Marco Domingos

Polycaprolactone Scaffolds Fabricated via Bioextrusion for Tissue Engineering Applications

The most promising approach in Tissue Engineering involves the seeding of porous, biocompatible/biodegradable scaffolds, with donor cells to promote tissue regeneration. Additive biomanufacturing processes are increasingly recognized as ideal techniques to produce 3D structures with optimal pore size and spatial distribution, providing an adequate mechanical support for tissue regeneration while shaping in-growing tissues. This paper presents a novel extrusion-based system to produce 3D scaffolds with controlled internal/external geometry for TE applications.The BioExtruder is a low-cost system that uses a proper fabrication code based on the ISO programming language enabling the fabrication of multimaterial scaffolds. Poly(ε-caprolactone) was the material chosen to produce porous scaffolds, made by layers of directionally aligned microfilaments. Chemical, morphological, and in vitro biological evaluation performed on the polymeric constructs revealed a high potential of the BioExtruder to produce 3D scaffolds with regular and reproducible macropore architecture, without inducing relevant chemical and biocompatibility alterations of the material.

Fabrication and characterisation of PCL and PCL/PLA scaffolds for tissue engineering

Purpose – The main purpose of this research work is to study the effect of poly lactic acid (PLA) addition into poly (e-caprolactone) (PCL) matrices, as well the influence of the mixing process on the morphological, thermal, chemical, mechanical and biological performance of the 3D constructs produced with a novel biomanufacturing device (BioCell Printing). Design/methodology/approach – Two mixing processes are used to prepare PCL/PLA blends, namely melt blending and solvent casting. PCL and PCL/PLA scaffolds are produced via BioCell Printing using a 300-μm nozzle, 0/90° lay down pattern and 350-μm pore size. Several techniques such as scanning electron microscopy (SEM), simultaneous thermal analyzer (STA), nuclear magnetic resonance (NMR), static compression analysis and Alamar BlueTM are used to evaluate scaffolds morphological, thermal, chemical, mechanical and biological properties. Findings – Results show that the addition of PLA to PCL scaffolds strongly improves the biomechanical performance of th...

Effect of process parameters on the morphological and mechanical properties of 3D Bioextruded poly(?????caprolactone) scaffolds

Purpose – This paper aims to report a detailed study regarding the influence of process parameters on the morphological/mechanical properties of poly(e‐caprolactone) (PCL) scaffolds manufactured by using a novel extrusion‐based system that is called BioExtruder.Design/methodology/approach – In this study the authors focused investigations on four parameters, namely the liquefier temperature (LT), screw rotation velocity (SRV), deposition velocity (DV) and slice thickness (ST). Scaffolds were fabricated by employing three different values of each parameter. Through a series of trials, scaffolds were manufactured varying iteratively one parameter while maintaining constant the other ones. The morphology of the structures was investigated using a scanning electron microscope (SEM), whilst the mechanical performance was assessed though compression tests.Findings – Experimental results highlight a direct influence of the process parameters on the PCL scaffolds properties. In particular, DV and SRV have the hig...

Three-dimensional printed bone scaffolds: The role of nano/micro-hydroxyapatite particles on the adhesion and differentiation of human mesenchymal stem cells

Bone tissue engineering is strongly dependent on the use of three-dimensional scaffolds that can act as templates to accommodate cells and support tissue ingrowth. Despite its wide application in tissue engineering research, polycaprolactone presents a very limited ability to induce adhesion, proliferation and osteogenic cell differentiation. To overcome some of these limitations, different calcium phosphates, such as hydroxyapatite and tricalcium phosphate, have been employed with relative success. This work investigates the influence of nano-hydroxyapatite and micro-hydroxyapatite (nHA and mHA, respectively) particles on the in vitro biomechanical performance of polycaprolactone/hydroxyapatite scaffolds. Morphological analysis performed with scanning electron microscopy allowed us to confirm the production of polycaprolactone/hydroxyapatite constructs with square interconnected pores of approximately 350 µm and to assess the distribution of hydroxyapatite particles within the polymer matrix. Compression mechanical tests showed an increase in polycaprolactone compressive modulus (E) from 105.5 ± 11.2 to 138.8 ± 12.9 MPa (PCL_nHA) and 217.2 ± 21.8 MPa (PCL_mHA). In comparison to PCL_mHA scaffolds, the addition of nano-hydroxyapatite enhanced the adhesion and viability of human mesenchymal stem cells as confirmed by Alamar Blue assay. In addition, after 14 days of incubation, PCL_nHA scaffolds showed higher levels of alkaline phosphatase activity compared to polycaprolactone or PCL_mHA structures.

Tissue Engineering 3D Neurovascular Units: A Biomaterials and Bioprinting Perspective

Neurovascular dysfunction is a central process in the pathogenesis of stroke and most neurodegenerative diseases, including Alzheimers disease. The multicellular neurovascular unit (NVU) combines the neural, vascular and extracellular matrix (ECM) components in an important interface whose correct functioning is critical to maintain brain health. Tissue engineering is now offering new tools and insights to advance our understanding of NVU function. Here, we review how the use of novel biomaterials to mimic the mechanical and functional cues of the ECM, coupled with precisely layered deposition of the different cells of the NVU through 3D bioprinting, is revolutionising the study of neurovascular function and dysfunction.

