Bioinks For Bioprinting Research Articles

Bioprinting PDLSC-Laden Collagen Scaffolds for Periodontal Ligament Regeneration.

Periodontitis and severe trauma are major causes of damage to the periodontal ligament (PDL). Repairing the native conditions of the PDL is essential for the stability of the tissue and its interfaces. Bioprinting periodontal ligament stem cells (PDLSCs) is an interesting approach to guide the regeneration of PDL and interfacial integration. Herein, a collagen-based bioink mimicking the native extracellular matrix conditions and carrying PDLSCs was tested to guide the periodontal ligament organization. The bioink was tested at two different concentrations (10 and 15 mg/mL) and characterized by swelling and degradation, microstructural organization, and rheological properties. The biological properties were assessed after loading PDLSCs into bioinks for bioprinting. The characterization was performed through cell viability, alizarin red assay, and expression for ALP, COL1A1, RUNX2, and OCN. The in vivo biocompatibility of the PDLSC-laden bioinks was verified using subcutaneous implantation in mice. Later, the ability of the bioprinted PDLSC-laden bioinks on dental root fragments to form PDL was also investigated in vivo in mice for 4 and 10 weeks. The bioinks demonstrated typical shear-thinning behavior, a porous microstructure, and stable swelling and degradation characteristics. Both concentrations were printable and provided suitable conditions for a high cell survival, proliferation, and differentiation. PDLSC-laden bioinks demonstrated biocompatibility in vivo, and the bioprinted scaffolds on the root surface evidenced PDLSC alignment, organization, and PDLSC migration to the root surface. The versatility of collagen-based bioinks provides native ECM conditions for PDLSC proliferation, alignment, organization, and differentiation, with translational applications in bioprinting scaffolds for PDL regeneration.

Collagen as Bioink for Bioprinting: A Comprehensive Review.

Biomaterials made using collagen are successfully used as a three-dimensional (3D) substrate for cell culture and considered to be promising scaffolds for creating artificial tissues. An important task that arises for engineering such materials is the simulation of physical and morphological properties of tissues, which must be restored or replaced. Modern additive technologies, including 3D bioprinting, can be applied to successfully solve this task. This review provides the latest evidence on advances of 3D bioprinting with collagen in the field of tissue engineering. It contains modern approaches for printing pure collagen bioinks consisting only of collagen and cells, as well as the obtained results from the use of pure collagen bioinks in different fields of tissue engineering.

Exploring the potential of supramolecular hydrogels as advanced bioinks for bioprinting and biomedical applications

Supramolecular hydrogels have emerged as versatile bioinks in tissue engineering, providing a promising avenue for constructing intricate and functional biological structures. This paper explores the significance of employing supramolecular hydrogels as advanced bioinks for three-dimensional bioprinting and various biomedical applications. Supramolecular hydrogels possess distinct and tunable characteristics attributed to the dynamic nature of supramolecular host–guest interactions alongside interactions based on DNA and peptides, which increases their significance in tissue engineering. These interactions are essential for enhancing the mechanical properties, injectability, printability, post-printing stability, and biocompatibility of hydrogels. Gelation kinetics and rheological properties can also be manipulated to suit specific printing techniques. Furthermore, these supramolecular interactions facilitate the incorporation of bioactive molecules to regulate cellular behavior and tissue development. These diverse interactions observed in supramolecular hydrogels underscore their ability to emulate the dynamic and responsive nature of the cell’s extracellular matrix, which fosters cell growth, adherence, and differentiation. This review specifically highlights the cucurbit[n]uril and cyclodextrin-based host–guest supramolecular hydrogels, as well as peptide and DNA-based supramolecular structures as advanced bioinks and brief examples of their applications in various biomedical fields. These advanced bioinks would drive the development of intricate tissue constructs with enhanced biomimicry and therapeutic potential in regenerative medicine.

Synthesis of Alginate/Collagen Bioink for Bioprinting Respiratory Tissue Models.

