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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Materials Science".

Deadline for manuscript submissions: closed (15 January 2022) | Viewed by 29897

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Prof. Dr. Agata Przekora

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Independent Unit of Tissue Engineering and Regenerative Medicine, Medical University of Lublin, 20-093 Lublin, Poland
Interests: biomaterials; bioceramics; biopolymers; tissue engineering; bone scaffolds; wound dressings; skin substitutes; stem cells; cell culture models; cell-biomaterial interactions
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Dear Colleagues,

Biomaterials for regenerative medicine applications should be non-toxic and biocompatible. Biocompatibility has a very broad meaning, including non-toxicity of the materials, their ability to support cell adhesion, proliferation, and differentiation, as well as their non-immunogenic properties. Therefore, the evaluation of major cell–biomaterial interactions is a key factor determining the biocompatibility and clinical usefulness of new biomaterials. It is important to know that in vitro tests can effectively replace animal models in the preliminary evaluation of (1) cytotoxicity, (2) cell adhesion, spreading, and proliferation on a biomaterial, (3) cell differentiation, and (4) immune response to a biomaterial. Despite the possibility of the use of in vitro cellular models for the evaluation of materials biocompatibility, researchers still preferentially choose in vivo animal tests for this purpose. Nevertheless, the use of animal models at a preliminary stage or for comparative purposes is against the principles of the 3Rs, aiming to Replace, Reduce, and Refine the use of animals wherever possible.

The main goal of this Special Issue is to highlight recent progress in molecular biology and biotechnological techniques that allow a better exploitation of the potential of in vitro cellular models for biocompatibility testing of novel biomaterials, suggesting molecular mechanisms of cell adhesion, proliferation, and biomaterial-induced activation of immune cells. All papers (reviews and original research articles) dealing with in vitro and ex vivo determination of cell–biomaterial interactions are welcome. Manuscripts presenting interactions of biomaterials with prokaryotic cells (e.g., antibiofilm or antibacterial activity of a material) are also encouraged; however, at least basic cytotoxicity tests with eukaryotic cells should be included.

Prof. Dr. Agata Przekora
Guest Editor

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Keywords

ex vivo tissue explant models
in vitro cellular models
cell adhesion and spreading on the biomaterial
cell proliferation and differentiation on the biomaterial
immune response to biomaterial
biomaterial interactions with mesenchymal stem cells
biomaterial-induced macrophage polarization and cytokine release
bacteria adhesion and biofilm formation on the biomaterials

Related Special Issues

Interactions of Cells with Biomaterials for Regenerative Medicine in International Journal of Molecular Sciences (18 articles)
Interactions of Cells with Biomaterials for Regenerative Medicine 3.0 in International Journal of Molecular Sciences (9 articles)
Interactions of Cells with Biomaterials for Regenerative Medicine 4.0 in International Journal of Molecular Sciences (2 articles)

Published Papers (11 papers)

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Research

Jump to: Review

12 pages, 2412 KiB

Open AccessArticle

Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation Protocol

by Serena Bianchi, Sara Bernardi, Antonella Mattei, Loredana Cristiano, Leonardo Mancini, Diana Torge, Giuseppe Varvara, Guido Macchiarelli and Enrico Marchetti

Int. J. Mol. Sci. 2022, 23(9), 5273; https://doi.org/10.3390/ijms23095273 - 09 May 2022

