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1.
Regen Biomater ; 11: rbae033, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38845855

RESUMEN

Biofabrication techniques allow for the construction of biocompatible and biofunctional structures composed from biomaterials, cells and biomolecules. Bioprinting is an emerging 3D printing method which utilizes biomaterial-based mixtures with cells and other biological constituents into printable suspensions known as bioinks. Coupled with automated design protocols and based on different modes for droplet deposition, 3D bioprinters are able to fabricate hydrogel-based objects with specific architecture and geometrical properties, providing the necessary environment that promotes cell growth and directs cell differentiation towards application-related lineages. For the preparation of such bioinks, various water-soluble biomaterials have been employed, including natural and synthetic biopolymers, and inorganic materials. Bioprinted constructs are considered to be one of the most promising avenues in regenerative medicine due to their native organ biomimicry. For a successful application, the bioprinted constructs should meet particular criteria such as optimal biological response, mechanical properties similar to the target tissue, high levels of reproducibility and printing fidelity, but also increased upscaling capability. In this review, we highlight the most recent advances in bioprinting, focusing on the regeneration of various tissues including bone, cartilage, cardiovascular, neural, skin and other organs such as liver, kidney, pancreas and lungs. We discuss the rapidly developing co-culture bioprinting systems used to resemble the complexity of tissues and organs and the crosstalk between various cell populations towards regeneration. Moreover, we report on the basic physical principles governing 3D bioprinting, and the ideal bioink properties based on the biomaterials' regenerative potential. We examine and critically discuss the present status of 3D bioprinting regarding its applicability and current limitations that need to be overcome to establish it at the forefront of artificial organ production and transplantation.

2.
Biomater Adv ; 157: 213737, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38211506

RESUMEN

Graphitic carbon nitride (g-C3N4) is explored as a novel sustainable visible light photoinitiator for the preparation of biomimetic 3D hydrogel scaffolds comprising gelatin methacrylamide (GelMA) and dopamine methacrylamide for use in tissue engineering. The initiator efficiency was assessed by comparing the swelling behavior and the stability of photopolymerized hydrogels prepared with GelMA of different degrees of functionalization and different comonomer compositions. Bioactive composite hydrogels with a 50 wt% nanohydroxyapatite (nHAp) content, to closely mimic the actual bone composition, were successfully obtained by the introduction of nHAp in the prepolymer solutions followed by photopolymerization. The composite hydrogels demonstrated enhanced mechanical properties and excellent stability in PBS verifying the preparation of robust 3D scaffolds for use in cancellous or pre-calcified bone tissue engineering applications. The in vitro cell response of the composite scaffolds exhibited high cell viability and enhanced differentiation of pre-osteoblasts to mature osteoblasts, demonstrating their osteogenic potential. This work establishes, for the first time, the excellent properties of g-C3N4 as a sustainable, visible light initiator, fully satisfying the principles of green chemistry, for the preparation of robust and biologically relevant hydrogels, and proposes a new approach to overcome the main challenges of conventional photoinitiators in cell scaffold fabrication, such as photobleaching, high cost and non-scalable synthesis employing toxic organic precursors and solvents.


Asunto(s)
Acrilamidas , Biomimética , Grafito , Compuestos de Nitrógeno , Pirenos , Ingeniería de Tejidos , Luz , Hidrogeles
3.
Gels ; 9(12)2023 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-38131970

RESUMEN

In this work, a sodium alginate-based copolymer grafted by thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) chains was used as gelator (Alg-g-PNIPAM) in combination with methylcellulose (MC). It was found that the mechanical properties of the resulting gel could be enhanced by the addition of MC and calcium ions (Ca2+). The proposed network is formed via a dual crosslinking mechanism including ionic interactions among Ca2+ and carboxyl groups and secondary hydrophobic associations of PNIPAM chains. MC was found to further reinforce the dynamic moduli of the resulting gels (i.e., a storage modulus of ca. 1500 Pa at physiological body and post-printing temperature), rendering them suitable for 3D printing in biomedical applications. The polymer networks were stable and retained their printed fidelity with minimum erosion as low as 6% for up to seven days. Furthermore, adhered pre-osteoblastic cells on Alg-g-PNIPAM/MC printed scaffolds presented 80% viability compared to tissue culture polystyrene control, and more importantly, they promoted the osteogenic potential, as indicated by the increased alkaline phosphatase activity, calcium, and collagen production relative to the Alg-g-PNIPAM control scaffolds. Specifically, ALP activity and collagen secreted by cells were significantly enhanced in Alg-g-PNIPAM/MC scaffolds compared to the Alg-g-PNIPAM counterparts, demonstrating their potential in bone tissue engineering.

