PLGA-Chitosan Encapsulated IL-10 Nanoparticles Modulate Chlamydia Inflammation in Mice

Abstract

Abebayehu N Yilma,1,2,* Rajnish Sahu,1,* Praseetha Subbarayan,1,* Francois Villinger,3 Mamie T Coats,4 Shree R Singh,1 Vida A Dennis1 1Center for NanoBiotechnology Research (CNBR), Alabama State University, Montgomery, AL, USA; 2Department of Public Health Sciences, Pennsylvania State University College of Medicine, Hershey, PA, USA; 3Department of Biology, University of Louisiana at Lafayette, New Iberia, LA, USA; 4Department of Clinical and Diagnostics Sciences, School of Health Professionals, The University at Alabama at Birmingham (UAB), Birmingham, AL, USA*These authors contributed equally to this workCorrespondence: Vida A Dennis, Center for NanoBiotechnology Research (CNBR), Alabama State University, 1627 Harris Way, Montgomery, AL, USA, Tel [phone], Email [email protected]: Interleukin-10 (IL-10) is a key anti-inflammatory mediator in protecting host from over-exuberant responses to pathogens and play important roles in wound healing, autoimmunity, cancer, and homeostasis. However, its application as a therapeutic agent for biomedical applications has been limited due to its short biological half-life. Therefore, it is important to prolong the half-life of IL-10 to replace the current therapeutic application, which relies on administering large and repeated dosages. Therefore, not a cost-effective approach. Thus, studies that aim to address this type of challenges are always in need.Methods: Recombinant IL-10 was encapsulated in biodegradable nanoparticles (Poly-(Lactic-co-Glycolic Acid) and Chitosan)) by the double emulsion method and then characterized for size, surface charge, thermal stability, cytotoxicity, in vitro release, UV–visible spectroscopy, and Fourier Transform-Infrared Spectroscopy as well as evaluated for its anti-inflammatory effects. Bioactivity of encapsulated IL-10 was evaluated in vitro using J774A.1 macrophage cell-line and in vivo using BALB/c mice. Inflammatory cytokines (IL-6 and TNF-α) were quantified from culture supernatants using specific enzyme-linked immunosorbent assay (ELISA), and significance was analyzed using ANOVA.Results: We obtained a high 96% encapsulation efficiency with smooth encapsulated IL-10 nanoparticles of ~100– 150 nm size and release from nanoparticles as measurable to 22 days. Our result demonstrated that encapsulated IL-10 was biocompatible and functional by reducing the inflammatory responses induced by LPS in macrophages. Of significance, we also proved the functionality of encapsulated IL-10 by its capacity to reduce inflammation in BALB/c mice as provoked by Chlamydia trachomatis, an inflammatory sexually transmitted infectious bacterium.Discussion: Collectively, our results show the successful IL-10 encapsulation, slow release to prolong its biological half-life and reduce inflammatory cytokines IL-6 and TNF production in vitro and in mice. Our results serve as proof of concept to further explore the therapeutic prospective of encapsulated IL-10 for biomedical applications, including inflammatory diseases.Keywords: Chlamydia, delivery system, chitosan, nanoparticles, IL-10, inflammation

Keywords

• chlamydia
• delivery system
• chitosan
• nanoparticles
• il-10
• inflammation.