Extraction of hyaluronic acid and chondroitin sulfate from marine biomass for their application in the treatment of the dry eye disease

Abstract

AbstractPurposeTear film instability, hyperosmolarity and neurosensory abnormality are the key etiological contributors to the pathophysiological process of the Dry Eye Disease (DED).1 Hyaluronic acid (HA) and Chondroitin sulfate (CS) have shown improvements in the corneal epithelial barrier in patients with DED.2 Moreover they are used in pharmaceutical applications due to their biocompatibility, viscoelasticity, lubricity and immunostimulatory effects. In this study, they are extracted from marine wastes and will be evaluated as possible DED treatment using natural, green and novel Deep Eutectic Solvents (DES) to replace time‐consuming and expensive conventional methods and maintaining the quality and purity of the isolated HA and CS. Natural compounds such as menthol and trehalose, are used in DES preparation and have shown to be effective in DED treatment.3,4MethodsHA and CS are isolated from marine wastes using conventional methods and DES as low‐cost alternative technique. DES are prepared by heating and mixing the components at certain ratios. They are mixed with the marine sources at specific extraction conditions. The extracts are then enzymatically hydrolyzed and analyzed by HPLC and capillary electrophoresis to quantify HA and CS.ResultsHA and CS are extracted from mussels, codfish bones and furtherly tuna eyeballs and shark fins using DES. Several DES were tested such as menthol:borneol, menthol:camphor, trehalose:glycerol, and significant amounts of HA and CS were extracted with high specificity. For example, 37.92 µg/gdry codfish bones CS disaccharide (ΔHexAβ1‐3GalNAc4S) were extracted using menthol:borneol.ConclusionDES are effective compounds for the extraction of valuable biopolymers HA and CS. The isolated HA and CS, as well as their therapeutic mixture with DES will be studied in vitro on conjunctival and corneal epithelial human cell lines to assess their toxicity, biocompatibility, muco‐adhesion and anti‐oxidant effects for the treatment of the DED.References Craig, Jennifer P., et al. “TFOS DEWS II report executive summary.” The Ocular Surface 15.4 (2017): 802‐812. Limberg, Michael B., et al. “Topical application of hyaluronic acid and chondroitin sulfate in the treatment of dry eyes.” American journal of ophthalmology 103.2 (1987): 194‐197. Arita, Reiko, et al. “Effects of a warm compress containing menthol on the tear film in healthy subjects and dry eye patients.” Scientific reports 7 (2017): 45848. Fondi, Klemens, et al. “Effect of Hyaluronic Acid/Trehalose in Two Different Formulations on Signs and Symptoms in Patients with Moderate to Severe Dry Eye Disease.” Journal of ophthalmology 2018 (2018).