Construction of 3D in vitro models by bioprinting human pluripotent stem cells: Challenges and opportunities

Three-dimensional (3D) printing of biological material, or 3D bioprinting, is a rapidly expanding field with interesting applications in tissue engineering and regenerative medicine. Bioprinters use cells and biocompatible materials as an ink (bioink) to build 3D structures representative of organs and tissues, in a controlled manner and with micrometric resolution. Human embryonic (hESCs) and induced (hiPSCs) pluripotent stem cells are ideally able to provide all cell types found in the human body. A limited, but growing, number of recent reports suggest that cells derived by differentiation of hESCs and hiPSCs can be used as building blocks in bioprinted human 3D models, reproducing the cellular variety and cytoarchitecture of real tissues. In this review we will illustrate these examples, which include hepatic, cardiac, vascular, corneal and cartilage tissues, and discuss challenges and opportunities of bioprinting more demanding cell types, such as neurons, obtained from human pluripotent stem cells

Biofabrication and Bone Tissue Regeneration: Cell Source, Approaches, and Challenges

The growing occurrence of bone disorders and the increase in aging population have resulted in the need for more effective therapies to meet this request. Bone tissue engineering strategies, by combining biomaterials, cells, and signaling factors, are seen as alternatives to conventional bone grafts for repairing or rebuilding bone defects. Indeed, skeletal tissue engineering has not yet achieved full translation into clinical practice because of several challenges. Bone biofabrication by additive manufacturing techniques may represent a possible solution, with its intrinsic capability for accuracy, reproducibility, and customization of scaffolds as well as cell and signaling molecule delivery. This review examines the existing research in bone biofabrication and the appropriate cells and factors selection for successful bone regeneration as well as limitations affecting these approaches. Challenges that need to be tackled with the highest priority are the obtainment of appropriate vascularized scaffolds with an accurate spatiotemporal biochemical and mechanical stimuli release, in order to improve osseointegration as well as osteogenesis

3D bioprinting for tissue engineering: Stem cells in hydrogels

Surgical limitations require alternative methods of repairing and replacing diseased and damaged tissue. Regenerative medicine is a growing area of research with engineered tissues already being used successfully in patients. However, the demand for such tissues greatly outweighs the supply and a fast and accurate method of production is still required. 3D bioprinting offers precision control as well as the ability to incorporate biological cues and cells directly into the material as it is being fabricated. Having precise control over scaffold morphology and chemistry is a significant step towards controlling cellular behaviour, particularly where undifferentiated cells, i.e., stem cells, are used. This level of control in the early stages of tissue development is crucial in building more complex systems that morphologically and functionally mimic in vivo tissue. Here we review 3D printing hydrogel materials for tissue engineering purposes and the incorporation of cells within them. Hydrogels are ideal materials for cell culture. They are structurally similar to native extracellular matrix, have a high nutrient retention capacity, allow cells to migrate and can be formed under mild conditions. The techniques used to produce these materials, as well as their benefits and limitations, are outlined

3D Cell Printed Tissue Analogues: A New Platform for Theranostics

Stem cell theranostics has received much attention for noninvasively monitoring and tracing transplanted therapeutic stem cells through imaging agents and imaging modalities. Despite the excellent regenerative capability of stem cells, their efficacy has been limited due to low cellular retention, low survival rate, and low engraftment after implantation. Three-dimensional (3D) cell printing provides stem cells with the similar architecture and microenvironment of the native tissue and facilitates the generation of a 3D tissue-like construct that exhibits remarkable regenerative capacity and functionality as well as enhanced cell viability. Thus, 3D cell printing can overcome the current concerns of stem cell therapy by delivering the 3D construct to the damaged site. Despite the advantages of 3D cell printing, the in vivo and in vitro tracking and monitoring of the performance of 3D cell printed tissue in a noninvasive and real-time manner have not been thoroughly studied. In this review, we explore the recent progress in 3D cell technology and its applications. Finally, we investigate their potential limitations and suggest future perspectives on 3D cell printing and stem cell theranostics.116Nsciescopu

Recent Applications of Three Dimensional Printing in Cardiovascular Medicine

Three dimensional (3D) printing, which consists in the conversion of digital images into a 3D physical model, is a promising and versatile field that, over the last decade, has experienced a rapid development in medicine. Cardiovascular medicine, in particular, is one of the fastest growing area for medical 3D printing. In this review, we firstly describe the major steps and the most common technologies used in the 3D printing process, then we present current applications of 3D printing with relevance to the cardiovascular field. The technology is more frequently used for the creation of anatomical 3D models useful for teaching, training, and procedural planning of complex surgical cases, as well as for facilitating communication with patients and their families. However, the most attractive and novel application of 3D printing in the last years is bioprinting, which holds the great potential to solve the ever-increasing crisis of organ shortage. In this review, we then present some of the 3D bioprinting strategies used for fabricating fully functional cardiovascular tissues, including myocardium, heart tissue patches, and heart valves. The implications of 3D bioprinting in drug discovery, development, and delivery systems are also briefly discussed, in terms of in vitro cardiovascular drug toxicity. Finally, we describe some applications of 3D printing in the development and testing of cardiovascular medical devices, and the current regulatory frameworks that apply to manufacturing and commercialization of 3D printed products

