3D bioprinting

Three dimensional (3D) bioprinting is the utilization of 3D printing–like techniques to combine cells, growth factors, and biomaterials to fabricate biomedical parts that maximally imitate natural tissue characteristics.[1] Generally, 3D bioprinting utilizes the layer-by-layer method to deposit materials known as bioinks to create tissue-like structures that are later used in medical and tissue engineering fields. Bioprinting covers a broad range of biomaterials.

Currently, bioprinting can be used to print tissues and organs to help research drugs and pills.[2] However, emerging innovations span from bioprinting of cells or extracellular matrix deposited into a 3D gel layer by layer to produce the desired tissue or organ. In addition, 3D bioprinting has begun to incorporate the printing of scaffolds.[3] These scaffolds can be used to regenerate joints and ligaments.[4]

Process

3D bioprinting generally follows three steps, pre-bioprinting, bioprinting, and post-bioprinting.[5][6]

Pre-bioprinting

Pre-bioprinting is the process of creating a model that the printer will later create and choosing the materials that will be used. One of the first steps is to obtain a biopsy of the organ. Common technologies used for bioprinting are computed tomography (CT) and magnetic resonance imaging (MRI). To print with a layer-by-layer approach, tomographic reconstruction is done on the images. The now-2D images are then sent to the printer to be made. Once the image is created, certain cells are isolated and multiplied.[5] These cells are then mixed with a special liquefied material that provides oxygen and other nutrients to keep them alive. In some processes, the cells are encapsulated in cellular spheroids 500μm in diameter. This aggregation of cells does not require a scaffold, and are required for placing in the tubular-like tissue fusion for processes such as extrusion.[7]:165

Bioprinting

In the second step, the liquid mixture of cells, matrix, and nutrients known as bioinks are placed in a printer cartridge and deposited using the patients' medical scans.[8] When a bioprinted pre-tissue is transferred to an incubator, this cell-based pre-tissue matures into a tissue.[3]

3D bioprinting for fabricating biological constructs typically involves dispensing cells onto a biocompatible scaffold using a successive layer-by-layer approach to generate tissue-like three-dimensional structures.[9] Artificial organs such as livers and kidneys made by 3D bioprinting have been shown to lack crucial elements that affect the body such as working blood vessels, tubules for collecting urine, and the growth of billions of cells required for these organs. Without these components the body has no way to get the essential nutrients and oxygen deep within their interiors.[9] Given that every tissue in the body is naturally composed of different cell types, many technologies for printing these cells vary in their ability to ensure stability and viability of the cells during the manufacturing process. Some of the methods that are used for 3D bioprinting of cells are photolithography, magnetic bioprinting, stereolithography, and direct cell extrusion.[7]:196

Post-bioprinting

The post-bioprinting process is necessary to create a stable structure from the biological material. If this process is not well-maintained, the mechanical integrity and function of the 3D printed object is at risk.[5] To maintain the object, both mechanical and chemical stimulations are needed. These stimulations send signals to the cells to control the remodeling and growth of tissues. In addition, in recent development, bioreactor technologies[1] have allowed the rapid maturation of tissues, vascularization of tissues and the ability to survive transplants.[6]

Bioreactors work in either providing convective nutrient transport, creating microgravity environments, changing the pressure causing solution to flow through the cells, or add compression for dynamic or static loading. Each type of bioreactor is ideal for different types of tissue, for example compression bioreactors are ideal for cartilage tissue.[7]:198

Bioprinting approach

Researchers in the field have developed approaches to produce living organs that are constructed with the appropriate biological and mechanical properties. 3D bioprinting is based on three main approaches: Biomimicry, autonomous self-assembly and mini-tissue building blocks.[10]

Biomimicry

The first approach of bioprinting is called biomimicry. The main goal of this approach is to create fabricated structures that are identical to the natural structure that are found in the tissues and organs in the human body. Biomimicry requires duplication of the shape, framework, and the microenvironment of the organs and tissues.[11] The application of biomimicry in bioprinting involves creating both identical cellular and extracellular parts of organs. For this approach to be successful, the tissues must be replicated on a micro scale. Therefore, it is necessary to understand the microenvironment, the nature of the biological forces in this microenvironment, the precise organization of functional and supporting cell types, solubility factors, and the composition of extracellular matrix.[10]