Cited by 11 | Viewed by 2072

Abstract

Several types of deproteinised bovine bone mineral (DBBM) are available on the market, and each one is obtained with a thermic and chemical process that can differ, achieving different results. Currently, several protocols using low temperature are suggested to reduce the possible particle crystallisation during the production process. This study aimed to evaluate the biomorphological reaction of periodontal fibroblast cultures in contact with different DBBM particles treated with a low-temperature protocol (Thermagen^®) and without exposure to sodium hydroxide (NaOH). Morphological evaluation was performed using light, confocal laser, and scanning electron microscopy, and the biological reaction in terms of proliferation was performed using an XTT proliferation assay at 24 h (T1), 72 h (T2), and 7 days (T3). The morphological analysis highlighted how the presence of the materials stimulated a change in the morphology of the cells into a polygonal shape, surface reactions with the thickening of the membrane, and expression of actin. In particular, the morphological changes were appreciable from T1, with a progressive increase in the considered morphological characteristics at T2 and T3 follow-ups. The proliferation assay showed a statistical significance between the different experimental materials and the negative control in T2 and T3 follow-ups. The post hoc analysis did not reveal any differences between the materials. In conclusion, the grafts obtained with the low-temperature extractions protocol and not exposed to NaOH solution showed positive morphological reactions with no differences in the sizes of particles. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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19 pages, 38450 KiB

Open AccessArticle

Laser Direct Writing of Dual-Scale 3D Structures for Cell Repelling at High Cellular Density

by Irina Alexandra Paun, Bogdan Stefanita Calin, Roxana Cristina Popescu, Eugenia Tanasa and Antoniu Moldovan

Int. J. Mol. Sci. 2022, 23(6), 3247; https://doi.org/10.3390/ijms23063247 - 17 Mar 2022

Cited by 2 | Viewed by 1598

Abstract

The fabrication of complex, reproducible, and accurate micro-and nanostructured interfaces that impede the interaction between material’s surface and different cell types represents an important objective in the development of medical devices. This can be achieved by topographical means such as dual-scale structures, mainly represented by microstructures with surface nanopatterning. Fabrication via laser irradiation of materials seems promising. However, laser-assisted fabrication of dual-scale structures, i.e., ripples relies on stochastic processes deriving from laser–matter interaction, limiting the control over the structures’ topography. In this paper, we report on laser fabrication of cell-repellent dual-scale 3D structures with fully reproducible and high spatial accuracy topographies. Structures were designed as micrometric “mushrooms” decorated with fingerprint-like nanometric features with heights and periodicities close to those of the calamistrum, i.e., 200–300 nm. They were fabricated by Laser Direct Writing via Two-Photon Polymerization of IP-Dip photoresist. Design and laser writing parameters were optimized for conferring cell-repellent properties to the structures, even for high cellular densities in the culture medium. The structures were most efficient in repelling the cells when the fingerprint-like features had periodicities and heights of ≅200 nm, fairly close to the repellent surfaces of the calamistrum. Laser power was the most important parameter for the optimization protocol. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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23 pages, 5038 KiB

Open AccessArticle

3D Printed Poly(𝜀-caprolactone)/Hydroxyapatite Scaffolds for Bone Tissue Engineering: A Comparative Study on a Composite Preparation by Melt Blending or Solvent Casting Techniques and the Influence of Bioceramic Content on Scaffold Properties

by Sara Biscaia, Mariana V. Branquinho, Rui D. Alvites, Rita Fonseca, Ana Catarina Sousa, Sílvia Santos Pedrosa, Ana R. Caseiro, Fernando Guedes, Tatiana Patrício, Tânia Viana, Artur Mateus, Ana C. Maurício and Nuno Alves

Int. J. Mol. Sci. 2022, 23(4), 2318; https://doi.org/10.3390/ijms23042318 - 19 Feb 2022

Cited by 15 | Viewed by 3308

Abstract

Bone tissue engineering has been developed in the past decades, with the engineering of bone substitutes on the vanguard of this regenerative approach. Polycaprolactone-based scaffolds are fairly applied for bone regeneration, and several composites have been incorporated so as to improve the scaffolds’ mechanical properties and tissue in-growth. In this study, hydroxyapatite is incorporated on polycaprolactone-based scaffolds at two different proportions, 80:20 and 60:40. Scaffolds are produced with two different blending methods, solvent casting and melt blending. The prepared composites are 3D printed through an extrusion-based technique and further investigated with regard to their chemical, thermal, morphological, and mechanical characteristics. In vitro cytocompatibility and osteogenic differentiation was also assessed with human dental pulp stem/stromal cells. The results show the melt-blending-derived scaffolds to present more promising mechanical properties, along with the incorporation of hydroxyapatite. The latter is also related to an increase in osteogenic activity and promotion. Overall, this study suggests polycaprolactone/hydroxyapatite scaffolds to be promising candidates for bone tissue engineering, particularly when produced by the MB method. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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15 pages, 5848 KiB

