition time groups, respectively. In general, CHX treatment did not significantly affect NIH-F3T3 cell viability (p > 0.05), whereas cell metabolism (MTT assay) was affected by CHX, with the 35 min of plasma deposition time group displaying the lowest values as compared to bulk cpTi (p < 0.05). Moreover, data analysis showed that films, with or without CHX, significantly affected the expression profile of inflammatory cytokines, including IL-4, IL-6, IL-17, IFN-y and TNF-α by NIH-F3T3 cells (p < 0.05). Co-culture demonstrated that CHX-doped film did not affect the metabolic activity, cytotoxicity and viability of fibroblasts cells (p > 0.05). Altogether, the findings of the current study support the conclusion that silicon films added with CHX can be successfully created on titanium discs and have the potential to affect bacterial growth and inflammatory markers without affecting cell viability/proliferation rates.

A novel combination of biallelic ALPL mutations associated with adult hypophosphatasia: A phenotype-genotype association and computational analysis study.

Hypophosphatasia (HPP) is an inherited metabolic disorder that causes defective skeletal and dental mineralization. HPP exhibits a markedly heterogeneous range of clinical manifestations caused by dysfunction of the tissue-nonspecific isozyme of alkaline phosphatase (TNSALP), resulting from loss-of-function mutations in the ALPL gene. HPP has been associated with predominantly missense mutations in ALPL, and a number of compound heterozygous genotypes have been identified. Here, we describe a case of a subject with adult-onset HPP caused by a novel combination of missense mutations p.Gly473Ser and p.Ala487Val, resulting in chronic musculoskeletal pain, myopathy, persistent fatigue, vomiting, and an uncommon dental phenotype of short-rooted permanent teeth. Pedigree and biochemical analysis indicated that severity of symptoms was correlated with levels of residual ALP activity, and co-segregated with the p.Gly473Ser missense mutation. Bioinformatic analysis to predict the structural and functional impact of each of the point mutations in the TNSALP molecule, and its potential contribution to the clinical symptoms, revealed that the affected Gly473 residue is localized in the homodimer interface and predicted to have a dominant negative effect. The affected Ala487 residue was predicted to bind to Tyr479, which is closely located the N-terminal α-helix of TNSALP monomer 2, suggesting that both changes may impair dimer stability and catalytic functions. In conclusion, these findings assist in defining genotype-phenotype associations for HPP, and further define specific sites within the TNSALP molecule potentially related to neuromuscular manifestations in adult HPP, allowing for a better understanding of HPP pathophysiology.