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Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds

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A special issue of Materials (ISSN 1996-1944).

Deadline for manuscript submissions: closed (30 June 2019) | Viewed by 82818

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Special Issue Editor

Dr. Iman Roohani

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Guest Editor

School of Chemistry, University of New South Wales, Sydney, Australia
Interests: biomaterials; stem cells; 3D printing; musculoskeletal tissue regeneration; bioceramics; hydrogels
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The design and fabrication of innovative scaffolds for tissue engineering applications are the most interesting topics in the biomaterials and tissue engineering community. In tissue engineering, scaffolds play a crucial role. A scaffold, which is a porous three-dimensional (3D) structure, is used to facilitate cell/tissue growth and the transportation of nutrients and wastes while interacting with biological environment. The emergence of new technology- additive manufacturing and 3D printing has enabled scientists to fine tune the internal and external structure of scaffolds and to incorporate various bioinstructive molecules such as genes, growth factors, and cytokines within the scaffolds to ultimately enhance rate of tissue regeneration. The present Special Issue of Materials will include the most recent and relevant contributions from materials scientists, biologists, and tissue engineers, focusing on novel 3D tissue scaffold fabrication; tissue and organ printing; the modelling of biofabrication processes and biofabricated constructs; architecture optimisation; and the fabrication of bioinstructive scaffolds using the volumetric incorporation of genes, growth factors, and cytokines.

Dr. Iman Roohani-Esfahani
Guest Editor

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Keywords

bioactive materials
3D printing
additive manufacturing
tissue regeneration
bioceramics
biodegradable polymers
hydrogels
porous scaffolds
tissue engineering

Published Papers (11 papers)

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Research

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16 pages, 2358 KiB

Open AccessFeature PaperArticle

3D Cultures of Salivary Gland Cells in Native or Gelled Egg Yolk Plasma, Combined with Egg White and 3D-Printing of Gelled Egg Yolk Plasma

by André M. Charbonneau, Joseph M. Kinsella and Simon D. Tran

Materials 2019, 12(21), 3480; https://doi.org/10.3390/ma12213480 - 24 Oct 2019

Cited by 16 | Viewed by 3097

Abstract

For salivary gland (SG) tissue engineering, we cultured acinar NS-SV-AC cell line or primary SG fibroblasts for 14 days in avian egg yolk plasma (EYP). Media or egg white (EW) supplemented the cultures as they grew in 3D-Cryo histology well inserts. In the second half of this manuscript, we measured EYP’s freeze-thaw gelation and freeze-thaw induced gelled EYP (_GEYP), and designed and tested further _GEYP tissue engineering applications. With a 3D-Cryo well insert, we tested _GEYP as a structural support for 3D cell culture or as a bio-ink for 3D-Bioprinting fluorescent cells. In non-printed EYP + EW or _GEYP + EW cultures, sagittal sections of the cultures showed cells remaining above the well’s base. Ki-67 expression was lacking for fibroblasts, contrasting NS-SV-AC’s constant expression. Rheological viscoelastic measurements of _GEYP at 37 °C on seven different freezing periods showed constant increase from 0 in mean storage and loss moduli, to 320 Pa and 120 Pa, respectively, after 30 days. We successfully 3D-printed _GEYP with controlled geometries. We manually extruded _GEYP bio-ink with fluorescence cells into a 3D-Cryo well insert and showed cell positioning. The 3D-Cryo well inserts reveal information on cells in EYP and we demonstrated _GEYP cell culture and 3D-printing applications. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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18 pages, 10784 KiB

Open AccessArticle

Medical-Grade PCL Based Polyurethane System for FDM 3D Printing—Characterization and Fabrication

by Agnieszka Haryńska, Justyna Kucinska-Lipka, Agnieszka Sulowska, Iga Gubanska, Marcin Kostrzewa and Helena Janik