Bioprinting with live cells

Tissue engineering is an emerging multidisciplinary field to regenerate damaged or diseased tissues and organs. Traditional tissue engineering strategies involve seeding cells into porous scaffolds to regenerate tissues or organs. However, there are still some challenges such as difficulty in seeding different type of cells spatially into a scaffold, limited oxygen and nutrient delivery and removal of metabolic waste from scaffold and weak cell-adhesion to scaffold material need to be overcome for clinically successful results. Because of those challenges, novel scaffold-free approaches based on cellular self-assembly or three-dimensional (3D) bioprinting have been recently pursued. Bioprinting is a relatively new technology where living cells with or without biomaterials are printed layer-by-layer in order to create 3D living structures. In 3D bioprinting, cell aggregates and hydrogels are termed as bioink used as building blocks that are placed by the bioprinter into precise architecture according to developed computer models. In this chapter, we focus on the scaffold-free, self-assembly based bioprinting approaches and some of the novel developments in this field. This chapter will also discuss the importance as well as the challenges for 3D bioprinting using stem cells. We aim to highlight the importance of the continuous cell printing in order to fabricate 3D biological structures with predefined shapes as being the building blocks of large and complex tissues

Toward personalized medicine: human adipose-derived stem cells (ASCs) and xenogenic-free media as enabling tools in tendon tissue engineering strategies.

L'abstract è presente nell'allegato / the abstract is in the attachmen

3D Engineered Peripheral Nerve: Towards A New Era of Patient-Specific Nerve Repair Solutions

Reconstruction of peripheral nerve injuries (PNIs) with substance loss remains challenging because of limited treatment solutions and unsatisfactory patient outcomes. Currently, nerve autografting is the first-line management choice for bridging critical-sized nerve defects. The procedure, however, is often complicated by donor site morbidity and paucity of nerve tissue, raising a quest for better alternatives. The application of other treatment surrogates, such as nerve guides remains questionable, and inefficient in irreducible nerve gaps. More importantly, these strategies lack customization for personalized patient therapy, which is a significant drawback of these nerve repair options. This negatively impacts the fascicle-to-fascicle regeneration process, critical to restoring the physiological axonal pathway of the disrupted nerve. Recently, the use of additive manufacturing (AM) technologies has offered major advancements to the bioengineering solutions for PNI therapy. These techniques aim to reinstate the native nerve fascicle pathway using biomimetic approaches, thereby augmenting end-organ innervation. AM-based approaches, such as 3D bioprinting, are capable of biofabricating 3D engineered nerve graft scaffolds in a patient-specific manner with high precision. Moreover, realistic in vitro models of peripheral nerve tissues that represent the physiologically and functionally relevant environment of human organs could also be developed. However, the technology is still nascent and faces major translational hurdles. In this review, we spotlighted the clinical burden of PNIs and most up-to-date treatment to address nerve gaps. Next, a summarized illustration of the nerve ultrastructure that guides research solutions is discussed. This is followed by a contrast of the existing bioengineering strategies used to repair peripheral nerve discontinuities. In addition, we elaborated on the most recent advances in 3D printing (3DP) and biofabrication applications in peripheral nerve modeling and engineering. Finally, the major challenges that limit the evolution of the field along with their possible solutions are also critically analyzed

Current standards and ethical landscape of engineered tissues—3D bioprinting perspective

Tissue engineering is an evolving multi-disciplinary field with cutting-edge technologies and innovative scientific perceptions that promise functional regeneration of damaged tissues/organs. Tissue engineered medical products (TEMPs) are biomaterial-cell products or a cell-drug combination which is injected, implanted or topically applied in the course of a therapeutic or diagnostic procedure. Current tissue engineering strategies aim at 3D printing/bioprinting that uses cells and polymers to construct living tissues/organs in a layer-by-layer fashion with high 3D precision. However, unlike conventional drugs or therapeutics, TEMPs and 3D bioprinted tissues are novel therapeutics and need different regulatory protocols for clinical trials and commercialization processes. Therefore, it is essential to understand the complexity of raw materials, cellular components, and manufacturing procedures to establish standards that can help to translate these products from bench to bedside. These complexities are reflected in the regulations and standards that are globally in practice to prevent any compromise or undue risks to patients. This review comprehensively describes the current legislations, standards for TEMPs with a special emphasis on 3D bioprinted tissues. Based on these overviews, challenges in the clinical translation of TEMPs & 3D bioprinted tissues/organs along with their ethical concerns and future perspectives are discussed