bims-biprem 2023-10-01 papers

bims-biprem

Biomed News

on Bioprinting for regenerative medicine

Issue of 2023–10–01
thirteen papers selected by
Seerat Maqsood, University of Teramo

Translational biomaterials of four-dimensional bioprinting for tissue regeneration.
Three-Dimensional Bioprinting of Human Organs and Tissues: Bioethical and Medico-Legal Implications Examined through a Scoping Review.
Alginate-waterborne polyurethane 3D bioprinted scaffolds for articular cartilage tissue engineering.
Development and Prospective Applications of 3D Membranes as a Sensor for Monitoring and Inducing Tissue Regeneration.
Progress in microsphere-based scaffolds in bone/cartilage tissue engineering.
Production of a bioink containing decellularized spinal cord tissue for 3D bioprinting.
Application of Advanced Technologies-Nanotechnology, Genomics Technology, and 3D Printing Technology-In Precision Anesthesia: A Comprehensive Narrative Review.
Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting.
3D Filaments Based on Polyhydroxy Butyrate-Micronized Bacterial Cellulose for Tissue Engineering Applications.
Bioinspired 3D-printed scaffold embedding DDAB-nano ZnO/nanofibrous microspheres for regenerative diabetic wound healing.
Widening the use of 3D printing.
Biocomposites containing poly(lactic acid) and chitosan for 3D printing - Assessment of mechanical, antibacterial and in vitro biodegradability properties.
Directly coaxial bioprinting of 3D vascularized tissue using novel bioink based on decellularized human amniotic membrane.

Biofabrication. 2023 Sep 27.

Translational biomaterials of four-dimensional bioprinting for tissue regeneration.

Leah Faber, Anne Yau, Yupeng Chen.

  Bioprinting is an additive manufacturing (AM) technique that combines living cells, biomaterials, and biological molecules to develop biologically functional constructs. Three-dimensional (3D) bioprinting is commonly used as an in vitro modeling system and is a more accurate representation of in vivo conditions in comparison to two-dimensional (2D) cell culture. Although 3D bioprinting has been utilized in various tissue engineering and clinical applications, it only takes into consideration the initial state of the printed scaffold or object. Four-dimensional (4D) bioprinting has emerged in recent years to incorporate the additional dimension of time within the printed 3D scaffolds. During the 4D bioprinting process, an external stimulus is exposed to the printed construct, which ultimately changes its shape or functionality. By studying how the structures and the embedded cells respond to various stimuli, researchers can gain a deeper understanding of the functionality of native tissues. This review paper will focus on the biomaterial breakthroughs in the newly advancing field of 4D bioprinting and their applications in tissue engineering and regeneration. In addition, the use of smart biomaterials and 4D printing mechanisms for tissue engineering applications is discussed to demonstrate potential insights for novel 4D bioprinting applications. To address the current challenges with this technology, we will conclude with future perspectives involving the incorporation of biological scaffolds and self-assembling nanomaterials in bioprinted tissue constructs.

Keywords:  4D bioprinting; biological scaffolds; self-assembling nanomaterials; stimuli-responsive biomaterials; tissue engineering

DOI:  https://doi.org/10.1088/1758-5090/acfdd0
Bioengineering (Basel). 2023 Sep 07. pii: 1052. [Epub ahead of print]10(9):

Three-Dimensional Bioprinting of Human Organs and Tissues: Bioethical and Medico-Legal Implications Examined through a Scoping Review.

Giovanna Ricci, Filippo Gibelli, Ascanio Sirignano.

  Three-dimensional bioprinting is a rapidly evolving technology that holds the promise of addressing the increasing demand for organs, tissues, and personalized medicine. By employing computer-aided design and manufacturing processes, 3D bioprinting allows for the precise deposition of living cells, biomaterials, and biochemicals to create functional human tissues and organs. The potential applications of this technology are vast, including drug testing and development, disease modeling, regenerative medicine, and ultimately, organ transplantation. However, as with any groundbreaking technology, 3D bioprinting presents several ethical, legal, and regulatory concerns that warrant careful consideration. As the technology progresses towards clinical applications, it is essential to address these challenges and establish appropriate frameworks to guide the responsible development of 3D bioprinting. This article, utilizing the Arksey and O'Malley scoping review model, is designed to scrutinize the bioethical implications, legal and regulatory challenges, and medico-legal issues that are intertwined with this rapidly evolving technology.

