Gene polA as a Suitable Reference for Studying Antibacterial Effect of Hydroxyapatite

Journal of Biomedical Nanotechnology
Yu-Tzu HuangAnkit Rawat

Abstract

Hydroxyapatite (HAP: Ca10(PO4)6(OH)2) is a widely used biocompatible material; however, information on the reaction mechanism between HAP and microorganisms is insufficient. This study aimed to identify a stable reference gene for studying the antibacterial activity of HAP. The half maximal inhibitory concentration (C50) and gene expression of a human symbiotic Escherichia coli strain TOP10, was investigated at various concentrations of HAP. Our uniformly sized HAP nanoparticles (HANPs) had a high surface area and an estimated IC50 of 75 mg/mL. The expressions of genes, including those of DNA polymerase I (poIA), DNA polymerase II (poIB), cytochrome d complex (cyd), glucan biosynthesis protein G (mdoG), D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and 16S ribosomal RNA (16S rRNA), were analyzed by performing quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and using reported and newly designed primers. The changes in the copy numbers compared to non-HANPs-treated conditions for polB, cyd, mdoG, GAPDH, and 16S rRNA were 50%-381%, 0.7%-66.3%, 1.1%-7.8%, 1.6%-86.3%, and 0.3%-8.1%, respectively. The expressions of polA remained stable under all conditions (62.8%-88.4%). Therefore, we identifie...Continue Reading

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