chondroitin-sulfates and Corneal-Ulcer

Corneal lesions cause significant pain and visual impairment and, in many cases, are unresponsive to conventional treatments. Platelet lysate (PL) is an haemoderivative rich in growth factors (GFs) that are released by platelets after freeze-thawing destruction of platelet rich plasma (PRP). The aim of the present work was to develop thermosensitive and mucoadhesive eyedrops to maintain and prolong the contact of platelet lysate (PL) with cornea ulcers. A sterile vehicle based on chondroitin sulphate sodium (CS) and hydroxypropylmethyl cellulose (HPMC) was developed. An extemporaneous loading of the vehicle with PL was performed and the obtained formulation was able to quickly thermogelify at about 32 °C and was characterized by good mucoadhesive properties. ELISA evidenced that the growth factor PDGF AB was compatible with the vehicle and stable in the formulation up to 15 days of storage at 2-8 °C. In vitro wound healing and proliferation test (performed using rabbit corneal epithelial cells (RCE)) showed that the formulation enhanced cell growth and put in evidence a synergistic effect of CS and PL in stimulating cell proliferation. The overall results indicate that PL loaded in thermosensitive and mucoadhesive eyedrops can be profitably employed to treat corneal lesions.

Topics: Adhesiveness; Animals; Biological Products; Blood Platelets; Cell Extracts; Cell Line; Cell Proliferation; Chemistry, Pharmaceutical; Chondroitin Sulfates; Corneal Ulcer; Drug Carriers; Enzyme-Linked Immunosorbent Assay; Epithelium, Corneal; Gels; Humans; Hypromellose Derivatives; Methylcellulose; Ophthalmic Solutions; Platelet-Derived Growth Factor; Rabbits; Rheology; Temperature; Wound Healing

To construct a suitable ex vivo model for the research of molecular mechanisms and the pharmacological screening of fungal adherence on the corneal surface.. Mouse eyes were divided into three groups as follows: a control group with normal corneal epithelium, a group with corneal epithelium that was needle-scarified, and a group with corneal epithelium that was completely debrided. All 96 corneas were placed in organ culture and inoculated with 5 μl spore suspensions of Candida albicans at 10⁹, 10⁸, or 10⁷ colony-forming units (CFU)/ml and incubated for 0, 30, 60, or 120 min. The corneas were homogenated and diluted for quantification by counting the CFU. The effects of amphotericin B or chondroitin sulfate on the adherence of the fungal spores were evaluated with the ex vivo organ culture model and were also compared with the human corneal epithelium monolayer model in vitro.. Compared with the normal corneas with intact epithelium, the corneas with scarified and debrided epithelium adhered more spores for above two and four folds. The spore adhesion on the corneal surface was in an inoculation concentration- and incubation time-dependent manner. Moreover, both amphotericin B and chondroitin sulfate inhibited the adhesion of C. albicans spores on the corneal surface, but the inhibitory rates were different between the ex vivo corneal organ culture model and the in vitro corneal epithelium monolayer model.. The corneal organ culture was a suitable ex vivo model for the research of fungal adhesion mechanisms and drug screening.

Topics: Amphotericin B; Animals; Bacterial Adhesion; Candida albicans; Candidiasis; Chondroitin Sulfates; Colony Count, Microbial; Cornea; Corneal Injuries; Corneal Ulcer; Disease Models, Animal; Eye Infections, Fungal; Mice; Mice, Inbred BALB C; Organ Culture Techniques; Time Factors

We used a synthetic defensin (rabbit neutrophil peptide-1; NP-1) as a microbicide in a corneal storage medium, Optisol without antimicrobial compounds. We established growth curves in Optisol for Staphylococcus aureus, Streptococcus pneumoniae, and Pseudomonas aeruginosa. Each organism was evaluated separately at 4 degrees C, 23 degrees C, and 37 degrees C in Optisol with NP-1 at each of four different concentrations including 1, 10, 100, and 200 micrograms/ml. When the three organisms were evaluated in Optisol containing NP-1, we found that a concentration of 200 micrograms/ml of NP-1 successfully killed 99.9% (the limits of our assay) of all three organisms at all temperatures tested. We conclude that NP-1 exhibits promise as a nonantibiotic preservative agent in corneal storage media, since it was effective in killing organisms at all temperatures, including 4 degrees C.

Topics: Antifungal Agents; Bacteria; Blood Proteins; Chondroitin Sulfates; Colony Count, Microbial; Complex Mixtures; Cornea; Corneal Ulcer; Culture Media, Serum-Free; Defensins; Dextrans; Gentamicins; Humans; Microbiological Techniques; Organ Preservation; Pseudomonas aeruginosa; Staphylococcus aureus; Streptococcus pneumoniae