Bioprinted Tissues Research Articles

Carbon-Based Nanoarchitectonics in Advancing Cardiac Tissue Bioprinting: A Review

Carbon-Based Nanoarchitectonics in Advancing Cardiac Tissue Bioprinting: A Review

Bioengineered skin-substitutes incorporating rete-ridges using a bioprinting process

Abstract Bioprinting is a widely used technique for creating three-dimensional, complex, and heterogeneous artificial tissue constructs that are biologically and biophysically similar to natural tissues. The skin is composed of several layers including the epidermis, basement membrane (BM), and dermis. However, the unique undulating structure of basement membranes (i.e., rete ridges) and the function of BM have not been extensively studied in the fabrication of engineered skin substitutes. In this study, a novel engineered skin substitute incorporating an artificially designed rete ridge (i.e., mogul-shape) was developed using bioprinting and bioinks prepared using collagen and fibrinogen. To mimic the structure of the rete ridges of skin tissue, we developed a modified bioprinting technique, controlling rheological property of bioink to create a mogul-shaped layer. In vitro cellular activities, including the expression of specific genes (those encoding vimentin, laminin-5, collagen IV, and cytokeratins), demonstrated that the engineered skin substitute exhibited more potent cellular responses than the normally bioprinted control owing to the favorable biophysical BM structure and the bioink microenvironment. Additionally, the feasibility of utilizing the bioprinted skin-structure was evaluated in a mouse model, and in vivo results demonstrated that the bioprinted skin substitutes effectively promoted wound healing capabilities. Based on these results, we suggest that bioprinted skin tissues and the bioprinting technique for mimicking rete ridges can be used not only as potential lab-chip models for testing cosmetic materials and drugs, but also as complex physiological models for understanding human skin.

3D Bioprinting of Thick Adipose Tissues with Integrated Vascular Hierarchies

AbstractEngineering functional, thick tissues with integrated vascular architectures is crucial for advancing regenerative medicine. This study applied a novel 3D bioprinting strategy to construct thick adipose tissues featuring complex, hierarchical vascular networks. Mature adipocytes and endothelial cells are encapsulated within a collagen matrix, with structural integrity and vascular functionality ensured through a combination of bioinks. An innovative approach is utilized to establish millimetric vascular channels that connect to microvascular networks, forming a fully perfusable, hierarchical vasculature throughout the 3D bioprinted tissue. The structural stability and endothelial functionality of the vascularized constructs are assessed under continuous flow. In‐vitro analyses confirmed the integrity of the vascular networks and demonstrated the functional characteristics of the printed mature adipose cells. Direct anastomosis of the bioprinted tissue to a rat femoral artery resulted in effective vascular integration and tissue viability, as evidenced by efficient blood perfusion and significant host vascular ingrowth within the implanted tissues. The findings highlighted the potential of the presented bioprinting technique in the development of fully functional, vascularized adipose tissues suitable for surgical reconstruction and regenerative medicine applications.

Remote-Controlled Gene Delivery in Coaxial 3D-Bioprinted Constructs using Ultrasound-Responsive Bioinks

IntroductionCoaxial 3D bioprinting has advanced the formation of tissue constructs that recapitulate key architectures and biophysical parameters for in-vitro disease modeling and tissue-engineered therapies. Controlling gene expression within these structures is critical for modulating cell signaling and probing cell behavior. However, current transfection strategies are limited in spatiotemporal control because dense 3D scaffolds hinder diffusion of traditional vectors. To address this, we developed a coaxial extrusion 3D bioprinting technique using ultrasound-responsive gene delivery bioinks. These bioink materials incorporate echogenic microbubble gene delivery particles that upon ultrasound exposure can sonoporate cells within the construct, facilitating controllable transfection.MethodsPhospholipid-coated gas-core microbubbles were electrostatically coupled to reporter transgene plasmid payloads and incorporated into cell-laden alginate bioinks at varying particle concentrations. These bioinks were loaded into the coaxial nozzle core for extrusion bioprinting with CaCl2 crosslinker in the outer sheath. Resulting bioprints were exposed to 2.25 MHz focused ultrasound and evaluated for microbubble activation and subsequent DNA delivery and transgene expression.ResultsCoaxial printing parameters were established that preserved the stability of ultrasound-responsive gene delivery particles for at least 48 h in bioprinted alginate filaments while maintaining high cell viability. Successful sonoporation of embedded cells resulted in DNA delivery and robust ultrasound-controlled transgene expression. The number of transfected cells was modulated by varying the number of focused ultrasound pulses applied. The size region over which DNA was delivered was modulated by varying the concentration of microbubbles in the printed filaments.ConclusionsOur results present a successful coaxial 3D bioprinting technique designed to facilitate ultrasound-controlled gene delivery. This platform enables remote, spatiotemporally-defined genetic manipulation in coaxially bioprinted tissue constructs with important applications for disease modeling and regenerative medicine.

