Escherichia coli e coli

The caffeic acid, keracyanin chloride, kuromanin chloride, Folin-Ciocalteau, and quercetin used in the experiment were purchased from Sigma-Aldrich (St. Louis, MO, USA). Aluminum chloride hexahydrate and potassium acetate were purchased from Junsei Chemical (Tokyo, Japan). Citric acid, sodium alginate, mannitol, and poloxamer 188 were purchased from Daejung (Seoul, Republic of Korea). Whey protein isolate, lecithin, and ascorbyl palmitate were purchased from ESfood (Gumpo, Republic of Korea). Lacticaseibacillus rhamnosus (L. rhamnosis), Pediococcus pentosaceus (P. pentosaceus), Enterococcus faecalis (E. faecalis). Escherichia coli (E. coli), and Streptococcus aureus (E. aureus) were purchased from ATCC (American Type Culture Collection). Man-Rogasa-shape (MRS) and brain heart infusion (BHI) medium was purchased from Solarbio (Shanghai, China).

The E. spiralis stem was collected from the forest in Tasik Chini, Pahang, Malaysia before being chopped and grounded to obtain E. spiralis stem powder. The silver nitrate was purchased from Acros organic, USA and was used without any purification. The deionized water from the ELGA Lab-Water/VWS (UK) purification system was used throughout the experiment. Four species of bacteria, including two Gram-positive species Staphylococcus aureus (S. aureus) (ATCC 25923) and Enterococcus faecalis (E. faecalis) (ATCC 29212) as well as two Gram-negative species Escherichia coli (E. coli) (ATCC 25922) and Proteus vulgaris (P. vulgaris) (ATCC 33420) were bought from Choice Care Sdn. Bhd, Kuala Lumpur, Malaysia. The stock culture was prepared in the Mueller Hinton broth (Difco, Malaysia) and incubated at 37°C overnight. For further usage, the stock culture was kept in the refrigerator at a temperature of 4–8°C.

S. aureus subspecies (MRSA, ATCC 43,300, methicillin-resistant, second-generation species) was purchased from BeNa Culture Collection (Beijing, China). S. aureus (ATCC 29,213), Streptococcus pneumoniae (S. pneumoniae, ATCC 49,619), Escherichia coli (E. Coli, ATCC 25,922), Pseudomonas aeruginosa (P. aeruginosa, ATCC 27,853) and Candida albicans (ATCC 10,231) were obtained from quality-control strains in microbiology laboratory of University-Town Hospital of Chongqing Medical University. Enterococcus faecalis (E. faecalis) Enterobacter cloacaeE. cloacaeKlebsiella oxytoca were obtained from clinical wild strain in microbiology laboratory. All strains including the resistance of MRSA to penicillin and cephalosporins were confirmed by the phoenix 100 bacterial identification instrument (BD, USA). All strains preserved in the original bacterial suspension (72 mM K₂HPO₄, 3.8 mM sodium citrate, 0.7 mM MgSO₄, 88% glycerol) were cultured on Blood, MacConkey or Sabouraud agar plate medium respectively in an incubator containing 5% CO₂ at 37 ℃ for 12–24 h. Bacterial suspensions with concentration of 0.5 MCF determined by a scattered light turbid meter according to the optical density measured at the wavelength of 600 nm in PBS were prepared for nucleic acid extraction or other experiments.

Freeze dried S. aureus (ATCC 25923), Staphylococcus epidermidis (S. epidermidis) (ATCC 12228), Escherichia coli (E. coli) (ATCC 13115), E. coli* (ATCC 15597), Enterococcus faecalis (E. faecalis) (ATCC 29212), and Enterococcus casseliflavus (E. casseliflavus) (ATCC 9199) were purchased from American Type Culture Collection (ATCC) and revived according to ATCC protocols. ATCC bacteria culture stocks were stored at −80 °C in a cryomedium to glycerol ratio of 3:2. When needed, bacteria cultures were inoculated into the brain hearth infusion (BHI) liquid media and incubated at 37 °C.

Chitosan (CTS) with 85% deacetylation (molecular weight < 5 kDa), type B gelatin (GEL), methacrylic anhydride (MA), fluorescamine (98%), dimethyl sulfoxide (DMSO), and lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP; 95%) were purchased from Sigma-Aldrich (Aldrich, St. Louis, MO, USA). Gallic acid (GA; 3,4,5-trihydroxybenzoic acid monohydrate, 98%), pyrogallol (PG; 1,2,3-trihydroxybenzene, 99%), and tannic acid (TA, 95%) were obtained from Alfa Aesar (Heysham, UK), and their chemical structures are shown in Figure 1a.
Escherichia coli (E. coli; ATCC 8739), Staphylococcus aureus (S. aureus; ATCC 25923), Porphyromonas gingivalis (P. gingivalis; A7436), Streptococcus mutans (S. mutans; ATCC 700610) strains, and L929 fibroblast cell line (RM60091) were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan). Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), and penicillin/streptomycin solution were purchased from Gibco (Langley, OK, USA). The alamarBlue reagent was from Invitrogen (Grand Island, NY, USA). Bacto tryptic soy broth and Wilkins-Chalgren anaerobe broth were purchased from Becton Dickinson (Sparks, MD, USA) and Oxoid (Hampshire, UK), respectively. All reagents were of analytical grade and used as received without further purification.

PVDF particles, dimethyl formamide and ZnO nanoparticles were purchased from Sigma Aldrich (China). Human osteoblasts were provided by ATCC(China). Escherichia coli (E.coli) and Staphylococcus aureus (SA) were provided by ATCC(China). Cell proliferation assay kit was provided by Colorimetric (China).