Keywords:  bioethics; bioprinting; precision medicine; tissue engineering

DOI:  https://doi.org/10.3390/bioengineering10091052
Int J Biol Macromol. 2023 Sep 23. pii: S0141-8130(23)03967-3. [Epub ahead of print] 127070

Alginate-waterborne polyurethane 3D bioprinted scaffolds for articular cartilage tissue engineering.

R Olmos-Juste, G Larrañaga-Jaurrieta, I Larraza, S Ramos-Diez, S Camarero-Espinosa, N Gabilondo, A Eceiza.

  Articular cartilage defects comprise a spectrum of diseases associated with degeneration or damage of the connective tissue present in particular joints, presenting progressive osteoarthritis if left untreated. In vitro tissue regeneration is an innovative treatment for articular cartilage injuries that is attracting not only clinical attention, but also great interest in the development of novel biomaterials, since this procedure involves the formation of a neotissue with the help of material support. In this work, functional alginate and waterborne polyurethane (WBPU) scaffolds have been developed for articular cartilage regeneration using 3D bioprinting technology. The particular properties of alginate-WBPU blends, like mechanical strength, elasticity and moistening, mimic the original cartilage tissue characteristics, being ideal for this application. To fabricate the scaffolds, mature chondrocytes were loaded into different alginate-WBPU inks with rheological properties suitable for 3D bioprinting. Bioinks with high alginate content showed better 3D printing performance, as well as structural integrity and cell viability, being most suitable for scaffolds fabrication. After 28 days of in vitro cartilage formation experiments, scaffolds containing 3.2 and 6.4 % alginate resulted in the maintenance of cell number in the range of 104 chondrocytes/scaffold in differentiated phenotypes, capable of synthesizing specialized extracellular matrix (ECM) up to 6 μg of glycosaminoglycans (GAG) and thus, showing a potential application of these scaffolds for in vitro regeneration of articular cartilage tissue.

Keywords:  3D bioprinting; Alginate; Bioink; Chondrocytes cartilage tissue engineering; Waterborne polyurethane

DOI:  https://doi.org/10.1016/j.ijbiomac.2023.127070
Membranes (Basel). 2023 Sep 18. pii: 802. [Epub ahead of print]13(9):

Development and Prospective Applications of 3D Membranes as a Sensor for Monitoring and Inducing Tissue Regeneration.

Hanning Wu, Jiawen Chen, Pengxiang Zhao, Mengyu Liu, Fei Xie, Xuemei Ma.

  For decades, tissue regeneration has been a challenging issue in scientific modeling and human practices. Although many conventional therapies are already used to treat burns, muscle injuries, bone defects, and hair follicle injuries, there remains an urgent need for better healing effects in skin, bone, and other unique tissues. Recent advances in three-dimensional (3D) printing and real-time monitoring technologies have enabled the creation of tissue-like membranes and the provision of an appropriate microenvironment. Using tissue engineering methods incorporating 3D printing technologies and biomaterials for the extracellular matrix (ECM) containing scaffolds can be used to construct a precisely distributed artificial membrane. Moreover, advances in smart sensors have facilitated the development of tissue regeneration. Various smart sensors may monitor the recovery of the wound process in different aspects, and some may spontaneously give feedback to the wound sites by releasing biological factors. The combination of the detection of smart sensors and individualized membrane design in the healing process shows enormous potential for wound dressings. Here, we provide an overview of the advantages of 3D printing and conventional therapies in tissue engineering. We also shed light on different types of 3D printing technology, biomaterials, and sensors to describe effective methods for use in skin and other tissue regeneration, highlighting their strengths and limitations. Finally, we highlight the value of 3D bioengineered membranes in various fields, including the modeling of disease, organ-on-a-chip, and drug development.

