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1.
ACS Nano ; 16(10): 16234-16248, 2022 10 25.
Article in English | MEDLINE | ID: mdl-36190461

ABSTRACT

Although hydrogel-based patches have shown promising therapeutic efficacy in myocardial infarction (MI), synergistic mechanical, electrical, and biological cues are required to restore cardiac electrical conduction and diastolic-systolic function. Here, an injectable mechanical-electrical coupling hydrogel patch (MEHP) is developed via dynamic covalent/noncovalent cross-linking, appropriate for cell encapsulation and minimally invasive implantation into the pericardial cavity. Pericardial fixation and hydrogel self-adhesiveness properties enable the MEHP to highly compliant interfacial coupling with cyclically deformed myocardium. The self-adaptive MEHP inhibits ventricular dilation while assisting cardiac pulsatile function. The MEHP with the electrical conductivity and sensitivity to match myocardial tissue improves electrical connectivity between healthy and infarcted areas and increases electrical conduction velocity and synchronization. Overall, the MEHP combined with cell therapy effectively prevents ventricular fibrosis and remodeling, promotes neovascularization, and restores electrical propagation and synchronized pulsation, facilitating the clinical translation of cardiac tissue engineering.


Subject(s)
Hydrogels , Myocardial Infarction , Humans , Hydrogels/pharmacology , Hydrogels/therapeutic use , Myocardium , Myocardial Infarction/drug therapy , Electric Conductivity , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology
2.
Int J Biol Macromol ; 165(Pt B): 2528-2540, 2020 Dec 15.
Article in English | MEDLINE | ID: mdl-33098901

ABSTRACT

A new sponge-type hydrogel was obtained by cross-linking hyaluronic acid (HA) and poly(methylvinylether-alt-maleic acid) P(MVE-alt-MA) through a solvent-free thermal method. The sponge-type hydrogel was characterized and checked as a support for cell growth. The influence of concentration and weight ratio of polymers on the morphology and hydrogel stability was investigated. The total polymers concentration of 3% (w/w) and the weight ratio of 1:1 were optimal for the synthesis of a stable hydrogel (HA3P50) and to promote cell proliferation. The swelling measurements revealed a high-water absorption capacity of the hydrogel in basic medium. Diphenhydramine (DPH), lidocaine (Lid) and propranolol (Prop) were loaded within the hydrogel as a model drugs to investigate the ability of drug transport and release. In vitro studies revealed that HA3P50 hydrogel promoted the adhesion and proliferation of human hepatocellular carcinoma cell line HepG2, providing a good support for 3D cell culture to obtain surrogate tumor scaffold suitable for preclinical anti-cancer drug screening.


Subject(s)
Cell Proliferation/drug effects , Hyaluronic Acid/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Hydrogels/pharmacology , Carcinoma, Hepatocellular/drug therapy , Cell Cycle/drug effects , Diphenhydramine/pharmacology , Hep G2 Cells , Humans , Hyaluronic Acid/pharmacology , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Hydrogels/chemistry , Lidocaine/pharmacology , Liver Neoplasms/drug therapy , Maleates/chemistry , Maleates/pharmacology , Propranolol/pharmacology
3.
Biotechnol Prog ; 36(4): e2986, 2020 07.
Article in English | MEDLINE | ID: mdl-32108999

ABSTRACT

Cardiovascular disease is the leading cause of death worldwide, and current treatments are ineffective or unavailable to majority of patients. Engineered cardiac tissue (ECT) is a promising treatment to restore function to the damaged myocardium; however, for these treatments to become a reality, tissue fabrication must be amenable to scalable production and be used in suspension culture. Here, we have developed a low-cost and scalable emulsion-based method for producing ECT microspheres from poly(ethylene glycol) (PEG)-fibrinogen encapsulated mouse embryonic stem cells (mESCs). Cell-laden microspheres were formed via water-in-oil emulsification; encapsulation occurred by suspending the cells in hydrogel precursor solution at cell densities from 5 to 60 million cells/ml, adding to mineral oil and vortexing. Microsphere diameters ranged from 30 to 570 µm; size variability was decreased by the addition of 2% poly(ethylene glycol) diacrylate. Initial cell encapsulation density impacted the ability for mESCs to grow and differentiate, with the greatest success occurring at higher cell densities. Microspheres differentiated into dense spheroidal ECTs with spontaneous contractions occurring as early as Day 10 of cardiac differentiation; furthermore, these ECT microspheres exhibited appropriate temporal changes in gene expression and response to pharmacological stimuli. These results demonstrate the ability to use an emulsion approach to encapsulate pluripotent stem cells for use in microsphere-based cardiac differentiation.