Materials 2019, 12(6), 887; https://doi.org/10.3390/ma12060887 - 16 Mar 2019

Cited by 83 | Viewed by 9029

Abstract

The widespread use of three-dimensional (3D) printing technologies in medicine has contributed to the increased demand for 3D printing materials. In addition, new printing materials that are appearing in the industry do not provide a detailed material characterization. In this paper, we present the synthesis and characterization of polycaprolactone (PCL) based medical-grade thermoplastic polyurethanes, which are suitable for forming in a filament that is dedicated to Fused Deposition Modeling 3D (FDM 3D)printers. For this purpose, we synthesized polyurethane that is based on PCL and 1,6-hexamethylene diisocyanate (HDI) with a different isocyanate index NCO:OH (0.9:1, 1.1:1). Particular characteristics of synthesized materials included, structural properties (FTIR, Raman), thermal (differential scanning calorimetry (DSC), thermogravimetric analysis (TGA)), mechanical and surfaces (contact angle) properties. Moreover, pre-biological tests in vitro and degradation studies were also performed. On the basis of the conducted tests, a material with more desirable properties S-TPU(PCL)0.9 was selected and the optimization of filament forming via melt-extrusion process was described. The initial biological test showed the biocompatibility of synthesized S-TPU(PCL)0.9 with respect to C2C12 cells. It was noticed that the process of thermoplastic polyurethanes (TPU) filaments forming by extrusion was significantly influenced by the appropriate ratio between the temperature profile, rotation speed, and dosage ratio. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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18 pages, 7978 KiB

Open AccessArticle

Improved Bioactivity of 3D Printed Porous Titanium Alloy Scaffold with Chitosan/Magnesium-Calcium Silicate Composite for Orthopaedic Applications

by Chun-Hao Tsai, Chih-Hung Hung, Che-Nan Kuo, Cheng-Yu Chen, Yu-Ning Peng and Ming-You Shie

Materials 2019, 12(2), 203; https://doi.org/10.3390/ma12020203 - 09 Jan 2019

Cited by 62 | Viewed by 5108

Abstract

Recently, cases of bone defects have been increasing incrementally. Thus, repair or replacement of bone defects is gradually becoming a huge problem for orthopaedic surgeons. Three-dimensional (3D) scaffolds have since emerged as a potential candidate for bone replacement, of which titanium (Ti) alloys are one of the most promising candidates among the metal alloys due to their low cytotoxicity and mechanical properties. However, bioactivity remains a problem for metal alloys, which can be enhanced using simple immersion techniques to coat bioactive compounds onto the surface of Ti–6Al–4V scaffolds. In our study, we fabricated magnesium-calcium silicate (Mg–CS) and chitosan (CH) compounds onto Ti–6Al–4V scaffolds. Characterization of these surface-modified scaffolds involved an assessment of physicochemical properties as well as mechanical testing. Adhesion, proliferation, and growth of human Wharton’s Jelly mesenchymal stem cells (WJMSCs) were assessed in vitro. In addition, the cell attachment morphology was examined using scanning electron microscopy to assess adhesion qualities. Osteogenic and mineralization assays were conducted to assess osteogenic expression. In conclusion, the Mg–CS/CH coated Ti–6Al–4V scaffolds were able to exhibit and retain pore sizes and their original morphologies and architectures, which significantly affected subsequent hard tissue regeneration. In addition, the surface was shown to be hydrophilic after modification and showed mechanical strength comparable to natural bone. Not only were our modified scaffolds able to match the mechanical properties of natural bone, it was also found that such modifications enhanced cellular behavior such as adhesion, proliferation, and differentiation, which led to enhanced osteogenesis and mineralization downstream. In vivo results indicated that Mg–CS/CH coated Ti–6Al–4V enhances the bone regeneration and ingrowth at the critical size bone defects of rabbits. These results indicated that the proposed Mg–CS/CH coated Ti–6Al–4V scaffolds exhibited a favorable, inducive micro-environment that could serve as a promising modification for future bone tissue engineering scaffolds. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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16 pages, 4103 KiB

