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1.
Biomacromolecules ; 25(6): 3449-3463, 2024 Jun 10.
Article in English | MEDLINE | ID: mdl-38739908

ABSTRACT

Using supramolecular self-assembled nanocomposite materials made from protein and polysaccharide components is becoming more popular because of their unique properties, such as biodegradability, hierarchical structures, and tunable multifunctionality. However, the fabrication of these materials in a reproducible way remains a challenge. This study presents a new evaporation-induced self-assembly method producing layered hydrogel membranes (LHMs) using tropocollagen grafted by partially deacetylated chitin nanocrystals (CO-g-ChNCs). ChNCs help stabilize tropocollagen's helical conformation and fibrillar structure by forming a hierarchical microstructure through chemical and physical interactions. The LHMs show improved mechanical properties, cytocompatibility, and the ability to control drug release using octenidine dihydrochloride (OCT) as a drug model. Because of the high synergetic performance between CO and ChNCs, the modulus, strength, and toughness increased significantly compared to native CO. The biocompatibility of LHM was tested using the normal human dermal fibroblast (NHDF) and the human osteosarcoma cell line (Saos-2). Cytocompatibility and cell adhesion improved with the introduction of ChNCs. The extracted ChNCs are used as a reinforcing nanofiller to enhance the performance properties of tropocollagen hydrogel membranes and provide new insights into the design of novel LHMs that could be used for various medical applications, such as control of drug release in the skin and bone tissue regeneration.


Subject(s)
Hydrogels , Humans , Hydrogels/chemistry , Hydrogels/pharmacology , Nanoparticles/chemistry , Chitin/chemistry , Cell Line, Tumor , Biocompatible Materials/chemistry , Biocompatible Materials/pharmacology , Fibroblasts/drug effects , Fibroblasts/cytology , Membranes, Artificial , Nanocomposites/chemistry , Cell Adhesion/drug effects
2.
J Periodontal Res ; 59(3): 468-479, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38311974

ABSTRACT

OBJECTIVE: The aim of this study was to: (1) evaluate the anti-inflammatory effects of cannabidiol (CBD) on primary cultures of human gingival fibroblasts (HGFs) and (2) to clinically monitor the effect of CBD in subjects with periodontitis. BACKGROUND: The use of phytocannabinoids is a new approach in the treatment of widely prevalent periodontal disease. MATERIALS AND METHODS: Cannabinoid receptors were analyzed by western blot and interleukin production detected using enzyme immunoassay. Activation of the Nrf2 pathway was studied via monitoring the mRNA level of heme oxygenase-1. Antimicrobial effects were determined by standard microdilution and 16S rRNA screening. In the clinical part, a placebo-control double-blind randomized study was conducted (56 days) in three groups (n = 90) using dental gel without CBD (group A) and with 1% (w/w) CBD (group B) and corresponding toothpaste (group A - no CBD, group B - with CBD) for home use to maintain oral health. Group C used dental gel containing 1% chlorhexidine digluconate (active comparator) and toothpaste without CBD. RESULTS: Human gingival fibroblasts were confirmed to express the cannabinoid receptor CB2. Lipopolysaccharide-induced cells exhibited increased production of pro-inflammatory IL-6 and IL-8, with deceasing levels upon exposure to CBD. CBD also exhibited antimicrobial activities against Porphyromonas gingivalis, with an MIC of 1.5 µg/mL. Activation of the Nrf2 pathway was also demonstrated. In the clinical part, statistically significant improvement was found for the gingival, gingival bleeding, and modified gingival indices between placebo group A and CBD group B after 56 days. CONCLUSIONS: Cannabidiol reduced inflammation and the growth of selected periodontal pathogenic bacteria. The clinical trial demonstrated a statistically significant improvement after CBD application. No adverse effects of CBD were reported by patients or observed upon clinical examination during the study. The results are a promising basis for a more comprehensive investigation of the application of non-psychotropic cannabinoids in dentistry.