Biofabrication of engineered tissues by 3D bioprinting of tissue specific high cell-density bioinks.

Bioprinting of high cell-density bioinks is a promising technique for cellular condensation-based tissue engineering and regeneration medicine. However, it remains difficult to create precisely controlled complex structures and organization of tissues with high cell-density bioink-based bioprinting for tissue specific condensation. In this study, we present newly biofabricated tissues from directly assembled, tissue specific, high cell-density bioinks which have been three-dimensionally printed into a photocrosslinkable and biodegradable hydrogel microparticle supporting bath. Three types of tissue specific high cell-density bioinks have been prepared with individual stem cells or stem cell aggregates by incorporation of growth factor-loaded gelatin microparticles. The bioprinted tissue specific high cell-density bioinks in the photocrosslinked microgel supporting bath condense together and differentiate down tissue-specific lineages to form multi-phase tissues (e.g., osteochondral tissues). By changing the growth factors and cell types, these tissue specific high cell-density bioinks enable engineering of various functional tissues with controlled architecture and organization of cells.

Innovations in Bioink Materials and 3D Bioprinting for Precision Tissue Engineering

The progress in bioink materials and 3D bioprinting techniques has created new opportunities in tissue engineering. The goal is to develop cellular implants that closely resemble native tissues in structure, function, and microenvironment, addressing challenges such as nutrient transport and mechanical support for tissue regeneration. This literature review evaluates current trends in bioink development and 3D bioprinting methodologies, assessing their effectiveness for enhancing tissue-engineered constructs for clinical applications. Recent studies were comprehensively analyzed, focusing on novel bioink formulations, optimization of 3D bioprinting processes, and evaluate on of printed constructs' mechanical and biological properties. Various fabrication techniques and their implications for tissue integration were examined. The review shows significant progress in bioink compositions that enhance cell viability and nutrient diffusion within printed scaffolds. Constructs demonstrated improved mechanical properties and biological functionality, enabling better integration with host tissues. In vivo studies highlighted the potential of these bioprinted tissues to support cellular activity and regeneration, signifying significant advancements in clinical viability. The findings emphasize the crucial role of bioink materials and bioprinting technology in advancing tissue engineering. Continual innovation in bioink formulation and printing techniques is essential to overcome current limitations and achieve widespread clinical application. Future research should focus on personalized bioink development and expanding the range of tissues that can be effectively engineered, ultimately improving patient outcomes in regenerative medicine.

A Futuristic Development in 3D Printing Technique Using Nanomaterials with a Step Toward 4D Printing.

3D bioprinting has shown great promise in tissue engineering and regenerative medicine for creating patient-specific tissue scaffolds and medicinal devices. The quickness, accurate imaging, and design targeting of this emerging technology have excited biomedical engineers and translational medicine researchers. Recently, scaffolds made from 3D bioprinted tissue have become more clinically effective due to nanomaterials and nanotechnology. Because of quantum confinement effects and high surface area/volume ratios, nanomaterials and nanotechnological techniques have unique physical, chemical, and biological features. The use of nanomaterials and 3D bioprinting has led to scaffolds with improved physicochemical and biological properties. Nanotechnology and nanomaterials affect 3D bioprinted tissue engineered scaffolds for regenerative medicine and tissue engineering. Biomaterials and cells that respond to stimuli change the structural shape in 4D bioprinting. With such dynamic designs, tissue architecture can change morphologically. New 4D bioprinting techniques will aid in bioactuation, biorobotics, and biosensing. The potential of 4D bioprinting in biomedical technologies is also discussed in this article.

Efficacy assessment of a novel pan-RAS inhibitor in KRAS-mutant and wild type colorectal 3D bioprinted organoid tumor tissue.