Subject(s)
Cell Differentiation/drug effects , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Mouse Embryonic Stem Cells/cytology , Pluripotent Stem Cells/cytology , Animals , Cell Differentiation/genetics , Cell Encapsulation/methods , Cell Proliferation/drug effects , Emulsions/chemistry , Emulsions/pharmacology , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Mice , Microspheres , Mouse Embryonic Stem Cells/drug effects , Pluripotent Stem Cells/drug effects , Tissue Engineering/trends
4.
J Biomed Mater Res B Appl Biomater ; 108(1): 14-21, 2020 01.
Article in English | MEDLINE | ID: mdl-30893515

ABSTRACT

The purpose is to estimate the oxygen diffusion coefficient and the relaxation time of the cornea with respect to the oxygen tension at the cornea-tears interface. Both findings are discussed. From the experimental data provided by Bonanno et al., the oxygen tension measurements in vivo for human cornea-tears-contact lens (CL), the relaxation time of the cornea, and their oxygen diffusion coefficient were obtained by numerical calculation using the Monod-kinetic model. Our results, considering the relaxation time of the cornea, observe a different behavior. At the time less than 8 s, the oxygen diffusivity process is upper-diffusive, and for the relaxation time greater than 8 s, the oxygen diffusivity process is lower-diffusive. Both cases depend on the partial pressure of oxygen at the entrance of the cornea. The oxygen tension distribution in the cornea-tears interface is separated into two different zones: one for conventional hydrogels, which is located between 6 and 75 mmHg, with a relaxation time included between 8 and 19 s, and the other zone for silicone hydrogel CLs, which is located at high oxygen tension, between 95 and 140 mmHg, with a relaxation time in the interval of 1.5-8 s. It is found that in each zone, the diffusion coefficient varies linearly with the oxygen concentration, presenting a discontinuity in the transition of 8 s. This could be interpreted as an aerobic-to-anaerobic transition. We attribute this behavior to the coupling formalism between oxygen diffusion and biochemical reactions to produce adenosine triphosphate. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 108B:14-21, 2020.


Subject(s)
Contact Lenses, Hydrophilic , Cornea/metabolism , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Models, Biological , Oxygen/metabolism , Cornea/pathology , Diffusion , Humans , Tears
5.
Molecules ; 24(9)2019 May 07.
Article in English | MEDLINE | ID: mdl-31067670

ABSTRACT

Three-dimensional (3D) tissue models replicating liver architectures and functions are increasingly being needed for regenerative medicine. However, traditional studies are focused on establishing 2D environments for hepatocytes culture since it is challenging to recreate biodegradable 3D tissue-like architecture at a micro scale by using hydrogels. In this paper, we utilized a gelatin methacryloyl (GelMA) hydrogel as a matrix to construct 3D lobule-like microtissues for co-culture of hepatocytes and fibroblasts. GelMA hydrogel with high cytocompatibility and high structural fidelity was determined to fabricate hepatocytes encapsulated micromodules with central radial-type hole by photo-crosslinking through a digital micromirror device (DMD)-based microfluidic channel. The cellular micromodules were assembled through non-contact pick-up strategy relying on local fluid-based micromanipulation. Then the assembled micromodules were coated with fibroblast-laden GelMA, subsequently irradiated by ultraviolet for integration of the 3D lobule-like microtissues encapsulating multiple cell types. With long-term co-culture, the 3D lobule-like microtissues encapsulating hepatocytes and fibroblasts maintained over 90% cell viability. The liver function of albumin secretion was enhanced for the co-cultured 3D microtissues compared to the 3D microtissues encapsulating only hepatocytes. Experimental results demonstrated that 3D lobule-like microtissues fabricated by GelMA hydrogels capable of multicellular co-culture with high cell viability and liver function, which have huge potential for liver tissue engineering and regenerative medicine applications.


Subject(s)
Gelatin/chemical synthesis , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Liver/drug effects , Tissue Engineering/methods , Cell Survival/drug effects , Coculture Techniques , Fibroblasts/chemistry , Fibroblasts/drug effects , Gelatin/chemistry , Gelatin/pharmacology , Hep G2 Cells , Hepatocytes/drug effects , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/chemical synthesis , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Liver/growth & development , Regenerative Medicine
6.
ACS Appl Mater Interfaces ; 11(18): 16320-16327, 2019 May 08.
Article in English | MEDLINE | ID: mdl-30985103