Subject(s)
Cannabidiol , Fibroblasts , Gingiva , Gingivitis , Humans , Cannabidiol/pharmacology , Cannabidiol/therapeutic use , Double-Blind Method , Fibroblasts/drug effects , Adult , Male , Female , Gingiva/drug effects , Gingivitis/drug therapy , Middle Aged , NF-E2-Related Factor 2 , Anti-Inflammatory Agents/therapeutic use , Anti-Inflammatory Agents/pharmacology , Chlorhexidine/therapeutic use , Chlorhexidine/pharmacology , Chlorhexidine/analogs & derivatives , Cells, Cultured , Interleukin-6/analysis , Periodontitis/drug therapy , Interleukin-8/drug effects , Heme Oxygenase-1
3.
Eur J Pharm Biopharm ; 185: 126-136, 2023 Apr.
Article in English | MEDLINE | ID: mdl-36871904

ABSTRACT

In this work oleuropein and lentisk oil have been co-loaded in different phospholipid vesicles (i.e., liposomes, transfersomes, hyalurosomes and hyalutransfersomes), to obtain a formulation capable of both inhibiting the production of different markers connected with inflammation and oxidative stress and promoting the skin repair. Liposomes were prepared using a mixture of phospholipids, oleuropein and lentisk oil. Tween 80, sodium hyaluronate or their combination have been added to the mixture to obtain transfersomes, hyalurosomes and hyalutransfersomes. Size, polydispersity index, surface charge and stability on storage was evaluated. The biocompatibility, anti-inflammatory activity and wound healing effect were tested using normal human dermal fibroblasts. Vesicles were small (mean diameter âˆ¼ 130 nm) and homogeneously dispersed (polydispersity index âˆ¼ 0.14), highly negatively charged (zeta potential 02053-64 mV) and capable of loading 20 mg/mL of oleuropein and 75 mg/mL of lentisk oil. The freeze-drying of dispersions with a cryoprotectant permitted to improve their stability on storage. The co-loading of oleuropein and lentisk oil in vesicles inhibited the overproduction of inflammatory markers, especially MMP-1 and IL-6, counteracted the oxidative stress induced in cells using hydrogen peroxide, and promoted the healing of a wounded area performed in vitro in a cell monolayer of fibroblasts. The proposed co-loading of oleuropein and lentisk oil in natural-based phospholipid vesicles may hold promising therapeutic value especially for the treatment of a wide variety of skin disorders.


Subject(s)
Liposomes , Phospholipids , Humans , Phospholipids/metabolism , Liposomes/metabolism , Skin/metabolism , Oxidative Stress , Wound Healing , Cytokines/metabolism
4.
Molecules ; 28(4)2023 Feb 11.
Article in English | MEDLINE | ID: mdl-36838716

ABSTRACT

Aging is a complex physiological process that can be accelerated by chemical (high blood glucose levels) or physical (solar exposure) factors. It is accompanied by the accumulation of altered molecules in the human body. The accumulation of oxidatively modified and glycated proteins is associated with inflammation and the progression of chronic diseases (aging). The use of antiglycating agents is one of the recent approaches in the preventive strategy of aging and natural compounds seem to be promising candidates. Our study focused on the anti-aging effect of the flavonoid hesperetin, its glycoside hesperidin and its carbohydrate moieties rutinose and rhamnose on young and physiologically aged normal human dermal fibroblasts (NHDFs). The anti-aging activity of the test compounds was evaluated by measuring matrix metalloproteinases (MMPs) and inflammatory interleukins by ELISA. The modulation of elastase, hyaluronidase, and collagenase activity by the tested substances was evaluated spectrophotometrically by tube tests. Rutinose and rhamnose inhibited the activity of pure elastase, hyaluronidase, and collagenase. Hesperidin and hesperetin inhibited elastase and hyaluronidase activity. In skin aging models, MMP-1 and MMP-2 levels were reduced after application of all tested substances. Collagen I production was increased after the application of rhamnose and rutinose.


Subject(s)
Hesperidin , Rhamnose , Skin Aging , Humans , Collagenases/metabolism , Hesperidin/pharmacology , Hyaluronoglucosaminidase , Pancreatic Elastase , Rhamnose/pharmacology , Skin Aging/drug effects
5.
Materials (Basel) ; 16(3)2023 Jan 25.
Article in English | MEDLINE | ID: mdl-36770074

ABSTRACT

In this work, composite filaments in the form of sticks and 3D-printed scaffolds were investigated as a future component of an osteochondral implant. The first part of the work focused on the development of a filament modified with bioglass (BG) and Zn-doped BG obtained by injection molding. The main outcome was the manufacture of bioactive, strong, and flexible filament sticks of the required length, diameter, and properties. Then, sticks were used for scaffold production. We investigated the effect of bioglass addition on the samples mechanical and biological properties. The samples were analyzed by scanning electron microscopy, optical microscopy, infrared spectroscopy, and microtomography. The effect of bioglass addition on changes in the SBF mineralization process and cell morphology was evaluated. The presence of a spatial microstructure within the scaffolds affects their mechanical properties by reducing them. The tensile strength of the scaffolds compared to filaments was lower by 58-61%. In vitro mineralization experiments showed that apatite formed on scaffolds modified with BG after 7 days of immersion in SBF. Scaffold with Zn-doped BG showed a retarded apatite formation. Innovative 3D-printing filaments containing bioglasses have been successfully applied to print bioactive scaffolds with the surface suitable for cell attachment and proliferation.

