Sanjairaj Vijayavenkataraman

3D bioprinting of skin: a state-of-the-art review on modelling, materials, and processes.

The skin is the largest organ of the body, having a complex multi-layered structure and guards the underlying muscles, bones, ligaments, and internal organs. It serves as the first line of defence to any external stimuli, hence it is the most vulnerable to injury and warrants the need for rapid and reliable regeneration methods. Tissue engineered skin substitutes help overcome the limitations of traditional skin treatment methods, in terms of technology, time, and cost. While there is commendable progress in the treating of superficial wounds and injuries with skin substitutes, treatment of full-thickness injuries, especially with third or fourth degree burns, still looks murkier. Engineering multi-layer skin architecture, conforming to the native skin structure is a tougher goal to achieve with the current tissue engineering methods, if not impossible, restoring all the functions of the native skin. The testing of drugs and cosmetics is another area, where engineered skins are very much needed, with bans being imposed on product testing on animals. Given this greater need, 3D bioprinting is a promising technology that can achieve rapid and reliable production of biomimetic cellular skin substitutes, satisfying both clinical and industrial needs. This paper reviews all aspects related to the 3D bioprinting of skin, right from imaging the injury site, 3D model creation, biomaterials that are used and their suitability, types of cells and their functions, actual bioprinting technologies, along with the challenges and future prospects.

An Overview of Scaffold Design and Fabrication Technology for Engineered Knee Meniscus

Current surgical treatments for meniscal tears suffer from subsequent degeneration of knee joints, limited donor organs and inconsistent post-treatment results. Three clinical scaffolds (Menaflex CMI, Actifit® scaffold and NUsurface® Meniscus Implant) are available on the market, but additional data are needed to properly evaluate their safety and effectiveness. Thus, many scaffold-based research activities have been done to develop new materials, structures and fabrication technologies to mimic native meniscus for cell attachment and subsequent tissue development, and restore functionalities of injured meniscus for long-term effects. This study begins with a synopsis of relevant structural features of meniscus and goes on to describe the critical considerations. Promising advances made in the field of meniscal scaffolding technology, in terms of biocompatible materials, fabrication methods, structure design and their impact on mechanical and biological properties are discussed in detail. Among all the scaffolding technologies, additive manufacturing (AM) is very promising because of its ability to precisely control fiber diameter, orientation, and pore network micro-architecture to mimic the native meniscus microenvironment.

Investigation of process parameters of electrohydro-dynamic jetting for 3D printed PCL fibrous scaffolds with complex geometries

Tissue engineering is a promising technology in the field of regenerative medicine, with the potential to create tissues de novo . Though there has been a good progress in this field so far, there still exists the challenge of providing a 3D micro-architecture to the artificial tissue construct, to mimic the native cell or tissue environment. 3D bioprinting is looked upon as a solution, with its capability of mimicking the native tissue architecture, layer by layer, with high precision and appreciable resolution. Electrohydrodynamic jetting (E-jetting) is one type of 3D bioprinting, where a high electric voltage is applied between the extruding nozzle and the substrate, to print highly controlled fibres. In this study, an E-jetting system developed in-house is used to 3D print fibrous scaffolds. Effect of various E-jetting parameters, namely the supply voltage, solution concentration, nozzle-to-substrate distance, stage (printing) speed and solution dispensing feed rate on the diameter of printed fibres is studied at the first stage. Optimized parameters are then used to print Polycaprolactone (PCL) scaffolds of highly complex geometries, like semi-lunar and spiral geometries, with the aim of demonstrating the flexibility and capability of our system to fabricate complex geometry scaffolds, to biomimic the complex 3D micro-architecture of native tissue environment. The spiral geometry may help in better cell migration during cell culture and tissue maturation.

Design of Three-Dimensional Scaffolds with Tunable Matrix Stiffness for Directing Stem Cell Lineage Specification: An In Silico Study