ABSTRACT

Bacterial infection is one of main causes that threaten global human health. Especially, antibiotic-resistant bacteria like methicillin-resistant Staphylococcus aureus (MRSA) lead to high mortality rate and more expensive treatment cost. Here, a novel amino-acid-modified conjugated oligomer OTE-d-Phe was synthesized by modifying the side chain of conjugated oligo(thiophene ethynylene) with d-phenylalanine. By mixing 9-fluorenylmethyloxycarbonyl-l-phenylalanin (Fmoc-l-Phe) with OTE-d-Phe, a new and biocompatible low-molecular weight hydrogel (HG-2) was prepared through self-assembly. In solution, HG-2 can effectively capture bacteria spontaneously, such as Escherichia coli and MRSA. Most importantly, the hydrogel has specific and strong antibacterial activity against MRSA over methicillin-susceptible S. aureus, Staphylococcus epidermidis, and E. coli. Interestingly, when the hydrogel was put on a model surface, a piece of cloth, it also is able to selectively kill MRSA with low cell cytotoxicity. The antibacterial mechanism was investigated, and it demonstrated that the HG-2 interacts with and physically breaks the cell wall and membrane, which leads to MRSA death. Therefore, this new conjugated oligomer-based hydrogel provides promising applications in disinfection and therapy of MRSA in hospital and in community.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Synergism , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Staphylococcal Infections/drug therapy , Amino Acids/drug effects , Anti-Bacterial Agents/chemistry , Escherichia coli/drug effects , Escherichia coli/pathogenicity , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/chemical synthesis , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Methicillin-Resistant Staphylococcus aureus/drug effects , Methicillin-Resistant Staphylococcus aureus/pathogenicity , Microbial Sensitivity Tests , Phenylalanine/chemistry , Staphylococcal Infections/microbiology , Staphylococcus epidermidis/drug effects , Staphylococcus epidermidis/pathogenicity , Thiophenes/chemical synthesis , Thiophenes/chemistry , Thiophenes/pharmacology
7.
Mater Sci Eng C Mater Biol Appl ; 99: 905-918, 2019 Jun.
Article in English | MEDLINE | ID: mdl-30889765

ABSTRACT

BACKGROUND: Due to unmet need for bone augmentation, our aim was to promote osteogenic differentiation of human adipose stem cells (hASCs) encapsulated in gellan gum (GG) or collagen type I (COL) hydrogels with bioactive glass (experimental glass 2-06 of composition [wt-%]: Na2O 12.1, K2O 14.0, CaO 19.8, P2O5 2.5, B2O3 1.6, SiO2 50.0) extract based osteogenic medium (BaG OM) for bone construct development. GG hydrogels were crosslinked with spermidine (GG-SPD) or BaG extract (GG-BaG). METHODS: Mechanical properties of cell-free GG-SPD, GG-BaG, and COL hydrogels were tested in osteogenic medium (OM) or BaG OM at 0, 14, and 21 d. Hydrogel embedded hASCs were cultured in OM or BaG OM for 3, 14, and 21 d, and analyzed for viability, cell number, osteogenic gene expression, osteocalcin production, and mineralization. Hydroxyapatite-stained GG-SPD samples were imaged with Optical Projection Tomography (OPT) and Selective Plane Illumination Microscopy (SPIM) in OM and BaG OM at 21 d. Furthermore, Raman spectroscopy was used to study the calcium phosphate (CaP) content of hASC-secreted ECM in GG-SPD, GG-BaG, and COL at 21 d in BaG OM. RESULTS: The results showed viable rounded cells in GG whereas hASCs were elongated in COL. Importantly, BaG OM induced significantly higher cell number and higher osteogenic gene expression in COL. In both hydrogels, BaG OM induced strong mineralization confirmed as CaP by Raman spectroscopy and significantly improved mechanical properties. GG-BaG hydrogels rescued hASC mineralization in OM. OPT and SPIM showed homogeneous 3D cell distribution with strong mineralization in BaG OM. Also, strong osteocalcin production was visible in COL. CONCLUSIONS: Overall, we showed efficacious osteogenesis of hASCs in 3D hydrogels with BaG OM with potential for bone-like grafts.


Subject(s)
Adipose Tissue/cytology , Cell Differentiation , Collagen Type I/pharmacology , Glass/chemistry , Osteogenesis , Polysaccharides, Bacterial/pharmacology , Stem Cells/cytology , Animals , Biomarkers/metabolism , Calcification, Physiologic/drug effects , Cell Count , Cell Differentiation/drug effects , Cell Survival/drug effects , Compressive Strength , Cross-Linking Reagents/chemistry , Durapatite/chemistry , Female , Gene Expression Regulation/drug effects , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Ions , Middle Aged , Minerals/chemistry , Osteocalcin/metabolism , Osteogenesis/drug effects , Osteogenesis/genetics , Rats , Serum/metabolism , Spectrum Analysis, Raman , Stem Cells/drug effects , Tissue Scaffolds/chemistry
8.
Colloids Surf B Biointerfaces ; 177: 370-376, 2019 May 01.
Article in English | MEDLINE | ID: mdl-30785034