6.
J Funct Biomater ; 13(4)2022 Nov 16.
Article in English | MEDLINE | ID: mdl-36412890

ABSTRACT

The cornea and the skin are two organs that form the outer barrier of the human body. When either is injured (e.g., from surgery, physical trauma, or chemical burns), wound healing is initiated to restore integrity. Many cells are activated during wound healing. In particular, fibroblasts that are stimulated often transition into repair fibroblasts or myofibroblasts that synthesize extracellular matrix (ECM) components into the wound area. Control of wound ECM deposition is critical, as a disorganized ECM can block restoration of function. One of the most abundant structural proteins in the mammalian ECM is collagen. Collagen type I is the main component in connective tissues. It can be readily obtained and purified, and short analogs have also been developed for tissue engineering applications, including modulating the wound healing response. This review discusses the effect of several current collagen implants on the stimulation of corneal and skin wound healing. These range from collagen sponges and hydrogels to films and membranes.

7.
Article in English | MEDLINE | ID: mdl-35332345

ABSTRACT

Cannabidiol (CBD), a non-psychotropic cannabinoid produced by the genus Cannabis, is a phytoceutical that activates the endocannabinoid system (ECS) through binding to CB1 and CB2 receptors. The ECS is involved in cellular homeostasis and regulates metabolic processes in virtually all mammalian tissues. Published studies on CBD focus, inter alia, on its use in prophylaxis and as an anti-inflammatory agent. Here the authors present a critical assessment of the effects of CBD on inflammatory periodontal diseases caused by bacterial virulence factors, and evaluate critically the possible benefits and drawbacks of CBD use in dentistry. Particular attention is paid to the interaction of CBD with microbially colonized oral tissues, the inflammatory response in relation to the immune response, and the destruction/regeneration of hard and soft tissues of the periodontium.


Subject(s)
Cannabidiol , Cannabinoids , Periodontal Diseases , Analgesics , Animals , Cannabidiol/metabolism , Cannabidiol/pharmacology , Cannabidiol/therapeutic use , Humans , Mammals/metabolism , Periodontal Diseases/drug therapy , Receptor, Cannabinoid, CB1
8.
J Appl Biomed ; 19(3): 149-158, 2021 09.
Article in English | MEDLINE | ID: mdl-34907758

ABSTRACT

Myricetin (MYR) and dihydromyricetin (DHM) are classified as natural flavonoids. Both substances are known for their anti-inflammatory and antioxidant properties. In this study, an in vitro model of inflammation was demonstrated on monolayers of scratched fibroblasts or keratinocytes exposed to LPS from Pseudomonas aeruginosa for six hours. MYR and DHM were subsequently applied to the cells for 24 hours at sub toxic concentrations (5-15 µM). Inflammatory parameters were analysed in collected cell medium and lysate after the incubation period using the Enzyme-Linked ImmuneSorbent Assay (ELISA) and Western blot. Both flavonoids inhibit the production of pro-inflammatory cytokines (IL-6, IL-8) in LPS-stimulated skin cells as well as the decreased level of MMP-1 in fibroblasts. However, the application of MYR and DHM dose dependently increased the level of MMP-1 in keratinocytes. In our experiments, we focused on the anti-glycation activity of MYR and DHM, where the higher concentration of MYR seems to be more effective.