Tissue engineering is a multi-disciplinary area of research bringing together the fields of engineering and life sciences with the aim of fabricating tissue constructs aiding in the regeneration of damaged tissues and organs. Scaffolds play a key role in tissue engineering, acting as the templates for tissue regeneration and guiding the growth of new tissue. The use of stem cells in tissue engineering and regenerative medicine becomes indispensable, especially for applications involving successful long-term restoration of continuously self-renewing tissues, such as skin. The differentiation of stem cells is controlled by a number of cues, of which the nature of the substrate and its innate stiffness plays a vital role in stem cell fate determination. By tuning the substrate stiffness, the differentiation of stem cells can be directed to the desired lineage. Many studies on the effect of substrate stiffness on stem cell differentiation has been reported, but most of those studies are conducted with two-dimensional (2D) substrates. However, the native in vivo tissue microenvironment is three-dimensional (3D) and life science researchers are moving towards 3D cell cultures. Porous 3D scaffolds are widely used by the researchers for 3D cell culture and the properties of such scaffolds affects the cell attachment, proliferation, and differentiation. To this end, the design of porous scaffolds directly influences the stem cell fate determination. There exists a need to have 3D scaffolds with tunable stiffness for directing the differentiation of stem cells into the desired lineage. Given the limited number of biomaterials with all the desired properties, the design of the scaffolds themselves could be used to tune the matrix stiffness. This paper is an in silico study, investigating the effect of various scaffold parameter, namely fiber width, porosity, number of unit cells per layer, number of layers, and material selection, on the matrix stiffness, thereby offering a guideline for design of porous tissue engineering scaffolds with tunable matrix stiffness for directing stem cell lineage specification.

Influence of electrohydrodynamic jetting parameters on the morphology of PCL scaffolds

One of the important constituents in tissue engineering is scaffold, which provides structural support and suitable microenvironment for the cell attachment, growth and proliferation. To fabricate micro/nano structures for soft tissue repair and three-dimensional (3D) cell culture, the key is to improve fibre-based scaffold fabrication. Electrohydrodynamic (EHD) jetting is capable of producing and orientating submicron fibres for 3D scaffold fabrication. In this work, an EHD-jetting system was developed to explore the relationship between vital processing parameters and fibre characteristics. In this study, polycaprolactone (PCL) solution prepared by dissolving PCL pellets in acetic acid was used to fabricate the scaffolds. The influence of voltage, motorized stage speed, solution feed rate, and solution concentration on fibre characteristics and scaffold pattern were studied. Morphology of the EHD-jetted PCL fibres and scaffolds were analysed using optical microscope images and scanning electron microscope (SEM) images. Multi-layer scaffolds with the varied coiled pattern were fabricated and analysed. Cell attachment and proliferation have to be investigated in the future by further cell culture studies on these multi-layer coiled scaffolds.

3D Printing and 3D Bioprinting in Pediatrics

Additive manufacturing, commonly referred to as 3D printing, is a technology that builds three-dimensional structures and components layer by layer. Bioprinting is the use of 3D printing technology to fabricate tissue constructs for regenerative medicine from cell-laden bio-inks. 3D printing and bioprinting have huge potential in revolutionizing the field of tissue engineering and regenerative medicine. This paper reviews the application of 3D printing and bioprinting in the field of pediatrics.

3D bioprinting of tissues and organs for regenerative medicine

&NA; 3D bioprinting is a pioneering technology that enables fabrication of biomimetic, multiscale, multi‐cellular tissues with highly complex tissue microenvironment, intricate cytoarchitecture, structure‐function hierarchy, and tissue‐specific compositional and mechanical heterogeneity. Given the huge demand for organ transplantation, coupled with limited organ donors, bioprinting is a potential technology that could solve this crisis of organ shortage by fabrication of fully‐functional whole organs. Though organ bioprinting is a far‐fetched goal, there has been a considerable and commendable progress in the field of bioprinting that could be used as transplantable tissues in regenerative medicine. This paper presents a first‐time review of 3D bioprinting in regenerative medicine, where the current status and contemporary issues of 3D bioprinting pertaining to the eleven organ systems of the human body including skeletal, muscular, nervous, lymphatic, endocrine, reproductive, integumentary, respiratory, digestive, urinary, and circulatory systems were critically reviewed. The implications of 3D bioprinting in drug discovery, development, and delivery systems are also briefly discussed, in terms of in vitro drug testing models, and personalized medicine. While there is a substantial progress in the field of bioprinting in the recent past, there is still a long way to go to fully realize the translational potential of this technology. Computational studies for study of tissue growth or tissue fusion post‐printing, improving the scalability of this technology to fabricate human‐scale tissues, development of hybrid systems with integration of different bioprinting modalities, formulation of new bioinks with tuneable mechanical and rheological properties, mechanobiological studies on cell‐bioink interaction, 4D bioprinting with smart (stimuli‐responsive) hydrogels, and addressing the ethical, social, and regulatory issues concerning bioprinting are potential futuristic focus areas that would aid in successful clinical translation of this technology.