Autonomous self-assembly

The second approach of bioprinting is autonomous self-assembly. This approach relies on the physical process of embryonic organ development as a model to replicate the tissues of interest.[11] When cells are in their early development, they create their own extracellular matrix building block, the proper cell signaling, and independent arrangement and patterning to provide the required biological functions and micro-architecture.[10] Autonomous self-assembly demands specific information about the developmental techniques of the tissues and organs of the embryo.[11] There is a "scaffold-free" model that uses self-assembling spheroids that subjects to fusion and cell arrangement to resemble evolving tissues. Autonomous self-assembly depends on the cell as the fundamental driver of histogenesis, guiding the building blocks, structural and functional properties of these tissues. It demands a deeper understanding of how embryonic tissues mechanisms develop as well as the microenvironment surrounded to create the bioprinted tissues.[10]

Mini-tissue

The third approach of bioprinting is a combination of both the biomimicry and self-assembly approaches, which is called mini tissues.[12] Organs and tissues are built from very small functional components. Mini-tissue approach takes these small pieces and manufacture and arrange them into larger framework.[11][10]

Printers
A 3D bioprinter

Akin to ordinary ink printers, bioprinters have three major components to them. These are the hardware used, the type of bio-ink, and the material it is printed on (biomaterials).[5] "Bio-ink is a material made from living cells that behaves much like a liquid, allowing people to 'print' it in order to create the desired shape. To make bio-ink, scientists create a slurry of cells that can be loaded into a cartridge and inserted into a specially designed printer, along with another cartridge containing a gel known as bio-paper."[13] test In bioprinting, there are three major types of printers that have been used. These are inkjet, laser-assisted, and extrusion printers. Inkjet printers are mainly used in bioprinting for fast and large-scale products. One type of inkjet printer, called drop-on-demand inkjet printer, prints materials in exact amounts, minimizing cost and waste.[14] Printers that utilize lasers provide high-resolution printing; however, these printers are often expensive. Extrusion printers print cells layer-by-layer, just like 3D printing to create 3D constructs. In addition to just cells, extrusion printers may also use hydrogels infused with cells.[5]

Applications

There are several applications for 3D bioprinting in the medical field. An infant patient with a rare respiratory disease known as Tracheobronchomalacia (TBM) was given a tracheal splint that was created with 3D printing.[15] 3D bioprinting can be used to reconstruct tissue from various regions of the body. Patients with end-stage bladder disease can be treated by using engineered bladder tissues to rebuild the damaged organ.[16] This technology can also potentially be applied to bone, skin, cartilage and muscle tissue.[17] Though one long-term goal of 3D bioprinting technology is to reconstruct an entire organ, there has been little success in printing fully functional organs.[18] Unlike implantable stents, organs have complex shapes and are significantly harder to bioprint. A bioprinted heart, for example, must not only meet structural requirements, but also vascularization, mechanical load, and electrical signal propagation requirements.[19] Israeli researchers constructed a rabbit-sized heart out of human cells in 2019.[20]

Impact

3D bioprinting contributes to significant advances in the medical field of tissue engineering by allowing for research to be done on innovative materials called biomaterials. Biomaterials are the materials adapted and used for printing three-dimensional objects. Some of the most notable bioengineered substances are usually stronger than the average bodily materials, including soft tissue and bone. These constituents can act as future substitutes, even improvements, for the original body materials. Alginate, for example, is an anionic polymer with many biomedical implications including feasibility, strong biocompatibility, low toxicity, and stronger structural ability in comparison to some of the body's structural material.[21] Synthetic hydrogels are also commonplace, including PV-based gels. The combination of acid with a UV-initiated PV-based cross-linker has been evaluated by the Wake Forest Institute of Medicine and determined to be a suitable biomaterial.[22] Engineers are also exploring other options such as printing micro-channels that can maximize the diffusion of nutrients and oxygen from neighboring tissues[8] In addition, the Defense Threat Reduction Agency aims to print mini organs such as hearts, livers, and lungs as the potential to test new drugs more accurately and perhaps eliminate the need for testing in animals.[8]

