A comprehensive comparison of cell seeding methods using highly porous melt electrowriting scaffolds

Blaudez, Fanny; Ivanovski, Sašo; Ipe, Deepak S.; Vaquette, Cédryck

doi:10.1016/j.msec.2020.111282

Cited by 19 publications

(18 citation statements)

References 61 publications

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“…3D PCL fibrous substrates were treated with 2 N sodium hydroxide (NaOH) for 1 h before coating with FBS in order to enhance cell seeding efficacy, as described previously [32].…”

Section: Human Primary Osteoblast Culture and Differentiationmentioning

confidence: 99%

The Mechanosensing and Global DNA Methylation of Human Osteoblasts on MEW Fibers

et al. 2021

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Cells interact with 3D fibrous platform topography via a nano-scaled focal adhesion complex, and more research is required on how osteoblasts sense and respond to random and aligned fibers through nano-sized focal adhesions and their downstream events. The present study assessed human primary osteoblast cells’ sensing and response to random and aligned medical-grade polycaprolactone (PCL) fibrous 3D scaffolds fabricated via the melt electrowriting (MEW) technique. Cells cultured on a tissue culture plate (TCP) were used as 2D controls. Compared to 2D TCP, 3D MEW fibrous substrates led to immature vinculin focal adhesion formation and significantly reduced nuclear localization of the mechanosensor-yes-associated protein (YAP). Notably, aligned MEW fibers induced elongated cell and nucleus shape and highly activated global DNA methylation of 5-methylcytosine, 5-hydroxymethylcytosine, and N-6 methylated deoxyadenosine compared to the random fibers. Furthermore, although osteogenic markers (osterix-OSX and bone sialoprotein-BSP) were significantly enhanced in PCL-R and PCL-A groups at seven days post-osteogenic differentiation, calcium deposits on all seeded samples did not show a difference after normalizing for DNA content after three weeks of osteogenic induction. Overall, our study linked 3D extracellular fiber alignment to nano-focal adhesion complex, nuclear mechanosensing, DNA epigenetics at an early point (24 h), and longer-term changes in osteoblast osteogenic differentiation.

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“…3D PCL fibrous substrates were treated with 2 N sodium hydroxide (NaOH) for 1 h before coating with FBS in order to enhance cell seeding efficacy, as described previously [32].…”

Section: Human Primary Osteoblast Culture and Differentiationmentioning

confidence: 99%

The Mechanosensing and Global DNA Methylation of Human Osteoblasts on MEW Fibers

et al. 2021

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“…SEM imaging demonstrated that the MEW technique successfully produced controlled layer-by-layer fibre stacking with high precision (pore size 499 ± 17.3 μm) and fibre resolution (diameter was 17.5 ± 1.3 μm), consistent with our previous work. 32 In addition, images taken pre- and post-immobilisations demonstrated that the morphology of the scaffold was not compromised by the S1-RBD protein immobilisation process (Fig. 1b).…”

Section: Resultsmentioning

confidence: 81%

“…All scaffolds were manufactured by melt electrowriting (MEW) using medical-grade PCL as previously described. 30,32 Briey, polymer pellets (PC12, Corbion, Amsterdam, The Netherlands) were heated to 74 C and 83 C at the cartridge and needle locations within a blunt 23 G 2 mL syringe, respectively. The polymer was then extruded onto a programmable x-y stage at a pressure of 1.2 bar, a voltage of 8.2 kV, a translational speed of 850 mm min À1 and a distance of 7.4 mm between the collector plate and the spinneret.…”

Section: Manufacture Of 3d Pcl Mew Scaffoldsmentioning

confidence: 99%

“…Several studies have shown that high-resolution melt electrowritten (MEW) poly(3-caprolactone) (PCL) scaffolds are suitable for cell growth, drug delivery and tissue regeneration. [30][31][32][33][34][35][36][37][38] Given their advantageous properties, including precise architecture, high porosity and greater surface area, the potential of MEW PCL scaffolds as 3D immunodetection platforms for COVID-19 antibodies warrants investigation.…”

Section: Introductionmentioning

confidence: 99%

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Salivary SARS-CoV-2 antibody detection using S1-RBD protein-immobilized 3D melt electrowritten poly(ε-caprolactone) scaffolds

Han

Staples

et al. 2022

RSC Adv.

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“…Indeed, 3D-printed scaffolds with highly ordered internal arrangements and macroscopic pores, required for proper in vivo vascularization, are notoriously difficult to efficiently and homogenously seed with cells . Several technological solutions have been implemented for enhancing initial cell adhesion; − however, cell seeding efficacy remains poor for 3D-printed scaffolds . In addition, the pore size of the 3D constructs has a direct influence on cell proliferation and differentiation, as previously demonstrated. − While there exits some contradictory results in the literature, it seems that a small pore size of around 100–300 μm is the most appropriate dimension for enhancing in vitro osteogenic differentiation. ,, …”

Section: Introductionmentioning

confidence: 99%

Effect of Dual Pore Size Architecture on In Vitro Osteogenic Differentiation in Additively Manufactured Hierarchical Scaffolds

Ratheesh

Shi

Lau

et al. 2021

ACS Biomater. Sci. Eng.

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The combination of macro-and microporosity is a potent manner of enhancing osteogenic potential, but the biological events leading to this increase in osteogenesis are not well understood. In this study, we investigated the effect of a dual pore size scaffold on the physical and biological properties, with the hypothesis that cell condensation is the determining factor for enhanced osteogenic differentiation. To this end, a hierarchical scaffold possessing a dual (large and small) pore size was fabricated by combining two additive manufacturing techniques: melt electrospinning writing (MEW) and fused deposition modeling (FDM). The scaffolds showed a mechanical stiffness of 23.2 ± 1.5 MPa similar to the FDM control scaffold, while the hybrid revealed an increased specific surface area of 1.4 ± 0.1 m 2 /g. The scaffold was cultured with primary human osteoblasts for 28 days, which showed enhanced cell adhesion and proliferation. The hierarchical structure was also beneficial for in vitro alkaline phosphate activity and mineralization and showed an increased expression of osteogenic protein and genes. Mesenchymal condensation markers related to osteoblastic differentiation (CDH2, RhoA, Rac1, and Cdc42) were upregulated in the hybrid construct, demonstrating that the MEW membrane provided an environment more suitable for the recapitulation of cell condensation, which in turn leads to higher osteogenic differentiation. In summary, this study demonstrated that the hierarchical scaffold developed in this paper leads to a significant improvement in the scaffold properties such as increased specific surface area, initial cell adhesion, cell proliferation, and in vitro osteogenesis.

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A comprehensive comparison of cell seeding methods using highly porous melt electrowriting scaffolds

Cited by 19 publications

References 61 publications

The Mechanosensing and Global DNA Methylation of Human Osteoblasts on MEW Fibers

The Mechanosensing and Global DNA Methylation of Human Osteoblasts on MEW Fibers

Salivary SARS-CoV-2 antibody detection using S1-RBD protein-immobilized 3D melt electrowritten poly(ε-caprolactone) scaffolds

Effect of Dual Pore Size Architecture on In Vitro Osteogenic Differentiation in Additively Manufactured Hierarchical Scaffolds

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