A spheroid is a complex, spherical cellular aggregate supporting cell-cell and cell-matrix interactions in an environment that mimics the real-world situation. In terms of tissue engineering, spheroids are important building blocks that replace two-dimensional cell cultures. Spheroids replicate tissue physiological activities. The use of spheroids with/without scaffolds yields structures that engage in desired activities and replicate the complicated geometry of three-dimensional tissues. In this mini-review, we describe conventional and novel methods by which scaffold-free and scaffolded spheroids may be fabricated and discuss their applications in tissue regeneration and future perspectives.
In the human body, there are numerous microtubular tissue structures, such as muscles, vessels, nerves, and tendons. Tissue engineering scaffolds have been regarded as a high-potential candidate for providing such aligned instructive niches to facilitate cell-recruitment and differentiation, and eventually, successful tissue regeneration. Moreover, scaffolds derived from the extracellular matrix (ECM) can provide excellent biocompatibility. However, the fabrication of such microtubular hierarchical scaffolds using ECM has proven to be difficult, and thus, innovative fabrication approaches are required. Herein, we have developed a biofabrication system involving a sequential removal of supporting materials (polycaprolactone (PCL) and poly(vinyl alcohol) (PVA)) to fabricate a uniaxially aligned microtubular collagen scaffold, a lotus-like structure. To generate the unique morphological structures of the scaffold, we manipulated various material-related and processing factors, such as the molecular weight of PVA and the weight fraction of collagen coating. Physical and biological activities of the aligned hierarchical microtubular collagen scaffolds were compared with those of the controls (conventional collagen struts and microtubular collagen scaffolds void of a uniaxial topographical cue). In conclusion, the instructive niche on the aligned hierarchical microtubular collagen structure induced high degrees of myoblast alignment and efficient myogenic differentiation.
The fabrication of biomimetic scaffolding is a challenging issue in tissue engineering. Scaffolds must be designed with micrometer precision to enable cell proliferation and tissue growth, requiring customization based on the type of tissue being developed. Biomimetic scaffolds have attracted interest for their potential in spinal fusion applications. By providing a structured environment to promote osteogenesis, these materials offer a robust and minimally invasive means to fuse vertebrae. The present study describes the successful preparation of a biomimetic collagen/hydroxyapatite hierarchical scaffold, with each strut having several microchannels via 3D printing, leaching, and coating processes (i.e., one-way shape morphing, 4D printing). The biophysical properties of the scaffold were analyzed, as were its various cellular activities, using human adipose stem cells. This biomimetic microchannel scaffold demonstrated great potential for osteogenic activities in vitro and significantly increased new bone formation and ingrowth of blood vessels in vivo in a mouse model of posterolateral lumbar spinal fusion. These in vitro and in vivo results suggest that the microchannel collagen/hydroxyapatite scaffold could act as a potential bone graft substitute to promote high rates of successful fusion.
In tissue engineering, biocompatible scaffolds are used as 3D cell niches to provide a similar environment to that of native tissue for seeded cells to regenerate the target tissue. When engineering bone tissue, high mechanical strength and calcium phosphate composition are essential factors to consider. In this study, we fabricated biocompatible composite scaffolds composed of synthetic polymers (polycaprolactone (PCL) and poly (vinyl alcohol) (PVA)), natural polymers (gelatin and collagen) and bioceramic (hydroxyapatite; HA) for bone tissue engineering. The synthetic polymers were used to enhance the mechanical properties of the composite scaffolds while the natural protein-based polymers were used to enhance various cellular activities, such as cell adhesion and proliferation. Meanwhile, the bioceramic was introduced to promote osteogenic differentiation. Composite scaffolds were evaluated for their physical characteristics, such as mechanical, swelling and protein absorbing properties as well as biological properties (cell proliferation, alkaline phosphatase (ALP) activities and calcium deposition) with human osteoblast-like cells (MG63). Consequently, incorporation of hydroxyapatite into the gelatin/PVA (C-GPH) scaffold showed 5-fold and 1.5-fold increase in calcium deposition and ALP activities, respectively compared to gelatin/PVA scaffold (C-GP). Moreover, compressive modulus also increased 1.8-fold. Integration of PCL core into gelatin/PVA/hydroxyapatite scaffold (C-PGPH) further amplified the compressive modulus 1.5-fold. In conclusion, the scaffold that is reinforced with HA particles and integrated with PCL core of the struts showed significant potential in field of bone tissue engineering.
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