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1.
ACS Appl Mater Interfaces ; 15(42): 48855-48870, 2023 Oct 25.
Article in English | MEDLINE | ID: mdl-37823476

ABSTRACT

By means of a "live-cell" template strategy, silica replicas displaying the same morphology and topography of the mammalian cells used as templates are fabricated. The replicas are used as substrates to direct the differentiation of mesenchymal stem cells (MSCs) to predefined cell lineages. Upregulation of specific genes shows how the silica replica-based substrates have the ability to induce the molecular characteristics of the mature cell types from which they have been derived from. Thus, MSCs cultured in the presence of silica replicas of human osteoblasts (HObs) differentiate into HObs-like cells, as shown by the upregulation of specific osteogenic genes. Likewise, when MSCs are incubated with silica replicas derived from human chondrocytes, an enhanced expression of chondrogenic markers is observed. Importantly, the effects of the silica replicas are cell type-specific since the incubation of MSCs with HObs silica replicas does not result in upregulation of chondrogenic markers and vice versa. What is more, for both cases, the differentiation rate is enhanced when the silica replicas are used in combination with growth factors, suggesting a potential synergistic effect. These results demonstrate the potential of this innovative method as an efficient and cheap approach with the potential to eliminate, or at least reduce, the use of biochemically soluble compounds in stem cells research.


Subject(s)
Mesenchymal Stem Cells , Animals , Humans , Cell Lineage , Cell Differentiation , Chondrocytes , Osteogenesis , Cells, Cultured , Chondrogenesis , Mammals
2.
ACS Appl Mater Interfaces ; 15(25): 29729-29742, 2023 Jun 28.
Article in English | MEDLINE | ID: mdl-37319328

ABSTRACT

Soft tissue defects or pathologies frequently necessitate the use of biomaterials that provide the volume required for subsequent vascularization and tissue formation as autrografts are not always a feasible alternative. Supramolecular hydrogels represent promising candidates because of their 3D structure, which resembles the native extracellular matrix, and their capacity to entrap and sustain living cells. Guanosine-based hydrogels have emerged as prime candidates in recent years since the nucleoside self-assembles into well-ordered structures like G-quadruplexes by coordinating K+ ions and π-π stacking, ultimately forming an extensive nanofibrillar network. However, such compositions were frequently inappropriate for 3D printing due to material spreading and low shape stability over time. Thus, the present work aimed to develop a binary cell-laden hydrogel capable of ensuring cell survival while providing enough stability to ensure scaffold biointegration during soft tissue reconstruction. For that purpose, a binary hydrogel made of guanosine and guanosine 5'-monophosphate was optimized, rat mesenchymal stem cells were entrapped, and the composition was bioprinted. To further increase stability, the printed structure was coated with hyperbranched polyethylenimine. Scanning electron microscopic studies demonstrated an extensive nanofibrillar network, indicating excellent G-quadruplex formation, and rheological analysis confirmed good printing and thixotropic qualities. Additionally, diffusion tests using fluorescein isothiocyanate labeled-dextran (70, 500, and 2000 kDa) showed that nutrients of various molecular weights may diffuse through the hydrogel scaffold. Finally, cells were evenly distributed throughout the printed scaffold, cell survival was 85% after 21 days, and lipid droplet formation was observed after 7 days under adipogenic conditions, indicating successful differentiation and proper cell functioning. To conclude, such hydrogels may enable the 3D bioprinting of customized scaffolds perfectly matching the respective soft tissue defect, thereby potentially improving the outcome of the tissue reconstruction intervention.


Subject(s)
Bioprinting , Hydrogels , Rats , Animals , Hydrogels/pharmacology , Hydrogels/chemistry , Guanosine Monophosphate , Guanosine , Biocompatible Materials , Tissue Engineering , Printing, Three-Dimensional , Tissue Scaffolds/chemistry
3.
Front Bioeng Biotechnol ; 11: 1147943, 2023.
Article in English | MEDLINE | ID: mdl-37020512

