Among the most difficult bacterial infections encountered in treating patients are wound infections, which may occur in burn victims, patients with traumatic wounds, necrotic lesions in people with diabetes, and patients with surgical wounds. Within a wound, infecting bacteria frequently develop biofilms. Many current wound dressings are impregnated with antimicrobial agents, such as silver or antibiotics. Diffusion of the agent(s) from the dressing may damage or destroy nearby healthy tissue as well as compromise the effectiveness of the dressing. In contrast, the antimicrobial agent selenium can be covalently attached to the surfaces of a dressing, prolonging its effectiveness. We examined the effectiveness of an organoselenium coating on cellulose discs in inhibitingPseudomonas aeruginosaandStaphylococcus aureusbiofilm formation. Colony biofilm assays revealed that cellulose discs coated with organoselenium completely inhibitedP. aeruginosaandS. aureusbiofilm formation. Scanning electron microscopy of the cellulose discs confirmed these results. Additionally, the coating on the cellulose discs was stable and effective after a week of incubation in phosphate-buffered saline. These results demonstrate that 0.2% selenium in a coating on cellulose discs effectively inhibits bacterial attachment and biofilm formation and that, unlike other antimicrobial agents, longer periods of exposure to an aqueous environment do not compromise the effectiveness of the coating.

Animals are commonly used for pharmacokinetic studies which are the most frequent events tested during ocular drug development and preclinical evaluation. Inaccuracy, cost, and ethical criticism in these tests have created a need to construct an in vitro model for studying corneal constraints. In this work, a porous membrane embedded microfluidic platform is fabricated that separates a chip into an apical and basal side. After functionalizing the membrane surface with fibronectin, the membrane's mechanical and surface properties are measured to ensure correct modeling of in vivo characteristics. Immortalized human corneal epithelial cells are cultured on the membrane to create a microengineered corneal epithelium-on-a-chip (cornea chip) that is validated with experiments designed to test the barrier properties of the human corneal epithelium construct using model drugs. A pulsatile flow model is used that closely mimics the ocular precorneal constraints and is reasonable for permeability analysis that models in vivo conditions. This model can be used for preclinical evaluations of potential therapeutic drugs and to mimic the environment of the human cornea.

Oral bacteria, including S treptococcus mutans and Streptococcus salivarius, contribute to tooth decay and plaque formation; therefore, it is essential to develop strategies to prevent dental caries and plaque formation. We recently showed that organo-selenium compounds covalently attached to different biomaterials inhibited bacterial biofilms. Our current study investigates the efficacy of an organo-selenium dental sealant (SeLECT-Defense^TM sealant) in inhibiting S. mutans and S. salivarius biofilm formation in vitro. The organo-selenium was synthesized and covalently attached to dental sealant material via standard polymer chemistry. By colony-forming unit (CFU) assay and confocal microscopy, SeLECT-Defense^TM sealant was found to completely inhibit the development of S. mutans and S. salivarius biofilms. To assess the durability of the anti-biofilm effect, we soaked the SeLECT-Defense^TM sealant in PBS for 2 mos at 37°C and found that the biofilm-inhibitory effect was not diminished after soaking. To determine if organo-selenium inhibits bacterial growth under the sealant, we placed SeLECT-Defense sealant over a lawn of S. mutans. In contrast to a control sealant, SeLECT-Defense^TM sealant completely inhibited the growth of S. mutans. These results suggest that the inhibitory effect of SeLECT-Defense^TM sealant against S. mutans and S. salivarius biofilms is very effective and durable.

Experiments were carried out in the early 1990s to investigate the cell types involved in a pterygium and to determine a possible mechanism of formation. Our first experiments used monoclonal antibodies to keratins and an associated protein (vimentin), to look at the cells that compose a pterygium. These experiments demonstrated that a pterygium is the result of an abnormal limbal basal epithelial stem cell that moves onto Bowman's layer and brings about the dissolution of this layer. More importantly, these data showed that the clear corneal epithelial cells in front of the pterygium also contained these abnormal limbal cells, which we named the pterygium cell. This demonstrated that when a pterygium is removed, a wide area of what appears to be normal epithelium must be removed to inhibit reoccurrence of the growth. Later experiments using expressed sequence tag analysis of an un-normalized unamplified complementary DNA library from surgically removed pterygia were compared with normal cornea and confirmed the role of the epithelial cells in this growth. The gene expression studies also showed that genes involved in cellular migration are stimulated, and this led to studies on polyamine analogs as inhibitors of pterygial migration. Immunohistochemical studies with antibodies to matrix metalloproteinases (MMPs) showed that it is the pterygium cell that produces the MMPs that dissolve Bowman's layer resulting in the growth stimulation of stromal fibroblasts. This led to experiments on the use of MMP inhibitors to inhibit the growth of pterygia.

Purpose: To determine whether topical Substance-P (SP) plus insulin-like growth factor-1 (IGF-1) can improve corneal healing after photorefractive surface ablation in a rabbit. Methods: After a 9.0-mm corneal de-epithelialization using a combination of chemical (18% alcohol) and mechanical debridement, excimer photorefractive surface ablation was performed bilaterally in eight rabbits (16 eyes) with an 8.0-mm ablation zone and 70-μm depth. The right eye was treated with SP (250 μg/mL) and IGF-1 (25 ng/mL) in hyaluronic acid, one drop twice a day, and the other eye treated with only hyaluronic acid. The epithelial healing process was documented photographically twice a day until healing was complete. Six rabbits were sacrificed 6 weeks after photorefractive keratectomy (PRK) and corneas examined histologically. Results: Seven of eight rabbit eyes treated with SP/IGF-1 healed in a shorter time than the untreated eye. For rabbit #6, both eyes healed at the same time. The average healing time (total time until wound closure) for the treated eyes was 99 hours, while the average healing time for the untreated eyes was 170 hours (P = 0.0490). A persistent epithelial defect was found in two of the nontreated eyes but none in the treated eyes. Corneal pathology showed some degree of epithelial separation in the central corneal wound in three out of six nontreated eyes and in just the treated eye of rabbit #6. Conclusion: Topical SP plus IGF-1 increases the epithelial healing rate after PRK. There may have been beneficial effects upon cell adhesion as well. Translational Relevance: Better and faster healing.