A multifunctional silk coating on additively manufactured porous titanium to prevent implant-associated infection and stimulate bone regeneration.

Despite tremendous progress in the design and manufacturing of metallic implants, they do not outlive the patient. To illustrate, more than half of hip replacements will fail, mainly due to implant infection and loosening. Surface engineering approaches and, in particular, coatings can facilitate implant bio-functionality via the recruitment of more host cells for new bone formation and inhibition of bacterial colonization. Here, we used electrophoretic deposition to apply a silk fibroin solution consisting of tricalcium phosphate (TCP) and vancomycin as a coating on the surface of additively-manufactured porous titanium. Furthermore, the surface properties of the coatings developed and the release kinetics of the vancomycin were studied to evaluate the applied coating. The in vitro antibacterial behavior of the multifunctional coating, as well as the cell viability and osteogenic differentiation of the MC3T3-E1 cell line were extensively studied. The biomaterials developed exhibited an antibacterial behavior with a reduction of up to four orders of magnitude in both planktonic and adherent bacteria for 6 h and 1 d. A live-dead assay, the Alamar Blue activity, the DNA content, and cytoskeleton staining demonstrated a significant increase in the cell density of the coated groups versus the as-manufactured ones. The significantly enhanced calcium deposition and the increase in mineralization for the groups with TCP after 21 and 28 d, respectively, demonstrate upregulation of the MC3T3 cells' osteogenic differentiation. Our results collectively show that the multifunctional coating studied here can be potentially used to develop a new generation of orthopedic implants.

Simultaneous Delivery of Multiple Antibacterial Agents from Additively Manufactured Porous Biomaterials to Fully Eradicate Planktonic and Adherent Staphylococcus aureus.

Implant-associated infections are notoriously difficult to treat and may even result in amputation and death. The first few days after surgery are the most critical time to prevent those infections, preferably through full eradication of the micro-organisms entering the body perioperatively. That is particularly important for patients with a compromised immune system such as orthopedic oncology patients, as they are at higher risk for infection and complications. Full eradication of bacteria is, especially in a biofilm, extremely challenging due to the toxicity barrier that prevents delivery of high doses of antibacterial agents. This study aimed to use the potential synergistic effects of multiple antibacterial agents to prevent the use of toxic levels of these agents and achieve full eradication of planktonic and adherent bacteria. Silver ions and vancomycin were therefore simultaneously delivered from additively manufactured highly porous titanium implants with an extremely high surface area incorporating a bactericidal coating made from chitosan and gelatin applied by electrophoretic deposition (EPD). The presence of the chitosan/gelatin (Ch+Gel) coating, Ag, and vancomycin (Vanco) was confirmed by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). The release of vancomycin and silver ions continued for at least 21 days as measured by inductively coupled plasma (ICP) and UV-spectroscopy. Antibacterial behavior against Staphylococcus aureus, both planktonic and in biofilm, was evaluated for up to 21 days. The Ch+Gel coating showed some bactericidal behavior on its own, while the loaded hydrogels (Ch+Gel+Ag and Ch+Gel+Vanco) achieved full eradication of both planktonic and adherent bacteria without causing significant levels of toxicity. Combining silver and vancomycin improved the release profiles of both agents and revealed a synergistic behavior that further increased the bactericidal effects.

Additively Manufactured and Surface Biofunctionalized Porous Nitinol.

Enhanced bone tissue regeneration and improved osseointegration are among the most important goals in design of multifunctional orthopedic biomaterials. In this study, we used additive manufacturing (selective laser melting) to develop multifunctional porous nitinol that combines superelasticity with a rationally designed microarchitecture and biofunctionalized surface. The rational design based on triply periodic minimal surfaces aimed to properly adjust the pore size, increase the surface area (thereby amplifying the effects of surface biofunctionalization), and resemble the curvature characteristics of trabecular bone. The surface of additively manufactured (AM) porous nitinol was biofunctionalized using polydopamine-immobilized rhBMP2 for better control of the release kinetics. The actual morphological properties of porous nitinol measured by microcomputed tomography (e.g., open/close porosity, and surface area) closely matched the design values. The superelasticity originated from the austenite phase formed in the nitinol porous structure at room temperature. Polydopamine and rhBMP2 signature peaks were confirmed by X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy tests. The release of rhBMP2 continued until 28 days. The early time and long-term release profiles were found to be adjustable independent of each other. In vitro cell culture showed improved cell attachment, cell proliferation, cell morphology (spreading, spindle-like shape), and cell coverage as well as elevated levels of ALP activity and increased calcium content for biofunctionalized surfaces as compared to as-manufactured specimens. The demonstrated functionalities of porous nitinol could be used as a basis for deployable orthopedic implants with rationally designed microarchitectures that maximize bone tissue regeneration performance by release of biomolecules with adjustable and well-controlled release profiles.