ABSTRACT

Tissue defects can lead to serious health problems and often require grafts or transplants to repair damaged soft tissues. However, these procedures can be complex and may not always be feasible due to a lack of available tissue. Hydrogels have shown potential as a replacement for tissue grafts due to their ability to support cell survival and encapsulate biomolecules such as growth factors. In particular, guanosine-based hydrogels have been explored as a potential solution, but they often exhibit limited stability which hampers their use in the biofabrication of complex grafts. To address this issue, we explored the use of borate ester chemistry and more complex boric acid derivatives to improve the stability and properties of guanosine-based hydrogels. We hypothesized that the aromatic rings in these derivatives would enhance the stability and printability of the hydrogels through added π-π stack interactions. After optimization, 13 compositions containing either 2-naphthylboronic acid or boric acid were selected. Morphology studies shows a well-defined nanofibrilar structure with good printable properties (thixotropic behaviour, print fidelity and printability). Moreover, the pH of all tested hydrogels was within the range suitable for cell viability (7.4-8.3). Nevertheless, only the boric acid-based formulations were stable for at least 7 days. Thus, our results clearly demonstrated that the presence of additional aromatic rings did actually impair the hydrogel properties. We speculate that this is due to steric hindrance caused by adjacent groups, which disrupt the correct orientation of the aromatic groups required for effective π-π stack interactions of the guanosine building block. Despite this drawback, the developed guanosine-boric acid hydrogel exhibited good thixotropic properties and was able to support cell survival, proliferation, and migration. For instance, SaOS-2 cells planted on these printed structures readily migrated into the hydrogel and showed nearly 100% cell viability after 7 days. In conclusion, our findings highlight the potential of guanosine-boric acid hydrogels as tissue engineering scaffolds that can be readily enhanced with living cells and bioactive molecules. Thus, our work represents a significant advancement towards the development of functionalized guanosine-based hydrogels.

4.
Int J Mol Sci ; 24(4)2023 Feb 20.
Article in English | MEDLINE | ID: mdl-36835636

ABSTRACT

Tissue engineering focuses on the development of materials as biosubstitutes that can be used to regenerate, repair, or replace damaged tissues. Alongside this, 3D printing has emerged as a promising technique for producing implants tailored to specific defects, which in turn increased the demand for new inks and bioinks. Especially supramolecular hydrogels based on nucleosides such as guanosine have gained increasing attention due to their biocompatibility, good mechanical characteristics, tunable and reversible properties, and intrinsic self-healing capabilities. However, most existing formulations exhibit insufficient stability, biological activity, or printability. To address these limitations, we incorporated polydopamine (PDA) into guanosine-borate (GB) hydrogels and developed a PGB hydrogel with maximal PDA incorporation and good thixotropic and printability qualities. The resulting PGB hydrogels exhibited a well-defined nanofibrillar network, and we found that PDA incorporation increased the hydrogel's osteogenic activity while having no negative effect on mammalian cell survival or migration. In contrast, antimicrobial activity was observed against the Gram-positive bacteria Staphylococcus aureus and Staphylococcus epidermidis. Thus, our findings suggest that our PGB hydrogel represents a significantly improved candidate as a 3D-printed scaffold capable of sustaining living cells, which may be further functionalized by incorporating other bioactive molecules for enhanced tissue integration.


Subject(s)
Borates , Hydrogels , Animals , Guanosine , Tissue Engineering , Cell Differentiation , Anti-Bacterial Agents/pharmacology , Printing, Three-Dimensional , Tissue Scaffolds , Mammals
5.
ACS Biomater Sci Eng ; 9(1): 40-61, 2023 01 09.
Article in English | MEDLINE | ID: mdl-36524860

ABSTRACT

Supramolecular hydrogels are of great interest in tissue scaffolding, diagnostics, and drug delivery due to their biocompatibility and stimuli-responsive properties. In particular, nucleosides are promising candidates as building blocks due to their manifold noncovalent interactions and ease of chemical modification. Significant progress in the field has been made over recent years to allow the use of nucleoside-based supramolecular hydrogels in the biomedical field, namely drug delivery and 3D bioprinting. For example, their long-term stability, printability, functionality, and bioactivity have been greatly improved by employing more than one gelator, incorporating different cations, including silver for antibacterial activity, or using additives such as boric acid or even biomolecules. This now permits their use as bioinks for 3D printing to produce cell-laden scaffolds with specified geometries and pore sizes as well as a homogeneous distribution of living cells and bioactive molecules. We have summarized the latest advances in nucleoside-based supramolecular hydrogels. Additionally, we discuss their synthesis, structural properties, and potential applications in tissue engineering and provide an outlook and future perspective on ongoing developments in the field.


