Vascularized cardiac tissue construction with orientation by layer-by-layer method and 3D printer

Abstract

Herein, we report the fabrication of native organ-like three-dimensional (3D) cardiac tissue with an oriented structure and vascular network using a layer-by-layer (LbL), cell accumulation and 3D printing technique for regenerative medicine and pharmaceutical applications. We firstly evaluated the 3D shaping ability of hydroxybutyl chitosan (HBC), a thermoresponsive polymer, by using a robotic dispensing 3D printer. Next, we tried to fabricate orientation-controlled 3D cardiac tissue using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) and normal human cardiac fibroblasts (NHCF) coated with extracellular matrix (ECM) nanofilms by layer-by-layer technique. these cells were seeded in the fabricated rectangular shape HBc gel frame. After cultivation of the fabricated tissue, fluorescence staining of the cytoskeleton revealed that hiPSC-CM and NHCF were aligned in one direction. Moreover, we were able to measure its contractile behavior using a video image analysis system. these results indicate that orientation-controlled cardiac tissue has more remarkable contractile function than uncontrolled cardiac tissue. finally, co-culture with human cardiac microvascular endothelial cells (HMVEC) successfully provided a vascular network in orientationcontrolled 3D cardiac tissue. The constructed 3D cardiac tissue with an oriented structure and vascular network would be a useful tool for regenerative medicine and pharmaceutical applications.

Advances in 3D bioprinting technology for cardiac tissue engineering and regeneration

“… Vascularization and contractility of the fabricated unit can be simultaneously possessed. [ 69 ] Needle-arrayed system None Sphere-like cell population composed of human iPSC-derived CM, fibroblast and umbilical vein endothelial cell (HUVEC) Wnt signaling activator The direct-write bioprinted cardiac patch was cultured in vitro with shaking for 72 h to obtain synchronized beating function, followed by being implanted to the infarcted area with pull-up omentum as a fixture of nude mice. Compared with the control group, the infarct area in the myocardial infarction area decreased, while its angiogenesis and cardiac ejection fraction increased.…”

“…Before the official printing operation, the preparation of active cardiac-bioink cells could be divided into three steps, namely, acquisition, cultivation or induction, and encapsulation. Currently, the practicable heart cell source can be human, mouse, or rat, and the first-hand genre covers the primary cells (such as primary rat ventricular CMs and human coronary artery endothelial cells), the cell lines (such as H9C2), or the iPSCs and other progenitor cells derived from iPSCs [ [67] , [68] , [69] ]. Because of the cell growth proficiency of the first-hand genre in vitro, the 2D culture systems for cell expansion are generally experienced, while the methods towards of stem cell differentiation and somatic cell trans -differentiation tend to be explorative and optimizable [ 70 , 71 ].…”

“…Another 3D-CTE bioink preparation method that can additionally increase the systematic mechanical property and/or bio-activity is the input of a hydrogel dECM [ 93 ], which is also a frontier hotspot in this field [ 94 , 95 ]. As a cofactor for the encapsulated living heart cells, a pre-printed cardiac dECM can be either processed solely with the biomaterial as a scaffold source [ 70 ] or mixed with the cells [ 69 , 85 ]. Similar to the source of instrumental cells, the preparation of a cardiac dECM reproducing the original physical and mechanical microenvironment of the heart can be different, which is mostly determined by the species and myocardial regions according to the specific requirements of 3D bioprinting applications [ 96 , 97 ].…”

Application of additive manufacturing in customized titanium mandibular implants for patients with oral tumors

“…Using digital medical imaging technology and metal 3D printing technology, according to CT scan data of patients, porous titanium alloy bone tissue substitute materials that are similar in shape, size and weight to the bone defect area can be designed and manufactured to achieve personalized repair and reconstruction of bone defects (30). At present, this technology has been applied in the field of hip, knee, spine and skull reconstruction (31,32); however, to the best of our knowledge, there are limited studies reporting its use in oral and maxillofacial defects (33). The present study included 4 patients with individualized titanium alloy implants and the outcomes were compared with 16 patients who received traditional reconstructed titanium plates and vascularized fibula free flaps through a retrospective controlled study.…”

In Vitro Modeling of Non-Solid Tumors: How Far Can Tissue Engineering Go?

“…In vivo, individual cells are harbored in specific niches where they integrate many external cues (including those that arise from extracellular matrix (ECM), mechanical stimulation and soluble signals from adjacent and distant cells) to generate a basal phenotype and respond to perturbations in their environment. The development of 3D platforms with well-defined architectures resembling native cellular environments has contributed to significant advances, among other tissues, in liver or heart modeling [ 6 , 7 , 8 ]. The integration of three dimensionality, multi-cellular interactions, patient-specific polymorphisms, fine control of chemical parameters (pH, oxygen level, biochemical gradients) and ECM composition are the main assets of this engineered tissues [ 4 , 9 , 10 ].…”