How to Use Protein-based Bioinks


Bioprinting, a cutting-edge technology in the field of regenerative medicine, holds tremendous promise for creating functional tissues and organs. Central to the success of bioprinting is the bioink, a substance that serves as the building material for 3D printing biological structures. In recent years, protein-based bioinks have emerged as a versatile and promising option for creating biocompatible and bioactive constructs. In this article, we will delve into the fundamentals and practical aspects of using protein-based bioinks in bioprinting.

Protein-Based Bioinks

Protein-based bioinks utilize proteins as the primary building blocks for the ink formulation. Proteins such as collagen, gelatin, fibrin, and silk have gained popularity due to their biocompatibility and ability to mimic the extracellular matrix (ECM) of natural tissues. The ECM is crucial for providing structural support and biochemical signals necessary for cell proliferation and differentiation.

Conventional cell types selected for cell embedding in protein-based composites according and biomedical application. Figure 1. Conventional cell types selected for cell embedding in protein-based composites according and biomedical application. (Veiga A, et al.; 2021)

Key Advantages of Protein-Based Bioinks

  • Biocompatibility

Protein-based bioinks closely resemble the natural environment of living tissues, promoting cell adhesion, proliferation, and differentiation. This biocompatibility is essential for the successful integration of printed structures into the host tissue.

  • Bioactivity

Proteins carry bioactive signals that play a crucial role in cellular behavior. By using protein-based bioinks, researchers can incorporate these signals into the printed structures, influencing cell behavior and tissue development. This is particularly important for applications in tissue engineering and regenerative medicine.

  • Printability

Protein-based bioinks exhibit favorable rheological properties, allowing them to be easily manipulated during the printing process. This printability ensures precise deposition of layers, contributing to the accuracy and reproducibility of the printed structures.

Practical Steps for Using Protein-Based Bioinks

  • Bioink Formulation

Begin by preparing the protein-based bioink formulation. This typically involves mixing the protein of choice with other components such as crosslinking agents, cells, and growth factors. The exact formulation will depend on the specific requirements of the intended application.

  • Rheological Characterization

Assess the rheological properties of the bioink to ensure optimal printability. This involves evaluating viscosity, shear-thinning behavior, and gelation kinetics. Adjustments to the formulation may be necessary to achieve the desired rheological properties.

  • Cell Integration

If the goal is to print living tissues, incorporate cells into the bioink. Cells should be evenly distributed to ensure uniform tissue formation. Optimizing cell viability and maintaining their functionality during the printing process are critical considerations.

  • Printing Process

Load the bioink into the bioprinter and set the printing parameters. The choice of printing technique, such as extrusion-based or inkjet printing, will depend on the bioink properties and the desired structure. Ensure proper calibration to achieve accurate layer deposition.

  • Crosslinking

Many protein-based bioinks require crosslinking to stabilize the printed structure. Common crosslinking methods include chemical crosslinking using agents like genipin or physical crosslinking through temperature or pH adjustments. Carefully select the crosslinking method that aligns with the bioink composition and application.

  • Post-Printing Processing

After printing, allow the structure to undergo post-printing processes such as maturation and culture. This step is crucial for tissue development and integration. Provide an appropriate culture environment to support cell viability and functionality.


Protein-based bioinks offer a promising avenue for advancing the capabilities of bioprinting in the realm of regenerative medicine. By harnessing the biocompatibility and bioactivity of proteins, researchers and clinicians can pave the way for the creation of functional tissues and organs. As the field continues to evolve, the practical implementation of protein-based bioinks will undoubtedly play a pivotal role in bringing bioprinting closer to realizing its transformative potential in healthcare and tissue engineering.

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  1. Mirzaei M, et al.; Protein-Based 3D Biofabrication of Biomaterials. Bioengineering (Basel). 2021, 8(4):48.
  2. Veiga A, et al.; Current Trends on Protein Driven Bioinks for 3D Printing. Pharmaceutics. 2021, 13(9):1444.
For research use only, not intended for any clinical use.