Open AccessArticle

Biocompatibility and Electrical Stimulation of Skeletal and Smooth Muscle Cells Cultured on Piezoelectric Nanogenerators

by Andreu Blanquer, Oriol Careta, Laura Anido-Varela, Aida Aranda, Elena Ibáñez, Jaume Esteve, Carme Nogués and Gonzalo Murillo

Int. J. Mol. Sci. 2022, 23(1), 432; https://doi.org/10.3390/ijms23010432 - 31 Dec 2021

Cited by 5 | Viewed by 2080

Abstract

Nanogenerators are interesting for biomedical applications, with a great potential for electrical stimulation of excitable cells. Piezoelectric ZnO nanosheets present unique properties for tissue engineering. In this study, nanogenerator arrays based on ZnO nanosheets are fabricated on transparent coverslips to analyse the biocompatibility and the electromechanical interaction with two types of muscle cells, smooth and skeletal. Both cell types adhere, proliferate and differentiate on the ZnO nanogenerators. Interestingly, the amount of Zn ions released over time from the nanogenerators does not interfere with cell viability and does not trigger the associated inflammatory response, which is not triggered by the nanogenerators themselves either. The local electric field generated by the electromechanical nanogenerator–cell interaction stimulates smooth muscle cells by increasing cytosolic calcium ions, whereas no stimulation effect is observed on skeletal muscle cells. The random orientation of the ZnO nanogenerators, avoiding an overall action potential aligned along the muscle fibre, is hypothesised to be the cause of the cell-type dependent response. This demonstrates the need of optimizing the nanogenerator morphology, orientation and distribution according to the potential biomedical use. Thus, this study demonstrates the cell-scale stimulation triggered by biocompatible piezoelectric nanogenerators without using an external source on smooth muscle cells, although it remarks the cell type-dependent response. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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20 pages, 3252 KiB

Open AccessArticle

A Fibrin Coating Method of Polypropylene Meshes Enables the Adhesion of Menstrual Blood-Derived Mesenchymal Stromal Cells: A New Delivery Strategy for Stem Cell-Based Therapies

by Federica Marinaro, Joana M. Silva, Alexandre A. Barros, Ivo M. Aroso, Juan C. Gómez-Blanco, Isaac Jardin, Jose J. Lopez, María Pulido, María Ángeles de Pedro, Rui L. Reis, Francisco Miguel Sánchez-Margallo, Javier G. Casado and Esther López

Int. J. Mol. Sci. 2021, 22(24), 13385; https://doi.org/10.3390/ijms222413385 - 13 Dec 2021

Cited by 7 | Viewed by 3724

Abstract

Polypropylene (PP) mesh is well-known as a gold standard of all prosthetic materials of choice for the reinforcement of soft tissues in case of hernia, organ prolapse, and urinary incontinence. The adverse effects that follow surgical mesh implantation remain an unmet medical challenge. Herein, it is outlined a new approach to allow viability and adhesion of human menstrual blood-derived mesenchymal stromal cells (MenSCs) on PP surgical meshes. A multilayered fibrin coating, based on fibrinogen and thrombin from a commercial fibrin sealant, was optimized to guarantee a homogeneous and stratified film on PP mesh. MenSCs were seeded on the optimized fibrin-coated meshes and their adhesion, viability, phenotype, gene expression, and immunomodulatory capacity were fully evaluated. This coating guaranteed MenSC viability, adhesion and did not trigger any change in their stemness and inflammatory profile. Additionally, MenSCs seeded on fibrin-coated meshes significantly decreased CD4+ and CD8+ T cell proliferation, compared to in vitro stimulated lymphocytes (p < 0.0001). Hence, the proposed fibrin coating for PP surgical meshes may allow the local administration of stromal cells and the reduction of the exacerbated inflammatory response following mesh implantation surgery. Reproducible and easy to adapt to other cell types, this method undoubtedly requires a multidisciplinary and translational approach to be improved for future clinical uses. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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14 pages, 5798 KiB

