The aim of this paper was to design and fabricate a novel composite scaffold based on the combination of 3D-printed polylactic acid-based triply periodic minimal surfaces (TPMSs) and cell-laden alginate hydrogel. This novel scaffold improves the low mechanical properties of alginate hydrogel and can also provide a scaffold with a suitable pore size, which can be used in bone regeneration applications. In this regard, an implicit function was used to generate some gyroid TPMS scaffolds. Then the fused deposition modeling process was employed to print the scaffolds. Moreover, the micro computed tomography technique was employed to assess the microstructure of 3D-printed TPMS scaffolds and obtain the real geometries of printed scaffolds. The mechanical properties of composite scaffolds were investigated under compression tests experimentally. It was shown that different mechanical behaviors could be obtained for different implicit function parameters. In this research, to assess the mechanical behavior of printed scaffolds in terms of the strain–stress curves on, two approaches were presented: equivalent volume and finite element-based volume. Results of strain–stress curves showed that the finite-element based approach predicts a higher level of stress. Moreover, the biological response of composite scaffolds in terms of cell viability, cell proliferation, and cell attachment was investigated. In this vein, a dynamic cell culture system was designed and fabricated, which improves mass transport through the composite scaffolds and applies mechanical loading to the cells, which helps cell proliferation. Moreover, the results of the novel composite scaffolds were compared to those without alginate, and it was shown that the composite scaffold could create more viability and cell proliferation in both dynamic and static cultures. Also, it was shown that scaffolds in dynamic cell culture have a better biological response than in static culture. In addition, scanning electron microscopy was employed to study the cell adhesion on the composite scaffolds, which showed excellent attachment between the scaffolds and cells.
The use of various chemical cross-linking agents for the improvement of scaffolds physical and mechanical properties is a common practical method, which is limited by cytotoxicity effects. Due to exerting contract type forces, chondrocytes are known to implement shrinkage on the tissue engineered constructs, which can be avoided by the scaffold cross-linking. In the this research, chitosan scaffolds are cross-linked with hydrothermal treatment with autoclave sterilization time of 0, 10, 20 and 30min, to avoid the application of the traditional chemical toxic materials. The optimization studies with gel content and crosslink density measurements indicate that for 20min sterilization time, the gel content approaches to ~80%. The scaffolds are fully characterized by the conventional techniques such as SEM, porosity and permeability, XRD, compression, thermal analysis and dynamic mechanical thermal analysis (DMTA). FT-IR studies shows that autoclave inter-chain cross-linking reduces the amine group absorption at 1560cm and increase the absorption of N-acetylated groups at 1629cm. It is anticipated, that this observation evidenced by chitosan scaffold browning upon autoclave cross-linking is an indication of the familiar maillard reaction between amine moieties and carbonyl groups. The biodegradation rate analysis shows that chitosan scaffolds with lower concentrations, possess suitable degradation rate for cartilage tissue engineering applications. In addition, cytotoxicity analysis shows that fabricated scaffolds are biocompatible. The human articular chondrocytes seeding into 3D cross-linked scaffolds shows a higher viability and proliferation in comparison with the uncross-linked samples and 2D controls. Investigation of cell morphology on the scaffolds by SEM, shows a more spherical morphology of chondrocytes on the cross-linked scaffolds for 21days of in vitro culture.
ABSTRACT:To prepare versatile polyurethane material suitable as insulating coatings, novel kinds of polyhydroxy compounds (PHCs) were prepared via transesterification reaction of poly(ethylene terphthalate), different molecular weight of poly(ethylene glycol)s (PEGs), and castor oil. The final networks were prepared via crosslinking of PHCs with a novel blocked isocyanate curing agent (BPI) made from trimethylol propane (TMP), toluene diisocyanate (TDI), and N-methyl aniline (NMA). Polyols and curing agent were characterized by conventional methods and the curing condition was optimized via gel content measurements. Curing kinetic of the polyurethane network formation was investigated by differential scanning calorimetric method and the kinetic parameters were derived. Crosslink density of the samples was determined via equilibrium swelling method and by using Flory-Rehner equations. Effects of crosslink density on electrical, physical, mechanical, and dynamic mechanical (DMTA) properties of the polyurethane coatings were investigated.
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