Insight into the Regulatory Function of Human Hair Keratins in Wound Healing Using Proteomics.

Keratins derived from human hair possess excellent wound healing qualities. However, their functional contribution to this process is poorly understood. In this study, the regulatory function of human hair keratins in wound healing is investigated using proteomic analysis by dividing keratins into different groups based on their molecular weight distributions: low molecular weight keratins (LMWK, 10-30 kDa), medium molecular weight keratins (MMWK, 30-50 kDa), and high molecular weight keratins (HMWK, >50 kDa). Keratin hydrogels with different molecular weights exhibit various morphologies, rheological properties, degradation rates, and wound healing activities. Using proteomic analysis, LMWK and HMWK hydrogels exhibit a stronger regulatory ability for wound healing at days 1 and 7, respectively. The major functions of LMWK during wound healing are regulation of cells communication and function. In contrast, proteins associated with energy metabolism are significantly expressed after HMWK hydrogel treatment at day 1, and these play an important role in cellular growth and reactive oxygen species scavenging at day 7. These results demonstrate that the wound healing qualities of human hair keratins are influenced by their molecular weight distribution, and the proteomic analysis sheds new light on the regulatory function of human hair keratins during wound healing.

Recombinant Human Hair Keratin Nanoparticles Accelerate Dermal Wound Healing.

In recent years, favorable enhanced wound-healing properties and excellent biocompatibility of keratin derived from human hair have attracted considerable attention. Recombinant keratin proteins can be produced by recombinant DNA technology and have higher purity than extracted keratin. However, the wound-healing properties of recombinant keratin proteins remain unclear. Herein, two recombinant trichocyte keratins including human type I hair keratin 37 and human type II hair keratin 81 were expressed using a bacterial expression system, and recombinant keratin nanoparticles (RKNPs) were prepared via an ultrasonic dispersion method. The molecular weight, purity, and physicochemical properties of the recombinant keratin proteins and nanoparticles were assessed using gel electrophoresis, circular dichroism, mass spectrometry, and scanning electron microscope analyses. The RKNPs significantly enhanced cell proliferation and migration in vitro, and the treatment of dermal wounds in vivo with RKNPs resulted in improved wound healing associated with improved epithelialization, vascularization, and collagen deposition and remodeling. In addition, the in vivo biocompatibility test revealed no systemic toxicity. Overall, this work demonstrates that RKNPs are a promising candidate for enhanced wound healing, and this study opens up new prospects for the development of keratin biomaterials.

Synthesis and fabrication of a keratin-conjugated insulin hydrogel for the enhancement of wound healing.

Accelerating and regulating collagen formation during wound healing repair is key issues for skin regeneration. Insulin can promote the healing of damaged skin by stimulating cellular migration and angiogenesis. Here, human hair keratin-conjugated insulin was synthesized to enhance full-thickness skin regeneration based on the excellent wound healing and hemostatic effects of keratin and the collagen deposition regulation ability of insulin. The insulin-conjugated keratin (Ins-K) was synthesized through the EDC/NHS reaction, which can supply a sustained release of insulin. The Ins-K hydrogel displayed similar water absorption, porosity and rheology properties to those of the keratin hydrogel. However, the Ins-K hydrogel shows a stronger hemostatic ability than the keratin hydrogel group, with a stronger wound healing effect found for the Ins-K hydrogel in the early regeneration stage (first 2 weeks) than for the keratin hydrogel treatment, resulting in smoother skin tissues at an excision section realized by regulating transforming growth factor ß1 (TGF-ß1) and hydroxyproline (HYP) expression. The results demonstrate that keratin promotes hemostasis and wound healing after insulin conjugation, which highlights the potential of keratin-based materials in tissue regeneration applications.