Open AccessArticle

Assessment of the Influence of Acetic Acid Residue on Type I Collagen during Isolation and Characterization

by Seon Young Bak, Sang Woo Lee, Chong Hyuk Choi and Hyun Woo Kim

Materials 2018, 11(12), 2518; https://doi.org/10.3390/ma11122518 - 11 Dec 2018

Cited by 17 | Viewed by 3436

Abstract

Various methods for isolation of type I collagen using acids, bases, enzymes, and their combinations have been applied. However, a lack of standardization exists among type I collagens isolated by various approaches. Consequently, in this study, we assessed the influence of acetic acid residue on type I collagen isolated by pepsin-acetic acid treatment, the fabrication of collagen-based porous scaffolds, and the seeded cells on collagen scaffolds. Unlike the isolated collagen dialyzed by deionized water (DDW), collagen dialyzed by 0.5 M acetic acid (DAC) exhibited structural and thermal denaturation. Both DDW- and DAC-based porous scaffolds at all collagen concentrations (0.5, 1 and 2% w/v) showed the high degree of porosity (>98%), and their pore morphologies were comparable at the same concentrations. However, the DDW- and DAC-based collagen scaffolds displayed significant differences in their physical properties (weight, thickness, and volume) and swelling behaviors. In particular, the weight losses induced by mechanical stimulation reflected the high degradation of DAC-collagen scaffolds. In cell culture experiments using adipose-derived stem cells (ADSCs), the characteristics of mesenchymal stem cell (MSC) did not change in both DDW- and DAC-collagen scaffolds for 10 days, although cells proliferated less in the DAC-collagen scaffolds. Our results suggest that the elimination of acetic acid residue from isolated collagen is recommended to produce collagen scaffolds that provide a stable environment for cells and cell therapy-related applications. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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14 pages, 10612 KiB

Open AccessArticle

Evaluation of Bone Sialoprotein Coating of Three-Dimensional Printed Calcium Phosphate Scaffolds in a Calvarial Defect Model in Mice

by Andreas Baranowski, Anja Klein, Ulrike Ritz, Hermann Götz, Stefan G. Mattyasovszky, Pol M. Rommens and Alexander Hofmann

Materials 2018, 11(11), 2336; https://doi.org/10.3390/ma11112336 - 21 Nov 2018

Cited by 11 | Viewed by 4235

Abstract

The bioactive coating of calcium phosphate cement (CPC) is a promising approach to enhance the bone-healing properties of bone substitutes. The purpose of this study was to evaluate whether coating CPCs with bone sialoprotein (BSP) results in increased bone formation. Forty-five female C57BL/6NRj mice with an average age of six weeks were divided into three groups. Either a BSP-coated or an uncoated three-dimensional plotted scaffold was implanted into a drilled 2.7-mm diameter calvarial defect, or the defect was left empty (control group; no CPC). Histological analyses revealed that BSP-coated scaffolds were better integrated into the local bone stock eight weeks after implantation. Bone volume/total volume (BV/TV) ratios and bone thickness at the bone–implant contact were analyzed via micro computed tomography (µCT) after eight weeks. BSP-coated scaffolds and uncoated CPC scaffolds increased bone thickness in comparison to the control (CPC + BSP: 691.1 ± 253.5 µm, CPC: 603.1 ± 164.4 µm, no CPC: 261.7 ± 37.8 µm, p < 0.01). Accordingly, BV/TV was enhanced in both scaffold groups (CPC + BSP: 1.3 ± 0.5%, CPC: 0.9 ± 0.5%, no CPC: 0.2 ± 0.3%, p < 0.01). The BSP coating showed a tendency towards an increased bone thickness (p = 0.18) and BV/TV (p = 0.18) in comparison to uncoated CPC scaffolds. However, a significant increase in bone formation through BSP coating was not found. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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Graphical abstract

16 pages, 4279 KiB

Open AccessArticle

The Physicochemical Properties of Decellularized Extracellular Matrix-Coated 3D Printed Poly(ε-caprolactone) Nerve Conduits for Promoting Schwann Cells Proliferation and Differentiation

by Chung-Chia Chen, Joyce Yu, Hooi-Yee Ng, Alvin Kai-Xing Lee, Chien-Chang Chen, Yueh-Sheng Chen and Ming-You Shie