Synthesis of bioinks for bioprinting of respiratory tissue requires considerations related to immunogenicity, mechanical properties, printability, and cellular compatibility. Biomaterials can be tailored to provide the appropriate combination of these properties through the synergy of materials with individual pros and cons. Sodium alginate, a water-soluble polymer derived from seaweed, is a cheap yet printable biomaterial with good structural properties; however, it lacks physiological relevance and cell binding sites. Collagen, a common component in the extra cellular matrix of many tissues, is expensive and lacks printability; however, it is highly biocompatible and exhibits sites for cellular binding. This paper presents our study on the synthesis of bioinks from alginate and collagen for use in bioprinting respiratory tissue models. Bioinks were synthesized from 40 mg/mL (4%) alginate and 3 mg/mL (0.3%) collagen in varying ratios (1:0, 4:1, 3:1, 2:1, and 1:1); then examined in terms of rheological properties, printability, compressive, and tensile properties and cellular compatibility. The results illustrate that the ratio of alginate to collagen has a profound impact on bioink performance and that, among the examined ratios, the 3:1 ratio is the most appropriate for use in bioprinting respiratory tissue scaffolds.

Extrusion bioprinting of elastin‐containing bioactive double‐network tough hydrogels for complex elastic tissue regeneration

AbstractDespite recent advances in extrusion bioprinting of cell‐laden hydrogels, using naturally derived bioinks to biofabricate complex elastic tissues with both satisfying biological functionalities and superior mechanical properties is hitherto an unmet challenge. Here, we address this challenge with precisely designed biological tough hydrogel bioinks featuring a double‐network structure. The tough hydrogels consisted of energy‐dissipative dynamically crosslinked glycosaminoglycan hyaluronic acid (o‐nitrobenzyl‐grafted hyaluronic acid) and elastin through Schiff's base reaction, and free‐radically polymerized gelatin methacryloyl. The incorporation of elastin further improved the elasticity, stretchability (∼170% strain), and toughness (∼45 kJ m−3) of the hydrogels due to the random coiling structure. We used this novel class of hydrogel bioinks to bioprint several complex elastic tissues with good shape retention. Furthermore, in vitro and in vivo experiments also demonstrated that the existence of elastin in the biocompatible bioinks facilitated improved cell behaviors and biological functions of bioprinted tissues, such as cell spreading and phenotype maintenance as well as tissue regeneration. The results confirmed the potential of the elastin‐containing tough hydrogel bioinks for bioprinting of 3D complex elastic tissues with biological functionalities, which may find widespread applications in elastic tissue regeneration.

Microgels for bioprinting: recent advancements and challenges.

Microgels have become a popular and powerful structural unit in the bioprinting field due to their advanced properties, ranging from the tiny size and well-connected hydrogel (nutrient) network to special rheological properties. Different microgels can be fabricated by a variety of fabrication methods including bulk crushing, auxiliary dripping, multiphase emulsion, and lithography technology. Traditionally, microgels can encapsulate specific cells and are used for in vitro disease models and in vivo organ regeneration. Furthermore, microgels can serve as a drug carrier to realize controlled release of drug molecules. Apart from being used as an independent application unit, recently, these microgels are widely applied as a specific bioink component in 3D bioprinting for in situ tissue repair or building special 3D structures. In this review, we introduce different methods used to generate microgels and the microgel-based bioink for bioprinting. Besides, the further tendency of microgel development in future is introduced and predicted to provide guidance for related researchers in exploring more effective ways to fabricate microgels and more potential bioprinting application cases as multifunctional bioink components.

Evaluating cells metabolic activity of bioinks for bioprinting: the role of cell-laden hydrogels and 3D printing on cell survival.

Tissue engineering has advanced significantly in recent years, owing primarily to additive manufacturing technology and the combination of biomaterials and cells known as 3D cell printing or Bioprinting. Nonetheless, various obstacles remain developing adequate 3D printed structures for biomedical applications, including bioinks optimization to meet biocompatibility and printability standards. Hydrogels are among the most intriguing bioinks because they mimic the natural extracellular matrix found in connective tissues and can create a highly hydrated environment that promotes cell attachment and proliferation; however, their mechanical properties are weak and difficult to control, making it difficult to print a proper 3D structure. In this research, hydrogels based on Alginate and Gelatin are tested to evaluate the metabolic activity, going beyond the qualitative evaluation of cell viability. The easy-to-make hydrogel has been chosen due to the osmotic requirements of the cells for their metabolism, and the possibility to combine temperature and chemical crosslinking. Different compositions (%w/v) are tested (8% gel-7% alg, 4% gel-4% alg, 4% gel-2% alg), in order to obtain a 3D structure up to 10.3 ± 1.4mm. The goal of this paper is to validate the obtained cell-laden 3D structures in terms of cell metabolic activity up to 7days, further highlighting the difference between printed and not printed cell-laden hydrogels. To this end, MS5 cells viability is determined by implementing the live/dead staining with the analysis of the cellular metabolic activity through ATP assay, enhancing the evaluation of the actual cells activity over cells number. The results of the two tests are not always comparable, indicating that they are not interchangeable but provide complementary pieces of information.