4.
Bioengineering (Basel) ; 10(5)2023 Apr 27.
Artículo en Inglés | MEDLINE | ID: mdl-37237602

RESUMEN

The application of mechanical stimulation on bone tissue engineering constructs aims to mimic the native dynamic nature of bone. Although many attempts have been made to evaluate the effect of applied mechanical stimuli on osteogenic differentiation, the conditions that govern this process have not yet been fully explored. In this study, pre-osteoblastic cells were seeded on PLLA/PCL/PHBV (90/5/5 wt.%) polymeric blend scaffolds. The constructs were subjected every day to cyclic uniaxial compression for 40 min at a displacement of 400 µm, using three frequency values, 0.5, 1, and 1.5 Hz, for up to 21 days, and their osteogenic response was compared to that of static cultures. Finite element simulation was performed to validate the scaffold design and the loading direction, and to assure that cells inside the scaffolds would be subjected to significant levels of strain during stimulation. None of the applied loading conditions negatively affected the cell viability. The alkaline phosphatase activity data indicated significantly higher values at all dynamic conditions compared to the static ones at day 7, with the highest response being observed at 0.5 Hz. Collagen and calcium production were significantly increased compared to static controls. These results indicate that all of the examined frequencies substantially promoted the osteogenic capacity.

5.
Int J Mol Sci ; 24(7)2023 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-37047438

RESUMEN

This study aims to investigate the impact of kappa-carrageenan on dental pulp stem cells (DPSCs) behavior in terms of biocompatibility and odontogenic differentiation potential when it is utilized as a component for the production of 3D sponge-like scaffolds. For this purpose, we prepared three types of scaffolds by freeze-drying (i) kappa-carrageenan/chitosan/gelatin enriched with KCl (KCG-KCl) as a physical crosslinker for the sulfate groups of kappa-carrageenan, (ii) kappa-carrageenan/chitosan/gelatin (KCG) and (iii) chitosan/gelatin (CG) scaffolds as a control. The mechanical analysis illustrated a significantly higher elastic modulus of the cell-laden scaffolds compared to the cell-free ones after 14 and 28 days with values ranging from 25 to 40 kPa, showing an increase of 27-36%, with the KCG-KCl scaffolds indicating the highest and CG the lowest values. Cell viability data showed a significant increase from days 3 to 7 and up to day 14 for all scaffold compositions. Significantly increasing alkaline phosphatase (ALP) activity has been observed over time in all three scaffold compositions, while the KCG-KCl scaffolds indicated significantly higher calcium production after 21 and 28 days compared to the CG control. The gene expression analysis of the odontogenic markers DSPP, ALP and RunX2 revealed a two-fold higher upregulation of DSPP in KCG-KCl scaffolds at day 14 compared to the other two compositions. A significant increase of the RunX2 expression between days 7 and 14 was observed for all scaffolds, with a significantly higher increase of at least twelve-fold for the kappa-carrageenan containing scaffolds, which exhibited an earlier ALP gene expression compared to the CG. Our results demonstrate that the integration of kappa-carrageenan in scaffolds significantly enhanced the odontogenic potential of DPSCs and supports dentin-pulp regeneration.