ABSTRACT

Although antibiotics have been widely used, the problem of bacterial infection in the medical field still faces many challenges. In this study, we designed a new lignin based antimicrobial hydrogel for antimicrobial application. First, we grafted the amino group onto sodium lignin sulfonate through Mannich reaction to obtain lignin amine (LA), which can cross-link with poly(vinyl alcohol) (PVA) to form hydrogel. Then, silver nitrate solution is added to the formed gel pre-solution to be in situ reduced to silver nanoparticles. The enhanced effect of antibacterial properties due to lignin and silver nanoparticles endows the hydrogel enhanced antibacterial properties. The modification of sodium lignosulfonate and the crosslinking reaction between LA and PVA are confirmed by FTIR, while the content of nitrogen in LA is characterized by XPS. The SEM image of the hydrogel after lyophilization illustrates its internal porous network structure. The rheological test of hydrogel demonstrates its good strength and elasticity. The hydrogel exhibits good antibacterial properties in in vitro antibacterial experiments towards both S. aureus and E. coli, while toxicity tests using L929 cells demonstrated good biocompatibility of the hydrogel.


Subject(s)
Anti-Bacterial Agents/pharmacology , Escherichia coli/drug effects , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Lignin/chemistry , Metal Nanoparticles/chemistry , Silver/chemistry , Silver/pharmacology , Staphylococcus aureus/drug effects , Animals , Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/chemistry , Biocompatible Materials/chemical synthesis , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Cell Line , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Lignin/pharmacology , Mice , Microbial Sensitivity Tests , Oxidation-Reduction , Particle Size , Surface Properties
9.
Mater Sci Eng C Mater Biol Appl ; 96: 205-217, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30606527

ABSTRACT

Although phenytoin is an antiepileptic drug used in the oral treatment of epilepsy, its off-label use as a cutaneous healing agent has been studied in recent years due to the frequent reports of gingival hyperplasia after oral administration. However, the cutaneous topical application of phenytoin should prevent percutaneous skin permeation. Therefore, the aim of this study was to evaluate the in vitro skin permeation/retention and in vivo effects of nanocapsules and nanoemulsions loaded with phenytoin and formulated as chitosan hydrogels on the healing process of cutaneous wounds in rats. The hydrogels had adequate pH values (4.9-5.6) for skin application, drug content of 0.025% (w/w), and non-Newtonian pseudoplastic rheological behaviour. Hydrogels containing nanocapsules and nanoemulsions enabled improved controlled release of phenytoin and adhesion to skin, compared with hydrogels containing non-encapsulated phenytoin. In vitro skin permeation studies showed that phenytoin permeation to the receptor compartment, and consequently the risk of systemic absorption, may be reduced by nanoencapsulation without any change in the in vivo performance of phenytoin in the wound healing process in rats.


Subject(s)
Chitosan , Hydrogel, Polyethylene Glycol Dimethacrylate , Nanocapsules , Phenytoin , Skin Absorption/drug effects , Wound Healing/drug effects , Wounds and Injuries/drug therapy , Administration, Topical , Animals , Chitosan/chemistry , Chitosan/pharmacokinetics , Chitosan/pharmacology , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacokinetics , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Male , Nanocapsules/chemistry , Nanocapsules/therapeutic use , Phenytoin/chemistry , Phenytoin/pharmacokinetics , Phenytoin/pharmacology , Rats , Rats, Wistar , Swine , Wounds and Injuries/metabolism
10.
Cell Tissue Bank ; 20(1): 25-34, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30673903

ABSTRACT

One of most important issue in the field of regenerative medicine is selection of appropriate cells, scaffolds and bioreactors. The present study aimed to investigate the appropriate method for the isolation of human UC-MSCs cells from explant cultured in alginate scaffold within novel perfusion bioreactor. MSCs were isolated with explant method and CD markers such CD73, CD31, CD90 and CD105 as were analyzed by flow cytometry. The culture chamber of the novel perfusion bioreactor was made from Plexiglas and housed the cell/scaffold constructs in the central part and the medium for the whole culture period. The flow behavior within the bioreactor chamber were performed for closed and open bypass systems. The shear stress profiles simulated using CFD modeling. The fluid flow distribution within the bioreactor chamber was performed in PBS solution containing a blue colorant. UC explants were resuspended in sodium alginate and were allowed to polymerize and placed in the perfusion bioreactor and cultured. MSCs were positive for mesenchymal markers such as CD73 and CD31. All 3D Perfusion bioreactor parts, except peristaltic pump was sterilizable by autoclaving. Results of CFD indicated very low wall shear stress on surface of culture chamber at flow rate 2 ml/min. The maximum wall shear stress was 1.10 × 10-3 m/s = 0.0110 dyne/cm2 (1 Pa = 10 dyne/cm2). The fluid flow distribution within the alginate gel initially exhibited oscillation. In comparison, when encapsulated explants were placed in the perfusion bioreactor, cell proliferation appeared faster (4.6 × 1011 ± 9.2 × 1011) than explants cultures in 2D conventional culture method (3.2 × 1011 ± 1 × 1011). Proliferated cell formed several colonies. Migration of chondrocytes to the periphery of the alginate bead was visible after 1 week of culture. Perfusion bioreactor with low shear stress and alginate hydrogel improve cell isolation and expansion and eliminate cell passaging and enhance colony forming unit of UC-MSCs.