Materials 2018, 11(9), 1665; https://doi.org/10.3390/ma11091665 - 09 Sep 2018

Cited by 36 | Viewed by 4241

Abstract

Although autologous nerve grafting remains the gold standard treatment for peripheral nerve injuries, alternative methods such as development of nerve guidance conduits have since emerged and evolved to counter the many disadvantages of nerve grafting. However, the efficacy and viability of current nerve conduits remain unclear in clinical trials. Here, we focused on a novel decellularized extracellular matrix (dECM) and polydopamine (PDA)-coated 3D-printed poly(ε-caprolactone) (PCL)-based conduits, whereby the PDA surface modification acts as an attachment platform for further dECM attachment. We demonstrated that dECM/PDA-coated PCL conduits possessed higher mechanical properties when compared to human or animal nerves. Such modifications were proved to affect cell behaviors. Cellular behaviors and neuronal differentiation of Schwann cells were assessed to determine for the efficacies of the conduits. There were some cell-specific neuronal markers, such as Nestin, neuron-specific class III beta-tubulin (TUJ-1), and microtubule-associated protein 2 (MAP2) analyzed by enzyme-linked immunosorbent assay, and Nestin expressions were found to be 0.65-fold up-regulated, while TUJ1 expressions were 2.3-fold up-regulated and MAP2 expressions were 2.5-fold up-regulated when compared to Ctl. The methodology of PDA coating employed in this study can be used as a simple model to immobilize dECM onto PCL conduits, and the results showed that dECM/PDA-coated PCL conduits can as a practical and clinically viable tool for promoting regenerative outcomes in larger peripheral nerve defects. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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Review

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42 pages, 11786 KiB

Open AccessReview

Bioprinting Vasculature: Materials, Cells and Emergent Techniques

by Clarissa Tomasina, Tristan Bodet, Carlos Mota, Lorenzo Moroni and Sandra Camarero-Espinosa

Materials 2019, 12(17), 2701; https://doi.org/10.3390/ma12172701 - 23 Aug 2019

Cited by 97 | Viewed by 8012

Abstract

Despite the great advances that the tissue engineering field has experienced over the last two decades, the amount of in vitro engineered tissues that have reached a stage of clinical trial is limited. While many challenges are still to be overcome, the lack of vascularization represents a major milestone if tissues bigger than approximately 200 µm are to be transplanted. Cell survival and homeostasis is to a large extent conditioned by the oxygen and nutrient transport (as well as waste removal) by blood vessels on their proximity and spontaneous vascularization in vivo is a relatively slow process, leading all together to necrosis of implanted tissues. Thus, in vitro vascularization appears to be a requirement for the advancement of the field. One of the main approaches to this end is the formation of vascular templates that will develop in vitro together with the targeted engineered tissue. Bioprinting, a fast and reliable method for the deposition of cells and materials on a precise manner, appears as an excellent fabrication technique. In this review, we provide a comprehensive background to the fields of vascularization and bioprinting, providing details on the current strategies, cell sources, materials and outcomes of these studies. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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20 pages, 1221 KiB

Open AccessReview

3D Printing of Bioceramic Scaffolds—Barriers to the Clinical Translation: From Promise to Reality, and Future Perspectives

by Kang Lin, Rakib Sheikh, Sara Romanazzo and Iman Roohani

Materials 2019, 12(17), 2660; https://doi.org/10.3390/ma12172660 - 21 Aug 2019

Cited by 95 | Viewed by 9712

Abstract

In this review, we summarize the challenges of the three-dimensional (3D) printing of porous bioceramics and their translational hurdles to clinical applications. The state-of-the-art of the major 3D printing techniques (powder-based and slurry-based), their limitations and key processing parameters are discussed in detail. The significant roadblocks that prevent implementation of 3D printed bioceramics in tissue engineering strategies, and medical applications are outlined, and the future directions where new research may overcome the limitations are proposed. In recent years, there has been an increasing demand for a nanoscale control in 3D fabrication of bioceramic scaffolds via emerging techniques such as digital light processing, two-photon polymerization, or large area maskless photopolymerization. However, these techniques are still in a developmental stage and not capable of fabrication of large-sized bioceramic scaffolds; thus, there is a lack of sufficient data to evaluate their contribution. This review will also not cover polymer matrix composites reinforced with particulate bioceramics, hydrogels reinforced with particulate bioceramics, polymers coated with bioceramics and non-porous bioceramics. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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18 pages, 1302 KiB

Open AccessFeature PaperReview

iPSC Bioprinting: Where are We at?

