Influence of the Tissue Collection Procedure on the Adipogenic Differentiation of Human Stem Cells: Ischemic versus Well-Vascularized Adipose Tissue.

Clinical and basic science applications using adipose-derived stem cells (ADSCs) are gaining popularity. The current adipose tissue harvesting procedures introduce nonphysiological conditions, which may affect the overall performance of the isolated ADSCs. In this study, we elucidate the differences between ADSCs isolated from adipose tissues harvested within the first 5 min of the initial surgical incision (well-vascularized, nonpremedicated condition) versus those isolated from adipose tissues subjected to medications and deprived of blood supply during elective free flap procedures (ischemic condition). ADSCs isolated from well-vascularized and ischemic tissues positively immunostained for several standard stem cell markers. Interestingly, the percent change in the CD36 expression for ADSCs isolated from ischemic versus well-vascularized tissue was significantly lower in males than females (p < 0.05). Upon differentiation and maturation to adipocytes, spheroids formed using ADSCs isolated from ischemic adipose tissue had lower triglyceride content compared to those formed using ADSCs isolated from the well-vascularized tissue (p < 0.05). These results indicate that ADSCs isolated from ischemic tissue either fail to uptake fatty acids or fail to efficiently convert those fatty acids into triglycerides. Therefore, more robust ADSCs suitable to establish in vitro adipose tissue models can be obtained by harvesting well-vascularized and nonpremedicated adipose tissues.

Antibacterial and biocompatible polyaniline-doped titanium oxide layers.

Titanium anodization has been shown to produce crystalline oxides exhibiting photocatalytic reactions that form reactive oxygen species (ROS) when exposed to UV light. The ROS subsequently attack bacteria cells, and thus reduce bacteria attachment on titanium implant surfaces. Polyaniline (PANI) is a conductive polymer that has shown antibacterial properties when electropolymerized onto titanium. Our research group hypothesized the addition of PANI to crystalline titanium oxide surfaces would increase the available free electrons and thus increase photocatalytic activity (PCA). This research led to the development of a novel single-step anodization approach for PANI doping crystalline titanium oxide layers. The objective of the present study was to determine the proper aniline electrolyte concentration needed to maximize the PCA and reduce bacterial attachment on the formed oxides. Aniline concentrations up to 1 M were added into a 1 M sulfuric acid electrolyte. The formed oxides exhibited increased PANI surface coverage but decreased anatase and rutile crystalline titanium oxide phase formation with increasing aniline electrolyte concentrations. Despite exhibiting the lowest levels of anatase and rutile formation, the 0.75 M and 1 M aniline oxides with the greatest PANI surface coverage also exhibited the highest PCA levels. 1 M aniline oxides showed significantly higher PCA under UVA irradiation compared to oxides formed from aniline concentrations up to 0.5 M (p < 0.001). 0.75 M aniline oxides exhibited significant reductions in Staphylococcus aureus attachment with or without UVA irradiation compared to control oxides without PANI. MTT and live/dead assays confirmed cytocompatibility and nearly 100% cell viability for the PANI doped oxides.

A novel single-step anodization approach for PANI-doping oxide surfaces to improve the photocatalytic activity of titanium implants.

Crystalline titanium oxides have shown photocatalytic activity (PCA) and the formation of antibacterial reactive oxygen species (ROS) when stimulated with UV light. Polyaniline (PANI) is a conductive polymer that has shown antibacterial effects. Previously, titanium oxides have been PANI-doped using a multi-step approach. In the present study, we compared PANI-doped specimens produced with a two-step method (ACV), to PANI-doped specimens produced by a novel single-step direct anodization (AAn) method, and a control group of anodized un-doped specimens. The surface morphology, oxide crystallinity, surface elemental composition, surface roughness, surface wettability, oxide adhesion, corrosion resistance, PCA, and ROS generation of each oxide group were evaluated. All groups exhibited mixed anatase and rutile phase oxides. The AAn group revealed less anatase and rutile, but more PANI-surface coverage. The AAn group exhibited significantly increased PCA after 60 minutes of direct UVA illumination compared to the ACV group, despite containing lower amounts of anatase and rutile. The ACV and AAn groups showed significant increases in ROS production after 4 hours UVA illumination while the control group showed similar ROS production. These findings suggested that PANI doping using the novel direct anodization technique significantly improved PCA even for oxides containing less crystallinity. The S. aureus attachment response to each oxide group was also compared under UVA pre-illumination, UVA direct illumination, and no illumination (dark) lighting conditions. Although no significant differences were shown in the bacterial response, both PANI-doped groups exhibited less average bacterial attachment compared to the control group. The response of MC3T3-E1 pre-osteoblast cells to each oxide group was evaluated using MTT and live/dead assays, and no evidence of cytotoxicity was found. Since many, if not most, titanium implant devices are routinely anodized as a part of the manufacturing processes, these study findings are applicable to a wide variety of implant applications.

Polydopamine Codoped BaTiO3-Functionalized Polyvinylidene Fluoride Coating as a Piezo-Biomaterial Platform for an Enhanced Cellular Response and Bioactivity.

For a number of clinical applications, Ti6Al4V implants with bioactive coatings are used. However, the deposition of a functional polymeric coating with desired physical properties, biocompatibility, and long-term stability remains largely unexplored. Among widely investigated synthetic biomaterials, polyvinylidene fluoride (PVDF) with ß-polymorph and barium titanate (BaTiO3, BT) are considered as good examples of piezo-biopolymers and bioceramics, respectively. In this work, an adherent PVDF-based nanocomposite coating is deposited onto a Ti6Al4V substrate to explore the impact of its functional characteristics (piezoactivity) on cellular behavior and bioactivity (apatite growth and mineralized matrix formation). The precursor solution was prepared by physically grafting PVDF with polydopamine (pDOPA), forming mPVDF. Subsequently, mPVDF was reinforced with BaTiO3 nanoparticles in dimethylformamide/acetone solution, and the resulting nanocomposite (mPVDF-BT) was then spray-coated onto a roughened Ti6Al4V substrate using an airbrush at 140 °C under a pressure of 2 bar. The reproducibility of this simple yet effective processing approach to deposit chemically stable and adherent coatings was established. Remarkably, the modification with pDOPA and reinforcement with BaTiO3 nanoparticles resulted in an enhanced ß-fraction of PVDF up to 96%. This nanocomposite encouraged cellular viability of preosteoblasts (∼158% at day 5) and characteristic spreading, in vitro. Our findings indicate that the mPVDF-BT coating facilitated faster nucleation and growth of the biomineralized apatite layer with ∼70% coverage within 3 days of incubation in the simulated body fluid. In addition, the coupling among surface polar energy (5.5 mN/m), fractional polarity (∼117%), roughness (8.7 µm), and fibrous morphology also endorsed better cellular behavior. Taken together, this coating deposition strategy will pave the pathway toward designing cell-instructive surface-modified Ti6Al4V biomaterials with tailored biomineralization and bioactivity properties for musculoskeletal reconstruction and regeneration applications.