Subject(s)
Hydrogels , Tissue Engineering , Hydrogels/chemistry , Nucleosides , Tissue Scaffolds , Printing, Three-Dimensional
6.
Bioact Mater ; 6(12): 4470-4490, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34027235

ABSTRACT

Bacterial infection of implanted scaffolds may have fatal consequences and, in combination with the emergence of multidrug bacterial resistance, the development of advanced antibacterial biomaterials and constructs is of great interest. Since decades ago, metals and their ions had been used to minimize bacterial infection risk and, more recently, metal-based nanomaterials, with improved antimicrobial properties, have been advocated as a novel and tunable alternative. A comprehensive review is provided on how metal ions and ion nanoparticles have the potential to decrease or eliminate unwanted bacteria. Antibacterial mechanisms such as oxidative stress induction, ion release and disruption of biomolecules are currently well accepted. However, the exact antimicrobial mechanisms of the discussed metal compounds remain poorly understood. The combination of different metal ions and surface decorations of nanoparticles will lead to synergistic effects and improved microbial killing, and allow to mitigate potential side effects to the host. Starting with a general overview of antibacterial mechanisms, we subsequently focus on specific metal ions such as silver, zinc, copper, iron and gold, and outline their distinct modes of action. Finally, we discuss the use of these metal ions and nanoparticles in tissue engineering to prevent implant failure.

7.
Int J Mol Sci ; 21(17)2020 Sep 03.
Article in English | MEDLINE | ID: mdl-32899269

ABSTRACT

We have previously reported the fabrication of a polycaprolactone and hydroxyapatite composite scaffold incorporating growth factors to be used for bone regeneration. Two growth factors were incorporated employing a multilayered coating based on polydopamine (PDA). In particular, Bone morphogenetic protein-2 (BMP-2) was bound onto the inner PDA layer while vascular endothelial growth factor (VEGF) was immobilized onto the outer one. Herein, the in vitro release of both growth factors is evaluated. A fastest VEGF delivery followed by a slow and more sustained release of BMP-2 was demonstrated, thus fitting the needs for bone tissue engineering applications. Due to the relevance of the crosstalk between bone-promoting and vessel-forming cells during bone healing, the functionalized scaffolds are further assessed on a co-culture setup of human mesenchymal stem cells and human endothelial progenitor cells. Osteogenic and angiogenic gene expression analysis indicates a synergistic effect between the growth factor-loaded scaffolds and the co-culture conditions. Taken together, these results indicate that the developed scaffolds hold great potential as an efficient platform for bone-tissue applications.


Subject(s)
Bone Morphogenetic Protein 2/metabolism , Endothelial Progenitor Cells/cytology , Indoles/chemistry , Mesenchymal Stem Cells/cytology , Neovascularization, Physiologic , Osteogenesis , Polymers/chemistry , Vascular Endothelial Growth Factor A/metabolism , Cells, Cultured , Coculture Techniques , Endothelial Progenitor Cells/metabolism , Humans , Mesenchymal Stem Cells/metabolism , Tissue Engineering/methods , Tissue Scaffolds/chemistry
8.
Mater Sci Eng C Mater Biol Appl ; 117: 111245, 2020 Dec.
Article in English | MEDLINE | ID: mdl-32919623

ABSTRACT

For bone tissue engineering applications, scaffolds that mimic the porous structure of the extracellular matrix are highly desirable. Herein, we employ a PCL/HA-based scaffold with a double-scaled architecture of small pores coupled to larger ones. To improve the osteoinductivity of the scaffold, we incorporate two different growth factors via polydopamine (PDA) coating. As a first step, we identify the maximum amount of PDA that can be deposited onto the scaffold. Next, to allow for the deposition of a second PDA layer which, in turn, will allow to increase the loading of growth factors, we incorporate a dithiol connecting layer. The thiol groups covalently react with the first PDA coating through Michael addition while also allowing for the incorporation of a second PDA layer. We load the first and second PDA layers with bone morphogenic protein-2 (BMP2) and vascular endothelial growth factor (VEGF), respectively, and evaluate the osteogenic potential of the functionalised scaffold by cell viability, alkaline phosphatase activity and the expression of three different osteogenesis-related genes of pre-seeded human mesenchymal stem cells. Through these studies, we demonstrate that the osteogenic activity of the scaffolds loaded with both BMP2 and VEGF is greater than scaffolds loaded only with BMP2. Importantly, the osteoinductivity is higher when the scaffolds are loaded with BMP2 and VEGF in two different PDA layers. Taken together, these results indicate that the as-prepared scaffolds could be a useful construct for bone-tissue applications.


