Targeted therapy of genital chlamydia infections: acting locally, efficiently and patient-friendly

Resveratrol-in-liposomes-in-hydrogel successfully inhibited Chlamydia infection in vitro.The advanced delivery system based on natural origin ingredients such as polyphenol resveratrol and polu,er chitosan can efficiently act on genital Chlamydia infection as confirmed in in vitro challange.Chlamydia trachomatis is the most common cause of bacterial sexually transmitted infections and causes serious reproductive tract complications among women. The limitations of existing oral antibiotics and treatment of antimicrobial resistance require alternative treatment options. We are proposing, for the first time, the natural polyphenol resveratrol (RES) in an advanced delivery system comprising liposomes incorporated in chitosan hydrogel, for the localized treatment of C. trachomatis infection. Both free RES and RES liposomes-in-hydrogel inhibited the propagation of C. trachomatis in a concentration-dependent manner, assessed by the commonly used in vitro model comprising McCoy cells. However, for lower concentrations, the anti-chlamydial eect of RES was enhanced when incorporated into a liposomes-in-hydrogel delivery system, with inhibition of 78% and 94% for 1.5 and 3 g/mL RES, respectively for RES liposomes-in-hydrogel, compared to 43% and 72%, respectively, for free RES. Furthermore, RES liposomes-in-hydrogel exhibited strong anti-inflammatory activity in vitro, in a concentration-dependent inhibition of nitric oxide production in the LPS-induced macrophages (RAW264.7). The combination of a natural substance exhibiting multi-targeted pharmacological properties, and a delivery system that provides enhanced activity as well as applicability for vaginal administration, could be a promising option for the localized treatment of C. trachomatis infection.

Resveratrol liposomes-in-hydrogel delivery system combats Chlamydia trachomatis in vitroIn the search for novel tools to tackle antimicrobial resistance, including sexually transmitted infections, natural substances have emerged as possible alternatives to antibiotics. We have utilized reseveratrol-in-liposomes-in hydrogel to treat Chøamydia trachomatis infections. The delivery system enhances the activity of resveratrol.Natural polyphenols such as resveratrol (RES) are attractive for treatments of various diseases, including vaginal infections and inflammation. RES exhibits strong anti-oxidative and anti-inflammatory properties. However, its low solubility and consequent poor bioavailability limit its wider therapeutic uses. To overcome these limitations, a vaginal delivery system comprising RES-in--liposomes-in-hydrogel was developed. This system permits dual therapeutic action of both liposomal polyphenol (RES) and chitosan-based hydrogel. Liposomes of around 200 nm and entrapment efficiency of 81 % were incorporated into chitosan hydrogel, respectively. Medium molecular weight chitosan (2.5 %, w/w) was found to have optimal texture properties and good mucoadhesiveness in ex vivo conditions. The in vitro release studies confirmed sustained prolonged and controlled release of RES from the system. Both liposomal polyphenol and polyphenol-in-liposomes-in-hydrogel were found to be non-toxic in in vitro conditions. Anti-inflammatory activity determined by measuring the inhibitory activity of formulations on the NO production in the LPS-induced macrophages (RAW 264.7) confirmed superiority of polyphenol-in-liposomes-in-hydrogel. Interestingly, The plain liposomes-in-hydrogel (free of RES) also exhibited potent anti-inflammatory activity, suggesting that the chitosan hydrogel also independently acts in synergy regarding anti-inflammatory effect of formulation. The formulation was challanged against Chlamydia trachomatis and confirmed its potential. Dr. Jøraholmen completed her stay at Karolinska Institute, Stockholm, Sweden. The stay focused on a safety evaluation of a novel delivery system for RES. The system was confirmed to be safe.

