Development of Bioinks for 3D Bioprinting

Project: “Development of Bioinks for 3D Bioprinting Based on Chemically Modified Porcine DECM, Enriched with Recombinant Hybrid Proteins, Nanomaterials, and Synthetic Polymers,” co-financed by the National Centre for Research and Development under the “Modern Material Technologies” TECHMATSTRATEG III program.

Implementation period: 01.01.2021 – 31.03.2024.

Total project cost: 22 444 594,83 PLN

NCBR funding: 21 962 857,33 PLN

The aim of the project was to develop a package of bioink modifications (including the production of recombinant hybrid protein) based on decellularized extracellular matrix (dECM) from porcine pancreas and meniscus, enhancing the efficiency of the 3D printing process and the durability of the obtained prints. As part of the project’s tasks, research was conducted to produce a ready-to-commercialize product in the fields of biotechnology, nanomaterial engineering, chemistry, and physiology.

It is worth noting that the products and technologies developed within this project have been secured by patents. Given the significant challenge posed by the lack of organs for transplantation in patients with type I diabetes and meniscus injuries, the implementation of this project carries substantial social and economic value.

3D bioprinting has become an extremely versatile tool in the fields of biological sciences and regenerative medicine over the past decade.

It is used in the production of bone defect implants, and increasingly in the construction of functional organs such as the pancreas, lungs, liver, heart, and skin. Moreover, tissue engineering is utilized in biotechnology research, in the creation of disease models for cancer, and in screening compounds in drug production. To achieve the desired tissue construct, a range of cells, methods, and materials can be used in 3D bioprinting, depending on the nature of the printed object.

The bioinks used in 3D bioprinting exhibit a very wide spectrum of biophysical characteristics: viscosity, printability, and degradation. Additionally, they must provide favorable conditions that allow for cell embedding and differentiation, without cytotoxicity.

The technology of 3D bioprinting is based on creating three-dimensional structures from living cells, which are suspended in a biocompatible hydrogel called bioink.

This hydrogel enables precise shaping of the printed object. Depending on the type of cells used, diverse research models can be obtained.

Bioinks should exhibit a wide range of biophysical properties, such as printability, viscosity, degradation rate, and the ability to gelate. These properties allow for the creation of precise and stable scaffolds, providing suitable conditions for cells to embed within the structure. We have successfully developed our own bioinks that meet all of these criteria.

Multidisciplinary Consortium

Multidisciplinary Consortium The project was carried out through the collaboration of a multidisciplinary consortium with significant experience in creating necessary materials for printing prototypes of bionic organs.

The consortium comprises the following centers and teams:

Foundation for Research and Development in Science (FBiRN), along with a team led by Dr. Michała Wszoły, who developed the technology for producing bioink based on the decellularization of porcine pancreas and printed a prototype of a bionic pancreas.

Team led by Prof. at the Center for Advanced Technologies, Adam Mickiewicz University (CZT UAM). Scientists from the team specialize in bioprinting meniscus using bioink based on porcine meniscus dECM and mesenchymal stem cells.

The consortium was supported by the Warsaw Medical University (WUM). Prof. Artur Kamiński – Director of the Biostructure Center, along with his team, has extensive experience in analyzing the toxicity of materials in animal models.

The business partner of the consortium was Polbionica, responsible for conducting part of the research and commercializing the obtained results.

As part of the implemented project, work was also carried out, resulting in the development of two unique recombinant hybrid proteins that combine functional domains of silk and elastin (p-SE17) and resilin and elastin (p-RE15).

The p-SE17 protein combines elastin and silk, exhibiting high solubility in aqueous buffers and providing flexibility to the final product. It significantly improves the rheological and mechanical properties of biomaterials, especially elasticity. Combinations of RGD and ZIP domains provide structural stability, creating a favorable microenvironment for seeded cells. With the potential for remodeling polymeric biomaterials, it enables high-resolution 3D printing of structures with complex spatial architecture.

The p-RE15 protein combines resilin and elastin to obtain unique mechanical, physicochemical, and biological properties. It contains RGD sequences and matrix metalloproteinase cleavage sites, allowing seeded cells to appropriately adjust the space by remodeling 3D structures. It strengthens the rheological properties of biomaterials, making them ideal for advanced medical 3D printing technologies.

KEY FEATURES

These innovative recombinant hybrid proteins are produced in an economical prokaryotic expression system, without the use of detergents or surfactants, making them ideal additives to bioinks. Resilin and silk sequences enhance plasticity, while resilin and elastin increase the elasticity of the printed product. These proteins create a favorable microenvironment, promoting cell colonization, migration, proliferation, and growth stimulation. Importantly, these proteins are characterized by high biocompatibility and biodegradability.

The new recombinant hybrid proteins represent innovative solutions that demonstrate promising potential and a wide range of applications. They can be used in the creation of new unique bioinks, tissue engineering, regenerative therapies, and as components used in the production of medical devices.