Subject(s)
Lipopolysaccharides , Matrix Metalloproteinase 1 , Flavonoids/pharmacology , Flavonols , Wound Healing
9.
Redox Biol ; 46: 102097, 2021 10.
Article in English | MEDLINE | ID: mdl-34418599

ABSTRACT

Lipid nitroalkenes - nitro-fatty acids (NO2-FAs) are formed in vivo via the interaction of reactive nitrogen species with unsaturated fatty acids. The resulting electrophilic NO2-FAs play an important role in redox homeostasis and cellular stress response. This study investigated the physicochemical properties and reactivity of two NO2-FAs: 9/10-nitrooleic acid (1) and its newly prepared 1-monoacyl ester, (E)-2,3-hydroxypropyl 9/10-nitrooctadec-9-enoate (2), both synthesized by a direct radical nitration approach. Compounds 1 and 2 were investigated in an aqueous medium and after incorporation into lipid nanoparticles prepared from 1-monoolein, cubosomes 1@CUB and 2@CUB. Using an electrochemical analysis and LC-MS, free 1 and 2 were found to be unstable under acidic conditions, and their degradation occurred in an aqueous environment within a few minutes or hours. This degradation was associated with the production of the NO radical, as confirmed by fluorescence assay. In contrast, preparations 1@CUB and 2@CUB exhibited a significant increase in the stability of the loaded 1 and 2 up to several days to weeks. In addition to experimental data, density functional theory-based calculation results on the electronic structure and structural variability (open and closed configuration) of 1 and 2 were obtained. Finally, experiments with a human HaCaT keratinocyte cell line demonstrated the ability of 1@CUB and 2@CUB to penetrate through the cytoplasmic membrane and modulate cellular pathways, which was exemplified by the Keap1 protein level monitoring. Free 1 and 2 and the cubosomes prepared from them showed cytotoxic effect on HaCaT cells with IC50 values ranging from 1 to 8 µM after 24 h. The further development of cubosomal preparations with embedded electrophilic NO2-FAs may not only contribute to the field of fundamental research, but also to their application using an optimized lipid delivery vehicle.


Subject(s)
Fatty Acids , Nitric Oxide , Humans , Kelch-Like ECH-Associated Protein 1 , NF-E2-Related Factor 2 , Nitric Oxide/metabolism , Nitro Compounds
10.
J Photochem Photobiol B ; 209: 111948, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32679512

ABSTRACT

The harmful effects of low energy UVA photons (315-400 nm) are associated with the massive production of reactive oxygen species resulting in oxidative stress. In response to oxidative damage, NF-E2-related factor 2 (Nrf2) is translocated to the nucleus and drives the expression of detoxication and antioxidant enzymes. UVA's effect on Nrf2 has been quite well characterised in dermal fibroblasts. However, there is a dearth of such information for keratinocytes. This study aimed to evaluate and compare the effect of UVA radiation on the Nrf2 pathway and oxidative stress related proteins in primary human dermal fibroblasts (NHDF), epidermal keratinocytes (NHEK) and human keratinocyte cell line HaCaT. NHDF were exposed to doses of 2.5-7.5 J/cm2, NHEK and HaCaT to 10-20 J/cm2 using a solar simulator. Effects on Nrf2 translocation were evaluated after 1, 3 and 6 h and Nrf2-controlled proteins (heme oxygenase 1 (HO-1), NAD(P)H:quinone oxidoreductase 1 (NQO1), glutathione reductase (GSR), glutathione-S-transferase (GST), interleukine-6 (IL-6), and matrix metalloproteinases (MMP-1, MMP-2)) after 3, 6 and 24 h. The results showed the fastest Nrf2 translocation was in UVA-irradiated HaCaT (1 h), persisting until the subsequent time interval (3 h), while in primary keratinocytes the effect of radiation was minimal. In NHDF, UVA-stimulated Nrf2 translocation was conspicuous 3 h after UVA treatment. In NHDF, most of the studied proteins (NQO1, HO-1, GSR, GSTM1 and MMP-1) showed the highest level 24 h after UVA exposure, except for MMP-2 and IL-6 which had their highest level at a shorter time incubation interval (3 h). In NHEK, NQO1, HO-1 and GST were increased 6 h after UVA exposure, GSR and MMP-2 level was slightly below or above the control level, and MMP-1 and IL-6 increased at shorter time intervals. When comparing NHEK and HaCaT, these cells displayed contrary responses in most of the Nrf2-controlled proteins. Thus, primary keratinocytes cannot be replaced with HaCaT when studying cell signalling such as the Nrf2 driven pathway and Nrf2-controlled proteins.


