Cellulose nanofibrillar bioink for 3D bioprinting for cell culturing, tissue engineering and regenerative medicine applications

Abstract

The present invention relates to biomaterial in the form of dispersion of cellulose nanofibrils with extraordinary shear thinning properties which can be converted into desire 3D shape using 3D Bioprinting technology. In this invention cellulose nanofibril dispersion, is processed through different mechanical, enzymatic and chemical steps to yield dispersion with desired morphological and rheological properties to be used as bioink in 3D Bioprinter. The processes are followed by purification, adjusting of osmolarity of the material and sterilization to yield biomaterial which has cytocompatibility and can be combined with living cells. Cellulose nanofibrils can be produced by microbial process but can also be isolated from plant secondary or primary cell wall, animals such as tunicates, algae and fungi. The present invention describes applications of this novel cellulose nanofibrillar bioink for 3D Bioprinting of tissue and organs with desired architecture.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]The present application is a Continuation application of U.S. application Ser. No. 15 / 537,154, filed Jun. 16, 2017. The '154 application is a National Stage application under 35 U.S.C. § 371 of International Application No. PCT / US15 / 66755 filed Dec. 18, 2015, which application relies on the disclosure of and claims priority to and the benefit of the filing date of U.S. Provisional Application No. 62 / 093,881, filed Dec. 18, 2014, the disclosures of each of which are hereby incorporated by reference herein in their entireties.BACKGROUND OF THE INVENTIONField of the Invention[0002]The present invention relates to novel bioink which is biomaterial in the form of water dispersion of cellulose nanofibrils and can be converted into desired 3D shapes using 3D Bioprinting technology.[0003]This novel bioink is suitable for 3D cell culturing and growing living tissues and organs. In this invention, cellulose nanofibrillar material is processed throu...

AI Technical Summary

Benefits of technology

The patent text describes a method to successfully print different types of mammalian cells in complex 3D shapes of human organs using a special ink called CELLINK™. These cells show good viability after printing and crosslinking, and long-term evaluation (up to 28 days) showed human cartilage regeneration in the 3D printed tissue. Additionally, CELLINK™ has been shown to be a great support material when printing complex tissues with collagen or decellularized matrix based bioinks. The biomimetic and biocompatibility characteristics of these novel nanocellulose fibrils based bioinks make them ideal candidates for applications in cell culture, tissue engineering, and regenerative medicine.

Problems solved by technology

The driving force has been a shortage of organs which are needed for transplantation.

These polymers have often fast degradation characteristics and / or produce an environment which causes inflammation.

While these materials can be fabricated into films, meshes, or more complex 3D structures, their successful use is limited by their physical and biochemical properties.

Fabrication of 3D structures with controlled architecture and interconnected porosity has been challenging.

The methods used, such as freeze drying, porogen removal or electrospinning, show poor reproducibility and lack of control of 3D architecture in micro scale.

As the consequence of that, there have been difficulties in cell seeding since cell migration requires good pore interconnectivity.

The disadvantage of 3D printing using thermoplastic materials is a difficulty in cell seeding due to limited cell migration into porous structures.

This technique can unfortunately not be combined with mammalian cells due to the differences in cultivation conditions at 37 degrees, which is required for mammalian cells, since bacterial cells are killed.

It is expected that a biosynthetic cellulose network cannot as such be used as a scaffold for tissue engineering because the relatively tight network of cellulose nanofibrils which make cell migration difficult to impossible.

None of the methods enable reproducible and scalable control of the architecture of the scaffolds or a convenient method to combine with the cells.

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