Electrohydrodynamic Jet 3D Printed Nerve Guide Conduits (NGCs) for Peripheral Nerve Injury Repair

The prevalence of peripheral nerve injuries resulting in loss of motor function, sensory function, or both, is on the rise. Artificial Nerve Guide Conduits (NGCs) are considered an effective alternative treatment for autologous nerve grafts, which is the current gold-standard for treating peripheral nerve injuries. In this study, Polycaprolactone-based three-dimensional porous NGCs are fabricated using Electrohydrodynamic jet 3D printing (EHD-jetting) for the first time. The main advantage of this technique is that all the scaffold properties, namely fibre diameter, pore size, porosity, and fibre alignment, can be controlled by tuning the process parameters. In addition, EHD-jetting has the advantages of customizability, repeatability, and scalability. Scaffolds with five different pore sizes (125 to 550 μm) and porosities (65 to 88%) are fabricated and the effect of pore size on the mechanical properties is evaluated. In vitro degradation studies are carried out to investigate the degradation profile of the scaffolds and determine the influence of pore size on the degradation rate and mechanical properties at various degradation time points. Scaffolds with a pore size of 125 ± 15 μm meet the requirements of an optimal NGC structure with a porosity greater than 60%, mechanical properties closer to those of the native peripheral nerves, and an optimal degradation rate matching the nerve regeneration rate post-injury. The in vitro neural differentiation studies also corroborate the same results. Cell proliferation was highest in the scaffolds with a pore size of 125 ± 15 μm assessed by the PrestoBlue assay. The Reverse Transcription-Polymerase Chain Reaction (RT-PCR) results involving the three most important genes concerning neural differentiation, namely β3-tubulin, NF-H, and GAP-43, confirm that the scaffolds with a pore size of 125 ± 15 μm have the highest gene expression of all the other pore sizes and also outperform the electrospun Polycaprolactone (PCL) scaffold. The immunocytochemistry results, expressing the two important nerve proteins β3-tubulin and NF200, showed directional alignment of the neurite growth along the fibre direction in EHD-jet 3D printed scaffolds.

Pluronic F127 blended polycaprolactone scaffolds via e-jetting for esophageal tissue engineering

AbstractSeveral attempts have been made to fabricate esophageal tissue engineering scaffolds. However, most of these scaffolds possess randomly oriented fibres and uncontrollable pore sizes. In order to mimic the native esophageal tissue structure, electro-hydrodynamic jetting (e-jetting) was used in this study to fabricate scaffolds with aligned fibres and controlled pore size. A hydrophilic additive, Pluronic F127 (F127), was blended with polycaprolactone (PCL) to improve the wettability of the scaffolds and hence the cell adhesion. PCL/F127 composite scaffolds with different weight ratios (0–12%) were fabricated. The wettability, phase composition, and the mechanical properties of the fabricated scaffolds were investigated. The results show that the e-jetted scaffolds have controllable fibres orientated in two perpendicular directions, which are similar to the human esophagus structure and suitable pore size for cell infiltration. In addition, the scaffolds with 8% F127 exhibited better wettability (contact angle of 14°) and an ultimate tensile strength (1.2 MPa) that mimics the native esophageal tissue. Furthermore, primary human esophageal fibroblasts were seeded on the e-jetted scaffolds. PCL/F127 scaffolds showed enhanced cell proliferation and expression of the vascular endothelial growth factor (VEGF) compared to pristine PCL scaffolds (1.5- and 25.8- fold increase, respectively; P < 0.001). An in vitro wound model made using the PCL/F127 scaffolds showed better cell migration than the PCL scaffolds. In summary, the PCL/F127 e-jetted scaffolds offer a promising strategy for the esophagus tissue repair.

A review on the use of computational methods to characterize, design, and optimize tissue engineering scaffolds, with a potential in 3D printing fabrication: COMPUTATIONAL METHOD FOR TE SCAFFOLD DESIGN AND 3D-PRINTING

The design and fabrication of tissue engineering scaffolds is a highly complex process. In order to provide a proper architecture for cells to grow, proliferate, and differentiate to form tissues, scaffolds have to be made with suitable properties. However, the limited structural designs and conventional fabrication techniques severely cripple the improvement of scaffold properties. To overcome these limitations, many researchers have recently adopted computational methods combined with 3D printing techniques as a new approach for scaffold design and fabrication. This approach allows scaffolds to be designed and fabricated with highly complex microstructures and good control and accuracy. Previous works have also shown this approach to be a very useful tool to predict the scaffold properties and to optimize the scaffold designs with a great reduction of experimental iterations. As this approach combining computational methods and 3D printing techniques for scaffold design and fabrication has many advantages over the conventional trial-and-error based approach, it is imperative to provide a state-of-the-art review on the topic. To this end, this article reviews the various applications of computational methods in scaffold design and simulation; it also briefly reviews the application of 3D printing techniques to fabricate the computationally designed scaffolds. Finally, the limitations and future trends of this approach are discussed. Overall, this review will enable readers to understand the benefits of using computational methods coupled with 3D printing to design and fabricate scaffolds, and thus help researchers to improve and optimize the scaffold properties for future tissue engineering research.