Subject(s)
NF-E2-Related Factor 2/metabolism , Signal Transduction/radiation effects , Skin/radiation effects , Ultraviolet Rays , Cell Survival/radiation effects , Cells, Cultured , Humans , Keratinocytes/metabolism , Keratinocytes/radiation effects , Protein Transport , Skin/cytology , Skin/metabolism
11.
Med Sci Monit ; 24: 511-522, 2018 Jan 26.
Article in English | MEDLINE | ID: mdl-29371587

ABSTRACT

BACKGROUND Paracrine factors secreted by adipose-derived stem cells can be captured, fractionated, and concentrated to produce therapeutic factor concentrate (TFC). The present study examined whether TFC effects could be enhanced by combining TFC with a biological matrix to provide sustained release of factors in the target region. MATERIAL AND METHODS Unilateral hind limb ischemia was induced in rabbits. Ischemic limbs were injected with either placebo control, TFC, micronized small intestinal submucosa tissue (SIS), or TFC absorbed to SIS. Blood flow in both limbs was assessed with laser Doppler perfusion imaging. Tissues harvested at Day 48 were assessed immunohistochemically for vessel density; in situ hybridization and quantitative real-time PCR were employed to determine miR-126 expression. RESULTS LDP ratios were significantly elevated, compared to placebo control, on day 28 in all treatment groups (p=0.0816, p=0.0543, p=0.0639, for groups 2-4, respectively) and on day 36 in the TFC group (p=0.0866). This effect correlated with capillary density in the SIS and TFC+SIS groups (p=0.0093 and p=0.0054, respectively, compared to placebo). A correlation was observed between miR-126 levels and LDP levels at 48 days in SIS and TFC+SIS groups. CONCLUSIONS A single bolus administration of TFC and SIS had early, transient effects on reperfusion and promotion of ischemia repair. The effects were not additive. We also discovered that TFC modulated miR-126 levels that were expressed in cell types other than endothelial cells. These data suggested that TFC, alone or in combination with SIS, may be a potent therapy for patients with CLI that are at risk of amputation.


Subject(s)
Adipose Tissue/cytology , Cell-Derived Microparticles/metabolism , Extracellular Matrix/metabolism , Extremities/blood supply , Ischemia/therapy , MicroRNAs/metabolism , Stem Cell Transplantation , Stem Cells/cytology , Animals , Disease Models, Animal , Extremities/pathology , Female , Gene Expression Regulation , Humans , Intestinal Mucosa/physiology , Intestine, Small/physiology , Ischemia/genetics , Ischemia/pathology , Laser-Doppler Flowmetry , MicroRNAs/genetics , Middle Aged , Perfusion , Rabbits , Reperfusion Injury/pathology , Reperfusion Injury/therapy , Skin/pathology
12.
Molecules ; 24(1)2018 Dec 30.
Article in English | MEDLINE | ID: mdl-30598040

ABSTRACT

In this study, we compared selected silymarin components, such as quercetin (QE), 2,3-dehydrosilybin (DHS) and silybin (SB), with the anti-inflammatory drug indomethacin (IND) in terms of their wound healing potential. In view of the fact that pathological cutaneous wound healing is associated with persistent inflammation, we studied their anti-inflammatory activity against inflammation induced by bacterial lipopolysaccharide (LPS). We investigated the regulation of crucial pro-inflammatory transcription factors-nuclear factor kappa-B (NF-κB) and activator protein 1 (AP-1)-as well as the expression of downstream inflammatory targets by Western blotting, real-time PCR (RT-PCR), electrophoretic mobility shift assay (EMSA), and/or enzyme-linked immunosorbent assay (ELISA) in vitro using primary normal human dermal fibroblasts (NHDF). We demonstrated the greater ability of DHS to modulate the pro-inflammatory cytokines production via the NF-κB and AP-1 signaling pathways when compared to other tested substances. The prolonged exposure of LPS-challenged human dermal fibroblasts to DHS had both beneficial and detrimental consequences. DHS diminished interleukin-6 (IL-6) and interleukin-8 (IL-8) secretion but induced the significant upregulation of IL-8 mRNA associated with NF-κB and AP-1 activation. The observed conflicting results may compromise the main expected benefit, which is the acceleration of the healing of the wound via a diminished inflammation.