91 Background: Colorectal cancer (CRC) is the third leading type of cancer worldwide, with ~150,000 new cases in the US annually and a grim 14% 5-year survival for patients diagnosed at a late stage. A lack of treatment options leads to persistently poor prognosis for patients with advanced stage disease. KRAS mutations are well known drivers of CRC and other GI cancers. Multiple KRAS mutations occur in CRC, including G12D (34%), G12V (21%), G13D (20%), G12C (8%), and others (18%). Existing KRAS-targeted therapies have limited use in CRC, underscoring the need for pan-RAS inhibitors in treating CRC and other RAS driven cancers. Objective: Assess activity of ADT-007, our pan-RAS inhibitor, on wild-type (WT) and KRAS-mutant 3D bioprinted organoid tumor (BOT) tissue using our high-throughput ex vivo platform. Methods: Using previously established bioprinting protocols, WT and mutant BOTs were printed with HT29 and HCT116 cells, respectively. HT29 is an established human WT CRC cell line with known sensitivity to proteosome and survivin inhibitors. HCT116 is a KRASG13D mutant human CRC cell line. 3 sets of BOTs were generated and acclimated for 24h. One set was treated for 72h with proteosome inhibitor Bortezomib, another with survivin inhibitor YM155, and the third with our novel pan-RAS inhibitor ADT-007. Dose response curves were generated from both conventional ATP luminescence readouts and high-content imaging. Results: BOT tissue microarchitecture was validated and >200 µm diffusion in BOTs was confirmed using high-content imaging. Differential response was quantified using Cell TiterGlo endpoint assay as well as advanced image processing of high-content live/dead nuclear stained images captured at multiple z-plains. ADT-007 IC50 was found to be substantially lower for mutant HCT116 compared to that for WT HT29 cell line BOTs, which was consistent with separately conducted in vitro and in vivo studies. Conclusions: A pan-RAS inhibitor, such as ADT-007 with high selectivity for cancer cells with activated RAS that is not limited to a specific KRAS mutant allele or RAS isozyme, could have broader use for CRC and other RAS-driven cancers. Further, due to their potential to replicate biophysical characteristics of a tumor and its microenvironment, BOT based precision and personalized medicine platforms can provide more accurate drug efficacy readout compared to in vitro cancer models.

Bioprinting of osteochondral tissues: A perspective on current gaps and future trends.

Osteochondral tissue regeneration has remained a critical challenge in orthopaedic surgery, especially due to complications of arthritic degeneration arising out of mechanical dislocations of joints. The common gold standard of autografting has several limitations in presenting tissue engineering strategies to solve the unmet clinical need. However, due to the complexity of joint anatomy, and tissue heterogeneity at the interface, the conventional tissue engineering strategies have certain limitations. The advent of bioprinting has now provided new opportunities for osteochondral tissue engineering. Bioprinting can uniquely mimic the heterogeneous cellular composition and anisotropic extra-cellular matrix (ECM) organization, while allowing for targeted gene delivery to achieve heterotypic differentiation. In this perspective, we discuss the current advances made towards bioprinting of composite osteochondral tissues and present an account of challenges—in terms of tissue integration, long-term survival, and mechanical strength at the time of implantation—required to be addressed for effective clinical translation of bioprinted tissues. Finally, we highlight some of the future trends related to osteochondral bioprinting with the hope of in-clinical translation.

Patterning of tissue spheroids biofabricated from human fibroblasts on the surface of electrospun polyurethane matrix using 3D bioprinter

Organ printing is a computer-aided additive biofabrication of functional three-dimensional human tissue and organ constructs according to digital model using the tissue spheroids as building blocks. The fundamental biological principle of organ printing technology is a phenomenon of tissue fusion. Closely placed tissue spheroids undergo tissue fusion driven by surface tension forces. In order to ensure tissue fusion in the course of post-printing, tissue spheroids must be placed and maintained close to each other. We report here that tissue spheroids biofabricated from primary human fibroblasts could be placed and maintained on the surface of biocompatible electrospun polyurethane matrix using 3D bioprinter according to desirable pattern. The patterned tissue spheroids attach to polyurethane matrix during several hours and became completely spread during several days. Tissue constructions biofabricated by spreading of patterned tissue spheroids on the biocompatible electrospun polyurethane matrix is a novel technological platform for 3D bioprinting of human tissue and organs.

Three-Dimensional Bioprinting: A Comprehensive Review for Applications in Tissue Engineering and Regenerative Medicine.

