# Nanostructured propolis ointment and platelet-rich plasma as novel biotherapeutics for cutaneous wound repair in an experimental canine model

## TL;DR

This study shows that combining nanostructured propolis ointment with platelet-rich plasma improves wound healing in dogs by boosting antioxidants and tissue repair.

## Contribution

The novel contribution is the development and evaluation of a bio-enhanced PRP formulation with nano-propolis for wound healing.

## Key findings

- PRP and PRP-nano-propolis reduced granulation tissue and improved collagen maturation compared to controls.
- The combination of PRP and nano-propolis enhanced antioxidant activity and TNF-α expression.
- PRP alone induces early cytokine activity, while propolis provides a delayed but sustained effect.

## Abstract

Effective management of cutaneous wounds is challenging in clinical practice. The present study aimed to investigate the relative efficacy and explore the potential differences of nanostructured propolis ointment, platelet rich plasma (PRP) and their combination in enhancing the healing of experimentally induced cutaneous defect in dog model. The study included 6 dogs with 6 skin wounds per dog. A 3-cm full-thickness skin wounds were surgically induced on the lateral thoracic walls. Wounds were randomly allocated to six treatment groups: control, lanolin (vehicle), nano-propolis, PRP, PRP-lanolin, and PRP-nano-propolis. Wound healing progression was evaluated clinically and histologically over 20 days using wound area measurements, epithelization, granulation tissue formation, and collagen deposition. The tumor necrosis factor-alpha (TNF-α) was immunohistochemically assessed. Biochemical markers including total antioxidant capacity (TAC), malondialdehyde (MDA), matrix extracellular phosphoglycoprotein (MEPE), transforming growth factor beta (TGF-β), platelet growth factor beta (PDGF-β) levels were also evaluated.

PRP and PRP-nano-propolis groups exhibited significantly decreased granulation tissue formation and enhanced collagen maturation compared to controls and lanolin-treated wounds (p < 0.05). The combination of PRP and nano-propolis resulted in superior modulation of oxidative stress, with marked elevation in TNF-α expression and antioxidant levels (p < 0.05). The temporal pattern of TGF-β expression suggests that PRP alone rapidly induces cytokine, while propolis exerts a delayed but sustained effect, and their combination yields both early and prolonged upregulation. These synergistic dynamics underscore the potential of bio-enhanced PRP formulations, particularly those incorporating nano-propolis, in accelerating and sustaining cutaneous wound healing. Histopathological and immunohistochemical findings supported these observations, indicating accelerated tissue regeneration and remodeling.

The application of nano-propolis and PRP, particularly in combination, significantly promotes cutaneous wound healing through enhanced antioxidant activity, and improved tissue repair mechanisms. These findings support their therapeutic potential and warrant further investigation in clinical settings.