Exploring nanofibrous self-assembling peptide hydrogels using mouse myoblast cells for three-dimensional bioprinting and tissue engineering applications.

Biomaterial-based 3D bioprinting strategy for orthopedic tissue engineering.

The ability of skeletal muscle to self-repair after a traumatic injury, tumor ablation, or muscular disease is slow and limited, and the capacity of skeletal muscle to self-regenerate declines steeply with age. Tissue engineering of functional skeletal muscle using 3D bioprinting technology is promising for creating tissue constructs that repair and promote regeneration of damaged tissue. Hydrogel scaffolds used as biomaterials for skeletal muscle tissue engineering can provide chemical, physical and mechanical cues to the cells in three dimensions thus promoting regeneration. Herein, we have developed two synthetically designed novel tetramer peptide biomaterials. These peptides are self-assembling into a nanofibrous 3D network, entrapping 99.9% water and mimicking the native collagen of an extracellular matrix. Different biocompatibility assays including MTT, 3D cell viability assay, cytotoxicity assay and live-dead assay confirm the biocompatibility of these peptide hydrogels for mouse myoblast cells (C2C12). Immunofluorescence analysis of cell-laden hydrogels revealed that the proliferation of C2C12 cells was well-aligned in the peptide hydrogels compared to the alginategelatin control. These results indicate that these peptide hydrogels are suitable for skeletal muscle tissue engineering. Finally, we tested the printability of the peptide bioinks using a commercially available 3D bioprinter. The ability to print these hydrogels will enable future development of 3D bioprinted scaffolds containing skeletal muscle myoblasts for tissue engineering applications.

The development of three-dimensional (3D) microenvironments that mimic the role of native extracellular matrix (ECM) components is very crucial. Self-assembling peptide (SAP) hydrogels have been utilized as scaffolds for encapsulation, 3D culture, proliferation, and differentiation of cells and also for repairing defects in cartilage tissue. SAPs can be biofunctionalized with short peptide epitopes to form biomimetic scaffolds. Herein, KLD (KLDLKLDLKLDL) SAP was functionalized with a short bioactive motif, EGDK, to fabricate KLD-HM (KLDLKLDLKLDL-EGDK) SAP hydrogel and self-assembled. In this study, it was aimed to investigate the effect of developed KLD-HM SAP hydrogels on the viability and proliferation of human mesenchymal stem cells (hMSCs). For ensuring the stability of SAPs, the rheological properties and degradation profile of produced SAP hydrogels were assessed. After the encapsulation of hMSCs in SAP hydrogels, MTT assay and Live and Dead staining assay were conducted. We showed that these biomimetic peptide hydrogel scaffolds provided a proper microenvironment for encapsulated hMSCs and the developed SAP hydrogels promoted the adhesion, viability, and proliferation of hMSCs. Our results suggest that designed bioactive SAP hydrogel scaffolds might be useful for promoting the regeneration of cartilage tissue.

PurposeTwo-dimensional (2D) cell culture is a valuable method for cell-based research but can provide unpredictable, misleading data about in vivo responses. In this study, we created a three-dimensional (3D) cell culture environment to mimic tumor characteristics and cell-cell interactions to better characterize the tumor formation response to chemotherapy.Materials and methodsWe fabricated the 3D cell culture samples using a 3D cell bio printer and the bladder cancer cell line 5637. T24 cells were used for 2D cell culture. Then, rapamycin and Bacillus Calmette-Guérin (BCG) were used to examine their cancer inhibition effects using the two bladder cancer cell lines. Cell-cell interaction was measured by measuring e-cadherin and n-cadherin secreted via the epithelial-mesenchymal transition (EMT).ResultsWe constructed a 3D cell scaffold using gelatin methacryloyl (GelMA) and compared cell survival in 3D and 2D cell cultures. 3D cell cultures showed higher cancer cell proliferation rates than 2D cell cultures, and the 3D cell culture environment showed higher cell-to-cell interactions through the secretion of E-cadherin and N-cadherin. Assessment of the effects of drugs for bladder cancer such as rapamycin and BCG showed that the effect in the 2D cell culture environment was more exaggerated than that in the 3D cell culture environment.ConclusionsWe fabricated 3D scaffolds with bladder cancer cells using a 3D bio printer, and the 3D scaffolds were similar to bladder cancer tissue. This technique can be used to create a cancer cell-like environment for a drug screening platform.

