Recolonization Dynamics of the Middle Ear Microbiota Following MESNA-Assisted Dissection in Paediatric Cholesteatomatous Chronic Otitis Media
This study characterizes the middle ear microbiome of paediatric patients with cholesteatomatous chronic otitis media (CCOM) and evaluates the ecological shifts induced by sodium 2-mercaptoethanesulphonate (MESNA)-assisted surgical dissection. 16S rRNA gene sequences (V3-V4 hypervariable regions) were generated from middle ear tissue biopsies collected from paediatric CCOM patients before (n = 13) and after (n = 13) MESNA application, and from healthy controls (n = 11). RNA was extracted using TRIzol reagent, reverse-transcribed to cDNA, and sequenced on the Illumina MiSeq platform (Zymo Research, Irvine, CA, USA). Bioinformatic processing was performed using QIIME2 and DADA2 for ASV inference, with taxonomic assignment against the SILVA 132 database. Downstream analyses employed a Compositional Data Analysis (CoDA) framework including Aitchison distance-based beta diversity (PERMANOVA), ALDEx2 differential abundance testing, consensus co-occurrence network reconstruction (SparCC, SPIEC-EASI, CLR-Pearson), Random Forest classification with Boruta feature selection, and PICRUSt2 functional inference. CCOM was associated with marked reduction in microbial network connectivity and taxonomic diversity compared to healthy controls, with network collapse from 185 edges in health to 2 edges in disease. MESNA disrupted the pathological community attractor and initiated secondary ecological succession, though full microbiome recovery was incomplete at the post-treatment sampling timepoint.
This study characterizes the middle ear microbiome of paediatric patients with cholesteatomatous chronic otitis media (CCOM) and evaluates the ecological shifts induced by sodium 2-mercaptoethanesulphonate (MESNA)-assisted surgical dissection. 16S rRNA gene sequences (V3-V4 hypervariable regions) were generated from middle ear tissue biopsies collected from paediatric CCOM patients before (n = 13) and after (n = 13) MESNA application, and from healthy controls (n = 11). RNA was extracted using TRIzol reagent, reverse-transcribed to cDNA, and sequenced on the Illumina MiSeq platform (Zymo Research, Irvine, CA, USA). Bioinformatic processing was performed using QIIME2 and DADA2 for ASV inference, with taxonomic assignment against the SILVA 132 database. Downstream analyses employed a Compositional Data Analysis (CoDA) framework including Aitchison distance-based beta diversity (PERMANOVA), ALDEx2 differential abundance testing, consensus co-occurrence network reconstruction (SparCC, SPIEC-EASI, CLR-Pearson), Random Forest classification with Boruta feature selection, and PICRUSt2 functional inference. CCOM was associated with marked reduction in microbial network connectivity and taxonomic diversity compared to healthy controls, with network collapse from 185 edges in health to 2 edges in disease. MESNA disrupted the pathological community attractor and initiated secondary ecological succession, though full microbiome recovery was incomplete at the post-treatment sampling timepoint.
hospital infantil de mexico federico gomez