ERA-NET: BlueBio - 3D Printed Biomarine Wound Healing Accelerant “AquaHeal3D”

AquaHeal3D, 3D Printed Biomarine Wound Healing Accelerant Aquaheal3D combines all renewable, marine sourced ingredients to develop a 3D-printed topical wound healing medical device. This is accomplished in collaboration between Regenics AS, RISE AB (Swe) and Scantox A/S (Dk). The wound dressing contains an extract with bioactive substances from salmon roe that accelerates wound healing. The goal was to develop a 3D-printed wound dressing based on renewable marine raw materials that is clinically safety-tested in animals and ready for a clinical study in patients. The matrix material is composed of alginate from seaweed and cellulose from tunicates. The final wound dressing will contain HTX, a patented extract originating from unfertilized salmon roe manufactured by Regenics AS. Unfertilized roe is a waste product, only used for additional fish farming but shows skin regenerative and anti-inflammatory activity and is therefore of value as starting material for marine bioactive. The challenge was to incorporate the HTX in the wound dressing materials in such a way that the wound healing ability is maintained during storage, and to control the release of the extract into the wound to accelerate wound healing. 3D printing is a manufacturing technology that enables high production reproducibility and offers unmatched freedom of design for a tailored dressing with a structure that is optimized to protect the extract during storage and provide controlled release during use. We have finalized a prototype of the patch with a formulation consisting of a defined concentration of cellulose, alginate and HTX. This formulation has been 3D printed into a flexible high-fidelity porous scaffold. This scaffold consists of HTX, which is readily released from the patch to exudate-like wound fluids. The initial phase of the release displays zero order kinetics and after 3 hours, ca 90% of the HTX content has been released from the patch. We have previously shown that one of the mechanisms causing HTX to accelerate wound healing is by enhancing Collagen 1 production. To determine the bioactivity of the patch, we have performed Collagen 1 ELISA assays from cell media containing HTX released from the patch. Data shows an increase in Collagen 1, even after 8 weeks of storage, indicating that the 3D printed all-marine patch is bioactive. We have further analyzed the pathc chemically, tested its stability and cytotoxicity and our data indicates that the patch is both safe, sterile, and non-toxic. It has further been analyzed on second degree burn wounds on minipig by Scantox A/S. On these wounds, the patch reduced the inflammatory response and accelerated wound closure in the wound compared to an inactive vaseline compress. The patch also created minimal irritation on the burn wounds and seems to be bioacompatible.

Responsible Innovation is needed in the EU maritime industry. Norway is the worlds second largest seafood exporter in 2017 with a 9.75 bn Euros export value. Aquatic biology and novel use of marine materials is underexplored in medical therapeutic contexts. We will polymerize seaweed alginate with nanocellulose from marine tunicates and combine with a skin regenerative bioactive from fish roe to safety testing in humans or equivalent safety model. A 3D-bioprinted medical patch for wound healing is contemplated. A medical device made of surplus marine waste materials is novel. Such a device can improve public attitudes towards bio-based marine products and services in markets as medical technology sector scores high on clean sterile regulatory demands for product approvals. The project may also carry policymaking in bio-based research, innovation and technology development by bringing marine biotechnology into e.g. consortia collaboration alongside non-marine sourcing and technology. During the project, a prototype have been developed and tested. Results indicate that the marine wound healing device is sterile, biocompatible, non-toxic and safe. In addition, studies on minipig indicate that it reduces inflammation and accelerates wound healing of burn wounds. Based on this, collaborations with the burn care unit has been established and clinical trials are currently being planned. According to the head of the Burn care unit at Haukeland University hospital, a dressing providing local anti-inflammation and anti-oxidation will cover an unmet need for the burn patients and has the potential to improve the burn care.

This project combines all renewable, marine sourced products to create a 3D printed wound healing medical device. We will incorporate Regenics’ bioactive substances into a topical wound healing dressing for chronic hard-to-heal wounds. Biorelease from the 3D patch and skin wound safety are important milestones of the project and will be explored using marine bioactives and enzymes. One bioactive ingredient, HTX, is a patented (EPO patent granted December 2018) purified heat-stable component of unfertilized salmon roe cytoplasm; HTX has strong skin regenerative and anti-inflammatory activity. Other marine derived bioactives may serve as biorelease markers. The marine protease LCE1 will be used as an antidote to HTX and has a potential in debridement of eschar. The project will deliver a 3D printed, bioactive, clinical safety-tested dressing, ready for clinical study in patients. No dressings with integrated skin regenerating and anti-inflammatory properties are available. PoC of marine bioactives of skin regenerative effects have been demonstrated in human explanted skin. Marine wound devices – may include, patches, 3D-printed patches, sprays and creams. The consortium consists of research and industry partners that are leaders in their respective fields; having a history of research and industrial success