In Vitro Evaluation of Rose Bengal Photoactivated by Custom-Built Green Light-Emitting Diode Source for Bacteria and Rapidly Growing Mycobacteria Inhibition.

Purpose: In vitro evaluation of rose bengal (RB) photoactivated by our custom-built green light-emitting diode (LED) source for the growth inhibition of bacterial strains and rapidly growing mycobacterial (RGM) isolates in infectious keratitis. Methods: Six corneal clinical bacteria isolates were included in this study: two Gram-positive bacteria (methicillin-resistant Staphylococcus aureus [MRSA] and Staphylococcus epidermidis), two Gram-negative bacteria (Pseudomonas aeruginosa and Serratia marcescens), and two RGM (Mycobacterium chelonae and Mycobacterium abscessus). Microorganisms were cultured and incubated at specific conditions and prepared in suspensions to adjust their concentration to 104 cells/mL. Different treatments were conducted in triplicates: Group I, no treatment; Group II, treated with 0.1% rose bengal alone (exposed to dark for 30 minutes); Group III, exposed to custom green LED for 30 minutes (12.87 J/cm2); and Group IV, treated with 0.1% rose bengal and exposed to custom green LED for 30 minutes. Agar plates were incubated at specific conditions and photographed after growth for pixel analyses. Results: Complete growth inhibition of all bacteria and RGM was observed in Group IV. MRSA and S. epidermidis in Group II also showed complete growth inhibition. Conclusions: The custom-built green LED presented good activity by photoactivating RB and inhibiting micro-organism growth. For the first time, we demonstrated the expressive growth inhibition effect of RB against S. epidermidis, RGM, and S. marcescens. Clinical treatment with RB may offer an alternate adjunct therapy for corneal surface infections. Translational Relevance: Validating in vitro the custom-built green LED encourages the clinical application for the treatment of infectious keratitis.

Titanium Powder 3D-Printing Technology for a Novel Keratoprosthesis in Alkali-Burned Rabbits.

Purpose: To evaluate the surgical technique, clinical performance, and biocompatibility of a novel keratoprosthesis (KPro) named KPro of Brazil (KoBra) in an alkali-burned rabbit model. Methods: Two-piece three-dimensional-printed titanium powder and polymethyl methacrylate KPros were implanted into 14 alkali-burned corneas of 14 rabbits using an autologous full-thickness corneal graft as the KPro carrier. Rabbits were examined weekly for 12 months to evaluate retention and postoperative complications. Anterior segment optical coherence tomography (AS-OCT) and scanning electron microscopy (SEM) were performed at the end of the experiment to evaluate the relationship between the KoBra and the carrier graft. Results: All surgeries were performed without intraoperative complications, and the immediate postoperative period was uneventful. In 12 eyes (85.7%), the implanted KPros integrated into the operated eyes and maintained clear optics without extrusion or further complications over 12 months. Two eyes presented late postoperative complications that progressed to KPro extrusion: one had a presumed infectious keratitis, and the other had sterile stromal necrosis. AS-OCT demonstrated the correct relationship of the device and carrier graft in all remaining animals at the final follow-up. SEM findings indicate the integration of the porous structure of the back plate into the surrounding tissue. Conclusions: Clinical evaluations, AS-OCT, and SEM findings indicate good biointegr-ation of the implanted device into the corneal carrier graft. KoBra has the advantage of using recipients' own corneas as the prosthesis supporter, and its surgical procedure is relatively simple and safe. Translational Relevance: Titanium three-dimensional-printed technology used in an animal limbal stem-cell deficiency model holds great promise for the treatment of corneal blindness in humans.