Subject(s)
Bioreactors , Cell Culture Techniques/methods , Cell Separation/methods , Perfusion , Stem Cells/cytology , Stress, Mechanical , Cell Proliferation , Cells, Cultured , Humans , Hydrodynamics , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Mesenchymal Stem Cells/cytology , Time Factors , Tissue Scaffolds/chemistry , Umbilical Cord/cytology
11.
J Colloid Interface Sci ; 535: 149-158, 2019 Feb 01.
Article in English | MEDLINE | ID: mdl-30292105

ABSTRACT

Graphene hydrogels hold great potential for the disinfection of bacteria-contaminated water. However, the intrinsic antibacterial activity of graphene hydrogels is not satisfactory, and the incorporation of other antibacterial agents often results in their unwanted releases. Here, we present a new strategy to improve the antibacterial activities of graphene hydrogels. We first synthesized a new pi-conjugated molecule containing five aromatic rings and two side-linked quaternary ammonium (QA) groups, denoted as piQA. Next, we fabricated composite gravity filters by assembling piQA with reduced graphene oxide (rGO) hydrogel. The rGO hydrogel helps to form a sponge-like physical sieve, contributes to the overall antibacterial activity, and provides abundant pi-rich surfaces. The large aromatic cores of piQA allow the formation of collectively strong pi-pi interactions with rGO, resulting in a high piQA mass loading of ∼31 wt%. Due to the sieving effect of rGO hydrogel and the synergistic antibacterial activity of rGO and piQA, the filters prepared based on piQA-rGO assemblies can remove over 99.5% of Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus) cells with a high-water treatment capacity of 10 L g-1. Furthermore, the piQA-rGO assemblies show low toxicity towards two different mammalian cell lines (L929 and macrophages), and the release of piQA is also negligible. Overall, the new piQA-rGO assembly demonstrates high potential for water disinfection applications.


Subject(s)
Anti-Bacterial Agents/pharmacology , Escherichia coli/drug effects , Graphite/pharmacology , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Quaternary Ammonium Compounds/pharmacology , Staphylococcus aureus/drug effects , Animals , Anti-Bacterial Agents/chemistry , Cell Line , Cell Survival/drug effects , Graphite/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Macrophages/drug effects , Mice , Microbial Sensitivity Tests , Particle Size , Quaternary Ammonium Compounds/chemistry , Surface Properties , Water Microbiology , Water Pollution , Water Supply
12.
ACS Appl Mater Interfaces ; 11(3): 2849-2859, 2019 Jan 23.
Article in English | MEDLINE | ID: mdl-30589529

ABSTRACT

The growing area of biomaterial sciences has attracted broad attention in recent years in the development of peptide-based biocompatible materials with inherent therapeutic potentials. Here, we developed an Amoc (9-anthracenemethoxycarbonyl)-capped dipeptide-based biocompatible, injectable, thixotropic, and self-healable hydrogel. In vitro cytotoxicity of the hydrogel was investigated with the human embryonic kidney cell (HEK293) line. We observed that the synthesized peptide is noncytotoxic. The hydrogel showed an antibacterial efficacy against Gram-positive and Gram-negative bacteria. In vivo anti-inflammatory activity of the hydrogel was investigated using the rat air pouch model of acute inflammation. The major parameters considered for the anti-inflammatory study were exudate volume, total and differential white blood cell count, tissue histology, and lipid peroxidation assay. These experimental data suggest biocompatibility and potential therapeutic applications of peptide hydrogel in inflammation.


Subject(s)
Biocompatible Materials/pharmacology , Cell Proliferation/drug effects , Inflammation/drug therapy , Peptides/pharmacology , Animals , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Biocompatible Materials/chemistry , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , HEK293 Cells , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Inflammation/microbiology , Inflammation/pathology , Lung/drug effects , Lung/microbiology , Lung/pathology , Peptides/chemistry , Rats
13.
Carbohydr Polym ; 202: 409-417, 2018 Dec 15.
Article in English | MEDLINE | ID: mdl-30287017

ABSTRACT

The study examines the use of macrobeads for the controlled-release of bacteria. Macrobeads were prepared by an easy dripping-technique using 20/80 wt/wt chitosan-starch blends and sodium tripolyphosphate as cross-linking agent. The resulting polymeric matrix was examined by SEM, XRD, TGA, and solid-RMN. The swelling-equilibrium, thermal behaviour, crystallinity, and size of macrobeads were affected by the autoclave-sterilization. The diameter of the sterilized xerogel was c.a. 1.6 mm. The results suggested that ionotropic-gelation and neutralization were the mechanisms underlying hydrogel formation. Plant growth-promoting bacteria (PGPB) were loaded into macrobeads separately or co-inoculated. Bacteria loaded macrobeads were dried and stored. Bacteria survived at least 12 months in orders of 109 CFU of A. brasilense/g and 108 CFU of P. fluorescens/g. Bacterial release in sterile saline solution tended to a super Case-II transport mechanism. Polymeric-matrix release efficiently both PGPB in natural soils, which uncovers their potential for the formulation of novel and improved biofertilizers.