Since three-dimensional (3D) bioprinting has emerged, it has continuously to evolved as a revolutionary technology in surgery, offering new paradigms for reconstructive and regenerative medical applications. This review highlights the integration of 3D printing, specifically bioprinting, across several surgical disciplines over the last five years. The methods employed encompass a review of recent literature focusing on innovations and applications of 3D-bioprinted tissues and/or organs. The findings reveal significant advances in the creation of complex, customized, multi-tissue constructs that mimic natural tissue characteristics, which are crucial for surgical interventions and patient-specific treatments. Despite the technological advances, the paper introduces and discusses several challenges that remain, such as the vascularization of bioprinted tissues, integration with the host tissue, and the long-term viability of bioprinted organs. The review concludes that while 3D bioprinting holds substantial promise for transforming surgical practices and enhancing patient outcomes, ongoing research, development, and a clear regulatory framework are essential to fully realize potential future clinical applications.

Optimizing Printhead Design for Enhanced Temperature Control in Extrusion-Based Bioprinting.

This study addresses critical challenges in the field of tissue engineering, specifically in the optimization of bioprinting technologies for the construction of complex, multicellular tissues. By utilizing a homemade piston-driven extrusion-based bioprinting (EBB) printhead, we performed detailed thermal and flow analyses to investigate the effects of temperature variations on the extrusion process of temperature-sensitive gelatin-alginate bioink. Through finite element method (FEM) simulations, we explored the temperature distribution within the printhead and its impact on bioink properties, such as viscosity, pressure, and shear stress. Key findings reveal significant temperature gradients from the printhead barrel to the nozzle tip, influencing bioink extrusion and filament morphology. This study further introduces an innovative hardware optimization with thermal insulators, designed to mitigate heat loss at the nozzle tip and ensure uniform temperature distribution. Both simulation and empirical printing experiments confirm the efficacy of thermal insulators in enhancing bioprinting fidelity and efficiency. This research contributes to the advancement of bioprinting technology by optimizing printhead design, with implications for improving the quality of bioprinted tissues and organs.

GPU-accelerated OCT imaging: Real-time data processing and artifact suppression for enhanced monitoring of 3D bioprinted tissues and vascular-like networks

Optical coherence tomography (OCT) imaging technology has significant advantages in in situ and noninvasive monitoring of biological tissues. However, it still faces the following challenges: including data processing speed, image quality, and improvements in three-dimensional (3D) visualization effects. OCT technology, especially functional imaging techniques like optical coherence tomography angiography (OCTA), requires a long acquisition time and a large data size. Despite the substantial increase in the acquisition speed of swept source optical coherence tomography (SS-OCT), it still poses significant challenges for data processing. Additionally, during in situ acquisition, image artifacts resulting from interface reflections or strong reflections from biological tissues and culturing containers present obstacles to data visualization and further analysis. Firstly, a customized frequency domain filter with anti-banding suppression parameters was designed to suppress artifact noises. Then, this study proposed a graphics processing unit (GPU)-based real-time data processing pipeline for SS-OCT, achieving a measured line-process rate of 800[Formula: see text]kHz for 3D fast and high-quality data visualization. Furthermore, a GPU-based real-time data processing for CC-OCTA was integrated to acquire dynamic information. Moreover, a vascular-like network chip was prepared using extrusion-based 3D printing and sacrificial materials, with sacrificial material being printed at the desired vascular network locations and then removed to form the vascular-like network. OCTA imaging technology was used to monitor the progression of sacrificial material removal and vascular-like network formation. Therefore, GPU-based OCT enables real-time processing and visualization with artifact suppression, making it particularly suitable for in situ noninvasive longitudinal monitoring of 3D bioprinting tissue and vascular-like networks in microfluidic chips.

