The effect of mesenchymal stem cell conditioned medium incorporated within chitosan nanostructure in clearance of common gastroenteritis bacteria in-vitro and in-vivo

• Published in: Scientific reports 2024-06, Vol.14 (1), p.14274-11, Article 14274
• Main Authors: Bagheri-Josheghani, Sareh, Saffari, Mahmood, Radaei, Tooba, Mirzaei, Hamed, Rashki, Somaye, Fatemi-Nasab, Zahra Sadat, Derakhshan-nezhad, Elahe, Bakhshi, Bita
• Format: Article
• Language: English
• Subjects: 631/326; 631/532; Animals; Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - pharmacology; Antibacterial; Antibacterial activity; Antibiofilm; Bacteria; Biofilms - drug effects; Campylobacter jejuni - drug effects; CD44 antigen; CD73 antigen; Chitosan; Chitosan - chemistry; Chitosan - pharmacology; Culture Media, Conditioned - pharmacology; Developing countries; E coli; Escherichia coli - drug effects; Gastroenteritis; Gastroenteritis - microbiology; Humanities and Social Sciences; Humans; LDCs; Light scattering; Mesenchymal stem cells; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - drug effects; Mesenchymal Stem Cells - metabolism; Mice; Microbial Sensitivity Tests; multidisciplinary; Nanoparticles; Nanoparticles - chemistry; Nanostructures - chemistry; Particle Size; Public health; Salmonella enteritidis - drug effects; Scanning electron microscopy; Science; Science (multidisciplinary); Shigella dysenteriae - drug effects; Stem cells; Synergistic effect; Zeta potential
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-64465-y

Gastroenteritis infection is a major public health concern worldwide, especially in developing countries due to the high annual mortality rate. The antimicrobial and antibiofilm activity of human mesenchymal stem cell-derived conditioned medium (hMSCsCM) encapsulated in chitosan nanoparticles (ChNPs) was studied in vitro and in vivo against common gastroenteritis bacteria. The synthesized ChNPs were characterized using Zeta potential, scanning electron microscopy (SEM), and dynamic light scattering (DLS) techniques. HMSC-derived conditioned medium incorporated into chitosan NPs (hMSCsCM-ChNPs) composite was fabricated by chitosan nanoparticles loaded with BM-MSCs (positive for CD73 and CD44 markers). The antimicrobial and antibiofilm activity of composite was investigated against four common gastroenteritis bacteria ( Campylobacter jejuni  ATCC29428, Salmonella enteritidis ATCC13076, Shigella dysenteriae  PTCC1188, and E. coli ATCC25922) in-vitro and in-vivo. Majority of ChNPs (96%) had an average particle size of 329 nm with zeta potential 7.08 mV. The SEM images confirmed the synthesis of spherical shape for ChNPs and a near-spherical shape for hMSCsCM-ChNPs. Entrapment efficiency of hMSCsCM-ChNPs was 75%. Kinetic profiling revealed that the release rate of mesenchymal stem cells was reduced following the pH reduction. The antibacterial activity of hMSCsCM-ChNPs was significantly greater than that of hMSCsCM and ChNPs at dilutions of 1:2 to 1:8 (P  10 6 CFU/ml to approximately (102 to 10) after 72 h. Both in vitro and in vivo assays demonstrated the antimicrobial and antibiofilm activities of ChNPs at a concentration of 0.1% and hMSCsCM at a concentration of 1000 μg/ml to be inferior to that of hMSCsCM-ChNPs (1000 μg/ml + 0.1%) composite. These results indicated the existence of a synergistic effect between ChNPs and hMSCsCM. The designed composite exhibited notable antibiofilm and antibacterial activities, demonstrating optimal release in simulated intestinal lumen conditions. The utilization of this composite is proposed as a novel treatment approach to combat gastroenteritis bacteria in the context of more challengin