Fabrication of single and bundled filament-like tissues using biodegradable hyaluronic acid-based hollow hydrogel fibers
Graphical abstract
Fabrication of single and bundled filament-like tissues using hyaluronic acid-based hollow hydrogel fibers.
Introduction
Hydrogels have emerged as leading matrices for engineered tissue scaffolds because of their hydrophilic characteristics and potential for biocompatibility [1], [2], [3], [4], [5]. Among hydrogels, extracellular matrix (ECM)-derived components have been recognized as ideal templates to prepare cell-laden hydrogels that can mimic the structural and biological properties of cellular environments in vivo [2], [4], [5], [6]. In particular, hyaluronic acid (HA), a major ECM component in various tissues, is an attractive template for the construction of hydrogels with desired morphology, mechanical strength, and bioactivity [2], [7], [8]. Until now, HA-based hydrogels have been used for tissue engineering applications in various shapes and architectures such as sheets, bulk constructs, microparticles, and microcapsules [9], [10], [11], [12], [13]. These hydrogels provide stable three-dimensional cultivation systems for the survival, expansion, and self-renewal of cells as well as differentiation of encapsulated cells [9], [11], [12], [13]. Among the various cell-laden hydrogels, fiber-shaped vehicles can be advantageous for fabrication of tissue-like biological constructs that mimic the structural and physical properties of native tissues [1], [3], [4], [5], [6]. In addition, fibers can be processed into woven, non-woven, and knitted constructs to fabricate complex tissue-like biological constructs [3], [4], [5], [6]. To the best of our knowledge, no report has adopted an HA-based hydrogel as a fiber-shaped vehicle for cell encapsulation.
In this study, we have developed an HA-based hydrogel as a tool to fabricate a cell-laden hollow fiber-shaped vehicle. Cell-laden hydrogel fibers have commonly been fabricated from unique or composite polymers, including gelatin, collagen, alginate, amylopectin, and chitosan [1], [3], [4], [5], [6], [14], [15], through techniques such as electrospinning, microfluidic spinning, wetspinning, and interfacial complexation [1], [3], [4], [5], [6], [13], [14], [15], [16], [17], [18]. Among the various existing fabrication methods, the microfluidic spinning technique allows precise control over the size, geometry, morphology, and chemical features of fibers [1], [3], [4], [5], [6], [13], [14], [15], [16], [17], [18]. Previously, we produced alginate-based hollow hydrogel fibers through enzymatic crosslinking and degradation using a coaxial double-orifice spinneret [14]. In this system, horseradish peroxidase (HRP) and catalase, both of which consume H2O2, were used to form a hollow in the hydrogel vehicles [14], [19], [20]. To this end, phenolic-substituted HA (HA-Ph) may be a useful bioactive and biodegradable material to create hollow hydrogel fibers that enable cell encapsulation while maintaining cell viability [9], [12], [21]. The HA-Ph fibers were made by a microfluidic spinning technique in which an aqueous solution of HA-Ph containing HRP and catalase was extruded into an ambient co-flowing aqueous solution containing H2O2 through a coaxial double-orifice spinneret. Subsequently, the fabricated fibers were simply bundled by rolling on a rotating cylindrical tube to prepare a higher-order fiber-shaped construct (Fig. 1B and C). The attractive point of using single and bundled HA-Ph hollow hydrogel fibers is that they provide a biomimetic environment for the growth of encapsulated cells while maintaining biodegradability.
In this study, we demonstrated the feasibility of single and bundled filament-like tissue fabrication using HA-Ph fiber-shaped hydrogel vehicles. We first investigated the diameter and membrane thickness of HA-Ph hydrogel fibers affected by changes in both the velocities of extruded solutions in the coaxial double-orifice spinneret and the concentration of reactants. Next, we examined the cytotoxicity of the fiber fabrication process and growth profiles of encapsulated cells in the HA-Ph hydrogel fibers. Finally, surface-coating with cells and the subsequent degradation of the HA-Ph hydrogel fibers were investigated to create a filament-like tissue construct covered with a heterogeneous cell layer. This study revealed the feasibility of HA-Ph cell-laden fiber-shaped vehicles for tissue engineering applications.
Section snippets
Materials
Sodium-HA (average molecular weight: 1.7 × 106 Da) was obtained from JNC Corp. (Tokyo, Japan). HRP (160 U/mg), H2O2 aqueous solution [30% (w/w)] and catalase (1.3 × 104 U/mg, from bovine liver) were purchased from Wako Pure Chemical Industries Ltd. (Osaka, Japan). Gelatin (type B; from porcine skin, 300 Bloom) and hyaluronidase from sheep testes (type II, ≥300 U/mg) were purchased from Sigma (St. Louis, MO, USA). HA-Ph and phenolic-substituted gelatin (gelatin-Ph) were prepared according to previously
Results and discussion
Hydrogels of HA-Ph obtained through an HRP-mediated reaction have been investigated as a promising biomaterial for various biomedical applications such as drug delivery, tissue engineering, and peritoneal adhesion prevention because of their excellent biocompatibility and biodegradability [8], [9], [10], [12], [22]. In addition, HA-Ph hydrogels have been used for encapsulation of cells in a variety of shapes and architectures [9], [12]. These studies illustrate that HA-Ph hydrogel is an
Conclusion
The current study reports the preparation of single and bundled cell-laden HA-Ph hollow hydrogel fibers as a template for filament-like tissues in tissue engineering applications. The HA-Ph hydrogel fibers were fabricated using a coaxial double-orifice spinneret through HRP- and catalase-mediated reactions. The fabricated cell-laden HA-Ph hydrogel fibers were bundled by rolling on a rotating cylindrical plastic tube to obtain a macro-scale fiber-shaped construct. The diameter and membrane
Acknowledgement
This work was supported by Japan Society for the Promotion of Science KAKENHI Grant Numbers 15H04194 and 16H02423.
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