Open AccessArticle

Polymeric Microspheres/Cells/Extracellular Matrix Constructs Produced by Auto-Assembly for Bone Modular Tissue Engineering

by Bartosz Mielan, Daniela M. Sousa, Małgorzata Krok-Borkowicz, Pierre Eloy, Christine Dupont, Meriem Lamghari and Elżbieta Pamuła

Int. J. Mol. Sci. 2021, 22(15), 7897; https://doi.org/10.3390/ijms22157897 - 23 Jul 2021

Cited by 6 | Viewed by 2710

Abstract

Modular tissue engineering (MTE) is a novel “bottom-up” approach to create engineered biological tissues from microscale repeating units. Our aim was to obtain microtissue constructs, based on polymer microspheres (MSs) populated with cells, which can be further assembled into larger tissue blocks and used in bone MTE. Poly(L-lactide-co-glycolide) MS of 165 ± 47 µm in diameter were produced by oil-in-water emulsification and treated with 0.1 M NaOH. To improve cell adhesion, MSs were coated with poly-L-lysine (PLL) or human recombinant collagen type I (COL). The presence of oxygenated functionalities and PLL/COL coating on MS was confirmed by X-ray photoelectron spectroscopy (XPS). To assess the influence of medium composition on adhesion, proliferation, and osteogenic differentiation, preosteoblast MC3T3-E1 cells were cultured on MS in minimal essential medium (MEM) and osteogenic differentiation medium (OSG). Moreover, to assess the potential osteoblast–osteoclast cross-talk phenomenon and the influence of signaling molecules released by osteoclasts on osteoblast cell culture, a medium obtained from osteoclast culture (OSC) was also used. To impel the cells to adhere and grow on the MS, anti-adhesive cell culture plates were utilized. The results show that MS coated with PLL and COL significantly favor the adhesion and growth of MC3T3-E1 cells on days 1 and 7, respectively, in all experimental conditions tested. On day 7, three-dimensional MS/cell/extracellular matrix constructs were created owing to auto-assembly. The cells grown in such constructs exhibited high activity of early osteogenic differentiation marker, namely, alkaline phosphatase. Superior cell growth on PLL- and COL-coated MS on day 14 was observed in the OSG medium. Interestingly, deposition of extracellular matrix and its mineralization was particularly enhanced on COL-coated MS in OSG medium on day 14. In our study, we developed a method of spontaneous formation of organoid-like MS-based cell/ECM constructs with a few millimeters in size. Such constructs may be regarded as building blocks in bone MTE. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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13 pages, 13629 KiB

Open AccessArticle

Effect of Endothelial Culture Medium Composition on Platelet Responses to Polymeric Biomaterials

by Skadi Lau, Anna Maier, Steffen Braune, Manfred Gossen and Andreas Lendlein

Int. J. Mol. Sci. 2021, 22(13), 7006; https://doi.org/10.3390/ijms22137006 - 29 Jun 2021

Cited by 1 | Viewed by 1767

Abstract

Near-physiological in vitro thrombogenicity test systems for the evaluation of blood-contacting endothelialized biomaterials requires co-cultivation with platelets (PLT). However, the addition of PLT has led to unphysiological endothelial cell (EC) detachment in such in vitro systems. A possible cause for this phenomenon may be PLT activation triggered by the applied endothelial cell medium, which typically consists of basal medium (BM) and nine different supplements. To verify this hypothesis, the influence of BM and its supplements was systematically analyzed regarding PLT responses. For this, human platelet rich plasma (PRP) was mixed with BM, BM containing one of nine supplements, or with BM containing all supplements together. PLT adherence analysis was carried out in six-channel slides with plasma-treated cyclic olefin copolymer (COC) and poly(tetrafluoro ethylene) (PTFE, as a positive control) substrates as part of the six-channel slides in the absence of EC and under static conditions. PLT activation and aggregation were analyzed using light transmission aggregometry and flow cytometry (CD62P). Medium supplements had no effect on PLT activation and aggregation. In contrast, supplements differentially affected PLT adherence, however, in a polymer- and donor-dependent manner. Thus, the use of standard endothelial growth medium (BM + all supplements) maintains functionality of PLT under EC compatible conditions without masking the differences of PLT adherence on different polymeric substrates. These findings are important prerequisites for the establishment of a near-physiological in vitro thrombogenicity test system assessing polymer-based cardiovascular implant materials in contact with EC and PLT. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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16 pages, 2571 KiB

