Table of Contents

Special Section: Bioprinting in Russia


by Peter Timashev, Vladimir Mironov

Bioprinting is a rapidly emerging biomedical research field. Three-dimensional bioprinting is defined as a robotic additive, layer-by-layer biofabrication of functional tissues and organs from living cells, and biomaterials according to a digital model. Bioprinting can revolutionize medicine by automated robotic production of human tissues and organs suitable for transplantation. Bioprinting is based on sophisticated high technology, and it is obvious that only technologically advanced countries can make a real contribution to this rapidly evolving multidisciplinary field. In this paper, we present main Russia’s achievements in bioprinting. Here, we also discuss challenges and perspectives of bioprinting research and development in Russia. Russian researchers already made some impressive contributions with long-lasting impact and they have capacities, potential, and ambitions to continue contribute to the advancements of bioprinting.

Perspective article

by Anastasia Kirillova, Stanislav Bushev, Aydar Abubakirov, Gennady Sukikh

Bioethical and legal issues of three-dimensional (3D) bioprinting as the emerging field of biotechnology have not yet been widely discussed among bioethicists around the world, including Russia. The scope of 3D bioprinting includes not only the issues of the advanced technologies of human tissues and organs printing but also raises a whole layer of interdisciplinary problems of modern science, technology, bioethics, and philosophy. This article addresses the ethical and legal issues of bioprinting of artificial human organs.

Review article

by Egor Olegovich Osidak, Vadim Igorevich Kozhukhov, Mariya Sergeevna Osidak, Sergey Petrovich Domogatskiy

Biomaterials made using collagen are successfully used as a three-dimensional (3D) substrate for cell culture and considered to be promising scaffolds for creating artificial tissues. An important task that arises for engineering such materials is the simulation of physical and morphological properties of tissues, which must be restored or replaced. Modern additive technologies, including 3D bioprinting, can be applied to successfully solve this task. This review provides the latest evidence on advances of 3D bioprinting with collagen in the field of tissue engineering. It contains modern approaches for printing pure collagen bioinks consisting only of collagen and cells, as well as the obtained results from the use of pure collagen bioinks in different fields of tissue engineering.

Review article

by Anastasia Shpichka, Daria Osipova, Yuri Efremov, Polina Bikmulina, Nastasia Kosheleva, Marina Lipina, Evgeny A. Bezrukov, Roman B. Sukhanov, Anna B. Solovieva, Massoud Vosough, Peter Timashev

For the past 10 years, the main efforts of most bioprinting research teams have focused on creating new bioink formulations, rather than inventing new printing set-up concepts. New tissue-specific bioinks with good printability, shape fidelity, and biocompatibility are based on “old” (well-known) biomaterials, particularly fibrin. While the interest in fibrin-based bioinks is constantly growing, it is essential to provide a framework of material’s properties and trends. This review aims to describe the fibrin properties and application in three-dimensional bioprinting and provide a view on further development of fibrin-based bioinks.

Review article

by Catherine Pakhomova, Dmitry Popov, Eugenii Maltsev, Iskander Akhatov, Alexander Pasko

The bioprinting of heterogeneous organs is a crucial issue. To reach the complexity of such organs, there is a need for highly specialized software that will meet all requirements such as accuracy, complexity, and others. The primary objective of this review is to consider various software tools that are used in bioprinting and to reveal their capabilities. The sub-objective was to consider different approaches for the model creation using these software tools. Related articles on this topic were analyzed. Software tools are classified based on control tools, general computer-aided design (CAD) tools, tools to convert medical data to CAD formats, and a few highly specialized research-project tools. Different geometry representations are considered, and their advantages and disadvantages are considered applicable to heterogeneous volume modeling and bioprinting. The primary factor for the analysis is suitability of the software for heterogeneous volume modeling and bioprinting or multimaterial three-dimensional printing due to the commonality of these technologies. A shortage of specialized suitable software tools is revealed. There is a need to develop a new application area such as computer science for bioprinting which can contribute significantly in future research work.

Review article

by Anton Elemoso, Grigoriy Shalunov, Yakov M. Balakhovsky, Alexander Yu. Ostrovskiy, Yusef D. Khesuani

Three-dimensional (3D) bioprinting as a technology is being researched and applied since 2003. It is actually several technologies (inkjet, extrusion, laser, magnetic bioprinting, etc.) under an umbrella term “3D bioprinting.” The versatility of this technology allows widespread applications in several; however, after almost 20 years of research, there is still a limited number of cases of commercialized applications. This article discusses the potential for 3D bioprinting in regenerative medicine, drug discovery, and food industry, as well as the existing cases of companies that create commercialized products and services in the aforementioned areas and even in fashion, including their go-to-market route and financing received. We also address the main barriers to creating practical applications of 3D bioprinting within each sphere the technology that is being studied for.