Keywords:  3D printing; artificial membrane application; biomaterials; sensors; tissue regeneration

DOI:  https://doi.org/10.3390/membranes13090802
Biomed Mater. 2023 Sep 26.

Progress in microsphere-based scaffolds in bone/cartilage tissue engineering.

Qian Pan, Weixian Su, Yongchang Yao.

  Bone/cartilage repair and regeneration have been popular and difficult issues in medical research. Tissue engineering is rapidly evolving to provide new solutions to this problem, and the key point is to design the appropriate scaffold biomaterial. In recent years，microsphere-based scaffolds have been considered suitable scaffold materials for bone/cartilage injury repair because microporous structures can form more internal space for better cell proliferation and other cellular activities, and these composite scaffolds can provide physical/chemical signals for neotissue formation with higher efficiency. This paper reviews the research progress regarding microsphere-based scaffolds in bone/chondral tissue engineering, briefly introduces types of microspheres made from organic and inorganic materials, discusses the preparation methods of microspheres and the exploration of suitable microsphere pore sizes in bone and cartilage tissue engineering, and finally details the application of microsphere-based scaffolds in biomimetic scaffolds, cell proliferation and drug delivery systems.&#xD.

Keywords:  cell proliferation; chondrogenesis; drug delivery; microsphere; osteogenesis; tissue engineering

DOI:  https://doi.org/10.1088/1748-605X/acfd78
Tissue Eng Part A. 2023 Sep 29.

Production of a bioink containing decellularized spinal cord tissue for 3D bioprinting.

Marcelo Garrido Dos Santos, Fernanda Stapenhorst França, João Pedro Prestes, Cristian Teixeira, Luiz Carlos Sommer, Laura Elena Sperling, Patricia Pranke.

Over the last few years, 3D bioprinting has emerged as a promising approach in the field of regenerative medicine. This technique allows for the production of three-dimensional scaffolds to support cell transplantation due to its ability to mimic the extracellular environment. One alternative to enhancing cell adhesion, survival and proliferation is the use of decellularized extracellular matrix as a bioink component. The aim of this study was to produce a bioink using lyophilized rat Decellularized Spinal Cord Tissue (DSCT) for 3D bioprinting of nervous tissue. DNA quantification, hematoxylin and eosin and DAPI staining indicated that 1% SDS and 9 hours processing were effective in removing the cells from the spinal cord samples. The cell viability assay showed that the decellularized matrix is not cytotoxic for PC12 cells. The hydrogel containing DSCT, alginate, and gelatine used as the base for the bioink has a shear thinning behavior and low G''/G' ratio, allowing for good printability without compromising cell viability after 3D bioprinting. The bioink supported long-term PC12 cell survival, with 93% of live cells 4 weeks after printing, and stimulated the production of laminin-1 and neurofilament-M. This bioink, therefore, represents an easily available biomaterial for central nervous system tissue engineering.

DOI: https://doi.org/10.1089/ten.TEA.2023.0078
Pharmaceutics. 2023 Sep 06. pii: 2289. [Epub ahead of print]15(9):

Application of Advanced Technologies-Nanotechnology, Genomics Technology, and 3D Printing Technology-In Precision Anesthesia: A Comprehensive Narrative Review.

Shiyao Gu, Qingyong Luo, Cen Wen, Yu Zhang, Li Liu, Liu Liu, Su Liu, Chunhua Chen, Qian Lei, Si Zeng.

  There has been increasing interest and rapid developments in precision medicine, which is a new medical concept and model based on individualized medicine with the joint application of genomics, bioinformatics engineering, and big data science. By applying numerous emerging medical frontier technologies, precision medicine could allow individualized and precise treatment for specific diseases and patients. This article reviews the application and progress of advanced technologies in the anesthesiology field, in which nanotechnology and genomics can provide more personalized anesthesia protocols, while 3D printing can yield more patient-friendly anesthesia supplies and technical training materials to improve the accuracy and efficiency of decision-making in anesthesiology. The objective of this manuscript is to analyze the recent scientific evidence on the application of nanotechnology in anesthesiology. It specifically focuses on nanomedicine, precision medicine, and clinical anesthesia. In addition, it also includes genomics and 3D printing. By studying the current research and advancements in these advanced technologies, this review aims to provide a deeper understanding of the potential impact of these advanced technologies on improving anesthesia techniques, personalized pain management, and advancing precision medicine in the field of anesthesia.