Subject(s)
Anti-Inflammatory Agents/pharmacology , Silymarin/pharmacology , Cell Proliferation/drug effects , Chemokines/metabolism , Cytokines/metabolism , Dermatitis/drug therapy , Dermatitis/genetics , Dermatitis/metabolism , Dermatitis/pathology , Fibroblasts/drug effects , Fibroblasts/metabolism , Gene Expression , Humans , Lipopolysaccharides/immunology , NF-kappa B/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Wound Healing/drug effects
13.
Interdiscip Toxicol ; 11(4): 289-293, 2018 Dec.
Article in English | MEDLINE | ID: mdl-31762680

ABSTRACT

Nanoparticles are utilized in a wide range of industries. The most studied silver nanoparticles (AgNPs) are used in medicine and also in several wound dressings due to their antimicrobial properties. The inflammatory response or potential morphological changes of skin cells after their application are not well known yet. In our study we used the model of human reconstructed epidermis (RHE), prepared in our laboratory, to evaluate whether the AgNPs penetrate through RHE, induce some morphological changes of keratinocytes or influence the production of pro-inflammatory cytokines (IL-6 and IL-8). After the application of three different concentrations (25 ppm, 2.5 ppm, 0.25 ppm) of AgNPs to of RHE for 24 hours we verified that AgNPs did not affect the production of pro-inflammatory cytokines (IL-6 and IL-8) and neither did they influence the expression of keratin K14 and loricrin. The morphology of the cells was likewise unchanged. Based on these results we conclude that AgNPs do not have any negative effect on the morphological changes and do not increase the production of pro-inflammatory cytokines.

14.
J Appl Biomater Funct Mater ; 14(2): e137-42, 2016 May 18.
Article in English | MEDLINE | ID: mdl-26952588

ABSTRACT

BACKGROUND: Nanoparticles are widely used in different technological fields, one of which is medicine. Because of their antibacterial properties, silver nanoparticles (AgNPs) are used in several types of wound dressings for the treatment of burns and nonhealing wounds, but their influence on each component of the wound-healing process remains unclear. In the present study, we evaluated the effects of AgNPs on normal human dermal fibroblasts (NHDFs) and normal human epidermal keratinocytes (NHEKs). Both cell types are important for wound healing, including with regard to inflammation, proliferation and tissue remodeling. Each phase of wound healing can be characterized by the secretion of cytokines, chemokines and growth factors. METHODS: The production of inflammatory parameters (tumor necrosis factor α [TNF-α], interleukin-6 [IL-6], IL-8 and IL-12 and cyclooxygenase-2 [COX-2]), angiogenesis parameters (vascular endothelial growth factor [VEGF], granulocyte macrophage colony-stimulating factor) and matrix metalloproteinases (MMP-1, MMP-2, MMP-3 and MMP-9) by NHDFs and NHEKs were examined by ELISA or Western blot after 24 and 48 hours of incubation with AgNPs. RESULTS: We found that AgNPs decreased some inflammatory cytokines (TNF-α and IL-12) and growth factors (VEGF) that were produced by NHDFs and NHEKs after 24 and 48 hours and decreased the expression of COX-2 after 24 hours but only at the highest concentration of AgNPs (25 parts per million). CONCLUSIONS: The results indicate that NHEKs are more susceptible to the application of AgNPs than NHDFs, and AgNPs may be useful for medical applications for the treatment of wounds.


Subject(s)
Fibroblasts/metabolism , Keratinocytes/metabolism , Metal Nanoparticles/chemistry , Models, Biological , Silver , Wound Healing/drug effects , Cells, Cultured , Female , Humans , Male , Primary Cell Culture , Silver/chemistry , Silver/pharmacology
15.
Article in English | MEDLINE | ID: mdl-29942372

ABSTRACT

Recently, due to their unique properties, gold nanoparticles (AuNPs) have been used in many biological applications. However, little is known about their toxicity when they come into contact with a biological system. Based on the proposal that AuNPs can have a positive effect on wound healing, the present study investigated the influence of negatively-charged-surface AuNPs (average diameter 25-50 nm) on the viability of normal human dermal fibroblasts (NHDF) and normal human epidermal keratinocytes (NHEK). Moreover, we evaluated the effect of AuNPs on the secretion of proteins involved in wound healing, such as interleukin-8 and - 12 (IL-8, IL-12), tumour necrosis factor-alpha (TNF-α), vascular endothelial growth factor (VEGF), basic fibroblast grow factor (bFGF), and granulocyte-macrophage colony-stimulating factor (GM-CSF). The results showed that AuNPs were not toxic to NHDF and NHEK. They showed a decrease in AuNPs' production of pro-inflammatory cytokines IL-6, IL-12 and TNF-α, as well as proteins involved in angiogenesis such as VEGF and bFGF. Thus, we suggest that AuNPs could have anti-inflammatory and anti-angiogenic activity.