Open AccessArticle

Vimentin Cytoskeleton Architecture Analysis on Polylactide and Polyhydroxyoctanoate Substrates for Cell Culturing

by Karolina Feliksiak, Daria Solarz, Maciej Guzik, Aneta Zima, Zenon Rajfur and Tomasz Witko

Int. J. Mol. Sci. 2021, 22(13), 6821; https://doi.org/10.3390/ijms22136821 - 25 Jun 2021

Cited by 2 | Viewed by 2282

Abstract

Polylactide (PLA), widely used in bioengineering and medicine, gained popularity due to its biocompatibility and biodegradability. Natural origin and eco-friendly background encourage the search of novel materials with such features, such as polyhydroxyoctanoate (P(3HO)), a polyester of bacterial origin. Physicochemical features of both P(3HO) and PLA have an impact on cellular response 32, i.e., adhesion, migration, and cell morphology, based on the signaling and changes in the architecture of the three cytoskeletal networks: microfilaments (F-actin), microtubules, and intermediate filaments (IF). To investigate the role of IF in the cellular response to the substrate, we focused on vimentin intermediate filaments (VIFs), present in mouse embryonic fibroblast cells (MEF). VIFs maintain cell integrity and protect it from external mechanical stress, and also take part in the transmission of signals from the exterior of the cell to its inner organelles, which is under constant investigation. Physiochemical properties of a substrate have an impact on cells’ morphology, and thus on cytoskeleton network signaling and assembly. In this work, we show how PLA and P(3HO) crystallinity and hydrophilicity influence VIFs, and we identify that two different types of vimentin cytoskeleton architecture: network “classic” and “nutshell-like” are expressed by MEFs in different numbers of cells depending on substrate features. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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19 pages, 20616 KiB

Open AccessArticle

Hydroxyapatite-Coated SPIONs and Their Influence on Cytokine Release

by Bernhard Friedrich, Jean-Philippe Auger, Silvio Dutz, Iwona Cicha, Eveline Schreiber, Julia Band, Aldo R. Boccacccini, Gerhard Krönke, Christoph Alexiou and Rainer Tietze

Int. J. Mol. Sci. 2021, 22(8), 4143; https://doi.org/10.3390/ijms22084143 - 16 Apr 2021

Cited by 8 | Viewed by 2614

Abstract

Hydroxyapatite- or calcium phosphate-coated iron oxide nanoparticles have a high potential for use in many biomedical applications. In this study, a co-precipitation method for the synthesis of hydroxyapatite-coated nanoparticles (SPION^HAp), was used. The produced nanoparticles have been characterized by dynamic light scattering, X-ray diffraction, vibrating sample magnetometry, Fourier transform infrared spectrometry, atomic emission spectroscopy, scanning electron microscopy, transmission electron microscopy, selected area diffraction, and energy-dispersive X-ray spectroscopy. The results showed a successful synthesis of 190 nm sized particles and their stable coating, resulting in SPION^HAp. Potential cytotoxic effects of SPION^HAp on EL4, THP-1, and Jurkat cells were tested, showing only a minor effect on cell viability at the highest tested concentration (400 µg Fe/mL). The results further showed that hydroxyapatite-coated SPIONs can induce minor TNF-α and IL-6 release by murine macrophages at a concentration of 100 µg Fe/mL. To investigate if and how such particles interact with other substances that modulate the immune response, SPION^HAp-treated macrophages were incubated with LPS (lipopolysaccharides) and dexamethasone. We found that cytokine release in response to these potent pro- and anti-inflammatory agents was modulated in the presence of SPION^HAp. Knowledge of this behavior is important for the management of inflammatory processes following in vivo applications of this type of SPIONs. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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20 pages, 2619 KiB