Subject(s)
Tissue Scaffolds , Vascular Endothelial Growth Factor A , Cell Differentiation , Humans , Indoles , Osteogenesis , Polymers , Tissue Engineering
9.
Polymers (Basel) ; 12(4)2020 Apr 01.
Article in English | MEDLINE | ID: mdl-32244655

ABSTRACT

Infections related to dental implants are a common complication that can ultimately lead to implant failure, and thereby carries significant health and economic costs. In order to ward off these infections, this paper explores the immobilization of triethoxysilylpropyl succinic anhydride (TESPSA, TSP) silane onto dental implants, and the interaction of two distinct monospecies biofilms and an oral plaque with the coated titanium samples. To this end, titanium disks from prior machining were first activated by a NaOH treatment and further functionalized with TESPSA silane. A porous sodium titanate surface was observed by scanning electron microscopy and X-ray photoelectron spectroscopy analyses confirmed the presence of TESPSA on the titanium samples (8.4% for Ti-N-TSP). Furthermore, a lactate dehydrogenase assay concluded that TESPSA did not have a negative effect on the viability of human fibroblasts. Importantly, the in vitro effect of modified surfaces against Streptococcus sanguinis, Lactobacillus salivarius and oral plaque were studied using a viable bacterial adhesion assay. A significant reduction was achieved in all cases but, as expected, with different effectiveness against simple mono-species biofilm (ratio dead/live of 0.4) and complete oral biofilm (ratio dead/live of 0.6). Nevertheless, this approach holds a great potential to provide dental implants with antimicrobial properties.

10.
Biomater Sci ; 7(11): 4813-4826, 2019 Nov 01.
Article in English | MEDLINE | ID: mdl-31535107

ABSTRACT

While ROS display crucial functions in many physiological processes, elevated ROS levels are also related to the initiation and progression of many severe diseases such as cancer, cardiovascular conditions or neurologic disorders. Research approaches to diminish ROS levels during disease progression are currently being focused on the therapeutic administration of antioxidant enzymes. However, enzyme administration suffers from several limitations including their fast elimination from blood upon administration, thus making crucial the development of enzyme encapsulating platforms. We have recently reported a multicompartment architecture constituted by two inherently different types of materials, i.e., polymeric microgels and liposomes. Poly(N-isopropylacrylamide-co-acrylic acid) microgels decorated with liposomes and subsequently coated by a protective poly(dopamine) shell (PDA) combine the benefits of both systems while minimizing some of their drawbacks. Herein, we exploit this dual-component platform as a microreactor for ROS depletion. We combine the intrinsic PDA's antioxidant properties with the encapsulation of the catalase enzyme within the liposomal compartments. The surface of the carrier is further functionalised with a poly(ethylene glycol) layer and the low fouling properties are demonstrated in terms of reduction of protein adsorption and cellular uptake. The potential of the carrier as an antioxidant microreactor is shown by its ability to deplete superoxide radicals and hydrogen peroxide, which can also take place in the presence of the two relevant cell lines.


Subject(s)
Acrylamides/metabolism , Antioxidants/metabolism , Catalase/metabolism , Indoles/metabolism , Polymers/metabolism , Reactive Oxygen Species/metabolism , Acrylamides/chemistry , Animals , Antioxidants/administration & dosage , Antioxidants/chemistry , Catalase/chemistry , Cells, Cultured , Indoles/chemistry , Liposomes/chemistry , Liposomes/metabolism , Mice , Particle Size , Polymers/chemistry , RAW 264.7 Cells , Surface Properties
11.
ACS Appl Mater Interfaces ; 11(6): 5862-5876, 2019 Feb 13.
Article in English | MEDLINE | ID: mdl-30605301

