Tarantula Cubensis Venom Theranekron-Embedded Hyaluronic Acid and Chitosan-Based Wound Dress Enhances Wound Healing by Ameliorating Histopathological Appearance and Enhancing Growth Factors in Low Doses: In Vitro and In Vivo Study.

Abstract

Background: This study aimed to produce wound dressing materials using the multilayer coating method(LbL) with varying doses of theranekron®(T) incorporated into hyaluronic acid (HA)/chitosan(Ch) multilayer films, demonstrating their proliferative effect through in vivo wound model experiments.

Material and methods: In vitro wound dressings were created by dipping 2 × 2 cm cotton fabrics into T solutions via the LbL method. Four circular wounds (1.5 cm diameter) were made on the dorsal area of 16 rats. Eight groups (control, HA-Ch, T₁₀, T₂₀, T₅₀, T₁₀₀, T₂₀₀, and T₅₀₀) used wound dressings impregnated with six doses of T µg. Dressing changes occurred every other day for 7 days, with rats killed on the 8th day. Skin samples underwent histopathological and histochemical examination, and growth factor expression levels were analyzed via qRT-PCR.

Results: SEM-EDS analysis of treated cotton fabric indicated compositions of 44.79% carbon, 48.56% oxygen, 6.19% nitrogen, and 0.46% calcium. Ch particles were evenly distributed on HA-Ch-coated fibers; however, crystalline structures diminished with increasing doses. Wound healing rates significantly increased in the T₂₀ and T₅₀ groups, showing reductions of 41.32% and 44.59% from day 1 to day 7. Masson trichrome staining revealed the most pronounced collagen formation in the T₁₀ group, with increased collagen also in T₂₀ and T₅₀ groups. Growth factor expression levels were notably higher in the T₁₀, T₂₀, and T₅₀ groups. In Picro Sirius Red-stained sections, enhanced collagen maturation was observed in T₁₀, T₂₀, and T₅₀ groups, while higher doses led to irregular and immature collagen deposition.

Conclusion: The study confirmed the in vivo and in vitro efficacy of the T + HA + Ch wound dressing material, identifying the optimal wound healing dose of T as 10-50 µg based on histopathological and genetic analyses.

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