In this study, we propose a new cell encapsulation method consisting of a dispensing method and an aerosol-spraying method. The aerosol spray using a cross-linking agent, calcium chloride (CaCl(2)), was used to control the surface gelation of dispensed alginate struts during dispensing. To show the feasibility of the method, we used preosteoblast (MC3T3-E1) cells. By changing the relationship between the various dispensing/aerosol-spraying conditions and cell viability, we could determine the optimal cell-dispensing process: a nozzle size (240 μm) and an aerosol spray flow rate (0.93 ± 0.12 mL min(-1)), 10 mm s(-1) nozzle moving speed, a 10 wt % concentration of CaCl(2) in the aerosol solution, and 2 wt % concentration of CaCl(2) in the second cross-linking process. Based on these optimized process conditions, we successfully fabricated a three-dimensional, pore-structured, cell-laden alginate scaffold of 20 × 20 × 4.6 mm(3) and 84% cell viability. During long cell culture periods (16, 25, 33, and 45 days), the preosteoblasts in the alginate scaffold survived and proliferated well.
A new cell-printed scaffold consisting of poly(ϵ-caprolactone) (PCL) and cell-embedded alginate struts is designed. The PCL and alginate struts are stacked in an interdigitated pattern in successive layers to acquire a three-dimensional (3D) shape. The hybrid scaffold exhibits a two-phase structure consisting of cell (MC3T3-E1)-laden alginate struts able to support biological activity and PCL struts able to provide controllable mechanical support of the cell-laden alginate struts. The hybrid scaffolds exhibit an impressive increase in tensile modulus and maximum strength compared to pure alginate scaffolds. Laden cells are homogeneously distributed throughout the alginate struts and the entire scaffold, resulting in cell viability of approximately 84%.
The goal of tissue engineering is to repair or regenerate damaged tissue using a combination of cellular biology and materials engineering techniques. One of the challenging problems in tissue engineering is the development of a reproducible three-dimensional (3D) scaffold to support cell migration and infiltration. Although natural polymers, such as dissolved collagen or alginate, are considered ideal for this purpose, their hydrophilic properties have hindered the fabrication of designed 3D scaffold structures. To overcome this problem, we developed a novel system for the cryogenic plotting of 3D scaffolds. Using this technique, we created various 3D collagen scaffolds with designed pore structures that exhibited desired properties. The diameter of the individual collagen strands, which varied from 250 mm to 500 mm, was reproducibly dependent on processing parameters, and the final collagen scaffold showed little shrinkage (less than 12%) relative to the initial design. To evaluate the fabricated scaffold, we adapted the scaffold to regenerate skin tissue. Immunohistochemical analysis demonstrated that co-cultured keratinocytes and fibroblasts completely migrated throughout the 3D collagen scaffold and keratinocytes were well differentiated on the surface of scaffold like a human skin.
Biomedical scaffolds used in bone tissue engineering should have various properties including appropriate bioactivity, mechanical strength, and morphologically optimized pore structures. Collagen has been well known as a good biomaterial for various types of tissue regeneration, but its usage has been limited due to its low mechanical property and rapid degradation. In this work, a new hybrid scaffold consisting of polycaprolactone (PCL) and collagen is proposed for bone tissue regeneration. The PCL enhances the mechanical properties of the hybrid scaffold and controls the pore structure. Layered collagen nanofibers were used to enhance the initial cell attachment and proliferation. The results showed that the hybrid scaffold yielded better mechanical properties of pure PCL scaffold as well as enhanced biological activity than the pure PCL scaffold did. The effect of pore size on bone regeneration was investigated using two hybrid scaffolds with pore sizes of 200 ± 20 and 300 ± 27 μm. After post-seeding for 7 days, the cell proliferation with pore size, 200 ± 20 μm, was greater than that with pore size, 300 ± 27 μm, due to the high surface area of the scaffold.
One of the challenges in tissue engineering is the development of a reproducible three-dimensional (3D) scaffold to support cell migration and infiltration. As a dermal substitute, 3D collagen scaffolds with precisely controlled pore structures were fabricated using an innovative cryogenic dispenser system. The scaffolds were composed of perpendicular, highly porous collagen strands in successive layers. The fabricated scaffolds were evaluated in an in vitro keratinocyte/fibroblast coculture test. Fibroblasts were well dispersed within the scaffold, and keratinocytes had completely migrated through the well-designed pore structure and differentiated on top of the scaffold surface. The differentiated keratinocytes generated a stratum corneum in the 3D dispensed scaffolds, similar to that in normal skin tissue.
Natural polymer-based drug carriers have been developed for antimicrobial applications but several problems remain with their poor controllability of drug loading and degradation. We introduce a novel method to produce improved antibiotic alginate fiber with high drug entrapment properties and a delayed degradation profile. A microfluidic spinning system with a low-polarity isopropyl alcohol (IPA) sheath flow was used to dehydrate an alginate/ampicillin aqueous solution and to form densely packed fiber with enhanced drug loading efficiency. The amounts of ampicillin initially loaded in the IPA-fiber were much higher than in the conventional water-based fiber and they released a more prolonged profile. The fibers were characterized by analyzing the morphology, mass loss and structural properties.The fibers were also used for an in vivo infected wound healing study. The results showed that the IPAbased fibrous alginate drug carrier possesses superior properties for loading drugs and potentials for wound healing applications with easy management.
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