Asunto(s)
Quitosano , Andamios del Tejido , Quitosano/metabolismo , Gelatina/metabolismo , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Carragenina/farmacología , Pulpa Dental/metabolismo , Células Cultivadas , Biomimética , Células Madre/metabolismo , Regeneración , Diferenciación Celular , Dentina/metabolismo
6.
Gels ; 9(2)2023 Jan 24.
Artículo en Inglés | MEDLINE | ID: mdl-36826273

RESUMEN

Bioprinting aims to provide new avenues for regenerating damaged human tissues through the controlled printing of live cells and biocompatible materials that can function therapeutically. Polymeric hydrogels are commonly investigated ink materials for 3D and 4D bioprinting applications, as they can contain intrinsic properties relative to those of the native tissue extracellular matrix and can be printed to produce scaffolds of hierarchical organization. The incorporation of nanoscale material additives, such as nanoparticles, to the bulk of inks, has allowed for significant tunability of the mechanical, biological, structural, and physicochemical material properties during and after printing. The modulatory and biological effects of nanoparticles as bioink additives can derive from their shape, size, surface chemistry, concentration, and/or material source, making many configurations of nanoparticle additives of high interest to be thoroughly investigated for the improved design of bioactive tissue engineering constructs. This paper aims to review the incorporation of nanoparticles, as well as other nanoscale additive materials, to printable bioinks for tissue engineering applications, specifically bone, cartilage, dental, and cardiovascular tissues. An overview of the various bioinks and their classifications will be discussed with emphasis on cellular and mechanical material interactions, as well the various bioink formulation methodologies for 3D and 4D bioprinting techniques. The current advances and limitations within the field will be highlighted.

7.
Int J Biol Macromol ; 209(Pt B): 1720-1730, 2022 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-35461864

RESUMEN

Kappa-carrageenan is a biocompatible natural polysaccharide able to form hydrogels for tissue regeneration. In bone tissue engineering, achieving a bioactive microenvironment with appropriate mechanical properties in polysaccharide-based scaffolds remains a challenge. This study aims to fabricate 3D scaffolds comprising kappa-carrageenan, chitosan and gelatin, crosslinked with KCl, and evaluate their mechanical and biological properties for bone tissue engineering. The produced scaffolds include kappa-carrageenan/chitosan (KC), kappa-carrageenan/chitosan/gelatin (KCG), kappa-carrageenan/chitosan/gelatin enriched with KCl (KCG-KCl), and chitosan/gelatin (CG). All scaffolds present degradation rates ranging from 30% weight loss on day 21, pore size distribution in the range of 100-160 µm and porosity above 80%. The Young modulus values range from 9 to 256 kPa, with the KCl-containing KCG scaffolds demonstrating the highest values, validating the role of KCl in the coil to helix transition of kappa-carrageenan leading to firmer structures. In vitro biological evaluation indicates that pre-osteoblasts proliferate significantly from day 3 up to day 14 on all scaffold compositions. The alkaline phosphatase activity shows a significant increase up to day 14. The calcium production displays a constant increase from day 14 up to day 28, proving that all scaffold compositions support the osteogenic differentiation potential.


Asunto(s)
Quitosano , Gelatina , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Carragenina/química , Carragenina/farmacología , Quitosano/química , Gelatina/química , Osteogénesis , Porosidad , Cloruro de Potasio/farmacología , Ingeniería de Tejidos , Andamios del Tejido/química
8.
Polymers (Basel) ; 13(10)2021 May 18.
Artículo en Inglés | MEDLINE | ID: mdl-34070123

RESUMEN

Main chain polyesters have been extensively used in the biomedical field. Despite their many advantages, including biocompatibility, biodegradability, and others, these materials are rather inert and lack specific functionalities which will endow them with additional biological and responsive properties. In this work, novel pH-responsive main chain polyesters have been prepared by a conventional condensation polymerization of a vinyl functionalized diol with a diacid chloride, followed by a photo-induced thiol-ene click reaction to attach functional carboxylic acid side-groups along the polymer chains. Two different mercaptocarboxylic acids were employed, allowing to vary the alkyl chain length of the polymer pendant groups. Moreover, the degree of modification, and as a result, the carboxylic acid content of the polymers, was easily tuned by varying the irradiation time during the click reaction. Both these parameters, were shown to strongly influence the responsive behavior of the polyesters, which presented adjustable pKα values and water solubilities. Finally, the difunctional polyesters bearing the alkene and carboxylic acid functionalities enabled the preparation of cross-linked polyester films by chemically linking the pendant vinyl bonds on the polymer side groups. The biocompatibility of the cross-linked polymers films was assessed in L929 fibroblast cultures and showed that the cell viability, proliferation, and attachment were greatly promoted on the polyester surface, bearing the shorter alkyl chain length side groups and the higher fraction of carboxylic acid functionalities.

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