ABSTRACT

Melanoma is malignant skin cancer occurring with increasing prevalence with no effective treatment. A unique feature of melanoma cells is that they require higher concentrations of ltyrosine (l-tyr) for expansion than normal cells. As such, it has been demonstrated that dietary l-tyr restriction lowers systemic l-tyr and suppresses melanoma advancement in mice. Unfortunately, this diet is not well tolerated by humans. An alternative approach to impede melanoma progression will be to administer the enzyme tyrosinase (TYR), which converts l-tyr into melanin. Herein, a multicompartment carrier consisting of a polymer shell entrapping thousands of liposomes is employed to act as a microreactor depleting l-tyr in the presence of melanoma cells. It is shown that the TYR enzyme can be incorporated within the liposomal subunits with preserved catalytic activity. Aiming to mimic the dynamic environment at the tumor site, l-tyr conversion is conducted by co-culturing melanoma cells and microreactors in a microfluidic setup with applied intratumor shear stress. It is demonstrated that the microreactors are concurrently depleting l-tyr, which translates into inhibited melanoma cell growth. Thus, the first microreactor where the depletion of a substrate translates into antitumor properties in vitro is reported.


Subject(s)
Monophenol Monooxygenase/metabolism , Animals , Bioreactors , Cell Line, Tumor , Cell Proliferation , Kinetics , Liposomes/chemistry , Melanins/metabolism , Melanoma, Experimental/metabolism , Melanoma, Experimental/pathology , Mice , Microscopy, Confocal , Microspheres , Monophenol Monooxygenase/chemistry , Polylactic Acid-Polyglycolic Acid Copolymer/chemistry , Quartz Crystal Microbalance Techniques , RAW 264.7 Cells , Tyrosine/metabolism
12.
Colloids Surf B Biointerfaces ; 171: 427-436, 2018 Nov 01.
Article in English | MEDLINE | ID: mdl-30075418

ABSTRACT

Advanced multicompartment drug delivery platforms ensure the co-localization of several drugs within the same carrier, thus making it possible to achieve a more effective and safe therapeutic outcome. Herein, we report a novel multicompartment architecture by combining two intrinsically different systems, i.e., polymeric microgels and liposomes, with the aim to achieve different release kinetics for model compounds. We assemble poly(N-isopropylacrylamide-co-acrylic acid) microgels decorated with liposomes which are subsequently coated with a protective poly(dopamine) shell and a poly(ethylene glycol) (PEG) layer. Since any intravenous administered drug delivery vehicle will get in contact with the dynamics of the blood flow, we evaluate the stealth properties of this novel multicompartment carrier towards protein adsorption and cellular uptake by three relevant cell lines (macrophages, endothelial and cancer cells) under physiological shear stress conditions. Our results demonstrate less protein adsorption for the PEGylated carriers and differences in the extent of internalized carriers depending on the presence of a PEG coating, the studied cell line and the intensity of the applied shear stress. Additionally, we demonstrate that, for all three tested cell lines, shear stress results in the activation of different cell entry pathways as compared to static conditions. All in all, we report a thorough study about the effect of shear stress on the cell association/uptake with a novel multicompartment carrier.


Subject(s)
Indoles/pharmacology , Polyethylene Glycols/pharmacology , Polymers/pharmacology , Shear Strength , Adsorption , Animals , Cell Line , Cell Survival , Drug Carriers/chemistry , Drug Delivery Systems , Gels/chemistry , Gels/pharmacology , Human Umbilical Vein Endothelial Cells/drug effects , Humans , Indoles/chemistry , Liposomes/chemistry , Macrophages/drug effects , Mice , Particle Size , Polyethylene Glycols/chemistry , Polymers/chemistry , RAW 264.7 Cells , Surface Properties
13.
ACS Appl Bio Mater ; 1(2): 452-461, 2018 Aug 20.
Article in English | MEDLINE | ID: mdl-35016368