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See also

References
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  2. Hinton TJ, Jallerat Q, Palchesko RN, Park JH, Grodzicki MS, Shue HJ, Ramadan MH, Hudson AR, Feinberg AW (23 October 2015). "Three-dimensional printing of complex biological structures by freeform reversible embedding of suspended hydrogels". Science Advances. 1 (9): e1500758. Bibcode:2015SciA....1E0758H. doi:10.1126/sciadv.1500758. PMC 4646826. PMID 26601312.
  3. Thomas, Daniel J. (August 2016). "Could 3D bioprinted tissues offer future hope for microtia treatment?". International Journal of Surgery. 32: 43–44. doi:10.1016/j.ijsu.2016.06.036. PMID 27353851.
  4. Nakashima, Yasuharu; Okazak, Ken; Nakayama, Koichiet; Okada, Seiji; Mizu-uchi, Hideki (January 2017). "Bone and Joint Diseases in Present and Future". Fukuoka Igaku Zasshi = Hukuoka Acta Medica. 108 (1): 1–7. PMID 29226660.
  5. Shafiee, Ashkan; Atala, Anthony (2016-03-01). "Printing Technologies for Medical Applications". Trends in Molecular Medicine. 22 (3): 254–265. doi:10.1016/j.molmed.2016.01.003. PMID 26856235.
  6. Ozbolat, Ibrahim T. (2015-07-01). "Bioprinting scale-up tissue and organ constructs for transplantation". Trends in Biotechnology. 33 (7): 395–400. doi:10.1016/j.tibtech.2015.04.005. PMID 25978871.
  7. Chua, C.K.; Yeong, W.Y. (2015). Bioprinting: Principles and Applications. Singapore: World Scientific Publishing Co. ISBN 9789814612104.
  8. Cooper-White, M. (1 March 2015). "How 3D Printing Could End The Deadly Shortage Of Donor Organs". Huffpost Science. TheHuffingtonPost.com, Inc. Retrieved 17 February 2016.
  9. Harmon, K. (2013). "A sweet solution for replacing organs" (PDF). Scientific American. 308 (4): 54–55. Bibcode:2013SciAm.308d..54H. doi:10.1038/scientificamerican0413-54. Archived from the original (PDF) on 2016-02-17. Retrieved 17 February 2016.
  10. Murphy, Sean; Atala, Anthony (August 5, 2014). "3D bioprinting of tissues and organs". Nature Biotechnology. 32 (8): 773–85. doi:10.1038/nbt.2958. PMID 25093879.
  11. Yoo, James; Atala, Anthony (2015). "Bio-printing: 3D printing comes to life". ProQuest 1678889578. Cite journal requires |journal= (help)
  12. Thomas, Daniel; Singh, Deepti (2018-06-12). "Novel techniques of engineering 3D vasculature tissue for surgical procedures". The American Journal of Surgery. 218 (1): 235–236. doi:10.1016/j.amjsurg.2018.06.004. PMID 29929908.
  13. Manappallil, John J (2015). Basic Dental Materials. JP Medical Ltd. ISBN 9789352500482.
  14. "3D Printing Technology At The Service Of Health". healthyeve. Retrieved 4 August 2016.
  15. Zopf, D. A.; Hollister, S. J.; Nelson, M. E.; Ohye, R. G.; Green, G. E. (2013). "Bioresorbable Airway Splint Created with a Three-Dimensional Printer". The New England Journal of Medicine. 368 (21): 2043–2045. doi:10.1056/NEJMc1206319. PMID 23697530.
  16. Atala, A.; Bauer, S. B.; Soker, S.; Yoo, J. J.; Retik, A. B. (2006). "Tissue-engineered autologous bladders for patients needing cystoplasty". Lancet. 367 (9518): 1241–1246. doi:10.1016/S0140-6736(06)68438-9. PMID 16631879.
  17. Hong, N.; Yang, G. H.; Lee, J.; Kim, G. (2017). "3D bioprinting and its in vivo applications". Journal of Biomedical Materials Research Part B: Applied Biomaterials. 106 (1): 444–459. doi:10.1002/jbm.b.33826. PMID 28106947.
  18. Sommer, A. C.; Blumenthal, E. Z. (2019). "Implementations of 3D printing in ophthalmology". Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht von Graefes Archiv Fur Klinische und Experimentelle Ophthalmologie. 257 (9): 1815–1822. doi:10.1007/s00417-019-04312-3. PMID 30993457.
  19. Haitao Cui, Shida Miao, Timothy Esworthy, Xuan Zhou, Se-jun Lee, Chengyu Liu, Zu-xi Yu, John P. Fisher, Muhammad Mohiuddin, Lijie Grace Zhang, "3D bioprinting for cardiovascular regeneration and pharmacology", Advanced Drug Delivery Reviews
  20. "Israeli scientists create world's first 3D-printed heart using human cells". NBC News. Retrieved 2019-04-20.
  21. Crawford, M. (May 2013). "Creating Valve Tissue Using 3-D Bioprinting". ASME.org. American Society of Mechanical Engineers. Retrieved 17 February 2016.
  22. Murphy, S.V.; Skardal, A.; Atala, A. (2013). "Evaluation of hydrogels for bio-printing applications". Journal of Biomedical Materials Research Part A. 101A (1): 272–84. doi:10.1002/jbm.a.34326. PMID 22941807.

Further reading
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