Hydrogels for 3D bioprinting in tissue engineering and regenerative medicine: Current progress and challenges.

Three-dimensional (3D) bioprinting is a promising and innovative biomanufacturing technology, which can achieve precise position controlling of cells and extracellular matrix components, and further create complex and functional multi-cellular tissues or organs in a 3D environment. Bioink in the form of the cell-loaded hydrogel is most commonly used in bioprinting, and it is vital to the process of bioprinting. The bionic scaffold should possess suitable mechanical strength, biocompatibility, cell proliferation, survival, and other biological characteristics. The disadvantages of natural polymer hydrogel materials include poor mechanical properties as well as low printing performance and shape fidelity. Over the past years, a series of synthetic, modified, and nanocomposite hydrogels have been developed, which can interact through physical interactions, chemical covalent bond crosslinking, and bioconjugation reactions to change the characteristics to satisfy the requirements. In this review, a comprehensive summary is provided on recent research regarding the unique properties of hydrogel bioinks for bioprinting, with optimized methods and technologies highlighted, which have both high-value research significance and potential clinical applications. A critical analysis of the strengths and weaknesses of each hydrogel-based biomaterial ink is presented at the beginning or end of each section, alongside the latest improvement strategies employed by current researchers to address their respective shortcomings. Furthermore, we propose potential repair sites for each hydrogel-based ink based on their distinctive repair features, while reflecting on current research limitations. Finally, we synthesize and analyze expert opinions on the future of these hydrogel-based bioinks in the broader context of tissue engineering and regenerative medicine, offering valuable insights for future investigations.

Molecular Tuning of a Benzene-1,3,5-Tricarboxamide Supramolecular Fibrous Hydrogel Enables Control over Viscoelasticity and Creates Tunable ECM-Mimetic Hydrogels and Bioinks.

Traditional synthetic covalent hydrogels lack the native tissue dynamics and hierarchical fibrous structure found in the extracellular matrix (ECM). These dynamics and fibrous nanostructures are imperative in obtaining the correct cell/material interactions. Consequently, the challenge to engineer functional dynamics in a fibrous hydrogel and recapitulate native ECM properties remains a bottle-neck to biomimetic hydrogel environments. Here,the molecular tuning of a supramolecular benzene-1,3,5-tricarboxamide (BTA) hydrogelator via simple modulation of hydrophobic substituents is reported. This tuning results in fibrous hydrogels with accessible viscoelasticity over 5 orders of magnitude, while maintaining a constant equilibrium storage modulus. BTA hydrogelators are created with systematic variations in the number ofhydrophobic carbon atoms, and this is observed to control the viscoelasticity and stress-relaxation timescales in a logarithmic fashion. Some of theseBTA hydrogels are shear-thinning, self-healing, extrudable, and injectable, and can be 3D printed into multiple layers. These hydrogels show high cell viability for chondrocytes and human mesenchymal stem cells, establishing their use in tissue engineering applications. This simple molecular tuning by changing hydrophobicity (with just a few carbon atoms) provides precise control over the viscoelasticity and 3D printability in fibrillar hydrogels and can be ported onto other 1D self-assembling structures. The molecular control and design of hydrogel network dynamics can push the field of supramolecular chemistry towardthe design of new ECM-mimicking hydrogelators for numerous cell-culture and tissue-engineering applications and give access towardhighly biomimetic bioinks for bioprinting.

Recent Advances in Decellularized Extracellular Matrix-Based Bioinks for 3D Bioprinting in Tissue Engineering.

In recent years, three-dimensional (3D) bioprinting has been widely utilized as a novel manufacturing technique by more and more researchers to construct various tissue substitutes with complex architectures and geometries. Different biomaterials, including natural and synthetic materials, have been manufactured into bioinks for tissue regeneration using 3D bioprinting. Among the natural biomaterials derived from various natural tissues or organs, the decellularized extracellular matrix (dECM) has a complex internal structure and a variety of bioactive factors that provide mechanistic, biophysical, and biochemical signals for tissue regeneration and remodeling. In recent years, more and more researchers have been developing the dECM as a novel bioink for the construction of tissue substitutes. Compared with other bioinks, the various ECM components in dECM-based bioink can regulate cellular functions, modulate the tissue regeneration process, and adjust tissue remodeling. Therefore, we conducted this review to discuss the current status of and perspectives on dECM-based bioinks for bioprinting in tissue engineering. In addition, the various bioprinting techniques and decellularization methods were also discussed in this study.