Keywords:  3D printing; algology; anesthesia; anesthetic safety; genomics; nanotechnology; precision medicine

DOI:  https://doi.org/10.3390/pharmaceutics15092289
J Funct Biomater. 2023 Sep 05. pii: 459. [Epub ahead of print]14(9):

Fibrinogen-Based Bioink for Application in Skin Equivalent 3D Bioprinting.

Aida Cavallo, Tamer Al Kayal, Angelica Mero, Andrea Mezzetta, Lorenzo Guazzelli, Giorgio Soldani, Paola Losi.

  Three-dimensional bioprinting has emerged as an attractive technology due to its ability to mimic native tissue architecture using different cell types and biomaterials. Nowadays, cell-laden bioink development or skin tissue equivalents are still at an early stage. The aim of the study is to propose a bioink to be used in skin bioprinting based on a blend of fibrinogen and alginate to form a hydrogel by enzymatic polymerization with thrombin and by ionic crosslinking with divalent calcium ions. The biomaterial ink formulation, composed of 30 mg/mL of fibrinogen, 6% of alginate, and 25 mM of CaCl2, was characterized in terms of homogeneity, rheological properties, printability, mechanical properties, degradation rate, water uptake, and biocompatibility by the indirect method using L929 mouse fibroblasts. The proposed bioink is a homogeneous blend with a shear thinning behavior, excellent printability, adequate mechanical stiffness, porosity, biodegradability, and water uptake, and it is in vitro biocompatible. The fibrinogen-based bioink was used for the 3D bioprinting of the dermal layer of the skin equivalent. Three different normal human dermal fibroblast (NHDF) densities were tested, and better results in terms of viability, spreading, and proliferation were obtained with 4 × 106 cell/mL. The skin equivalent was bioprinted, adding human keratinocytes (HaCaT) through bioprinting on the top surface of the dermal layer. A skin equivalent stained by live/dead and histological analysis immediately after printing and at days 7 and 14 of culture showed a tissuelike structure with two distinct layers characterized by the presence of viable and proliferating cells. This bioprinted skin equivalent showed a similar native skin architecture, paving the way for its use as a skin substitute for wound healing applications.

Keywords:  3D bioprinting; alginate; bioink; fibrinogen; skin equivalent

DOI:  https://doi.org/10.3390/jfb14090459
J Funct Biomater. 2023 Sep 09. pii: 464. [Epub ahead of print]14(9):

3D Filaments Based on Polyhydroxy Butyrate-Micronized Bacterial Cellulose for Tissue Engineering Applications.

Matheus F Celestino, Lais R Lima, Marina Fontes, Igor T S Batista, Daniella R Mulinari, Alessandra Dametto, Raphael A Rattes, André C Amaral, Rosana M N Assunção, Clovis A Ribeiro, Guillermo R Castro, Hernane S Barud.

  In this work, scaffolds based on poly(hydroxybutyrate) (PHB) and micronized bacterial cellulose (BC) were produced through 3D printing. Filaments for the printing were obtained by varying the percentage of micronized BC (0.25, 0.50, 1.00, and 2.00%) inserted in relation to the PHB matrix. Despite the varying concentrations of BC, the biocomposite filaments predominantly contained PHB functional groups, as Fourier transform infrared spectroscopy (FTIR) demonstrated. Thermogravimetric analyses (i.e., TG and DTG) of the filaments showed that the peak temperature (Tpeak) of PHB degradation decreased as the concentration of BC increased, with the lowest being 248 °C, referring to the biocomposite filament PHB/2.0% BC, which has the highest concentration of BC. Although there was a variation in the thermal behavior of the filaments, it was not significant enough to make printing impossible, considering that the PHB melting temperature was 170 °C. Biological assays indicated the non-cytotoxicity of scaffolds and the provision of cell anchorage sites. The results obtained in this research open up new paths for the application of this innovation in tissue engineering.

