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Bioprinting Towards Organ Fabrication: Challenges and Future Trends Ibrahim Ozbolat and Yin Yu

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Bioprinting Towards Organ Fabrication: Challenges and Future Trends Ibrahim Ozbolat and Yin Yu This article has been accepted for publication in a future issue of this journal, but has not been fully edited. Content may change prior to final publication. TBME-01840-2012 1 Bioprinting towards Organ Fabrication: Challenges and Future Trends Ibrahim Ozbolat and Yin Yu proliferation, and direct cell differentiation toward certain Abstract— Tissue engineering has been a promising field of lineages. Although significant success has been achieved in the research, offering hope for bridging the gap between organ past decades both in research and clinical applications [7], it is shortage and transplantation needs. However, building three- obvious that complex 3D organs require more precise multi- dimensional (3D) vascularized organs remains the main technological barrier to be overcome. Organ printing, which is cellular structures with vascular network integration, which defined as computer-aided additive biofabrication of 3D cellular cannot be fulfilled by traditional methods. tissue constructs, has shed light on advancing this field into a new era. Organ printing takes advantage of rapid prototyping (RP) technology to print cells, biomaterials, and cell-laden biomaterials individually or in tandem, layer by layer, directly creating 3D tissue-like structures. Here, we overview RP-based bioprinting approaches and discuss the current challenges and trends towards fabricating living organs for transplant in the near future. Index Terms— Bioadditive manufacturing, bioprinting, organ fabrication, tissue engineering I. INTRODUCTION RGAN shortage has become more problematic in spite of O an increase in willing donors. From July 2000 to July 2001, for example, approximately 80,000 people in the United States awaited an organ transplant, with less than a third receiving it [1]. The solution to this problem, as with the solutions to other grand engineering challenges, requires long- Fig. 1: Organ printing concept: 3D functional organs are printed from term solutions by building or manufacturing living organs from the bottom up using living cells in a supportive medium (image a person’s own cells. For the past three decades, tissue courtesy of Christopher Barnatt [8]). engineering has emerged as a multidisciplinary field involving A computer-aided bioadditive manufacturing process has scientists, engineers, and physicians, for the purpose of emerged to deposit living cells together with hydrogel-base creating biological substitutes mimicking native tissue to scaffolds for 3D tissue and organ fabrication. Bioprinting or replace damaged tissues or restore malfunctioning organs [2]. direct cell printing is an extension of tissue engineering, as it The traditional tissue engineering strategy is to seed cells onto intends to create de novo organs. It uses bioadditive scaffolds, which can then direct cell proliferation and manufacturing technologies, including laser-based writing [9], differentiation into three-dimensional (3D) functioning tissues. inkjet-based printing [10] and extrusion-based deposition [11]. Both synthetic and natural polymers have been used to Bioprinting offers great precision on spatial placement of the engineer various tissue grafts like skin, cartilage, bone, and cells themselves, rather than providing scaffold support alone bladder [3-6]. To be successfully used for tissue engineering, [12]. Although still in its infancy, this technology appears to be these materials must be biocompatible and biodegradable, with more promising for advancing tissue engineering towards the mechanical strength to support cell attachment, organ fabrication, ultimately mitigating organ shortage and saving lives. Figure 1 demonstrates the concept of futuristic Manuscript received December 12, 2012. This research was supported by 3D direct organ printing technology, where multiple living National Institutes of Health (NIH) and the Institute for Clinical and Translational Science (ICTS) grant number ULIRR024979. cells with the supportive media stored in cartridges are printed I.T. Ozbolat is with The University of Iowa Mechanical and Industrial layer by layer using inkjet printing technology. It offers a Engineering and Biomanufacturing Laboratory, Iowa City, IA, 52242 USA controllable fabrication process, which allows precise (corresponding author to provide phone: 319-384-0810; fax: 319-335-5669; e-mail: [email protected]). placement of various biomaterial and/or cell types Y. Yin is with The University of Iowa Biomedical Engineering simultaneously according to the natural compartments of the Department and Biomanufacturing Laboratory, Iowa City, IA 52242 USA (e- target tissue or organs. Multiple