Targeting genital chlamydia infections: localized delivery by nanocarriersIncreased antimicrobial resistance (AMR) and high recurrence/re-infection rates of genital Chlamydia are an increasing health hazard. Localized vaginal therapy is non-invasive and direct delivery of drugs to the site of action. Biodegradable advanced delivery systems represent a novel approach in targeting chlamydia infections and combating AMR.Chlamydia trachomatis is a major cause of sexually transmitted bacterial diseases with more than 130 million new infections reported globally each year. Localized vaginal therapy is the non-invasive and direct delivery of drugs to the site of action at lower doses than via systemic route, yet reducing the adverse effects associated with systemic drug administration. To achieve the efficient, safe and targeted vaginal therapy of C. trachomatis infections we are proposing development of a biodegradable and biocompatible synergy-based advanced delivery system comprising liposomal polyphenols, curcumin or resveratrol, incorporated into chitosan hydrogel. Such formulation permits dual antimicrobial action of liposomal polyphenol and chitosan-based hydrogel. Incorporation of polyphenols into liposomes allows fusion with bacterial cells, resulting in improved localized effect at lower doses. Incorporation of liposomes into the chitosan hydrogel enhances the residence time on the vaginal mucosa and further increases the antimicrobial action against bacteria. The project represents a novel approach in targeting chlamydia infections and combating AMR. The use of alternative active ingredients preserves available antibiotics as efficient drugs in the future.The combination of two delivery systems, one enabling the incorporation of poorly soluble substances (liposomes) and another assuring the prolonged contact time at the site of drug action (hydrogels), represents a novel and promising approach in optimization of localized drug therapy. The novel formulation has been tested and its promise confirmed regarding anti-inflmmatory activity of polyphenols (Manuscript I, currently under revision). We are completing the study confirming the ability of the formulation to suppress the growth of C. trachomatis and assure system’s safety. To establish the method not previously availabe at UiT/UNN, Dr. Jøraholmen spent 3 weeks in early 2018 at University of Helsinki, Finland, to learn the in vitro infection model for Chlamydia. She successfully transfered the methodology to UiT and very promising results have been obtain, soon to be published (Manuscripy II). Original research articles: M.W. Jøraholmen, P. Basnet, M.J. Tostrup, S. Moueffaq, N. Škalko-Basnet (2018) Localized therapy of vaginal inflammation: Liposomes-in-hydrogel delivery system for polyphenols, Pharmaceutics, Pharmaceutics (open acces journal). under revision Conference contributions: M.W. Jøraholmen, M. Tostrup, S. Moueffaq, P. Basnet, N. Škalko-Basnet, 2018: Liposome-in-hydrogel delivery system for polyphenols in the local therapy of vaginal infections, Controlled Release Society Annual Meeting & Exposition, New York, USA. Poster. M.W. Jøraholmen, K. Gravningen, M. Johannessen, P. Basnet, G. Acharya, N. Škalko-Basnet, 2018: Liposomes-in-hydrogel for the local treatment of chlamydia: Controlled delivery of natural substances, 14th International Symposium on Human Chlamydial Infections, Zeist, The Netherlands. Poster. Mobility: Started as a visiting researcher (October 2018 – June 2019, 9 months), Division of Obstetrics and Gynecology, Department of Clinical Science, Intervention and Technology, Karolinska University Hospital, Stockholm, Sweden. Helse Nord postdoctoral travelling grant received from January 2019. Visiting researcher (January 2018 – February 2018, 3 weeks), Department of Virology, Hartman Institute, University of Helsinki, Helsinki, Finland.

Local, efficient and patient-friendly therapy of genital Chlamydia: overcoming antibiotic resistanceIncreased antimicrobial resistance (AMR) and high recurrence/re-infection rates of genital Chlamydia are an increasing health hazard. Localized vaginal therapy is non-invasive and direct delivery of drugs to the site of action. Biodegradable advanced delivery systems represent a novel approach in targeting chlamydia infections and combating AMR.Chlamydia trachomatis is a major cause of sexually transmitted bacterial diseases with more than 130 million new infections reported globally each year. Currently, only oral (rarely intravenous) antibiotics are available for its treatment; however, increased antimicrobial resistance (AMR) and high recurrence/re-infection rates represent an increasing health hazard. Localized vaginal therapy is the non-invasive and direct delivery of drugs to the site of action at lower doses than via systemic route, yet reducing the adverse effects associated with systemic drug administration. To achieve the efficient, safe and targeted vaginal therapy of C. trachomatis infections we are proposing development of a biodegradable and biocompatible synergy-based advanced delivery system comprising liposomal polyphenols, curcumin or resveratrol, incorporated into chitosan hydrogel. Such formulation permits dual antimicrobial action of liposomal polyphenol and chitosan-based hydrogel. Incorporation of polyphenols into liposomes allows fusion with bacterial cells, resulting in improved localized effect at lower doses. Incorporation of liposomes into the chitosan hydrogel enhances the residence time on the vaginal mucosa and further increases the antimicrobial action against bacteria. The project represents a novel approach in targeting chlamydia infections and combating AMR. The use of alternative active ingredients preserves available antibiotics as efficient drugs in the future.The combination of two delivery systems, one enabling the incorporation of poorly soluble substances (liposomes) and another assuring the prolonged contact time at the site of drug action (hydrogels), represents a novel and promising approach in optimization of localized drug therapy. Newly developed liposomes were found to be in the desired size range (200 nm) and contained sufficient load of active ingredient to assure effective concentrations at the vaginal site. Texture properties of the chitosan hydrogels indicate that the system may exhibit prolonged residence time at vaginal site. The in vitro release studies confirmed a prolonged and controlled release of epicatechin and resveratrol from both liposomal and liposomes-in-hydrogel formulations compared to the controls. The next focuses will be to study the ability of the delivery system to suppress the growth of C. trachomatis and assure system’s safety.