by Sara Romanazzo, Stephanie Nemec and Iman Roohani

Materials 2019, 12(15), 2453; https://doi.org/10.3390/ma12152453 - 01 Aug 2019

Cited by 31 | Viewed by 8545

Abstract

Here, we present a concise review of current 3D bioprinting technologies applied to induced pluripotent stem cells (iPSC). iPSC have recently received a great deal of attention from the scientific and clinical communities for their unique properties, which include abundant adult cell sources, ability to indefinitely self-renew and differentiate into any tissue of the body. Bioprinting of iPSC and iPSC derived cells combined with natural or synthetic biomaterials to fabricate tissue mimicked constructs, has emerged as a technology that might revolutionize regenerative medicine and patient-specific treatment. This review covers the advantages and disadvantages of bioprinting techniques, influence of bioprinting parameters and printing condition on cell viability, and commonly used iPSC sources, and bioinks. A clear distinction is made for bioprinting techniques used for iPSC at their undifferentiated stage or when used as adult stem cells or terminally differentiated cells. This review presents state of the art data obtained from major searching engines, including Pubmed/MEDLINE, Google Scholar, and Scopus, concerning iPSC generation, undifferentiated iPSC, iPSC bioprinting, bioprinting techniques, cartilage, bone, heart, neural tissue, skin, and hepatic tissue cells derived from iPSC. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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25 pages, 4194 KiB

Open AccessReview

Integrated Design Approaches for 3D Printed Tissue Scaffolds: Review and Outlook

by Paul F. Egan

Materials 2019, 12(15), 2355; https://doi.org/10.3390/ma12152355 - 24 Jul 2019

Cited by 74 | Viewed by 6728

Abstract

Emerging 3D printing technologies are enabling the fabrication of complex scaffold structures for diverse medical applications. 3D printing allows controlled material placement for configuring porous tissue scaffolds with tailored properties for desired mechanical stiffness, nutrient transport, and biological growth. However, tuning tissue scaffold functionality requires navigation of a complex design space with numerous trade-offs that require multidisciplinary assessment. Integrated design approaches that encourage iteration and consideration of diverse processes including design configuration, material selection, and simulation models provide a basis for improving design performance. In this review, recent advances in design, fabrication, and assessment of 3D printed tissue scaffolds are investigated with a focus on bone tissue engineering. Bone healing and fusion are examples that demonstrate the needs of integrated design approaches in leveraging new materials and 3D printing processes for specified clinical applications. Current challenges for integrated design are outlined and emphasize directions where new research may lead to significant improvements in personalized medicine and emerging areas in healthcare. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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21 pages, 1596 KiB

Open AccessReview

An Introduction to 3D Bioprinting: Possibilities, Challenges and Future Aspects

by Željka P. Kačarević, Patrick M. Rider, Said Alkildani, Sujith Retnasingh, Ralf Smeets, Ole Jung, Zrinka Ivanišević and Mike Barbeck

Materials 2018, 11(11), 2199; https://doi.org/10.3390/ma11112199 - 06 Nov 2018

Cited by 268 | Viewed by 19839

Abstract

Bioprinting is an emerging field in regenerative medicine. Producing cell-laden, three-dimensional structures to mimic bodily tissues has an important role not only in tissue engineering, but also in drug delivery and cancer studies. Bioprinting can provide patient-specific spatial geometry, controlled microstructures and the positioning of different cell types for the fabrication of tissue engineering scaffolds. In this brief review, the different fabrication techniques: laser-based, extrusion-based and inkjet-based bioprinting, are defined, elaborated and compared. Advantages and challenges of each technique are addressed as well as the current research status of each technique towards various tissue types. Nozzle-based techniques, like inkjet and extrusion printing, and laser-based techniques, like stereolithography and laser-assisted bioprinting, are all capable of producing successful bioprinted scaffolds. These four techniques were found to have diverse effects on cell viability, resolution and print fidelity. Additionally, the choice of materials and their concentrations were also found to impact the printing characteristics. Each technique has demonstrated individual advantages and disadvantages with more recent research conduct involving multiple techniques to combine the advantages of each technique. Full article

(This article belongs to the Special Issue Additive manufacturing and Biofabrication of Tissue Engineering Scaffolds)

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