ABSTRACT

Graphene-based quantum dots (GQDs) are attractive fluorophores due to their excellent photoluminescence properties, water solubility, low cost, and low toxicity. However, the lack of simple, efficient, and environmental-friendly synthesis methods often limits their biological applications. Herein, we explore a novel, one-pot, green synthesis approach for the fabrication of fluorescent GQDs without involving any harsh reagents. Graphene oxide is used as a precursor, and a 2 h hydrothermal synthesis is carried out with assistance of hydrogen peroxide; no further post purification steps are required. The effects of reaction conditions on the characteristics of GQDs are comprehensively investigated. The as-synthesized GQDs show a high photostability and excellent biocompatibility as revealed by cell viability assays for three different cell lines, namely, macrophages, endothelial cells, and a model cancer cell line. The detailed studies of cellular uptake mechanisms suggest that for all of the three cell lines the major internalization route for GQDs is caveolae-mediated endocytosis followed by clathrin-mediated endocytosis at a less extent. Our results demonstrate the great potential of the as-synthesized GQDs as fluorescent nanoprobes. The study also provides unique insight into the cell-GQDs interactions, which is highly valuable for bioimaging and other related applications such as diagnostics and drug delivery.

14.
Adv Healthc Mater ; 7(5)2018 03.
Article in English | MEDLINE | ID: mdl-29205928

ABSTRACT

Artificial organelles created from a bottom up approach are a new type of engineered materials, which are not designed to be living but, instead, to mimic some specific functions inside cells. By doing so, artificial organelles are expected to become a powerful tool in biomedicine. They can act as nanoreactors to convert a prodrug into a drug inside the cells or as carriers encapsulating therapeutic enzymes to replace malfunctioning organelles in pathological conditions. For the design of artificial organelles, several requirements need to be fulfilled: a compartmentalized structure that can encapsulate the synthetic machinery to perform an enzymatic function, as well as a means to allow for communication between the interior of the artificial organelle and the external environment, so that substrates and products can diffuse in and out the carrier allowing for continuous enzymatic reactions. The most recent and exciting advances in architectures that fulfill the aforementioned requirements are featured in this review. Artificial organelles are classified depending on their constituting materials, being lipid and polymer-based systems the most prominent ones. Finally, special emphasis will be put on the intracellular response of these newly emerging systems.


Subject(s)
Artificial Cells , Nanotechnology/trends , Organelles , Animals , Artificial Cells/chemistry , Artificial Cells/classification , Artificial Cells/metabolism , Humans , Nanotechnology/methods , Organelles/chemistry , Organelles/metabolism
15.
ACS Appl Mater Interfaces ; 9(19): 15907-15921, 2017 May 17.
Article in English | MEDLINE | ID: mdl-28117959

ABSTRACT

Cell organelles are subcellular structures entrapping a set of enzymes to achieve a specific functionality. The incorporation of artificial organelles into cells is a novel medical paradigm which might contribute to the treatment of various cell disorders by replacing malfunctioning organelles. In particular, artificial organelles are expected to be a powerful solution in the context of enzyme replacement therapy since enzymatic malfunction is the primary cause of organelle dysfunction. Although several attempts have been made to encapsulate enzymes within a carrier vehicle, only few intracellularly active artificial organelles have been reported to date and they all consist of single-compartment carriers. However, it is noted that biological organelles consist of multicompartment architectures where enzymatic reactions are executed within distinct subcompartments. Compartmentalization allows for multiple processes to take place in close vicinity and in a parallel manner without the risk of interference or degradation. Here, we report on a subcompartmentalized and intracellularly active carrier, a crucial step for advancing artificial organelles. In particular, we develop and characterize a novel capsosome system, which consists of multiple liposomes and fluorescent gold nanoclusters embedded within a polymer carrier capsule. We subsequently demonstrate that encapsulated enzymes preserve their activity intracellularly, allowing for controlled enzymatic cascade reaction within a host cell.