Subject(s)
Azospirillum brasilense/drug effects , Chitosan/pharmacology , Crops, Agricultural/growth & development , Drug Delivery Systems , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Pseudomonas fluorescens/drug effects , Starch/pharmacology , Azospirillum brasilense/metabolism , Chitosan/chemistry , Crops, Agricultural/microbiology , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Particle Size , Pseudomonas fluorescens/metabolism , Starch/chemistry
14.
Biomolecules ; 8(4)2018 09 28.
Article in English | MEDLINE | ID: mdl-30274232

ABSTRACT

Nanoparticles can be used as a smart drug delivery system, when they release the drug only upon degradation by specific enzymes. A method to create such responsive materials is the formation of hydrogel nanoparticles, which have enzymatically degradable crosslinkers. Such hydrogel nanoparticles were prepared by ionotropic gelation sodium alginate with lysine-rich peptide sequences-either α-poly-L-lysine (PLL) or the aggrecanase-labile sequence KKKK-GRD-ARGSV↓NITEGE-DRG-KKKK. The nanoparticle suspensions obtained were analyzed by means of dynamic light scattering and nanoparticle tracking analysis. Degradation experiments carried out with the nanoparticles in suspension revealed enzyme-induced lability. Drugs present in the polymer solution during the ionotropic gelation can be encapsulated in the nanoparticles. Drug loading was investigated for interferon-ß (IFN-ß) as a model, using a bioluminescence assay with MX2Luc2 cells. The encapsulation efficiency for IFN-ß was found to be approximately 25%. The nanoparticles suspension can be used to spray-coat titanium alloys (Ti-6Al-4V) as a common implant material. The coatings were proven by ellipsometry, reflection-absorption infrared spectroscopy, and X-ray photoelectron spectroscopy. An enzyme-responsive decrease in layer thickness is observed due to the degradation of the coatings. The Alg/peptide coatings were cytocompatible for human gingival fibroblasts (HGFIB), which was investigated by CellTiterBlue and lactate dehydrogenase (LDH) assay. However, HGFIBs showed poor adhesion and proliferation on the Alg/peptide coatings, but these could be improved by modification of the alginate with a RGD-peptide sequence. The smart drug release system presented can be further tailored to have the right release kinetics and cell adhesion properties.


Subject(s)
Biopolymers/pharmacology , Coated Materials, Biocompatible/therapeutic use , Drug Liberation , Interferon-beta/pharmacology , Biopolymers/chemistry , Chitosan/chemistry , Chitosan/pharmacology , Coated Materials, Biocompatible/chemistry , Fibroblasts/drug effects , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Interferon-beta/chemistry , Nanoparticles/administration & dosage , Nanoparticles/chemistry , Polylysine/chemistry , Polylysine/pharmacology , Surface Properties , Titanium/chemistry
15.
Sci Rep ; 8(1): 16061, 2018 10 30.
Article in English | MEDLINE | ID: mdl-30375445

ABSTRACT

Since the oral mucosa is continuously exposed to abundant microbes, one of its most important defense features is a highly proliferative, thick, stratified epithelium. The cellular mechanisms responsible for this are still unknown. The aim of this study was to determine whether multi-species oral biofilm contribute to the extensive stratification and primed antimicrobial defense in epithelium. Two in vitro models were used: 3D reconstructed human gingiva (RHG) and oral bacteria representative of multi-species commensal biofilm. The organotypic RHG consists of a reconstructed stratified gingiva epithelium on a gingiva fibroblast populated hydrogel (lamina propria). Biofilm was cultured from healthy human saliva, and consists of typical commensal genera Granulicatella and major oral microbiota genera Veillonella and Streptococcus. Biofilm was applied topically to RHG and host-microbiome interactions were studied over 7 days. Compared to unexposed RHG, biofilm exposed RHG showed increased epithelial thickness, more organized stratification and increased keratinocyte proliferation. Furthermore biofilm exposure increased production of RHG anti-microbial proteins Elafin, HBD2 and HBD3 but not HBD1, adrenomedullin or cathelicidin LL-37. Inflammatory and antimicrobial cytokine secretion (IL-6, CXCL8, CXCL1, CCL20) showed an immediate and sustained increase. In conclusion, exposure of RHG to commensal oral biofilm actively contributes to RHG epithelial barrier function.