Open AccessArticle

Differential Responses to Bioink-Induced Oxidative Stress in Endothelial Cells and Fibroblasts

by Hatice Genç, Jonas Hazur, Emine Karakaya, Barbara Dietel, Faina Bider, Jürgen Groll, Christoph Alexiou, Aldo R. Boccaccini, Rainer Detsch and Iwona Cicha

Int. J. Mol. Sci. 2021, 22(5), 2358; https://doi.org/10.3390/ijms22052358 - 26 Feb 2021

Cited by 14 | Viewed by 3004

Abstract

A hydrogel system based on oxidized alginate covalently crosslinked with gelatin (ADA-GEL) has been utilized for different biofabrication approaches to design constructs, in which cell growth, proliferation and migration have been observed. However, cell–bioink interactions are not completely understood and the potential effects of free aldehyde groups on the living cells have not been investigated. In this study, alginate, ADA and ADA-GEL were characterized via FTIR and NMR, and their effect on cell viability was investigated. In the tested cell lines, there was a concentration-dependent effect of oxidation degree on cell viability, with the strongest cytotoxicity observed after 72 h of culture. Subsequently, primary human cells, namely fibroblasts and endothelial cells (ECs) were grown in ADA and ADA-GEL hydrogels to investigate the molecular effects of oxidized material. In ADA, an extremely strong ROS generation resulting in a rapid depletion of cellular thiols was observed in ECs, leading to rapid necrotic cell death. In contrast, less pronounced cytotoxic effects of ADA were noted on human fibroblasts. Human fibroblasts had higher cellular thiol content than primary ECs and entered apoptosis under strong oxidative stress. The presence of gelatin in the hydrogel improved the primary cell survival, likely by reducing the oxidative stress via binding to the CHO groups. Consequently, ADA-GEL was better tolerated than ADA alone. Fibroblasts were able to survive the oxidative stress in ADA-GEL and re-entered the proliferative phase. To the best of our knowledge, this is the first report that shows in detail the relationship between oxidative stress-induced intracellular processes and alginate di-aldehyde-based bioinks. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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Review

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22 pages, 22975 KiB

Open AccessReview

Bioengineered Living Bone Grafts—A Concise Review on Bioreactors and Production Techniques In Vitro

by Paulina Kazimierczak and Agata Przekora

Int. J. Mol. Sci. 2022, 23(3), 1765; https://doi.org/10.3390/ijms23031765 - 03 Feb 2022

Cited by 13 | Viewed by 3356

Abstract

It has been observed that bone fractures carry a risk of high mortality and morbidity. The deployment of a proper bone healing method is essential to achieve the desired success. Over the years, bone tissue engineering (BTE) has appeared to be a very promising approach aimed at restoring bone defects. The main role of the BTE is to apply new, efficient, and functional bone regeneration therapy via a combination of bone scaffolds with cells and/or healing promotive factors (e.g., growth factors and bioactive agents). The modern approach involves also the production of living bone grafts in vitro by long-term culture of cell-seeded biomaterials, often with the use of bioreactors. This review presents the most recent findings concerning biomaterials, cells, and techniques used for the production of living bone grafts under in vitro conditions. Particular attention has been given to features of known bioreactor systems currently used in BTE: perfusion bioreactors, rotating bioreactors, and spinner flask bioreactors. Although bioreactor systems are still characterized by some limitations, they are excellent platforms to form bioengineered living bone grafts in vitro for bone fracture regeneration. Moreover, the review article also describes the types of biomaterials and sources of cells that can be used in BTE as well as the role of three-dimensional bioprinting and pulsed electromagnetic fields in both bone healing and BTE. Full article

(This article belongs to the Special Issue Interactions of Cells with Biomaterials for Regenerative Medicine 2.0)

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