Subject(s)
Artificial Cells , Capsules , Gold , Liposomes , Polymers
16.
Adv Healthc Mater ; 6(4)2017 Feb.
Article in English | MEDLINE | ID: mdl-28004530

ABSTRACT

The creation of artificial organelles is a new paradigm in medical therapy that aims to substitute for missing cellular function by replenishing a specific cellular task. Artificial organelles tackle the challenge of mimicking metabolism, which is the set of chemical reactions that occur within a cell, mainly catalyzed by enzymes. So far, the few reported carriers able to conduct enzymatic reactions intracellularly are based on single-compartment carriers. However, cell organelles outperform by conducting multiple reactions simultaneously within confined sub-compartments. Here, the field of artificial organelles is advanced by reporting the assembly of a microreactor consisting of polymer capsules entrapping gold nanoclusters (AuNCs) and liposomes as sub-compartments. The fluorescence properties of AuNCs are employed to monitor the microreactors uptake by macrophages. Encapsulation is demonstrated and functionality of microreactors with trypsin (TRP) and horseradish peroxidase (HRP)-loaded liposomes is preserved. Multiple enzymatic reactions taking place simultaneously is demonstrated by exposing macrophages with the internalized microreactors to bis-(benzyloxycarbonyl-Ile-Pro-Arg)-Rho-110 and Amplex Red substrates, which are specific for TRP and HRP, respectively. Conversion of the substrates into the respective fluorescent products is observed. This report on the first microreactor conducting multiple enzymatic reactions simultaneously inside a cell is a considerable step in the field of artificial organelles.


Subject(s)
Gold/chemistry , Liposomes/chemistry , Metal Nanoparticles/chemistry , Organelles , Trypsin/chemistry , Animals , Cattle , Horseradish Peroxidase/chemistry , Organelles/chemistry , Organelles/enzymology
17.
Mater Sci Eng C Mater Biol Appl ; 69: 538-45, 2016 Dec 01.
Article in English | MEDLINE | ID: mdl-27612745

ABSTRACT

The aim of this study was to evaluate the in vivo effect of antibacterial modified dental implants in the first stages of peri-implantitis. Thirty dental implants were inserted in the mandibular premolar sites of 5 beagle dogs. Sites were randomly assigned to Ti (untreated implants, 10units), Ti_Ag (silver electrodeposition treatment, 10units), and Ti_TSP (silanization treatment, 10units). Coated implants were characterized by scanning electron microscopy, interferometry and X-ray photoelectron spectroscopy. Two months after implant insertion, experimental peri-implantitis was initiated by ligature placement. Ligatures were removed 2months later, and plaque formation was allowed for 2 additional months. Clinical and radiographic analyses were performed during the study. Implant-tissue samples were prepared for micro computed tomography, backscattered scanning electron microscopy, histomorphometric and histological analyses and ion release measurements. X-ray, SEM and histology images showed that vertical bone resorption in treated implants was lower than in the control group (P<0.05). This effect is likely due to the capacity of the treatments to reduce bacteria colonization on the implant surface. Histological analysis suggested an increase of peri-implant bone formation on silanized implants. However, the short post-ligature period was not enough to detect differences in clinical parameters among implant groups. Within the limits of this study, antibacterial surface treatments have a positive effect against bone resorption induced by peri-implantitis.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Bone Resorption/drug therapy , Bone Resorption/etiology , Coated Materials, Biocompatible/therapeutic use , Dental Implants/adverse effects , Animals , Anti-Bacterial Agents/pharmacology , Bone Resorption/diagnostic imaging , Bone Resorption/pathology , Coated Materials, Biocompatible/pharmacology , Dogs , Female , Gingiva/diagnostic imaging , Gingiva/drug effects , Gingiva/pathology , Ions , Surface Properties
18.
Mater Sci Eng C Mater Biol Appl ; 59: 524-532, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26652404

ABSTRACT

Bacterial infection in dental implants along with osseointegration failure usually leads to loss of the device. Bioactive molecules with antibacterial properties can be attached to titanium surfaces with anchoring molecules such as silanes, preventing biofilm formation and improving osseointegration. Properties of silanes as molecular binders have been thoroughly studied, but research on the biological effects of these coatings is scarce. The aim of the present study was to determine the in vitro cell response and antibacterial effects of triethoxysilypropyl succinic anhydride (TESPSA) silane anchored on titanium surfaces. X-ray photoelectron spectroscopy confirmed a successful silanization. The silanized surfaces showed no cytotoxic effects. Gene expression analyses of Sarcoma Osteogenic (SaOS-2) osteoblast-like cells cultured on TESPSA silanized surfaces reported a remarkable increase of biochemical markers related to induction of osteoblastic cell differentiation. A manifest decrease of bacterial adhesion and biofilm formation at early stages was observed on treated substrates, while favoring cell adhesion and spreading in bacteria-cell co-cultures. Surfaces treated with TESPSA could enhance a biological sealing on implant surfaces against bacteria colonization of underlying tissues. Furthermore, it can be an effective anchoring platform of biomolecules on titanium surfaces with improved osteoblastic differentiation and antibacterial properties.