Keywords:  3D printing; micronized bacterial cellulose; poly(hydroxybutyrate); scaffolds; tissue engineering

DOI:  https://doi.org/10.3390/jfb14090464
Biofabrication. 2023 Sep 26.

Bioinspired 3D-printed scaffold embedding DDAB-nano ZnO/nanofibrous microspheres for regenerative diabetic wound healing.

Walaa M Metwally, Salma E El-Habashy, Lobna S El-Hosseiny, Marwa M Essawy, Hoda M Eltaher, Labiba El-Khordagui.

  There is a constant demand for novel materials/biomedical devices to accelerate the healing of hard-to-heal wounds. Herein, an innovative 3D-printed bioinspired construct was developed as an antibacterial/regenerative scaffold for diabetic wound healing. Hyaluronic/chitosan ink was used to fabricate a bilayer scaffold comprising a dense plain hydrogel layer topping an antibacterial/regenerative nanofibrous layer obtained by incorporating the hydrogel with polylactic acid nanofibrous microspheres (MS). These were embedded with nano ZnO (ZNP) or didecyldimethylammonium bromide (DDAB)-treated ZNP (D-ZNP) to generate the antibacterial/healing nano/micro hybrid biomaterials, Z-MS@scaffold and DZ-MS@scaffold. Plain and composite scaffolds incorporating blank MS (blank MS@scaffold) or MS-free ZNP@scaffold and D-ZNP@scaffold were used for comparison. 3D printed bilayer constructs with customizable porosity were obtained as verified by SEM. The DZ-MS@scaffold exhibited the largest total pore area as well as the highest water-uptake capacity and in vitro antibacterial activity. Treatment of Staphylococcus aureus-infected full thickness diabetic wounds in rats indicated superiority of DZ-MS@scaffold as evidenced by multiple assessments. The scaffold afforded 95% wound-closure, infection suppression, effective regulation of healing-associated biomarkers as well as regeneration of skin structure in 14 days. On the other hand, healing of non-diabetic acute wounds was effectively accelerated by the simpler less porous Z-MS@scaffold. Information is provided for the first-time on the 3D printing of nanofibrous scaffolds using non-electrospun injectable bioactive nano/micro particulate constructs, an innovative ZNP-functionalized 3D-printed formulation and the distinct bioactivity of D-ZNP as a powerful antibacterial/wound healing promotor. In addition, findings underscored the crucial role of nanofibrous-MS carrier in enhancing the physicochemical, antibacterial, and wound regenerative properties of DDAB-nano ZnO. In conclusion, innovative 3D-printed DZ-MS@scaffold merging the MS-boosted multiple functionalities of ZNP and DDAB, the structural characteristics of nanofibrous MS in addition to those of the 3D-printed bilayer scaffold, provide a versatile bioactive material platform for diabetic wound healing and other biomedical applications.

Keywords:  Hyaluronic acid-chitosan ink; bioinspired hydrogel scaffolds; customization; infected-wound healing; multilevel porosity; nanofibrous microspheres; polylactic acid

DOI:  https://doi.org/10.1088/1758-5090/acfd60
Science. 2023 Sep 29. 381(6665): 1413-1414

Widening the use of 3D printing.

Daniel M Balazs, Maria Ibáñez.

A light-triggered fabrication method extends the functionality of printable nanomaterials.

DOI: https://doi.org/10.1126/science.adk3070
J Mech Behav Biomed Mater. 2023 Sep 21. pii: S1751-6161(23)00489-7. [Epub ahead of print]147 106136

Biocomposites containing poly(lactic acid) and chitosan for 3D printing - Assessment of mechanical, antibacterial and in vitro biodegradability properties.

Isabel Hui, Eva Pasquier, Amalie Solberg, Karin Agrenius, Joakim Håkansson, Gary Chinga-Carrasco.