Prospects of emerging 3D bioprinting technologies: major startup companies and regulatory issues for human use—part II

Abstract In its highly developed form, the evolving three-dimensional (3D) bioprinting technology aims to create 3D structures with living cells to mimic real tissue and organ functions. It would offer significant benefits across research, personalized medicine, and multiple other applications when adequately developed for human medicine. Presently, more technological activities are witnessed in North America, followed by Europe, Asia Pacific countries, Israel among the Middle East countries, and some South American countries. Around 75 commercial companies are active in 3D bioprinting, with only about a dozen making significant commercial progress. This number is expected to rise phenomenally as breakthroughs in manufacturing and the safe use of 3D bioprinted tissues and organs emerge. Legal frameworks for 3D bioprinting will likely be established, incorporating additions to existing drug laws once countries like the United States of America authorize using 3D bioprinted products in personalized medicine. The demand for 3D bioprinting products is rising based on expectations of future benefits. Therefore, intense research and development activities are ongoing, resulting in demands for the supply of research materials. The legal framework still needs to be put in place for the commercial use of 3D bioprinted tissues and organs in personalized medicine; therefore, laws are to be created for their safe use. This review provides a flavor of the evolution of 3D bioprinting startup companies globally using these technologies.

Bridging the Gap: Integrating 3D Bioprinting and Microfluidics for Advanced Multi-Organ Models in Biomedical Research.

Recent advancements in 3D bioprinting and microfluidic lab-on-chip systems offer promising solutions to the limitations of traditional animal models in biomedical research. Three-dimensional bioprinting enables the creation of complex, patient-specific tissue models that mimic human physiology more accurately than animal models. These 3D bioprinted tissues, when integrated with microfluidic systems, can replicate the dynamic environment of the human body, allowing for the development of multi-organ models. This integration facilitates more precise drug screening and personalized therapy development by simulating interactions between different organ systems. Such innovations not only improve predictive accuracy but also address ethical concerns associated with animal testing, aligning with the three Rs principle. Future directions include enhancing bioprinting resolution, developing advanced bioinks, and incorporating AI for optimized system design. These technologies hold the potential to revolutionize drug development, regenerative medicine, and disease modeling, leading to more effective, personalized, and humane treatments.

Growing Role of 3D In Vitro Cell Cultures in the Study of Cellular and Molecular Mechanisms: Short Focus on Breast Cancer, Endometriosis, Liver and Infectious Diseases.

Over the past decade, the development of three-dimensional (3D) models has increased exponentially, facilitating the unravelling of fundamental and essential cellular mechanisms by which cells communicate with each other, assemble into tissues and organs and respond to biochemical and biophysical stimuli under both physiological and pathological conditions. This section presents a concise overview of the most recent updates on the significant contribution of different types of 3D cell cultures including spheroids, organoids and organ-on-chip and bio-printed tissues in advancing our understanding of cellular and molecular mechanisms. The case studies presented include the 3D cultures of breast cancer (BC), endometriosis, the liver microenvironment and infections. In BC, the establishment of 3D culture models has permitted the visualization of the role of cancer-associated fibroblasts in the delivery of exosomes, as well as the significance of the physical properties of the extracellular matrix in promoting cell proliferation and invasion. This approach has also become a valuable tool in gaining insight into general and specific mechanisms of drug resistance. Given the considerable heterogeneity of endometriosis, 3D models offer a more accurate representation of the in vivo microenvironment, thereby facilitating the identification and translation of novel targeted therapeutic strategies. The advantages provided by 3D models of the hepatic environment, in conjunction with the high throughput characterizing various platforms, have enabled the elucidation of complex molecular mechanisms underlying various threatening hepatic diseases. A limited number of 3D models for gut and skin infections have been developed. However, a more profound comprehension of the spatial and temporal interactions between microbes, the host and their environment may facilitate the advancement of in vitro, ex vivo and in vivo disease models. Additionally, it may pave the way for the development of novel therapeutic approaches in diverse research fields. The interested reader will also find concluding remarks on the challenges and prospects of using 3D cell cultures for discovering cellular and molecular mechanisms in the research areas covered in this review.

Nanomaterials-Based Hybrid Bioink Platforms in Advancing 3D Bioprinting Technologies for Regenerative Medicine.

3D bioprinting is recognized as the ultimate additive biomanufacturing technology in tissue engineering and regeneration, augmented with intelligent bioinks and bioprinters to construct tissues or organs, thereby eliminating the stipulation for artificial organs. For 3D bioprinting of soft tissues, such as kidneys, hearts, and other human body parts, formulations of bioink with enhanced bioinspired rheological and mechanical properties were essential. Nanomaterials-based hybrid bioinks have the potential to overcome the above-mentioned problem and require much attention among researchers. Natural and synthetic nanomaterials such as carbon nanotubes, graphene oxides, titanium oxides, nanosilicates, nanoclay, nanocellulose, etc. and their blended have been used in various 3D bioprinters as bioinks and benefitted enhanced bioprintability, biocompatibility, and biodegradability. A limited number of articles were published, and the above-mentioned requirement pushed us to write this review. We reviewed, explored, and discussed the nanomaterials and nanocomposite-based hybrid bioinks for the 3D bioprinting technology, 3D bioprinters properties, natural, synthetic, and nanomaterial-based hybrid bioinks, including applications with challenges, limitations, ethical considerations, potential solution for future perspective, and technological advancement of efficient and cost-effective 3D bioprinting methods in tissue regeneration and healthcare.