16.
Int J Oral Maxillofac Implants ; 28(5): 1386-94, 2013.
Article in English | MEDLINE | ID: mdl-24066332

ABSTRACT

PURPOSE: To study the effects of different chemically modified titanium surfaces on the proliferation, differentiation, adhesion, and apoptosis of osteoblast-like SaOS-2 cells. MATERIALS AND METHODS: In this work, six different titanium materials were tested and compared to each other: (1) glazed; (2) unglazed; (3) unglazed and alkali-etched; (4) unglazed, sandblasted, acid- and alkali-etched; (5) unglazed and coated with zirconium nitride; and (6) unglazed, sandblasted, and acid-etched. The production of alkaline phosphatase (ALP), tumor necrosis factor alpha (TNF-α), matrix metalloproteinase-2, and the expression of adhesion proteins (integrin α3ß1, vinculin) were evaluated using ELISA. Finally, the apoptosis of cells was analyzed by flow cytometry. RESULTS: The most significant differences were found for unglazed sandblasted acid- and alkali-etched titanium discs compared with unglazed titanium discs. The production of TNF-α was decreased after 24 hours, as was the production of ALP after 72 hours. In contrast, the expression of integrin α3ß1 was increased after 6 hours. None of the titanium discs showed an apoptotic effect on cells. CONCLUSIONS: This study has shown that physical surface treatments (such as surface roughness) play a more important role than chemical modifications. Generally, chemical modifications such as acid- and alkali-etching can affect the wettability of titanium surfaces, making a surface hydrophilic or hydrophobic according to the modification. The cell attachment is better on hydrophilic surfaces, while hydrophilic surfaces may slightly decrease the expression of ALP activity.


Subject(s)
Dental Implants , Osteoblasts/physiology , Titanium , Alkaline Phosphatase/biosynthesis , Apoptosis , Cell Adhesion/physiology , Cell Cycle , Cell Differentiation/physiology , Cell Line, Tumor , Cell Survival/physiology , Dental Etching , Humans , Integrin alpha3beta1/metabolism , Matrix Metalloproteinase 2/biosynthesis , Osteoblasts/cytology , Surface Properties , Time Factors , Tumor Necrosis Factor-alpha/biosynthesis , Vinculin/metabolism , Wettability , Zirconium
17.
J Biomed Mater Res A ; 101(10): 2915-24, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23529774

ABSTRACT

The modification of implant surface situated in the area of peri-implant sulcus has important role in bacterial and cell adhesion. Six different chemically and physically modified titanium discs were prepared: glazed (Tis-MALP), unglazed (Tis-O), unglazed and alkali-etched (Tis-OA), unglazed and coated with ZrN (Tis-OZ), unglazed, sand blasted, and acid etched (Tis-OPAE), and unglazed, sand blasted, acid, and alkali etched (Tis-OPAAE). Analysis of surface topography was determined using scanning electron microscopy and atomic force microscopy (AFM). Biocompatibility of gingival fibroblasts was characterized by the production of tumor necrosis factor alpha, collagen I, matrix metalloproteinase 2 (MMP-2) after 24 and 72 h and expression of α3 ß1 integrin and vinculin using enzyme-linked immunosorbent assay (ELISA) or modified ELISA after 6 and 24 h. Microorganism adhesion (five bacterial strains) and biofilm formation was also evaluated. The adhesion of bacteria and gingival fibroblasts was significantly higher on titanium disc Tis-OPAAE and biofilm formation on the same surface for Streptococcus mutans, Streptococcus gordonii, and Streptococcus intermedius. The gingival fibroblasts on Tis-OPAAE disc had also significantly lower production of MMP-2. The collagen production was significantly lower on all surfaces with roughness higher than 0.2 µm. This study confirmed that the titanium disc with the surface roughness 3.39 µm (Tis-OPAAE) supported the adhesion of bacterial strains as well as gingival fibroblasts.


Subject(s)
Biocompatible Materials/pharmacology , Fibroblasts/cytology , Fibroblasts/microbiology , Gingiva/cytology , Materials Testing , Streptococcus/cytology , Titanium/pharmacology , Bacterial Adhesion/drug effects , Biofilms/drug effects , Biofilms/growth & development , Cell Adhesion/drug effects , Collagen Type I/biosynthesis , Dental Implants/microbiology , Fibroblasts/drug effects , Focal Adhesions/drug effects , Focal Adhesions/metabolism , Humans , Integrin alpha3beta1/metabolism , Matrix Metalloproteinase 2/biosynthesis , Microscopy, Atomic Force , Microscopy, Electron, Scanning , Spectrometry, X-Ray Emission , Streptococcus/drug effects , Streptococcus/physiology , Tumor Necrosis Factor-alpha/biosynthesis , Vinculin/metabolism
18.
Acta Dermatovenerol Croat ; 21(4): 219-23, 2013.
Article in English | MEDLINE | ID: mdl-24476607