Advanced mechanical and thermal characterization of 3D Bioextruded Poly(ε-caprolactone)-based composites

The purpose of this paper is to study the effect of nano hydroxyapatite (HA) and graphene oxide (GO) particles on thermal and mechanical performances of 3D printed poly(e-caprolactone) (PCL) filaments used in bone tissue engineering (BTE).,Raw materials were prepared by melt blending, followed by 3D printing via 3D Discovery (regenHU Ltd., CH) with all fabricating parameters kept constant. Filaments, including pure PCL, PCL/HA and PCL/GO, were tested under the same conditions. Several techniques were used to mechanically, thermally and microstructurally evaluate properties of these filaments, including differential scanning calorimetry, tensile test, nano indentation and scanning electron microscope.,Results show that both HA and GO nano particles are capable of improving mechanical performance of PCL. Enhanced mechanical properties of PCL/HA result from reinforcing effect of HA, while a different mechanism is observed in PCL/GO, where degree of crystallinity plays an important role. In addition, GO is more efficient at enhancing mechanical performance of PCL compared with HA.,For the first time, a systematic study about effects of nano HA and GO particles on bioactive scaffolds produced by additive manufacturing for BTE applications is conducted in this work. Mechanical and thermal behaviors of each sample, pure PCL, PCL/HA and PCL/GO, are reported, correlated and compared with literature.

An experimental study to investigate the micro-stereolithography tools for micro injection molding

Purpose The use of microstereolithography (μSL) parts as micro-injection molding (μIM) tools greatly reduces the time and cost to product and offers unique solutions for complex design issues. However, they present challenges to designers because of their strength, thermal characteristics and shorter lifetimes as compared to other mold materials. The purpose of this study is to use SL to build rapid injection mold tools directly combining micro features for short-run production. Design/methodology/approach In total, three tool inserts were produced. Two different μSL mold inserts were produced and evaluated in terms of different build approaches of micro features. One of the inserts includes micro features built horizontally, while the other one collaborates features built vertically, both having same geometrical dimensions. To evaluate the replication capability of prototype tools, two different thicknesses were set for micro features, that is, 30 and 120 μm. The mold inserts were assembled on a metallic mold frame and tested with polypropylene (PP). Findings It was observed that using inappropriate resin to fabricate the mold inserts can lead to inaccurate geometrical dimensions of the tool. Therefore, the material with high glass transition temperature (Tg) and low thermal conductivity is preferred. Also, the results of this research work showed that the processed material and technology play an important role both on part quality and tool life. After investigating the tool failure mechanisms during the injection, it was observed that tool failure occurred mainly because of excessive flexural stresses and ejection forces during the cavity filling and part ejection phases, respectively. Originality/value The paper describes the capability of μSL mold inserts for the production of small batches of micro-cantilevers which are used in MEMS relays.

Effect of in vitro enzymatic degradation on 3D printed poly(ε-caprolactone) scaffolds: morphological, chemical and mechanical properties

Background In recent years, the tissue engineering (TE) field has significantly benefited from advanced techniques such as additive manufacturing (AM), for the design of customized 3D scaffolds with the aim of guided tissue repair. Among the wide range of materials available to biomanufacture 3D scaffolds, poly(ε-caprolactone) (PCL) clearly arises as the synthetic polymer with the greatest potential, due to its unique properties – namely, biocompatibility, biodegradability, thermal and chemical stability and processability. This study aimed for the first time to investigate the effect of pore geometry on the in vitro enzymatic chain cleavage mechanism of PCL scaffolds manufactured by the AM extrusion process. Methods Methods: Morphological properties of 3D printed PCL scaffolds before and after degradation were evaluated using Scanning Electron Microscopy (SEM) and micro-computed tomography (μ-CT). Differential Scanning Calorimetry (DSC) was employed to determine possible variations in the crystallinity of the scaffolds during the degradation period. The molecular weight was assessed using Size Exclusion Chromatography (SEC) while the mechanical properties were investigated under static compression conditions. Results Morphological results suggested a uniform reduction of filament diameter, while increasing the scaffolds’ porosity. DSC analysis revealed and increment in the crystallinity degree while the molecular weight, evaluated through SEC, remained almost constant during the incubation period (25 days). Mechanical analysis highlighted a decrease in the compressive modulus and maximum stress over time, probably related to the significant weight loss of the scaffolds. Conclusions All of these results suggest that PCL scaffolds undergo enzymatic degradation through a surface erosion mechanism, which leads to significant variations in mechanical, physical and chemical properties, but which has little influence on pore geometry.

EFFECT OF PROCESS PARAMETERS ON THE PROPERTIES OF 3D BIOEXTRUDED POLY(?-CAPROLACTONE) SCAFFOLDS

Purpose – This paper aims to report a detailed study regarding the influence of process parameters on the morphological/mechanical properties of poly(e‐caprolactone) (PCL) scaffolds manufactured by using a novel extrusion‐based system that is called BioExtruder.Design/methodology/approach – In this study the authors focused investigations on four parameters, namely the liquefier temperature (LT), screw rotation velocity (SRV), deposition velocity (DV) and slice thickness (ST). Scaffolds were fabricated by employing three different values of each parameter. Through a series of trials, scaffolds were manufactured varying iteratively one parameter while maintaining constant the other ones. The morphology of the structures was investigated using a scanning electron microscope (SEM), whilst the mechanical performance was assessed though compression tests.Findings – Experimental results highlight a direct influence of the process parameters on the PCL scaffolds properties. In particular, DV and SRV have the hig...