Hyaluronic Acid‐Recombinant Collagen Type III Electrospun Nanomembranes: A Stratum Corneum‐Navigated Cutaneous Delivery Platform for Skin Rejuvenation
Xinyu Peng, Songhua He, Xiaofeng Qiu, Zhanhua Fan, Qiudong LiaoABSTRACT
Objective
This study tests the hypothesis that electrospun nanomembranes composed of recombinant human collagen type III (rhCol III, 35–50 kDa) and hyaluronic acid (HA, 20–40 kDa) enable stratum corneum–navigated cutaneous delivery of these two substances, thereby synergistically enhancing dermal physiology.
Methods
RhCol III (35–50 kDa) and HA (20–40 kDa) were dissolved in ultrapure water (Milli‐Q, 18.2 MΩ·cm) at a mass ratio of 5.25:1 (w/w) to yield a 25% (w/v) total polymer concentration. Electrospinning was performed using an electrospinning device (YT‐MS‐III) at 8–9 kV voltage and a 15–18 cm tip‐to‐collector distance, yielding nanomembranes with fiber diameters of 187 ± 42 nm ( n = 30). Control serum was prepared by dissolving rhCol III/HA powder in water; the composition and concentration are identical with the nanomembranes rehydrated by 7× the dry membrane weight (simulating clinical reconstitution). Ex vivo penetration was quantified using Franz diffusion cells (3 cm 2 area) on porcine dorsal skin (TEWL < 15 g·m −2 ·h −1 ); HA was measured via HPLC (Agilent 1260, MCI GEL column, 1% phosphoric acid mobile phase, 232 nm detection); rhCol III was quantified by immunofluorescence using anti‐His‐tag antibody (Abcam ab9108, 1:500) and Alexa Fluor 488–conjugated secondary antibody (Thermo Fisher A11001, 1:1000). A randomized, split‐face, 4‐week clinical trial enrolled 35 healthy Asian volunteers (45 ± 1.0 years; wrinkle grade 2–4) to assess hydration (Corneometer CM825), barrier function (Tewameter TM300), elasticity (Cutometer MPA580), wrinkles (VISIA‐CR + Image‐Pro Plus 7.0), and erythema (Mexameter MX18).
Results
At 24 h, rhCol III‐HA nanomembranes achieved a cumulative HA permeation of 0.136 ± 0.009 mg·cm −2 (mean ± SD), representing a 72.15% increase over control serum (0.079 ± 0.001 mg·cm −2 ; p < 0.001). Apparent permeability coefficient (K p ) was 4.90 × 10 −3 ± 1.2 × 10 −4 mg·cm‐2·h −1 for nanomembranes vs. 2.50 × 10 −3 ± 8.5 × 10 −5 mg·cm‐2·h −1 for serum ( p = 0.003). rhCol III immunofluorescence signal in the dermis was 3.86 ± 0.31‐fold higher in nanomembrane‐treated skin versus serum controls at 24 h ( p < 0.001). Clinically, 4‐week electrospun nanomembrane treatment increased stratum corneum hydration by 47.82% ( p < 0.001), reduced TEWL by 41.0% ( p < 0.001), improved elasticity (R2) by 16.58% ( p < 0.001), decreased periocular wrinkle area ratio by 36.88% ( p = 0.001), and lowered erythema index by 20.34% ( p < 0.001) vs. control (without treatment).
Conclusion
Electrospun rhCol III‐HA nanomembranes function as a stratum corneum–navigated cutaneous delivery platform that overcomes classical molecular weight barriers (20–50 kDa), enabling co‐delivery of bioactive macromolecules (generally molecular weight > 10 kDa, e.g., rhCol III, HA) with statistically significant enhancements in skin hydration, barrier integrity, biomechanics, and appearance.