Dynamic Alginate Hydrogel as an Antioxidative Bioink for Bioprinting.

3D bioprinting holds great potential for use in tissue engineering to treat degenerative joint disorders, such as osteoarthritis. However, there is a lack of multifunctional bioinks that can not only support cell growth and differentiation, but also offer protection to cells against injuries caused by the elevated oxidative stress; this conditions is a common characteristic of the microenvironment of the osteoarthritis disease. To mitigate oxidative stress-induced cellular phenotype change and malfunction, an anti-oxidative bioink derived from an alginate dynamic hydrogel was developed in this study. The alginate dynamic hydrogel gelated quickly via the dynamic covalent bond between the phenylboronic acid modified alginate (Alg-PBA) and poly (vinyl alcohol) (PVA). It presented good self-healing and shear-thinning abilities because of the dynamic feature. The dynamic hydrogel supported long-term growth of mouse fibroblasts after stabilization with a secondary ionic crosslinking between introduced calcium ions and the carboxylate group in the alginate backbone. In addition, the dynamic hydrogel showed good printability, resulting in the fabrication of scaffolds with cylindrical and grid structures with good structural fidelity. Encapsulated mouse chondrocytes maintained high viability for at least 7 days in the bioprinted hydrogel after ionic crosslinking. Most importantly, in vitro studies implied that the bioprinted scaffold could reduce the intracellular oxidative stress for embedded chondrocytes under H2O2 exposure; it could also protect the chondrocytes from H2O2-induced downregulation of extracellular matrix (ECM) relevant anabolic genes (ACAN and COL2) and upregulation of a catabolic gene (MMP13). In summary, the results suggest that the dynamic alginate hydrogel can be applied as a versatile bioink for the fabrication of 3D bioprinted scaffolds with an innate antioxidative ability; this technique is expected to improve the regenerative efficacy of cartilage tissues for the treatment of joint disorders.

Tunable metacrylated silk fibroin-based hybrid bioinks for the bioprinting of tissue engineering scaffolds.

Three-dimensional (3D) bioprinting is a powerful technique for the production of tissue-like structures to study cell behavior and tissue properties. A major challenge in 3D extrusion bioprinting is the limited diversity of bioinks, which fulfills the requirements of shear-thinning and strain recovery behaviors and can be solidified by a crosslinking process to retain their shape after printing. Herein, we aimed to develop a natural biopolymer-based formula with dual crosslinking performance to formulate a cell-laden bioink. In this study, methacrylate gelatin (GelMA) and methacrylated silk fibroin (SFMA) with different degrees of methacrylation were fabricated into hybrid bioinks. The GelMA/SFMA bioink of an optimal degree provides excellent rheological properties for extrusion bioprinting, and its hydrogel precursor polymer can form a polymer network at a low temperature and the high shape fidelity of the printed construct through photocrosslinking. Moreover, the hydrogel bioink can encapsulate different types of cells together to create 3D printed constructs that mimic the cellular microenvironment at a microscale level. Human umbilical vein endothelial cells (HUVECs) and rat pheochromocytoma (PC12) cells encapsulated in the 3D printed constructs can maintain high viability and proliferation ability for a long time. Furthermore, the GelMA/SFMA hydrogels were implanted in the subcutaneous tissue of SD rats for the evaluation of biocompatibility and degradability in vivo. Thus, the proposed GelMA/SFMA bioink expands the palette of available bioinks and offers opportunities for biomedical applications such as tissue engineering and soft robotics in clinical applications.

Formulation and characterization of gelatin methacrylamide-hydroxypropyl methacrylate based bioink for bioprinting applications