Original research article

by Vladimir Yusupov, Semyon Churbanov, Ekaterina Churbanova, Ksenia Bardakova, Artem Antoshin, Stanislav Evlashin, Peter Timashev, Nikita Minaev
Laser-induced forward transfer is a versatile, non-contact, and nozzle-free printing technique which has demonstrated high potential for different printing applications with high resolution. In this article, three most widely used hydrogels in bioprinting (2% hyaluronic acid sodium salt, 1% methylcellulose, and 1% sodium alginate) were used to study laser printing processes. For this purpose, the authors applied a laser system based on a pulsed infrared laser (1064 nm wavelength, 8 ns pulse duration, 1 – 5 J/cm2 laser fluence, and 30 μm laser spot size). A high-speed shooting showed that the increase in fluence caused a sequential change in the transfer regimes: No transfer regime, optimal jetting regime with a single droplet transfer, high speed regime, turbulent regime, and plume regime. It was demonstrated that in the optimal jetting regime, which led to printing with single droplets, the size and volume of droplets transferred to the acceptor slide increased almost linearly with the increase of laser fluence. It was also shown that the maintenance of a stable temperature (±2°C) allowed for neglecting the temperature-induced viscosity change of hydrogels. It was determined that under room conditions (20°C, humidity 50%), the hydrogel layer, due to drying processes, decreased with a speed of about 8 μm/min, which could lead to a temporal variation of the transfer process parameters. The authors developed a practical algorithm that allowed quick configuration of the laser printing process on an applied experimental setup. The configuration is provided by the change of the easily tunable parameters: Laser pulse energy, laser spot size, the distance between the donor ribbon and acceptor plate, as well as the thickness of the hydrogel layer on the donor ribbon slide.

Original research article

by Ilya I Bozo, Roman V. Deev, Igor V. Smirnov, Alexander Yu Fedotov, Vladimir K. Popov, Anton V. Mironov, Olga A. Mironova, Alexander Yu. Gerasimenko, Vladimir S Komlev

The aim of the study was the development of three-dimensional (3D) printed gene-activated implants based on octacalcium phosphate (OCP) and plasmid DNA encoding VEGFA. The first objective of the present work involved design and fabrication of gene-activated bone substitutes based on the OCP and plasmid DNA with VEGFА gene using 3D printing approach of ceramic constructs, providing the control of its architectonics compliance to the initial digital models. X-ray diffraction, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy, and compressive strength analyses were applied to investigate the chemical composition, microstructure, and mechanical properties of the experimental samples. The biodegradation rate and the efficacy of plasmid DNA delivery in vivo were assessed during standard tests with subcutaneous implantation to rodents in the next stage. The final part of the study involved substitution of segmental tibia and mandibular defects in adult pigs with 3D printed gene-activated implants. Biodegradation, osteointegration, and effectiveness of a reparative osteogenesis were evaluated with computerized tomography, SEM, and a histological examination. The combination of gene therapy and 3D printed implants manifested the significant clinical potential for effective bone regeneration in large/critical size defect cases.

Original research article

by Vladislav A. Parfenov, Stanislav V. Petrov, Frederico D. A. S. Pereira, Aleksandr A. Levin, Elizaveta V. Koudan, Pavel A. Karalkin, Mikhail M. Vasiliev, Oleg F. Petrov, Vladimir S Komlev, Yusef D. Khesuani, Vladimir Mironov

Scaffolding is the conceptual framework of conventional tissue engineering. Over the past decade, scaffold-free approaches as a potential alternative to classic scaffold-based methods have emerged, and scaffold-free magnetic levitational tissue engineering (magnetic force-based tissue engineering [Mag-TE]) is a type of this novel tissue engineering strategy. However, Mag-TE is often based on the use of potentially toxic magnetic nanoparticles. Scaffold-free and label-free magnetic levitational bioassembly do not employ magnetic nanoparticles and thus, the potential toxicity of magnetic nanoparticles can be avoided. In this short review, we describe the conceptual foundation of scaffold-free, label-free, and nozzle-free formative biofabrication using magnetic fields as “scaffields.” The design and implementation of “Organ.Aut,” the first commercial magnetic levitational bioassembler, and the potential applications of magnetic bioassembler are discussed as well.