In this study, chitosan oligosaccharide (COS) Nanoparticles (NPs) are loaded with Apocynin (APO), a bioactive phytochemical with prominent anti-inflammatory and anti-oxidant activities.  It targets potential antiulcerogenic activity against ketoprofen-induced gastric ulceration in rats.

Chitosan oligosaccharide (COS) is a low molecular weight chitosan and its oral nanoparticulate system for potentiating the antiulcerogenic activity of the loaded APO has been described here.

Methods: COS-nanoparticles (NPs) loaded with APO (using tripolyphosphate [TPP] as a cross-linker) were prepared by ionic gelation method and fully characterized. The chosen formula was extensively evaluated regarding in vitro release profile, kinetic analysis, and stability at refrigerated and room temperatures. Ultimately, the in vivo antiulcerogenic activity against ketoprofen (KP)-induced gastric ulceration in rats was assessed by macroscopic parameters including Paul’s index and antiulcerogenic activity, histopathological examination, immunohistochemical (IHC) evaluation of cyclooxygenase-2 (COX-2) expression level in ulcerated gastric tissue, and biochemical measurement of oxidative stress markers and nitric oxide (NO) levels.

Results: The selected NPs formula with COS (0.5 % w/v) and TPP (0.1% w/v) was the most appropriate one with drug entrapment efficiency percentage of 35.06%, particle size of 436.20 nm, zeta potential of +38.20 mV, and mucoadhesive strength of 51.22%. It exhibited a biphasic in vitro release pattern as well as high stability at refrigerated temperature for a 6-month storage period. APO-loaded COS-NPs provoked marvelous antiulcerogenic activity against KP-induced gastric ulceration in rats compared with free APO treated group, which was emphasized by histopathological, IHC, and biochemical studies.

Conclusion:  APO-loaded COS-NPs could be considered as a promising oral phytopharmaceutical nanoparticulate system for management of gastric ulceration.

chitosan oligosaccharides chemical structure
chitosan oligosaccharides (C0S) chemical structure

Advances in nanotechnology involving bioactive phytochemicals have provided numerous innovative delivery systems; including polymeric nanoparticles (PNPs). Oral PNPs have gained much attention as drug carriers because of their nanoscopic size, bioadhesion, targetability, and controlled release of drugs in the GIT, hence conferring enhanced bioavailability.1  Their ability to cross directly and/or adhere to the mucosa represents a prerequisite step prior to the translocation process of particles.2

From this perspective, bioadhesion plays a substantial role in delivering the drugs across the epithelia and subsequently averting hepatic first pass metabolism and enzymatic degradation in the GIT.1   Among natural mucoadhesive polymers, chitosan (CS) is the most extensively studied cationic polysaccharide for construction of PNPs owing to its characteristic features.3  Ionic gelation technique, a physical cross-linking process, is an efficient method adopted to prepare CS-NPs based on ionic interaction between positively charged primary amino groups of CS and negatively charged groups of polyanion like tripolyphosphate (TPP) (the most preferable cross-linker with safety and multivalent properties).4  Such positively charged CS-NPs possess mucoadhesive and permeation enhancing properties.5  Despite the advantages of CS, its poor aqueous solubility in physiological pH is considered a major limitation (readily soluble in acidic medium only). Besides, the degree of deacetylation, molecular weight (MW), and type of CS can influence its solubility.6,7  Therefore, CS derivatives have emerged in recent years to circumvent such limitation.  Among them; CS oligosaccharide (COS), an oligomer of β-(1–4)-linked D-glucosamine, is a low MW CS, typically below 10 kDa.  Its superior merits such as high water solubility, low viscosity, biocompatibility, biodegradability, mucoadhesiveness, and permeation enhancing capability boost its potential application in pharmaceutical and biomedical fields.6,7  COS has been reported as a biopolymer possessing versatile biological activities.8 The anti-inflammatory activity of COS was verified both in vitro and in vivo.9–11 Gastric ulcer has long been rated as one of the most prevalent gastrointestinal inflammatory disorders affecting approximately 10% of the world’s population. The pathogenesis of gastric ulcer is related to the disruption of the homeostasis between offensive factors and defensive factors.12 Despite the availability of different drug categories for treatment of gastric ulcer such as proton pump inhibitors and H2 receptor antagonists, their clinical evaluation has demonstrated numerous side effects and high incidence of relapse.13  Apocynin (APO) (4-hydroxy-3-methoxyacetophenone) is a bioactive phytochemical extracted from the roots of Apocynum cannabinum (Canadian hemp) or Picrorhiza kurroa native to the Himalayas. It possesses eminent anti-inflammatory and anti-oxidant activities that have been substantiated on a diversity of cell lines and animal models.14–16 APO’s mechanism is renowned through specific inhibition of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase as well as suppression of a series of inflammatory mediators.17,18 However, for successful prospective administration, the literatures recently published APO as nanoparticulate systems to overcome its poor oral bioavailability as a prime challenge.19–22  COS has been proven a promising candidate for preparation of several nano-delivery systems targeted to potentiate the therapeutic efficacy of the loaded drugs.23–26  However, very few articles have been published, e.g.,  Ye et al which stated the preparation of COSNPs using TPP as a cross-linker and evaluated the efficiency of such COS-TPP NPs as a non-viral vector for DNA delivery.7 Likewise, a sole pharmacological study manifested the gastroprotective activity of APO, as a free drug, against ethanol-induced gastric ulcer in rats.27 Nevertheless, designing a nano-delivery system targeted to potentiate the gastric antiulcerogenic activity (AA) of APO has not been investigated yet. This context paves the way to devote the current study to fabricate and extensively evaluate a novel oral phytopharmaceutical nanoparticulate system composed of COSNPs loaded with APO (using TPP as a cross-linker) in order to reconnoiter its potential for effective gastric AA.

Conclusion:  APO-loaded COS-NPs were successfully prepared by chitosan powder and the ionic gelation method. Maximum DEE % was achieved owing to the electrostatic interaction between the polymer and the drug. The results of FT-IR, DSC, and XRD of the chosen formula confirmed the drug entrapment in the matrix of the polymeric nanoparticulate system. Furthermore, the TEM displayed its nanosized and spherical shape. The in vitro release pattern of APO from COS-NPs exhibited a biphasic pattern (initial burst phase and sustained release phase) that can quickly confer therapeutic concentration of the drug which will be sustained over a long period of time. The prominent in vivo AA of APO-loaded COS-NPs was proven by histopathological, IHC, and biochemical studies. One might look to the prepared NPs as a “scaffold” for APO sustained release in the gastric mucosa. Actually, APO-loaded COS-NPs deserve profound attention for their prospective application as a promising phytopharmaceutical nanoparticulate system for potentiated gastric AA.

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Study paper by;  Anter HM, Abu Hashim II, Awadin W, Meshali MM.  Novel chitosan oligosaccharide-based nanoparticles for gastric mucosal administration of the phytochemical “apocynin”.  Int J Nanomedicine. 2019 Jul 5;14:4911-4929. doi: 10.2147/IJN.S209987. eCollection 2019.