Three-dimensional (3D) bioprinting has emerged as a revolutionary technology for constructing functional tissue equivalents/scaffolds for tissue engineering applications. Bioink design is a crucial element in 3D bioprinting, which typically comprises a mixture of biomaterials, biological molecules or cells followed by its printing and tissue maturation. An ideal bioink should possess suitable physicochemical, mechanical, rheological, and biological features of the target tissue. However, mimicking multifaceted compositions similar to native extracellular matrix (ECM) with bioactive milieu of soluble and non-soluble factors is challenging. Herein, we report the formulation and characterization of a bioink system, comprising methacrylamide modified gelatin (GelMA) and 2-hydroxylpropyl methacrylate (HPMA) with a cost-effective redox initiators based cross-linking. GelMA was synthesized by reacting gelatin with methacrylic anhydride (MA) and subsequently, copolymerized with HPMA at room temperature by redox mechanism. Various hydrogel formulations by varying GelMA: HPMA w/v% ratios (G:HP) were studied as 10:0 (G100HP0), 9.5:0.5 (G95HP05), 9:1 (G90HP10), 8:2 (G80HP20), and 6:4 (G60HP40), to identify the best bioink composition. The formulations were characterized for its opacity, chemical, rheological, mechanical, porosity and swelling properties and cytocompatibility as per ISO-10993 standards. Cell encapsulation studies using live/dead assay analyzed cell viability inside the handprinted and 3D printed constructs. The preliminary results indicate successful formulation of cytocompatible bioink for potential 3D bioprinting and biofabrication applications.

Properties of Collagen/Sodium Alginate Hydrogels for Bioprinting of Skin Models

3D printing technology has great potential for the reconstruction of human skin. However, the reconstructed skin has some differences from natural skin, largely because the hydrogel used does not have the appropriate biological and physical properties to allow healing and regeneration. This study examines the swelling, degradability, microstructure and biological properties of Collagen/Sodium Alginate (Col/SA) hydrogels of differing compositions for the purposes of skin printing. Increasing the content of sodium alginate causes the hydrogel to exhibit stronger mechanical and swelling properties, a faster degradation ratio, smaller pore size, and less favorable biological properties. An optimal 1% collagen hydrogel was used to print bi-layer skin in which fibroblasts and keratinocytes showed improved spreading and proliferation as compared to other developed formulations. The Col/SA hydrogels presented suitable tunability and properties to be used as a bioink for bioprinting of skin aiming at finding applications as 3D models for wound healing research.

3D Bio-printing For Skin Tissue Regeneration: Hopes and Hurdles.

For many years, discovering the appropriate methods for the treatment of skin irritation has been challenging for specialists and researchers. Bio-printing can be extensively applied to address the demand for proper skin substitutes to improve skin damage. Nowadays, to make more effective biomimicry of natural skin, many research teams have developed cell-seeded bio-inks for bioprinting of skin substitutes. These loaded cells can be single or co-cultured in these structures. The present review gives a comprehensive overview of the methods, substantial parameters of skin bioprinting, examples of in vitro and in vivo studies, and current advances and challenges in skin tissue engineering.

3D-bioprinted Recombination Structure of Hertwig's Epithelial Root Sheath Cells and Dental Papilla Cells for Alveolar Bone Regeneration.

Three-dimensional (3D) bioprinting is an emerging method for tissue regeneration. However, promoting the epithelial-mesenchymal interaction (EMI), while maintaining the characteristics of epithelial cells has always been a challenge in tissue engineering. Since EMI acts as a critical factor in bone regeneration, this study aims to promote EMI by recombining epithelial and mesenchymal cells through 3D bioprinting. Hertwig’s epithelial root sheath (HERS) is a transient structure appeared in the process of tooth root formation. Its epithelial characteristics are easy to attenuate under appropriate culture environment. We recombined HERS cells and dental papilla cells (DPCs) through 3D bioprinting to simulate the micro-environment of cell-cell interaction in vivo. HERS cells and DPCs were mixed with gelatin methacrylate (GelMA) separately to prepare bio-inks for bioprinting. The cells/GelMA constructs were transplanted into the alveolar socket of Sprague-Dawley rats and then observed for 8 weeks. Hematoxylin and eosin staining, Masson staining, and immunohistochemical analysis showed that dimensional cultural pattern provided ideal environment for HERS cells and DPCs to generate mineralization texture and promote alveolar bone regeneration through their interactions. 3D bioprinting technology provides a new way for the co-culture of HERS cells and DPCs and this study is inspiring for future research on EMI model.

