Keratin-collagen Matrices For Wound Healing Applications
Angana Kharge, PhD1, Ankur Gandhi, PhD1, Sita Damaraju, PhD1, Heli Modi, MS1, Luke Burnett, PhD2, Sunil Saini, PhD3.
1INTEGRA LIFESCIENCES, Plainsboro, NJ, USA, 2Keratin Biosciences, Winston-Salem, NC, USA, 3INTEGRA LIFESCIENCES, Plainsboro Center, NJ, USA.
Keratin is a fibrous structural protein that provides mechanical stability to cells of the epithelium. These proteins can be used in wound healing applications in the form of scaffolds, hydrogels, and wound dressings. Here, we evaluated the in vitro and in vivo biological response to different presentations of keratin within a bioengineered collagen matrix. Keratin was extracted from end-cut human hair through a proprietary oxidative process. Two different configurations of keratin-collagen matrices were fabricated: keratin hydrogel was superficially layered on one side of a collagen sponge, lyophilized, and sterilized (bilayer keratin-collagen matrix); keratin powder was directly incorporated into solubilized collagen, lyophilized, crosslinked, and sterilized (Keratin-collagen matrix). In in vitro cell culture, adult fibroblasts were seeded on matrices and cultured up to 14 days. Samples were analyzed for DNA, collagen type I (ELISA) and imaged using confocal microscope. In vivo, partial-thickness wounds (2.5cm square) were created on the dorsum of Yorkshire pigs. Wounds were treated with the bilayer keratin-collagen matrix, the keratin-collagen matrix or the collagen matrix. Dressing changes were performed at 3 or 4-day intervals up to sacrifice at Day 7 or 21. In vitro cell culture studies demonstrated that the keratin-based matrices supported fibroblast growth, collagen type I deposition and migration of dermal fibroblasts. There were no adverse effects. The keratin prototypes demonstrated complete matrix take. Histologically, at Day 7 keratin-collagen matrix demonstrated enhanced epithelial burrowing suggesting the incorporation of keratin into collagen resulted in greater keratinocyte activity. Further, there was less contraction in both keratin prototypes compared to collagen-only matrices suggesting greater mechanical stability. In conclusion, two different configurations of keratin-collagen scaffolds were successfully fabricated. Pilot results demonstrate enhanced keratinocyte activity in the keratin-collagen matrix in early stages of wound healing. Future studies evaluating the impact of keratin on long term healing are warranted.
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