Three-dimensional (3D) bioprinting is a promising technology which utilises additive manufacturing technologies to produce functional tissues and organs. A bioink is the term used to describe any biomaterial, containing cells or other biologics, which is compatible with 3D bioprinting processes. The ideal bioink is one which is biocompatible, biodegradable, can achieve high shape fidelity during and after the bioprinting process and can provide a microenvironment which can accommodate the formation of a functional biological tissue. This chapter will discuss the composition of multiple naturally derived bioinks and synthetic bioinks which are commonly used in 3D bioprinting. It will also discuss how bioink stiffness and composition can influence and direct cell fate. Finally, this chapter will demonstrate how bioinks have advanced the field of tissue engineering by highlighting some key strategies involving the 3D bioprinting of tissues such as bone, vascular tissue and liver.

Three-dimensional (3D) bioprinting technology has emerged as an ideal approach to address the challenges in regenerative dentistry by fabricating 3D tissue constructs with customized complex architecture. The dilemma with current dental treatments has led to the exploration of this technology in restoring and maintaining the function of teeth. This scoping review aims to explore 3D bioprinting technology together with the type of biomaterials and cells used for dental applications. Based on PRISMA-ScR guidelines, this systematic search was conducted by using the following databases: Ovid, PubMed, EBSCOhost and Web of Science. The inclusion criteria were (i) cell-laden 3D-bioprinted construct; (ii) intervention to regenerate dental tissue using bioink, which incorporates living cells or in combination with biomaterial; and (iii) 3D bioprinting for dental applications. A total of 31 studies were included in this review. The main 3D bioprinting technique was extrusion-based approach. Novel bioinks in use consist of different types of natural and synthetic polymers, decellularized extracellular matrix and spheroids with encapsulated mesenchymal stem cells, and have shown promising results for periodontal ligament, dentin, dental pulp and bone regeneration application. However, 3D bioprinting in dental applications, regrettably, is not yet close to being a clinical reality. Therefore, further research in fabricating ideal bioinks with implantation into larger animal models in the oral environment is very much needed for clinical translation.

Organoids provide more realistic in vitro models that closely mimic their corresponding tissues or organs. Three-dimensional (3D) bioprinting allows for precise control over cell distribution, arrangement, and the regulation of cell behavior and interactions, offering flexibility and repeatability. By combining 3D bioprinting with organoid culture, optimal extracellular matrix conditions can be created, better replicating the complex interactions between cells and their environment. The integration of 3D bioprinting amplifies the scalability of organoids, facilitating the development of large-scale organ models that significantly enhance research capabilities. This review explores the crucial role of 3D bioprinting in organoid development, emphasizing its contributions to improving organoid structure and functionality. We discuss how this innovative approach supports the creation of well-defined physiological microenvironments, aids in the development of functional organoids, and enables high-throughput cultivation. Furthermore, we highlight the advantages of optimizing bioprinting strategies and bioinks to advance organoid applications. In addition, we examine the importance of refining 3D bioprinting devices to enhance organoid fabrication and application. Finally, we outline strategies for leveraging 3D bioprinting to further advance organoid research and its implications for regenerative medicine and disease modeling.

Three-dimensional (3D) bioprinting offers great potential for the fabrication of complex 3D cell-laden constructs for clinical and research applications. The droplet formation process is the important first step in droplet-based 3D bioprinting, affecting the positional accuracy and printing fidelity. In this paper, the drop ejection behavior, thresholds for stable droplet generation, and formation of satellite drops are studied, under various ink properties, printing conditions, and input cell concentrations using a micro-valve-based 3D bioprinter. Three droplet ejection behaviors are identified under different conditions: an isolated stable droplet, satellites coalescing into a single droplet, and the presence of one/multiple satellites. The droplet state is represented by a phase diagram bounded by a dimensionless Z number (the inverse of the Ohnesorge number) and a jet Weber number, Wej, to define the printability of the utilized bioprinter. The printability range is defined as 2 < Z < 15 and 10 < Wej < 25 by considering characteristics, such as stable single droplet formability and sufficient drop falling velocity. There is no fatal damage on cells within this printability range. The results show there is no strong influence of an actuation system on droplet-based bioprinting printability. As the input cell concentration increases, the bioink's density and viscosity increases, and surface tension decreases, which, in turn, causes the Z number to slightly decrease. The change in the cell concentration (from 0 to 1×107 cells/ml), within a Newtonian bioink, has negligible impact on the droplet volume, falling velocity, drop ejection behavior, breakup time, and ligament length in microvalve-based bioprinting.

Three-dimensional (3D) bioprinting technology has emerged as a powerful biofabrication platform for tissue engineering because of its ability to engineer living cells and biomaterial-based 3D objects. Over the last few decades, droplet-based, extrusion-based, and laser-assisted bioprinters have been developed to fulfill certain requirements in terms of resolution, cell viability, cell density, etc. Simultaneously, various bio-inks based on natural–synthetic biomaterials have been developed and applied for successful tissue regeneration. To engineer more realistic artificial tissues/organs, mixtures of bio-inks with various recipes have also been developed. Taken together, this review describes the fundamental characteristics of the existing bioprinters and bio-inks that have been currently developed, followed by their advantages and disadvantages. Finally, various tissue engineering applications using 3D bioprinting are briefly introduced.

