Bulk RNA sequencing
Granulation tissues were collected on the day of treatment, placed in 100 μl of RNAlater, flash frozen, and stored in − 80 °C. Total RNA was obtained using the “Quick- RNA MicroPrep” kit (Zymo Research) according to manufacturer's instructions. Following this, total RNA was sent to the Oxford Genomics Centre (Oxford, United Kingdom) for quality control, library preparation and sequencing using Illumina NovaSeq6000 (150 bp, pair-ended) platform.The fastq reads where aligned to the human genome (GRCh38.p14/hg38) using an RNAseq specific aligner. To quantify gene expression levels, count tables were generated using the human hg38 transcriptome as reference. The count tables of each sample group were then compared and subjected to expression testing to identify differentially expressed genes between the two conditions. Raw reads were mapped to GRCh38.p14/hg38 using Hisat2, and EdgeR together with the Cufflinks pipeline were used to identify genes with differential gene expression. A gene was classified as being differentially expressed if it had a q < 0.05.Gene Ontology (GO) functional annotation and KEGG pathway enrichment analysis for the up and down-regulated expressed genes in the granulation tissues was performed using Enrichr database. Three categories, including biological process (BP), molecular function (MF), and KEGG pathway were included in GO annotation. The top 10-enriched GO items and KEGG pathways were displayed on the webpage and downloaded as images.