Kaempferia parviflora Extracellular Vesicle Loaded with Clarithromycin for the Treatment of Helicobacter pylori Infection

Abstract

Kaempferia parviflora extracellular vesicles (KPEVs) have been reported as promising nanovesicles for drug delivery. This study aimed to load clarithromycin (CLA) into KPEVs (KPEVS-CLA) and determine the physical properties, drug-releasing efficiency, gastric cell uptake, anti-H. pylori activities, and anti-inflammatory responses in comparison with free CLA and KPEVs. Methods: The size and surface charge of KPEVs-CLA were evaluated using dynamic light scattering and visualized using a transmission electron microscope. The encapsulation efficiency (EE%), loading capacity (LC%), and drug release of KPEVs-CLA were examined using HPLC. Anti-H. pylori growth and anti-adhesion were evaluated. IL-8 gene expression, NF-κB signaling proteins, and anti-inflammatory profiles were examined using qRT-PCR, Western blotting, and Bio-Plex immunoassay, respectively. Antichemotaxis was then examined using a Transwell assay. Results: KPEVs-CLA were intact and showed a negative surface charge similar to that of KPEVs. However, slightly enlarged KPEVs were observed. CLA was successfully loaded into KPEVs with EE of 93.45% ± 2.43%, LC of 9.3% ± 3.02%. CLA release in the PBS and gastric mimic buffer with Fickian diffusion (n ≤ 0.43) according to Korsmeyer-Peppas kinetic model (R 2 =0.98). KPEVs-CLA was localized in the gastric cells' cytoplasm and perinuclear region. Anti-H. pylori growth and anti-H. pylori adhesion of KPEVs-CLA were compared with those of free CLA with no cytotoxicity to adenocarcinoma gastric cells. KPEVs-CLA significantly reduced IL-8, G-CSF, MIP-1α, and MIP-1β levels. Moreover, KPEVs-CLA showed a superior effect over CLA in reducing G-CSF, MIP-1α, and NF-κB phosphorylation and monocyte chemotactic activities. Conclusion: KPEVs serve as potential carriers of CLA. They exhibited a higher efficiency in inhibiting gastric cell inflammation mediated by H. pylori infection than free CLA. The establishment of KPEVs-CLA as a nanodrug delivery model for H. pylori treatment could be applied to other plant extracellular vesicles or loaded with other cancer drugs for gastric cancer treatment.

Response to Article “Kaempferia parviflora Extracellular Vesicle Loaded with Clarithromycin for the Treatment of Helicobacter pylori Infection” [Letter]

“…2 In this study, clarithromycin was designed to be loaded into Kaempferia parviflora extracellular vesicles (KPEVs), while the drug delivery and final antibiotic activity of clarithromycin in KPEVs were examined and compared to the free form of clarithromycin in this in vitro study by using adenocarcinoma gastric cells. 1 Since the growing number of cases of antimicrobial resistance (AMR) worldwide are still rapidly reported, the results of this study showed promising novelty as the effort to find a way out of this global health issue. KPEVs were shown and proven to be promising drug delivery vesicles.…”

“…We felt really encouraged to read and review the recent research report reported by Nemidkanam et al since the study provides interesting ideas in providing solutions for antimicrobial resistance in bacterial infection. 1 Clarithromycin is a second-generation broad-spectrum macrolide antibiotic and was generally used in the treatment of Helicobacter pylori infection. 2 In this study, clarithromycin was designed to be loaded into Kaempferia parviflora extracellular vesicles (KPEVs), while the drug delivery and final antibiotic activity of clarithromycin in KPEVs were examined and compared to the free form of clarithromycin in this in vitro study by using adenocarcinoma gastric cells.…”

“… 2 In this study, clarithromycin was designed to be loaded into Kaempferia parviflora extracellular vesicles (KPEVs), while the drug delivery and final antibiotic activity of clarithromycin in KPEVs were examined and compared to the free form of clarithromycin in this in vitro study by using adenocarcinoma gastric cells. 1 …”

“…However, some effects shown by clarithromycin in KPEVs were only compared to the free-form clarithromycin in this study. 1 It could be better and convincing to compare to the KPEVs without clarithromycin as well. For example, the results of anti-inflammatory effects and cytotoxic effects of KPEVs alone on gastric cells were not shown in the results section of this study, even though a previous study of this research group had been showing the observation results of biochemical properties and bioactive compounds in KPEVs alone.…”