ABSTRACT

Levels of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and tumor necrosis factor-α (TNF-α) may influence wound healing and wound closure in non-healing wounds. The aim of the study was to test the hypothesis that hydrogencalcium salts of oxidized cellulose change the production of matrix metalloproteinases (MMPs) and TNF-α, wound size and number of bacterial strains in non-healing wounds. We analyzed MMP-2, MMP-9 and TNF-α in the wound fluid from 20 patients by ELISA every fourteen days over six weeks. Wound size, pain, wound closure and bacterial strains in the wound were also investigated. The wound size was reduced in 14 patients and pain in 16 patients. Bacterial contamination of the wound decreased significantly after treatment. The level of MMP-2 correlated with TNF-α production. The level of MMP-9 was unchanged during the healing period. We conclude that hydrogencalcium salts of oxidized cellulose have a favorable effect on the reduction of bacterial contamination, wound size and pain.


Subject(s)
Cellulose, Oxidized/pharmacology , Diabetic Foot/metabolism , Diabetic Foot/therapy , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Tumor Necrosis Factor-alpha/metabolism , Varicose Ulcer/metabolism , Varicose Ulcer/therapy , Wound Healing/physiology , Aged , Diabetic Foot/microbiology , Female , Humans , Male , Middle Aged , Varicose Ulcer/microbiology
19.
Article in English | MEDLINE | ID: mdl-21804619

ABSTRACT

BACKGROUND: Dental implants are a suitable option for the replacement of some or all missing teeth. Their main function is to secure the stability of the artificial tooth. The implant material interacts with several cell types including osteoblasts, gingival fibroblasts, periodontal ligament fibroblasts and monocytes. The most common material used is pure titanium which is corrosion resistant and has an elasticity modulus similar to that of bone. In recent years, diverse modified titanium surfaces have also been developed. The wound healing around the implant is a complex process that determines how well the host can heal and accept the implanted material. For this reason, search for markers of the biocompatibility of these new materials is paramount. To identify markers found to be suitable for studying the biocompatibility of dental implants. METHODS: Review of Pubmed and Web of Science databases for the years 1958-2010. CONCLUSIONS: The surface of dental implant material should enhance firm attachment of the implant to junctional epithelium, soft connective tissue and bone. For the purposes of dental implant biocompatibility studies, a number of markers produced by osteoblasts or by cells of periodontal ligament have been proposed. In general, the most typical markers for osteoblasts and fibroblasts are alkaline phosphatase and collagen I, respectively. The involvement of both cell types in the inflammatory response is primarily evaluated by determination of tumour necrosis factor α and proinflammatory interleukins.


Subject(s)
Biocompatible Materials , Dental Implantation, Endosseous , Dental Implants , Gingiva/metabolism , Osteoblasts/metabolism , Periodontal Ligament/metabolism , Alkaline Phosphatase/metabolism , Collagen/metabolism , Dental Materials , Fibroblasts/metabolism , Humans , Titanium , Wound Healing
20.
J Mater Sci Mater Med ; 17(10): 891-8, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16977386

ABSTRACT

Hyiodine (high molecular weight hyaluronan combined with KI3 complex) is a new non-adhesive wound dressing which significantly improves the healing process. The aim of the study was to investigate the effects of Hyoidine on functional properties of isolated human keratinocytes and leukocytes, and on those of U937 and HL60 cell lines. While KI3 complex inhibited the viability and proliferation of the cells tested, Hyiodine did not have any significant effect. The expression of CD11b, CD62L and CD69 on PMNL, monocytes and lymphocytes, as well as the oxidative burst of blood neutrophils, were not changed. On the contrary, Hyiodine inhibited the PMA-activated oxidative burst and significantly increased the production of IL-6 and TNF-alpha by lymphocytes. It was concluded that hyaluronan content of Hyiodine reduces the toxic effect of KI3 complex on cells and speeds up the wound healing process by increasing the production of inflammatory cytokines.


Subject(s)
Hyaluronic Acid/pharmacology , Immune System/drug effects , Iodine/pharmacology , Keratinocytes/drug effects , HL-60 Cells , Hemostatics/pharmacology , Humans , Immune System/cytology , Iodides/pharmacology , U937 Cells
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