Melt electrowriting (MEW) enables precise scaffold fabrication for biomedical applications. With a limited number of processable materials with short and tunable degradation times, polyhydroxyalkanoates (PHAs) present an interesting option. Here, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and a blend of PHBV and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (PHBV+P34HB) are successfully melt electrowritten into scaffolds with various architectures. PHBV+P34HB exhibits greater thermal stability, making it a superior printing material compared to PHBV in MEW. The PHBV+P34HB scaffolds subjected to enzymatic degradation show tunable degradation times, governed by enzyme dilution, incubation time, and scaffold surface area. PHBV+P34HB scaffolds seeded with human dermal fibroblasts (HDFs), demonstrate enhanced cell adherence, proliferation, and spreading. The HDFs, when exposed to the enzyme solutions and enzymatic degradation residues, show good viability and proliferation rates. Additionally, HDFs grown on enzymatically pre-incubated scaffolds do not show any difference in behavior compared those grown on control scaffolds. It is concluded that PHAs, as biobased materials with enzymatically tunable degradability rates, are an important addition to the already limited set of materials available for MEW technology.
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Polyhydroxyalkanoates (PHAs) are biodegradable biopolymers (polyesters), produced by a wide range of bacterial strains. They are gaining increasing interest in different research fields, due to their sustainability and environmental-friendly properties. Additionally, PHAs are also biocompatible, which makes them interesting for tissue engineering and regenerative medicine. At the same time, they are characterized by properties ideal for 3D printing processing, such as high tensile strength, easy processability and thermoplasticity. To date, the techniques employed in PHAs printing mostly include fused deposition modeling (FDM), selective laser sintering (SLS), electrospinning (ES), and melt electrospinning (MES). In this review, we provide a comprehensive summary of the versatile and sustainably sourced bacterial PHAs, also modified by blending with natural and synthetic polymers (e.g., PLA, PGA) or combining them with inorganic fillers (e.g., nanoparticles, glass), used for 3D printing in biomedical applications. We specify focus on the printing conditions and the properties of the obtained scaffolds with a focus on the print resolution and scaffolds mechanical and biological properties. New perspectives in the emerging field of PHAs biofabrication process, characterized by sustainability and efficiency of the scaffold production, are demonstrated. The use of alternative printing techniques, i.e. melt electrowriting (MEW), and producing smart and functional materials degrading on demand under in vitro and in vivo conditions is proposed.
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Multi-layer cell constructs produced in vitro are an innovative treatment option to support the growing demand for therapy in regenerative medicine. Our research introduces a novel construct integrating organ-derived decellularised extracellular matrix (dECM) hydrogels and 3D-printed biodegradable polymer meshes composed of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB) to support and maintain multiple layers of different cell types. We achieved that by integrating the mechanical stability of PHBV+P34HB, commonly used in the food storage industry, with a dECM hydrogel, which replicates organ stiffness and supports cellular survival and function. The construct was customised by adjusting the fibre arrangement and pore sizes, making it a suitable candidate for a personalised design. We showed that the polymer is degradable after precoating it with PHB depolymerase (PhaZ), with complete degradation achieved in 3–5 days and delayed by adding the hydrogel to 10 days, enabling tuneable degradation for regenerative medicine applications. Finally, as a proof of concept, we composed a three-layered tissue in vitro; each layer represented a different tissue type: epidermal, vascular, and subcutaneous layers. Possible future applications include wound healing and diabetic ulcer paths, personalised drug delivery systems, and personalised tissue implants.
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