New skin regeneration and wound healing products from eggshell membrane

Biovotec is finalizing the validation and optimization of the eggshell/eggshell membrane separation process in collaboration with Nortura and Adigo. This work will then qualify the process line for production of both fractions: the eggshell membrane (ESM) and the eggshell (ES). At the same time, it will allow Nortura to enter the market with two new products (ESM and ES) derived from fresh eggshell which they presently discard as byproduct. However, an extensive optimization is presently ongoing both to increase the operational parameters but also to ensure that the membrane produced can be used in a Medical device. As we expected by the end of 2017 both process optimization and validation will be completed and by this the products will be fully qualified according to specifications and will meet one of the main objectives of this project. During the entire project period, all BIA partners have dedicated contributed to the development, starting from the raw material until the development of a novel wound care products. The studies conducted during the last 4 years have clearly demonstrated the potential of this new bio material (ESM). Overall, the BIA project will have produced 6 scientific publications and 4 patents. During 2017, the following tasks and results have been achieved: - Biovotec DAC in Ireland has been focusing on the optimization of the ESM purification and milling as we in the beginning of 2017 through pig wound trials revealed that we had an overstimulation of granulation tissue formation. Additional animal trials were then performed to confirm the efficiency of the newly processed ESM. The tests gave us encouraging results and the obtained ESM product was further investigated by all BIA partners as well as introduced into new formulations for wound healing. In addition, new product development such as the fabrication of scaffold has been done and is currently tested by our BIA partners. This new product opens up new opportunities in the field of tissue engineering. - Biovotec is currently testing the most promising medical device formulation using different animal models with both newly processed ESM and the final medical device. The first results have shown that both newly processed ESM and it final formulation form promote wound closure, tissue granulation and tissue re-ephitiliziation within few days. - At Nofima, final experiments were carried out and confirm that processed ESM induces cell migration. Further studies confirmed that MMP activity enabling cell migration is increased in wound edges (epidermal) and granulation tissue during healing. All studies confirmed that process ESM has a significant effect on early phase of wound healing. In addition, processed ESM increased keratinocyte cell proliferation in the wound edges. Altogether, our data suggest that processed ESM is a stimulator of MMP activity and other early cellular events during wound healing. Nofima is also working with different type of scaffolds and the results indicate that ESM scaffolds could be a new product for tissue engineering. In parallel with the above Nofima is writing a new publication on MMPs. - The University of Ottawa has confirmed that processed ESM is a wound healing accelerator by using in vivo mouse model. The last proteomic analysis of processed ESM identified 110 proteins, including structural proteins such as collagen and cysteine-rich eggshell membrane proteins (CREMPs) which together constitute about 40% of processed ESM. Functional annotation clustering showed various predicted functionalities related to wound healing including response to external stimulus, defense response, inflammatory response, and cell-substrate adhesion. Processed ESM was found to significantly accelerate wound closure at days 3, 7, and 10 on animal models. Histological assessment showed higher levels of collagen deposition at day 10 in wounds treated with processed ESM, with limited inflammatory reaction. Therefore, PEP is a biocompatible and non-cytotoxic biomaterial that has great potential for development into a cost-effective wound healing product. - Our Partner Innovent has been working on the fabrication of various cryogels from methacrylated biopolymer-derived hyaluronan and dextran as well as from synthetic macromer polyethylene glycol diacrylate (PEGDA) with and without addition of processed ESM as potential mediator of the cell regeneration capacity of the final scaffolds. Among the studied cross linkable macromers, PEGDA has been found to provide cryogels excellent mechanical stability, inter-connecting porosity and in vitro cytocompatibility. Furthermore, the addition of processed ESM to the cryogels considerably increased the ability of cells to adhere onto the cryogel surfaces or even migrate into the porous structure. Overall, the synthesized cryogels are promising candidates for wound dressings that would support tissue regeneration.

Biovotec AS has an exclusive license to use eggshell membrane, produced from Agroplas AS newly developed process technology , as a raw material for biomedical applications included for wound healing. This allows for the first time ever to establish a complete supply chain from industrial egg/breaker facilities to final commercial wound healing product.