Harnessing electrostatic interactions for enhanced printability of alginate-based bioinks

3D bioprinting has received great attention in recent years because of its remarkable advantage in the fabrication of personalized biomaterial designs. In order to meet the full potential of bioprinting, it is necessary to develop new or improved bioinks with appropriate physicochemical properties supporting biological function. One potential bioink is sodium alginate (SA), an anionic polymer which can form an “egg-box” three-dimensional network complex with the introduction of cations. The rapid network formation allows for stabilization of extrudable alginate, making it suitable for use as a bioink in bioprinting applications. In this study, the complexation of positively charged chitosan (CS) and laponite (Lap) with negatively charged SA was investigated. 1-layer, 2-layer, and multi-layer models were established to evaluate and compare the printability of SA, SA-CS, and SA-Lap bioinks, including metrics of extrudability, filament stability, pore circularity, and structure fidelity. In addition, material properties of each bioink, including viscosity, compressive modulus, swelling rate, and biocompatibility, were assessed. The results showed the printability of SA was improved with the addition of CS or Lap. Moreover, the swelling rate of composite bioinks was reduced to half of that of sodium alginate counterparts. In addition, cell compatibility studies demonstrated that all bioinks were capable of maintaining cell proliferation over five days. As a whole, this work highlights the importance of electrostatic interactions of alginate-based bioinks for bioprinting.

Tunable Microgel‐Templated Porogel (MTP) Bioink for 3D Bioprinting Applications (Adv. Healthcare Mater. 8/2022)

Porous Bioinks for Bioprinting In article number 2200027 by Liliang Ouyang, Molly M. Stevens, and co-workers, a microgel-templated porogel bioink strategy is introduced to present micropores (10–100 µm) in the cell-laden building blocks for 3D bioprinting applications. It mediates the viscoelastic properties of a series of existing hydrogels and enhances embedded cell activity via better mass transport conditions.

Bioprinting of bioactive tissue scaffolds from ecologically-destructive fouling tunicates

Urochordates are the closest invertebrate relative to humans and commonly referred to as tunicates, a name ascribed to their leathery outer “tunic”. The tunic is the outer covering of the organism which functions as the exoskeleton and is rich in carbohydrates and proteins. Invasive or fouling tunicates pose a great threat to the indigenous marine ecosystem and governments spend several hundred thousand dollars for tunicate management, considering the huge adverse economic impact it has on the shipping and fishing industries. In this work, the environmentally destructive colonizing tunicate species of Polyclinum constellatum was successfully identified in the coast of Abu Dhabi and methods of sustainably using it as wound-dressing materials, decellularized extra-cellular matrix (dECM) scaffolds for tissue engineering applications and bioinks for bioprinting of tissue constructs for regenerative medicine are proposed. The intricate three-dimensional nanofibrous cellulosic networks in the tunic remain intact even after the multi-step process of decellularization and lyophilization. The lyophilized dECM tunics possess excellent biocompatibility and remarkable tensile modulus of 3.85 ± 0.93 MPa compared to ∼0.1–1 MPa of other hydrogel systems. This work demonstrates the use of lyophilized tunics as wound-dressing materials, having outperformed the commercial dressing materials with a capacity of absorbing 20 times its weight in the dry state. This work also demonstrates the biocompatibility of dECM scaffold and dECM-derived bioink (3D bioprinting with Mouse Embryonic Fibroblasts (MEFs)). Both dECM scaffolds and bioprinted dECM-based tissue constructs show enhanced metabolic activity and cell proliferation over time. Sustainable utilization of dECM-based biomaterials from ecologically-destructive fouling tunicates proposed in this work helps preserve the marine ecosystem, shipping and fishing industries worldwide, and mitigate the huge cost spent for tunicate management.

Collagen Bioinks for Bioprinting: A Systematic Review of Hydrogel Properties, Bioprinting Parameters, Protocols, and Bioprinted Structure Characteristics.

Bioprinting is a modern tool suitable for creating cell scaffolds and tissue or organ carriers from polymers that mimic tissue properties and create a natural environment for cell development. A wide range of polymers, both natural and synthetic, are used, including extracellular matrix and collagen-based polymers. Bioprinting technologies, based on syringe deposition or laser technologies, are optimal tools for creating precise constructs precisely from the combination of collagen hydrogel and cells. This review describes the different stages of bioprinting, from the extraction of collagen hydrogels and bioink preparation, over the parameters of the printing itself, to the final testing of the constructs. This study mainly focuses on the use of physically crosslinked high-concentrated collagen hydrogels, which represents the optimal way to create a biocompatible 3D construct with sufficient stiffness. The cell viability in these gels is mainly influenced by the composition of the bioink and the parameters of the bioprinting process itself (temperature, pressure, cell density, etc.). In addition, a detailed table is included that lists the bioprinting parameters and composition of custom bioinks from current studies focusing on printing collagen gels without the addition of other polymers. Last but not least, our work also tries to refute the often-mentioned fact that highly concentrated collagen hydrogel is not suitable for 3D bioprinting and cell growth and development.