RESUMEN
Encapsulation of live cells in protective, semipermeable microcapsules is one of the kernel techniques for in vitro tissue regeneration, cell therapies, and pharmaceutical screening. Advanced fabrication techniques for cell encapsulation have been developed to meet different requirements. Existing cell encapsulation techniques place substantial constraints on the spatial patterning of live cells as well as on the compartmentalization of heterotypic cells. Alginate-Poly-L-lysine-alginate (APA) microcapsules that use sodium alginate as the polyanion and poly-L-lysine (PLL) as the polycation have been extensively employed for cell microencapsulation due to their excellent biocompatibility and biodegradability. This study proposes a novel method for developing programmable Janus APA microcapsules with variable shapes and sizes by using electrodeposition. By the versatile design of the microelectrode device, sequential electrodeposition is triggered to electro-address the cells at specific locations immobilized within a Janus APA microcapsule. The osteogenesis is evaluated by resembling cell compartmentalized and vascularized osteoblast-laden constructs. This technique allows precise spatial patterning of heterotypic cells inside the APA microcapsule, enabling the observation of cellular growth, interactions, and differentiation in a well-controlled chemical and mechanical microenvironment.
Asunto(s)
Galvanoplastia , Polilisina , Alginatos , Cápsulas , Polilisina/análogos & derivadosRESUMEN
Tissue-engineered periosteum substitutes (TEPSs) incorporating hierarchical architecture with osteoprogenitor and vascular niches are drawing much attention as a promising tool to support functional cells in defined zones and nourish the cortical bone. Current TEPSs usually lack technologies to closely observe cell performance, especially at the cell contact interface between distinct compartments containing defined biological configurations and functions. Here, an electrodeposition strategy is reported, which enables the selective formation of TEPSs with osteoprogenitor and vascular niches in a multiphasic scaffold in combination with different human cell types for cartilage regeneration in an in vivo osteochondral defect model. Human umbilical vein endothelial cells (HUVECs), dermal fibroblasts (HDFs), and bone marrow mesenchymal stem cells (hMSCs) are used to mirror both the vascular and osteogenic niches, respectively. It is observed that the intrinsic viscoelastic nature of the porous solid matrix is essential to successfully induce angiogenesis. Coculture of hMSCs with functional cells (HUVECs/HDFs) in TEPSs also effectively promoted periosteal regeneration, including osteogenic and angiogenic processes. The osteoarthritis cartilage histopathology assessment and histologic/histochemical grading system data indicate that the TEPSs containing hMSCs/HUVECs/HDFs exhibit superior potential for cartilage regeneration.
Asunto(s)
Osteogénesis , Ingeniería de Tejidos , Humanos , Células Endoteliales de la Vena Umbilical Humana , Técnicas de Cocultivo , Diferenciación Celular , Cartílago , Periostio , Andamios del Tejido , Regeneración ÓseaRESUMEN
Heterogeneous spheroids that mimic the complex three-dimensional environment of natural tissues are needed in various biomedical applications. Geometric cues from cellular matrix play invaluable roles in governing cell behavior and phenotype. However, the structural complexity of interior morphologies of spheroids is currently limited due to poor spatial resolution of positioning/orientation of cellular constructs. Here, a coaxial capillary microfluidic device is developed to generate gelatin methacrylate (GelMA) microspheres with tunable dimensions and interior morphologies, such as core-shell, or microspheres with interior undulated wavy, or spiral canals, by manipulating the two-phase flow of hydrogel precursor solution and methylcellulose solution. The formation of diverse and exquisite interior morphologies is caused by the interacting viscous instabilities of the two-phase flow in the microfluidic system, followed by water-in-oil emulsion and photo-initiated polymerization. Polyethylene glycol diacrylate (PEGDA) is incorporated into the GelMA solution to tune the mechanical properties of the fabricated microspheres, and an optimized concentration of PEGDA is confirmed by evaluating thein vitroproliferation and vascularization of human umbilical vein endothelial cells. Further, a heterogeneous spheroid with spiral blood vessel lumen is constructed to demonstrate the versatility and potential of the proposed droplet-based microfluidic approach for building functional tissue constructs.
Asunto(s)
Gelatina , Microfluídica , Gelatina/química , Células Endoteliales de la Vena Umbilical Humana , Humanos , Hidrogeles/química , Microesferas , Esferoides CelularesRESUMEN
Internal gelation has been an important sol-gel route for the preparation of spherical microgel for drug delivery, cell therapy, or tissue regeneration. Despite high homogeneity and permeability, the internal gelated microgels often result in weak mechanical stability, unregular interface morphology and low cell survival rate. In this work, we have extensively improved the existing internal gelation approach and core-shell hydrogel microcapsules (200-600µm) with a smooth surface, high mechanical stability and cell survival rate, are successfully prepared by using internal gelation. A coaxial flow-focusing capillary-assembled microfluidic device was developed for the gelation. Rapid gelling behavior of alginate in the internal gelation makes it suitable for producing well-defined and homogenous alginate hydrogel microstructures that serve as the shell of the microcapsules. 2-[4-(2-Hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) was used in the shell stream during the internal gelation. Thus, a high concentration of acid in the oil solution can be used for better crosslinking the alginate while maintaining high cell viability. We further demonstrated that the gelation conditions in our approach were mild enough for encapsulating HepG2 cells and 3T3 fibroblasts without losing their viability and functionality in a co-culture environment.