Antimicrobial and Antibiofilm Effects of Human Amniotic/Chorionic Membrane Extract on Streptococcus pneumoniae

Abstract

Background: Streptococcus pneumoniae colonize the human nasopharynx in the form of biofilms. The biofilms act as bacterial reservoirs and planktonic bacteria from these biofilms can migrate to other sterile anatomical sites to cause pneumonia, otitis media (OM), bacteremia and meningitis. Human amniotic membrane contains numerous growth factors and antimicrobial activity; however, these have not been studied in detail. In this study, we prepared amniotic membrane extract and chorionic membrane extract (AME/CME) and evaluated their antibacterial and antibiofilm activities against S. pneumoniae using an in vitro biofilm model and in vivo OM rat model.Materials and Methods: The AME/CME were prepared and protein was quantified using DCTM (detergent compatible) method. The minimum inhibitory concentrations were determined using broth dilution method, and the synergistic effect of AME/CME with Penicillin-streptomycin was detected checkerboard. The in vitro biofilm and in vivo colonization of S. pneumoniae were studied using microtiter plate assay and OM rat model, respectively. The AME/CME-treated biofilms were examined using scanning electron microscope and confocal microscopy. To examine the constituents of AME/CME, we determined the proteins and peptides of AME/CME using tandem mass tag-based quantitative mass spectrometry.Results: AME/CME treatment significantly (p < 0.05) inhibited S. pneumoniae growth in planktonic form and in biofilms. Combined application of AME/CME and Penicillin-streptomycin solution had a synergistic effect against S. pneumoniae. Biofilms grown with AME/CME were thin, scattered, and unorganized. AME/CME effectively eradicated pre-established pneumococci biofilms and has a bactericidal effect. AME treatment significantly (p < 0.05) reduced bacterial colonization in the rat middle ear. The proteomics analysis revealed that the AME/CME contains hydrolase, ribonuclease, protease, and other antimicrobial proteins and peptides.Conclusion: AME/CME inhibits S. pneumoniae growth in the planktonic and biofilm states via its antimicrobial proteins and peptides. AME/CME are non-cytotoxic, natural human product; therefore, they may be used alone or with antibiotics to treat S. pneumoniae infections.

The use of irradiated amnion dressing for the treatment of antibiotic‐disinfected skin ulcer

“…These types of pathogenic microorganisms secrete toxins, adhesion molecules, and surface complexes, causing cytotoxicity to the wound and subsequently affecting the healing (Makrantonaki, Wlaschek, & Scharffetter‐Kochanek, 2017). The synergistic effect of AM with antibiotics against microbial growth and biofilm formation was proved previously (Yadav, Go, Kim, Chae, & Song, 2017). Hence, this study recommends vigorously washing the wound with antibiotic‐contained saline, before applying iHAM, for eliminating microbial colonization and their biofilms into wounds and to protect the wound from external infection.…”

<p>Different Effects Of Amniotic Membrane Homogenate On The Growth Of Uropathogenic <em>Escherichia coli</em>, <em>Staphylococcus aureus</em> And <em>Serratia marcescens</em></p>

“…[2][3][4] AM has been utilized in medicine for some time [4][5][6] and antimicrobial activity of AM has been previously reported. [7][8][9][10][11][12][13][14][15][16] However, due do the broad range of AM preparation techniques (e.g. AM patches, AM conditioned medium, AM extract) and various testing methods used, reported results are in some cases inconsistent and unclear.…”

Effect of Lead on Human Middle Ear Epithelial Cells

“…Therefore, this study evaluated the effects and mechanism of lead toxicity on human middle ear epithelial cells (HMEECs). In a previous study, we showed that amniotic membrane extract (AME) and chorionic membrane extract (CME) have anti-inflammatory effects on HMEECs; therefore, we evaluated the protective effect of AME and CME on lead toxicity [13]. …”