Honey-Propolis-Engineered Collagen Peptides as Promising Wound-Healing Matrix in Mouse Model

Abstract

In this study, collagen hydrolysates (CHDs) were fabricated with honey-propolis wax (HPW), structurally modified as a sponge matrix, and experimentalized on wound healing in a mouse model. The scaffold was characterized by means of in vitro enzymatic degradation; in vitro HPW release; and in vivo wound-healing mouse model, wound-healing-specific RNA, transcripts, and protein markers. The functional activity of the HPW extracted from raw propolis was determined using total flavonoids, antioxidant scavenging assays, and anti-hemolytic principles. The results indicated that HPW had a high flavonoid content (20 μg/mL of wax) and antioxidant activities. The effective concentration (EC50) of HPW was estimated (28 mg/mL) and was then used in the subsequent in vivo experiments. Additionally, the dopped mixture of CHDs and HPW substantially enhanced the wound-healing process and regulated wound biochemical markers such as hexoseamine and melondialdehyde. CHDs- HPW upregulated the expression of growth factors including vascular endothelial growth factor (VEGF) (2.3-fold), fibroblast growth factor (FGF) and epidermal growth factor (EGF) (1.7-fold), and transforming growth factor-beta (TGF-β) (3.1-fold), indicating their potential capacity to perform wound re-epithelialization and the loading of ground tissue. Pro-inflammatory markers IL-1 β (51 pg/mL) and TNF-α (220 pg/mL) were significantly reduced in the CHD-HPW-treated wound. These interesting results were further confirmed using mRNA and protein growth factors from the wound, which enhanced the load of collagen-I in the wound site. In conclusion, CHDs-HPW exhibited a significant reduction in inflammation and inflammatory markers and helped to obtain a faster wound-healing process in a mouse model. The newly engineered biosponge could be developed as a promising therapeutic approach for the regeneration and repair of damaged human skin in the future.

Honey and propolis for wound healing

“…1 produce honey with excellent nutritional and medicinal value with potent antimicrobial action 5,6 and enhance wound healing by reducing the level of inflammation, edema, and pain through suppression the activities of cyclooxygenases 1 and 2 (COX1 and COX2). 7 Correct treatment of chronic wounds and ulcers is a complex process, which requires de-trauma, drainage and anti-inflammatory therapies, 8 so honey is a favorable as it promotes tissue repair 9 re-epithelization, 10 collagen deposition, 2 debridement action, 11 plays a role in tissue regeneration since honey-induced extracellular Ca2+ entry results in wound healing, which is consistent with the role played by Ca2+ signaling in tissue regeneration. 12 Rather than it has durable adhesive characteristics for skin graft fixation.…”

“…The biological activities of propolis involved in wound healing and tissue regeneration correlates with its antimicrobial, anti-inflammatory and immunomodulatory properties. 74 The antimicrobial synergistic action of both honey and propolis is recommended in vitro 5,75 and in vivo 10 since their combination stimulates the monocytes/macrophages to release cytokines important for wound healing, and promote neutrophils 54 rather than B-lymphocytes (antibody production) 67 protecting the healing tissue from pathogenic infection. 54 A study 76 concluded that honey supplemented with propolis extract can be an excellent source of antioxidant compounds as significant increase in the content of polyphenolic compounds, including flavonoids and phenolic acids, of which chrysin, pinocembrin, p-coumaric acid, and ferulic acid were present in the highest amounts.…”

“…2 Otherwise, the release of honeypropolis-engineered collagen peptides in the wound site increases the absorption and re-epithelization of keratinocytes. 10 Reducing bacterial biofilm production: Biofilm production is a passive virulence factor facilitating many infectious diseases 79 where certain bacterial genes produce extracellular polysaccharides to protect these pathogens from antimicrobial agents and cellular immune elements 80 resulting in long standing chronic wounds. 65 So, fighting biofilm production through down-regulation of the biofilm genes may prevent colonization of host tissues including damaged tissues in wounds 81 is of great concern.…”

“…88 Another advantages of electro-spun nanofibers, sponge, foam that including a small diameter, high porosity, narrow diameter distribution, gas permeation, and high-specific surface to area ratio. 6,89 Various nanoscale strategies were explored for targeting different phases of wound repair 66 where these nanomaterials exhibit intrinsic properties beneficial therapeutic materials for wound treatment (natural as honey, propolis) 10,20,69,90 or employed as delivery vehicles (e.g., silver, gold, zinc) 88 for effective therapeutic agents. Nanoparticle properties increase the probability of biological interaction and injury penetration in films, foams, hydrogels, hydrocolloids, contact layers, and multilayer form dressings.…”

“…20 Honey propolis wax collagen hydrolysates exhibit a significant reduction in inflammation and inflammatory markers and helps to obtain a faster woundhealing. 10 Honey-processed Astragalus (bioactive peptide amphiphile nanofiber-based hydrogel biomaterial) may stimulate burn wound healing. 48 Propolis nanoparticles have anti-inflammatory and antiapoptotic effects as decreased expression of TGF-β and increased the expression of Bcl-2 in liver and kidney.…”