Nanoencapsulated Cannabidiol–Cannabigerol Using Eudragit L100: In Vitro and In Vivo Evidence in Murine Colitis Model

Abstract

Background: Phytocannabinoids such as cannabidiol (CBD) and cannabigerol (CBG) have received increasing attention in the context of inflammatory and intestinal disorders. However, direct comparisons between their individual and combined effects, as well as the influence of delivery systems, remain limited.

Objectives: This study evaluated the biological effects of free and nanoencapsulated CBD and CBG, including a cannabinoid–Eudragit L100 formulation, in an in vitro TNBS-treated intestinal cell model and an in vivo murine model of TNBS-induced colitis.

Methods: Cytotoxicity and treatment-associated effects of CBD, CBG, their 1:1 combination, and a nanoencapsulated formulation were assessed in TNBS-exposed Caco-2 cells. In parallel, BALB/c mice with TNBS-induced colitis were evaluated for colonic damage and inflammatory markers.

Results: CBD and CBG individually showed dose-dependent effects in Caco-2 cells, while their combined administration produced a greater effect than either compound alone at higher concentrations. The nanoencapsulated formulation preserved cellular metabolic activity following TNBS exposure. In vivo, both free combined and nanoencapsulated cannabinoids were associated with reduced epithelial damage and inflammatory alterations.

Conclusions: Nanoencapsulation using Eudragit L100 modulated the biological effects of CBD and CBG in experimental models of TNBS-induced intestinal injury.

Introduction

Inflammatory bowel diseases (IBDs), including Crohn’s disease (CD) and ulcerative colitis (UC), are chronic, relapsing inflammatory disorders of the gastrointestinal tract with heterogeneous clinical presentation and uncertain etiology [1]. Although their precise cause remains unclear, IBDs are widely considered a result of dysregulation in immune response to intestinal microbiota in genetically susceptible individuals [2]. Clinical manifestations range from different grades of pain, blood in stools or diarrhea, severe complications such as hemorrhage, growth impairment in pediatric patients and damage to other organs such as eyes, skin, liver and joints, particularly in CD [3].

Accurate diagnosis of IBD relies on the integration of clinical evidence such as endoscopy, laboratory and imaging assessments [4]. Current pharmacological management includes aminosalicylates, immunomodulators, biologic agents, and antibiotics. Despite their clinical utility, these therapies are often limited by incomplete efficacy, adverse effects, or loss of response over time, underscoring the need for alternative of complementary therapeutic strategies [5].

Components of the endocannabinoid system and endocannabinoidome are expressed throughout the gastrointestinal tract and participate in the regulation of intestinal permeability, motility, immune responses and inflammatory processes [6].

Experimental evidence indicates that modulation of these systems influences intestinal inflammation, supporting the investigation of cannabinoids in models of inflammatory bowel disease. In this context, non-psychotropic phytocannabinoids such as cannabidiol (CBD) and cannabigerol (CBG) have been evaluated for their anti-inflammatory and immunomodulatory effects in experimental models of intestinal inflammation [7]. Preclinical studies suggest that these compounds can modulate inflammatory pathways through interactions with defined molecular targets, including cannabinoid receptors (CB1/CB2), peroxisome proliferator activated receptors (PPARs), NF-κβ and inhibition of endocannabinoid uptake/degradation [8,9,10,11,12].

While cannabinoids have shown favorable safety profiles in experimental settings [13,14,15] a major limitation in the pharmacological use of phytocannabinoids is their poor aqueous solubility, chemical instability and low oral bioavailability, largely due to their lipophilic nature and extensive first-pass metabolism [16,17,18].

Nanotechnology-based delivery systems (NDSs) have been explored to address these limitations by improving compound stability and modulating drug release. Eudragit L100, a pH responsive methacrylic acid-methyl methacrylate copolymer soluble at pH above 6, has been widely used in enteric formulations to enable colon-targeted drug delivery [19,20]. Although Eudragit-based systems have been applied to various therapeutic agents, limited studies have investigated their use for the delivery of cannabinoids, and available reports lack biological evaluation in cellular or animal models [21].

This study evaluates the effects of cannabidiol and cannabigerol administered individually and in combination, using both in vitro and in vivo models of intestinal inflammation. First, cytotoxic and protective effects of free cannabinoids were evaluated in Caco-2 cells under basal conditions and following inflammatory injury induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS). Subsequently, cannabinoids were nanoencapsulated with the polymer Eudragit L100 to assess its impact on cellular metabolic activity. Finally, we examined the biological effects of free and nanoencapsulated cannabinoid formulation in a TNBS-induced murine model of colitis by evaluating histopathological alterations and inflammatory markers. This experimental approach was designed to determine whether Eudragit L100 nanoencapsulation modifies in vitro and in vivo biological response to cannabinoid administration under inflammatory conditions.

Download the full article as PDF here: Nanoencapsulated Cannabidiol–Cannabigerol Using Eudragit L100

or continue reading here

4. Materials and Methods

4.1. Materials and Reagents

The Eudragit® L100 polymer was purchased from Helm México (©HELM AG, Hamburg, Germany). Cannabidiol (CBD) and cannabigerol (CBG) were obtained from Botican (ICAN Connect to Cannabis, Mexico City, Mexico) in powder form, along with their respective certificates of analysis (100% CBD and 99.08% CBG) (Supplementary Material). Antibiotic/antimycotic solution (1% v/v of a commercial mixture of 100 U/mL penicillin and streptomycin), Dulbecco’s Modified Eagle Medium (DMEM culture medium), fetal bovine serum (FBS), and sodium bicarbonate (NaHCO3) were purchased from Gibco™ (Thermo Fisher Scientific Inc. Waltham, MA, USA). TRIzol™ Reagent was purchased from Invitrogen™ (Thermo Fisher Scientific Inc. Waltham, MA, USA). 2,4,6-trinitrobenzenesulfonic acid (TNBS), 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT), dimethylsulfoxide (DMSO), polyethylene glicol (PEG), sodium hydroxide (NaOH), and Zein powder were procured from Sigma-Aldrich® (Merck KGaA, Darmstadt, Germany). N-(2,6-Dimethylphenyl)-5,6-dihydro-4H-1,3-thiazin-2-amine (xylazine) was purchased from Supelco® (Merck KGaA, Darmstadt, Germany). All other products, solvents, and reagents were of analytical grade and purchased from CTRScientific® (Monterrey, NL, Mexico).

Cárdenas-Noriega, K.A.; Elizondo-Luévano, J.H.; Chávez-Montes, A.; Rodríguez-Tovar, L.E.; Franco-Molina, M.A.; Zárate-Triviño, D.G.; Pérez-Hernández, R.A.; Soto-Domínguez, A.; Castillo-Velázquez, U. Nanoencapsulated Cannabidiol–Cannabigerol Using Eudragit L100: In Vitro and In Vivo Evidence in Murine Colitis Model. Drugs Drug Candidates 2026, 5, 10. https://doi.org/10.3390/ddc5010010

Read more on Cannabidiol (CBD) here:

The Role of Excipients in CBD Products