Subject(s)
Biofilms/growth & development , Gingiva/growth & development , Host-Pathogen Interactions/genetics , Microbiota/genetics , Coculture Techniques , Elafin/genetics , Epithelial Cells/microbiology , Epithelial Cells/pathology , Fibroblasts/microbiology , Gene Expression Regulation/genetics , Gingiva/microbiology , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Mouth Mucosa/microbiology , Primary Cell Culture/methods , Saliva/microbiology , Streptococcus/growth & development , Streptococcus/pathogenicity , Veillonella/growth & development , Veillonella/pathogenicity , beta-Defensins/genetics
16.
Carbohydr Polym ; 202: 545-553, 2018 Dec 15.
Article in English | MEDLINE | ID: mdl-30287034

ABSTRACT

Here, we present a novel strategy to fabricate hyaluronic acid (HA) hydrogels with excellent physical and biological properties. The cross-linking of HA hydrogel by butanediol diglycidyle ether (BDDE) was characterized under different reaction temperatures, and the resulting physical properties (i.e., the storage modulus and swelling ratio) were measured. The ratio between the cross-linking rate (a strengthening effect) and the hydrolysis rate (a weakening effect) was much greater with lower cross-linking temperatures after sufficient cross-linking time, resulting in a noticeably higher storage modulus. As the cross-linking temperature decreased, the formed HA hydrogel structure became denser with smaller pores. Moreover, the introduction of low-temperature HA cross-linking strategy also resulted in an enhanced several important characteristics of HA hydrogels including its enzymatic resistivity and its ability to elicit a cellular response. These results indicate the performance of HA hydrogels can be markedly enhanced without further additives or modifications, which is expected to contribute to the advancement of applications of HA hydrogels in all industrial fields.


Subject(s)
Cross-Linking Reagents/chemistry , Hyaluronic Acid/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Temperature , Animals , Cell Line , Cell Proliferation/drug effects , Cell Survival/drug effects , Cross-Linking Reagents/pharmacology , Female , Fibroblasts/drug effects , Fibroblasts/metabolism , Hyaluronic Acid/pharmacology , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Hydrolysis , Mice , Mice, Inbred BALB C , Mice, Nude , Particle Size , Surface Properties
17.
Anal Chim Acta ; 1036: 115-120, 2018 Dec 07.
Article in English | MEDLINE | ID: mdl-30253821

ABSTRACT

In biosensors development, alginate hydrogels are a first choice for enabling stable biomolecules entrapment in biocompatible membranes obtained under soft physiological conditions. Although widely exploited, most alginate membranes are isolating and poorly repetitive, which limit their application in biosensing. Significant steps forward on improving repeatability and conductivity have been performed, but to date there is no single protocol for controlled deposition of live cells in replicable conductive alginate layers. Here, cell electrotrapping in conductive alginate hydrogels is examined in order to overcome these limitations. Conductive alginate-coated electrodes are obtained after potentiostatic electrodeposition of graphite-doped alginate samples (up to 4% graphite). The presence of graphite reduces electrode passivation and improves the electrochemical response of the sensor, although still significantly lower than that recorded with the naked electrode. Bacterial electrotrapping in the conductive matrix is highly efficient (4.4 × 107 cells per gel) and repetitive (CV < 0.5%), and does not compromise bacterial integrity or activity (cell viability = 56%). Biosensing based on ferricyanide respirometry yielded a four times increase in biosensor response with respect to non-conductive alginate membrane, providing toxicity values completely comparable to those reported. Cell electrotrapping in conductive hydrogels represents a step forward towards in high-sensitive cell-based biosensors development with important influence in environmental analysis, food and beverage industry as well as clinical diagnosis.


Subject(s)
Escherichia coli/drug effects , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Alginates/chemistry , Alginates/pharmacology , Biosensing Techniques , Electric Conductivity , Electrochemical Techniques , Electrodes , Escherichia coli/cytology , Ferrocyanides/metabolism , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry
18.
Stem Cells Dev ; 27(23): 1605-1620, 2018 12 01.
Article in English | MEDLINE | ID: mdl-30215325