cell types, including organ- mail: yin-yu@ uiowa.edu). specific cells and blood vessel cells, i.e., smooth muscle and Copyright (c) 2013 IEEE. Personal use is permitted. For any other purposes, permission must be obtained from the IEEE by emailing [email protected]. This article has been accepted for publication in a future issue of this journal, but has not been fully edited. Content may change prior to final publication. TBME-01840-2012 2 endothelial cells (ECs), constitute the entire organ. Although a certain pattern onto a substrate by a laser beam. This is the concept seems to be trivial considering the complexity and followed by deposition of hydrogel on top of each layer of functionality of the parts that can be manufactured using cells, and the process is repeated for multiple cycles to get a contemporary rapid prototyping (RP) technology [13], several 3D structure. Nahmias et al. successfully performed challenges impede the evolution of organ printing. This paper hepatocytes patterning in collagen and MatrigelTM using laser- discusses the current state of the art in bioprinting technology, guided 3D cell writing [16]. In their study, three layers of cells its advantages and limitations, and grand challenges and future and hydrogels were alternately deposited on top of each other, trends towards fabrication of living organs in both research forming a 3D cellular structure. Cell viability and proliferation and clinical scenarios. was well-maintained post-deposition. Inkjet-based bioprinting was introduced in the early 2000s II. BIOPRINTING: CURRENT STATE OF THE ART and built a great foundation for future organ printing Bioprinting, where living cells are precisely printed in a technologies. In this technique, living cells are printed in the certain pattern, has great potential and promise for fabricating form of droplets through cartridges instead of seeding them on engineered living organs. Based on their working principles, scaffolds (see Fig. 2(B)). It uses a non-contact reprographic bioprinting systems can be primarily classified as: (1) laser- technique that takes digital data from a computer representing based, (2) inkjet-based, or (3) extrusion-based. tissue or organs, and reproduces it onto a substrate using “bio- ink” made of cells and biomaterials [10]. Boland et al. used a thermal inkjet printer to successfully fabricate 3D cellular assemblies of bovine aortal ECs with thermosensitive gels [10]. Post-incubation, printed structures showed high cell viability and maintained cell phenotype. Cui and his coworkers [17] applied inkjet printing technology to repair human articular cartilage, showing its promising potential for high- efficiency direct tissue regeneration. Huang and his coworkers [18] developed a bipolar wave-based drop-on-demand jetting. In their studies, cell-encapsulated alginate microspheres were jetted and assembled to create vertically oriented, short, tubular structures [19]. Inkjet-based system allows printing single cells or cell aggregates [20, 21] by controlling process parameters such as cell concentration, drop volume, resolution, nozzle diameter and average diameter of printed cells [22]. Weiss and his coworkers [23] developed a multi-head inkjet- based bioprinting platform for fabricating heterogeneous structures with a concentration gradient changing from the Fig. 2: Bioprinting techniques including (A) laser-based writing of bottom up. Multiple growth factors such as fibrinogen and cells, (B) ink-jet based systems and (C) extrusion-based deposition. thrombin and cells were printed with spatial precision in a functionally graded manner into rat calvarial defect in-situ Laser technology has recently been applied in the cell [24]. They demonstrated the feasibility of in-situ printing; printing process, in which laser energy is used to excite the however, this technique did not seem to be a practical cells and give patterns to control spatially the cellular approach for clinicians due to complex nature of the process in environment. A laser-based system was first introduced in their study [24]. 1999 by Odde et al. to process 2D cell patterning [14]. Laser Another bioprinting technique has been introduced for direct-write (LDW) is a biofabrication method capable of printing living cells and is based on the extrusion of rapidly creating precise patterns of viable cells on petri dishes. continuous filaments made of biomaterials. It is a combination In LDW, cells suspended in a solution in donor slides are of a fluid-dispensing system and an automated three-axis transferred to a collector slide using laser energy. A laser pulse robotic system for extrusion and printing, respectively [11]. creates a bubble, and shock waves are generated by the bubble During printing, biomaterial is dispensed by a pressure- formation, which eventually propel cells towards the collector assisted system, under
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