Subject(s)
Bacterial Adhesion/drug effects , Biofilms/drug effects , Cell Differentiation/drug effects , Osteoblasts/drug effects , Silanes/chemistry , Titanium/chemistry , Bacteria/drug effects , Cell Line , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Humans , Silanes/pharmacology , Surface Properties , Titanium/pharmacology
19.
J Biomater Appl ; 30(3): 279-89, 2015 Sep.
Article in English | MEDLINE | ID: mdl-25876889

ABSTRACT

Acrylic bone cements have an elastic modulus several times higher than the surrounding trabecular bone. This has been hypothesized to contribute to certain clinical complications. There are indications that the addition of specific fatty acids and triglyceride oils may reduce the elastic modulus of these types of cements. Some of these additives also appear to have inherent antibiotic properties, although this has never been evaluated in bone cements. In this study, several types of fatty acids and triglyceride oils were evaluated for use in acrylic bone cements. Their mechanical properties were evaluated under uniaxial compression testing and selected cements were then further characterized in terms of microstructure, handling and antibacterial properties using scanning electron microscopy, polymerization temperature measurements, agar diffusion tests and bactericidal activity assays of cement extracts. It was found that any of the evaluated fatty acids or triglyceride oils could be used to tailor the stiffness of acrylic bone cements, although at varying concentrations, which also depended on the type of commercial base cement used. In particular, the addition of very small amounts of linoleic acid (<2.0 wt%) resulted in Young's moduli and compressive strengths in the range of human trabecular bone, while maintaining a similar setting time. Further, the addition of 12.6 wt% ricinoleic acid to Osteopal V cement was found to have a significant antibacterial effect, inhibiting growth of Staphylococcus aureus in an agar diffusion test as well as demonstrating 100% bactericidal activity against the same strain.


Subject(s)
Acrylates/pharmacology , Anti-Bacterial Agents/pharmacology , Bone Cements/pharmacology , Fatty Acids, Unsaturated/pharmacology , Triglycerides/administration & dosage , Acrylates/chemistry , Anti-Bacterial Agents/chemistry , Bone Cements/chemistry , Gas Chromatography-Mass Spectrometry , Materials Testing , Microbial Sensitivity Tests , Microscopy, Electron, Scanning , Staphylococcus aureus/drug effects
20.
ACS Appl Mater Interfaces ; 7(10): 5992-6001, 2015 Mar 18.
Article in English | MEDLINE | ID: mdl-25734758

ABSTRACT

Dental plaque is a biofilm that causes dental caries, gingivitis, and periodontitis. Most of the studies in antibacterial coatings have been conducted by in vitro single-species biofilm formation, but oral biofilm involves more than 700 different bacterial species that are able to interact. Therefore, new studies are focused on in vitro multispecies biofilm models that mimic in vivo biofilms. The aim of the present work was to study different antibacterial coatings onto titanium surfaces and evaluate the in vitro antimicrobial properties of the surfaces on two different bacterial species and an oral biofilm. The lactate dehydrogenase assay determined that treated samples did not affect fibroblast viability. In addition, the viability of microorganisms on modified samples was evaluated by a LIVE/DEAD BacLight bacterial viability kit. Although a decrease in viable bacteria onto treated samples was obtained, the results showed differences in effectiveness when single-biofilm and oral plaque were tested. It confirms, as we expected, the distinct sensitivities that bacterial strains have. Thus, this multispecies biofilms model holds a great potential to assess antibacterial properties onto samples for dental purposes.


Subject(s)
Anti-Bacterial Agents/pharmacology , Biofilms/drug effects , Biofilms/growth & development , Coated Materials, Biocompatible/pharmacology , Microbial Consortia/physiology , Titanium/chemistry , Anti-Bacterial Agents/chemistry , Cell Survival/drug effects , Cell Survival/physiology , Coated Materials, Biocompatible/chemical synthesis , Materials Testing , Microbial Consortia/drug effects , Surface Properties
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