ABSTRACT

Severe burns are some of the most challenging problems in clinics and still lack ideal modalities. Mesenchymal stem cells (MSCs) incorporated with biomaterial coverage of burn wounds may offer a viable solution. In this report, we seeded MSCs to a biodegradable hybrid hydrogel, namely ACgel, that was synthesized from unsaturated arginine-based poly(ester amide) (UArg-PEA) and chitosan derivative. MSC adhered to ACgels. ACgels maintained a high viability of MSCs in culture for 6 days. MSC seeded to ACgels presented well in third-degree burn wounds of mice at 8 days postburn (dpb) after the necrotic full-thickness skin of burn wounds was debrided and filled and covered by MSC-carrying ACgels. MSC-seeded ACgels promoted the closure, reepithelialization, granulation tissue formation, and vascularization of the burn wounds. ACgels alone can also promote vascularization but less effectively compared with MSC-seeded ACgels. The actions of MSC-seeded ACgels or ACgels alone involve the induction of reparative, anti-inflammatory interleukin-10, and M2-like macrophages, as well as the reduction of inflammatory cytokine TNFα and M1-like macrophages at the late inflammatory phase of burn wound healing, which provided the mechanistic insights associated with inflammation and macrophages in burn wounds. For the studied regimens of these treatments, no toxicity was identified to MSCs or mice. Our results indicate that MSC-seeded ACgels have potential use as a novel adjuvant therapy for severe burns to complement commonly used skin grafting and, thus, minimize the downsides of grafting.


Subject(s)
Burns/drug therapy , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/cytology , Amides/chemistry , Animals , Arginine/chemistry , Biodegradable Plastics/pharmacology , Burns/pathology , Chitosan/chemistry , Chitosan/pharmacology , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Mice , Polyesters/chemistry , Polyesters/pharmacology , Tissue Scaffolds/chemistry , Wound Healing
19.
Plast Surg Nurs ; 38(3): 133-138, 2018.
Article in English | MEDLINE | ID: mdl-30157127

ABSTRACT

Objective of this study was to evaluate the efficacy of the autolytic debridement promoted by hydrogel with sodium alginate enriched with fatty acids and vitamins A and E in the healing of foot wounds in diabetic patients. A clinical study was conducted at an outpatient clinic of medical specialties. The sample comprised 8 patients supervised for a 3-month period, from April to July 2017, by means of a clinical history, photographic record, planimetry, and classification of the wound severity by the Pressure Ulcer Scale for Healing (PUSH) system. Of the 8 patients supervised, 1 dropped out and 7 were followed up for 12 weeks. Only 2 had complete wound healing, but all presented a reduction of the lesion area of approximately 22.2% and PUSH score of 9.8 to 6.6. This study found that hydrogel showed good results for the treatment of diabetic feet, reducing the area and overall PUSH score of the wounds.


Subject(s)
Alginates/pharmacology , Diabetes Complications/therapy , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Wound Healing/drug effects , Aged , Aged, 80 and over , Alginates/therapeutic use , Bandages , Diabetes Mellitus/physiopathology , Female , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/therapeutic use , Male , Middle Aged
20.
Tissue Eng Part A ; 24(19-20): 1432-1443, 2018 10.
Article in English | MEDLINE | ID: mdl-30036140

ABSTRACT

Post-traumatic and focal cartilage defects of the knee affect over 3 million Americans annually. Autologous cell-based cartilage repair, for example, autologous chondrocyte implantation, is limited by the need for ex vivo chondrocyte expansion and donor site morbidity. Mesenchymal stem cells (MSCs), owing to their relative ease of isolation, higher replication activity, and chondrogenic potential, represent an alternative reparative cell type. Platelet-rich plasma (PRP) is an autologous, growth factor-rich biologic preparation that has recently received increasing attention and use as a therapeutic adjunct for the treatment of degenerative joint diseases, and there is evidence suggesting that PRP acts by promoting stem cell proliferation and tissue healing. In this study, we have examined the effect of PRP treatment on chondrogenic differentiation of adult human MSCs derived from infrapatellar fat pad-adipose stem cells (IFP-ASCs) and bone marrow (BM-MSCs). Both cell types were placed in high-density pellet culture and hydrogel-encapsulated culture under chondrogenic conditions. Our results showed that PRP did not improve IFP-ASC or BM-MSC chondrogenesis. In general, chondrogenesis was inhibited with increasing PRP concentrations and duration of exposure, on the basis of histological, biochemical, and gene expression analyses. Taken together, these findings suggest that although PRP is reported to be beneficial in terms of pain relief and joint function improvement, its mechanism of action is unlikely to directly involve enhancement of MSC-mediated hyaline cartilage formation.


Subject(s)
Adipose Tissue/cytology , Bone Marrow Cells/cytology , Cell Differentiation , Chondrogenesis , Mesenchymal Stem Cells/cytology , Platelet-Rich Plasma/metabolism , Adult , Bone Marrow Cells/metabolism , Cell Differentiation/drug effects , Cell Differentiation/genetics , Cells, Cultured , Chondrogenesis/drug effects , Chondrogenesis/genetics , Gene Expression Regulation/drug effects , Humans , Hydrogel, Polyethylene Glycol Dimethacrylate/pharmacology , Mesenchymal Stem Cells/metabolism
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