# Effects of Piper betle Leaf Extract-Coated Suture Material on Clinical Strains of Staphylococcus aureus and Staphylococcus pseudintermedius Isolated from Skin-Infected Dogs

## TL;DR

This study shows that sutures coated with Piper betle leaf extract can reduce bacterial infections in dogs by inhibiting bacteria, biofilms, and adhesion.

## Contribution

The novel contribution is the development of a biocidal suture coating using Piper betle leaf extract to combat staphylococcal infections in veterinary surgery.

## Key findings

- Piper betle extract-coated sutures inhibited Staphylococcus aureus and Staphylococcus pseudintermedius adhesion and biofilm formation.
- Hydroxychavicol, the main phytochemical in the extract, remained stable on the sutures during the experiment.
- Low-dose extract-coated sutures showed minimal cytotoxicity with high cell survival rates.

## Abstract

Non-absorbable sutures are predisposed to bacterial adhesion, increasing the risk of surgical site infections. Alternative prevention, such as using an extract from Piper betle leaves as a biocidal coating agent on suture materials, is an option. This study assessed the effectiveness of P. betle leaf extract-coated sutures against staphylococci. The extract was obtained from ethanolic extraction and analyzed for phytochemicals. Four treatments, including uncoated, antibiotic/extract-free-coated, P. betle leaf extract-coated, and gentamicin-coated sutures, were tested. Analysis of P. betle leaf extract revealed hydroxychavicol as the main phytochemical. The P. betle leaf extract-coated suture showed a stable hydroxychavicol amount on the coated suture during the experiment period. The P. betle leaf extract-coated sutures showed antibacterial, antibiofilm, and anti-adhesion activities to tested staphylococci. Furthermore, a low dose of the P. betle leaf extract-coating agent on the suture had less of a cytotoxicity effect.

Non-absorbable sutures provide a site for bacterial attachment and increase the risk of surgical site infections. An alternative prevention of infections requires plant-extract coatings on sutures. The objectives of this study were to develop P. betle leaf extract-coated non-absorbable sutures and to investigate their activities on staphylococci. P. betle leaves were extracted and analyzed for the phytochemicals. P. betle extract was coated on sutures, including polyester and polypropylene. The stability of hydroxychavicol on coated sutures was evaluated. Four treatments were designed, including (1) uncoated, (2) antibiotic/extract-free-coated, (3) extract-coated, and (4) gentamicin-coated sutures. Each treatment was tested for antibacterial, antibiofilm, and anti-adhesion activities on Staphylococcus aureus and Staphylococcus pseudintermedius. In addition, the cytotoxicity of extract-coated sutures was tested. Analysis of the extract identified hydroxychavicol (40.07%) as the primary phytochemical. Stability tests indicated higher hydroxychavicol on Day 1 of extract-coated polyester compared to polypropylene, and the levels decreased on the subsequent days (p < 0.05). Antibacterial activity of extract-coated polyester showed antibacterial effects during the experiment period (5.16 ± 2.35 mm), while polypropylene showed no effectiveness. Additionally, biofilm inhibition was found to be 36.63 ± 27.08% and 37.34 ± 26.98% in tested staphylococci for extract-coated polyester and polypropylene, respectively. Anti-adhesion showed that the extract-coated sutures had a higher ability to decrease tested bacteria attachment (56.25–60.42% living cell reduction). The cytotoxicity study revealed that extract-coated sutures of ≤1.5 mg/1.5 cm had a 99% survival rate. The findings indicate that the coated sutures showed antibacterial, antibiofilm, and anti-adhesion effects against staphylococci causing canine skin infections and might lead to alternative surgical use in veterinary medicine.