Unlocking the Future: Bioprinting Salivary Glands-From Possibility to Reality.

Salivary gland biofabrication represents a promising avenue in regenerative medicine, aiming to address the challenges of salivary gland dysfunction caused by various factors such as autoimmune diseases and radiotherapy. This review examines the current state of bioprinting technology, biomaterials, and tissue engineering strategies in the context of creating functional, implantable salivary gland constructs. Key considerations include achieving vascularization for proper nutrient supply, maintaining cell viability and functionality during printing, and promoting tissue maturation and integration with surrounding tissues. Despite the existing challenges, recent advancements offer significant potential for the development of personalized therapeutic options to treat salivary gland disorders. Continued research and innovation in this field hold the potential to revolutionize the management of salivary gland conditions, improving patient outcomes and quality of life. This systematic review covers publications from 2018 to April 2024 and was conducted on four databases: Google Scholar, PubMed, EBSCOhost, and Web of Science. The key features necessary for the successful creation, implantation and functioning of bioprinted salivary glands are addressed.

Magnetically Actuated GelMA‐Based Scaffolds as a Strategy to Generate Complex Bioprinted Tissues

AbstractThe 3D bioprinting of complex structures has attracted particular attention in recent years and has been explored in several fields, including dentistry, pharmaceutical technology, medical devices, and tissue/organ engineering. However, it still possesses major challenges, such as decreased cell viability due to the prolongation of the printing time, along with difficulties in preserving the print shape. The 4D bioprinting approach, which is based on controlled shape transformation upon stimulation after 3D bioprinting, is a promising innovative method to overcome these difficulties. Herein, the generation of skeletal muscle tissue‐like complex structures is demonstrated by 3D bioprinting of GelMA‐based C2C12 mouse myoblast‐laden bio‐ink on a polymeric magnetic actuator that enables on‐demand shape transformation (i.e., rolling motion) under a magnetic field. Bioprinted scaffolds are used in both unrolled (open as control) and rolled forms. The results indicate that C2C12s remain viable upon controlled shape transformation, and functional myotube formation is initiated by the 7th day within bioprinted platforms. Moreover, when the rolled and open groups are compared regarding MyoD1 staining intensity, the rolled one enhanced MyoD1 expression. These results provide a promising methodology for generating complex structures with a simple magnetic actuation procedure for the bioprinting of tissue‐engineered constructs with enhanced cell viability and functionality.

Cellular Patterning Alone Using Bioprinting Regenerates Articular Cartilage Through Native-Like Cartilagenesis.

Few studies have proved that bioprinting itself helps recapitulate native tissue functions mainly because the bioprinted macro shape can rarely, if ever, influence cell function. This can be more problematic in bioprinting cartilage, generally considered more challenging to engineer. Here a new method is shown to micro-pattern chondrocytes within bioprinted sub-millimeter micro tissues, denoted as patterned micro-articular-cartilages tissues (PA-MCTs). Under the sole influence of bioprinted cellular patterns. A pattern scoring system is developed after over 600 bioprinted cellular patterns are analyzed. The top-scored pattern mimics that of the isogenous group in native articular cartilage. Under the sole influence of this pattern during PA-MCTs bio-assembling into macro-cartilage and repairing cartilage defects, chondrogenic cell phenotype is preserved, and cartilagenesis is initiated and maintained. Neocartilage tissues from individual and assembled PA-MCTs are comparable to native articular cartilage and superior to cartilage bioprinted with homogeneously distributed cells in morphology, biochemical components, cartilage-specific protein and gene expression, mechanical properties, integration with host tissues, zonation forming and stem cell chondrogenesis. PA-MCTs can also be used as osteoarthritic and healthy cartilage models for therapeutic drug screening and cartilage development studies. This cellular patterning technique can pave a new way for bioprinting to recapitulate native tissue functions via tissue genesis.