  New bone repair materials are needed for treatment of trauma- and disease-related skeletal defects as they still represent a major challenge in clinical practice. Additionally, new strategies are required to combat orthopedic device-related infections (ODRI), given the rising incidence of total joint replacement and fracture fixation surgeries in increasingly elderly populations. Recently, the convergence of additive manufacturing (AM) and bone tissue engineering (BTE) has facilitated the development of bone healthcare to achieve personalized three-dimensional (3D) scaffolds. This study focused on the development of a 3D printable bone repair material, based on the biopolymers poly(lactic acid) (PLA) and chitosan. Two different types of PLA and chitosan differing in their molecular weight (MW) were explored. The novel feature of this research was the successful 3D printing using biocomposite filaments composed of PLA and 10 wt% chitosan, with clear chitosan entrapment within the PLA matrix confirmed by Scanning Electron Microscopy (SEM) images. Tensile testing of injection molded samples indicated an increase in stiffness, compared to pure PLA scaffolds, suggesting potential for improved load-bearing characteristics in bone scaffolds. However, the potential benefit of chitosan on the biocomposite stiffness could not be reproduced in compression testing of 3D printed cylinders. The antibacterial assays confirmed antibacterial activity of chitosan when dissolved in acetic acid. The study also verified the biodegradability of the scaffolds, with a process producing an acidic environment that could potentially be neutralized by chitosan. In conclusion, the study indicated the feasibility of the proposed PLA/chitosan biocomposite for 3D printing, demonstrating adequate mechanical strength, antibacterial properties and biodegradability, which could serve as a new material for bone repair.

Keywords:  3D printing; Antibacterial; Biocomposites; Biodegradability; Chitosan; Poly(lactic acid)

DOI:  https://doi.org/10.1016/j.jmbbm.2023.106136
Int J Biol Macromol. 2023 Sep 22. pii: S0141-8130(23)03938-7. [Epub ahead of print]253(Pt 4): 127041

Directly coaxial bioprinting of 3D vascularized tissue using novel bioink based on decellularized human amniotic membrane.

Faranak Heidari, Maryam Saadatmand, Sara Simorgh.

  Despite several progressions in the biofabrication of large-scale engineered tissues, direct biopri nting of perfusable three-dimensional (3D) vasculature remained unaddressed. Developing a feasible method to generate cell-laden thick tissue with an effective vasculature network to deliver oxygen and nutrient is crucial for preventing the formation of necrotic spots and tissue death. In this study, we developed a novel technique to directly bioprint 3D cell-laden prevascularized construct. We developed a novel bioink by mixing decellularized human amniotic membrane (dHAM) and alginate (Alg) in various ratios. The bioink with encapsulated human vein endothelial cells (HUVECs) and a crosslinker, CaCl2, were extruded via sheath and core nozzle respectively to directly bioprint a perfusable 3D vasculature construct. The various concentration of bioink was assessed from several aspects like biocompatibility, porosity, swelling, degradation, and mechanical characteristics, and accordingly, optimized concentration was selected (Alg 4 %w/v - dHAM 0.6 %w/v). Then, the crosslinked bioink without microchannel and the 3D bioprinted construct with various microchannel distances (0, 1.5 mm, 3 mm) were compared. The 3D bioprinted construct with a 1.5 mm microchannels distance demonstrated superiority owing to its 492 ± 18.8 % cell viability within 14 days, excellent tubulogenesis, remarkable expression of VEGFR-2 which play a crucial role in endothelial cell proliferation, migration, and more importantly angiogenesis, and neovascularization. This perfusable bioprinted construct also possess appropriate mechanical stability (32.35 ± 5 kPa Young's modulus) for soft tissue. Taking these advantages into the account, our new bioprinting method possesses a prominent potential for the fabrication of large-scale prevascularized tissue to serve for regenerative medicine applications like implantation, drug-screening platform, and the study of mutation disease.

Keywords:  3D-bioprinting; Alginate; Angiogenesis; Coaxial nozzle; Decellularized human amniotic membrane; HUVECs; Vascularization

DOI:  https://doi.org/10.1016/j.ijbiomac.2023.127041