An insight on advances and applications of 3d bioprinting: A review

Cancer vasculogenesis is a pivotal focus of cancer research and treatment given its critical role in tumor development, metastasis, and the formation of vasculogenic microenvironments. Traditional approaches to investigating cancer vasculogenesis face significant challenges in accurately modeling intricate microenvironments. Recent advancements in three-dimensional (3D) bioprinting technology present promising solutions to these challenges. This review provides an overview of cancer vasculogenesis and underscores the importance of precise modeling. It juxtaposes traditional techniques with 3D bioprinting technologies, elucidating the advantages of the latter in developing cancer vasculogenesis models. Furthermore, it explores applications in pathological investigations, preclinical medication screening for personalized treatment and cancer diagnostics, and envisages future prospects for 3D bioprinted cancer vasculogenesis models. Despite notable advancements, current 3D bioprinting techniques for cancer vasculogenesis modeling have several limitations. Nonetheless, by overcoming these challenges and with technological advances, 3D bioprinting exhibits immense potential for revolutionizing the understanding of cancer vasculogenesis and augmenting treatment modalities.

In the era of personalised medicine, novel therapeutic approaches raise increasing hopes to address currently unmet medical needs by developing patient-customised treatments. Three-dimensional (3D) bioprinting is rapidly evolving and has the potential to obtain personalised tissue constructs and overcome some limitations of standard tissue engineering approaches. Bioprinting could support a wide range of biomedical applications, such as drug testing, tissue repair or organ transplantation. There is a growing interest for 3D bioprinting in the orthopaedic field, with remarkable scientific and technical advances. However, the full exploitation of 3D bioprinting in medical applications still requires efforts to anticipate the upcoming challenges in translating bioprinted products from bench to bedside. In this review we summarised current trends, advances and challenges in the application of 3D bioprinting for bone and cartilage tissue engineering. Moreover, we provided a detailed analysis of the applicable regulations through the 3D bioprinting process and an overview of available standards covering bioprinting and additive manufacturing.

3D bioprinting is a disruptive, computer-aided, and additive manufacturing technology that allows the obtention, layer-by-layer, of 3D complex structures. This technology is believed to offer tremendous opportunities in several fields including biomedical, pharmaceutical, and food industries. Several bioprinting processes and bio-ink materials have emerged recently. However, there is still a pressing need to develop low-cost sustainable bio-ink materials with superior qualities (excellent mechanical, viscoelastic and thermal properties, biocompatibility, and biodegradability). Marine-derived biomaterials, including polysaccharides and proteins, represent a viable and renewable source for bio-ink formulations. Therefore, the focus of this review centers around the use of marine-derived biomaterials in the formulations of bio-ink. It starts with a general overview of 3D bioprinting processes followed by a description of the most commonly used marine-derived biomaterials for 3D bioprinting, with a special attention paid to chitosan, glycosaminoglycans, alginate, carrageenan, collagen, and gelatin. The challenges facing the application of marine-derived biomaterials in 3D bioprinting within the biomedical and pharmaceutical fields along with future directions are also discussed.

Background: An autologous minimally manipulated homologous adipose tissue (AMHAT) fabricated using three-dimensional (3D) bioprinting has shown potential in the treatment of diabetic foot ulcers and other chronic wounds. Methods: This study assessed the efficacy of AMHAT fabricated using 3D bioprinting for treating skin defects after a wide excision of skin cancer lesions where primary closure was not feasible. A total of 10 consecutive patients were included. The wounds were photographed and analyzed using AI, and the fat tissue harvested from the lower abdomen was processed into an AMHAT patch using a 3D bioprinter. The wound area was measured weekly until complete wound closure was achieved. Results: The area of the skin defects ranged from 1.77-6.22 cm2, averaging 2.72 cm2. Complete closure was achieved in 3-5 (average, 4.2) weeks. The residual area decreased from 52% after 1 week to 3% after 4 weeks. The scar appearance was esthetically favorable, with one patient showing mild nostril asymmetry. Furthermore, patient-reported outcome evaluation using the Japanese version of SCAR-Q for postoperative scars showed a very high level of satisfaction. Conclusions: The 3D bioprinted AMHAT accelerates wound healing with minimal scarring, offering an important option for skin defects where esthetics are a priority.

Three-dimensional bioprinting adipose tissue and mammary Organoids feasible for artificial breast structure regeneration