Inflammation-suppressing cornea-in-a-syringe with anti-viral GF19 peptide promotes regeneration in HSV-1 infected rabbit corneas.

Pathophysiologic inflammation, e.g., from HSV-1 viral infection, can cause tissue destruction resulting in ulceration, perforation, and ultimately blindness. We developed an injectable Cornea-in-a-Syringe (CIS) sealant-filler to treat damaged corneas. CIS comprises linear carboxylated polymers of inflammation-suppressing 2-methacryloyloxyethyl phosphorylcholine, regeneration-promoting collagen-like peptide, and adhesive collagen-citrate glue. We also incorporated GF19, a modified anti-viral host defense peptide that blocked HSV-1 activity in vitro when released from silica nanoparticles (SiNP-GF19). CIS alone suppressed inflammation when tested in a surgically perforated and HSV-1-infected rabbit corneal model, allowing tissue and nerve regeneration. However, at six months post-operation, only regenerated neocorneas previously treated with CIS with SiNP-GF19 had structural and functional features approaching those of normal healthy corneas and were HSV-1 virus-free. We showed that composite injectable biomaterials can be designed to allow regeneration by modulating inflammation and blocking viral activity in an infected tissue. Future iterations could be optimized for clinical application.

Synthesis and characterization of UV curable biocompatible hydrophilic copolymers containing siloxane units.

Tissues are highly three-dimensional structure complexes composed of different cell types and their interactions. One of the main challenges in tissue engineering is the inability to produce large, highly perfused scaffolds in which cells can grow at a high cell density and viability. Poly(dimethyl siloxane) (PDMS) is used as a flexible, biocompatible cell culture substrate with tunable mechanical properties. However, its fragility and hydrophobicity still pose a challenge. Here, we present a new strategy for the three-step one-pot synthesis of novel biocompatible hydrophilic copolymers containing siloxane units. In the first step, free radical copolymerization of acrylic acid (AA), butyl methacrylate (BMA), and 2-hydroxyethyl methacrylate (HEMA) was carried out in dioxane (DO) solution in the presence of 2,2'-azodiisobutyronitrile (AIBN). In the second step, the copolymers were modified with diepoxypropoxypropyl-terminated polydimethylsiloxane (DE-PDMS), and in the third step, the copolymers were additionally modified with glycidyl methacrylate (GMA). The modified copolymers were characterized by FTIR, NMR spectroscopy and elemental analysis. Films of modified copolymers were prepared by UV curing. SEM studies revealed microphase separated morphology with distribution of PDMS domains. The mechanical properties of the films depended on the amount of incorporated silicone modifier. The films were more hydrophilic than PDMS films. All novel copolymers showed high biocompatibility.

Genetic Ablation of Ankrd1 Mitigates Cardiac Damage during Experimental Autoimmune Myocarditis in Mice.

Myocarditis (MC) is an inflammatory disease of the myocardium that can cause sudden death in the acute phase, and dilated cardiomyopathy (DCM) with chronic heart failure as its major long-term outcome. However, the molecular mechanisms beyond the acute MC phase remain poorly understood. The ankyrin repeat domain 1 (ANKRD1) is a functionally pleiotropic stress/stretch-inducible protein, which can modulate cardiac stress response during various forms of pathological stimuli; however, its involvement in post-MC cardiac remodeling leading to DCM is not known. To address this, we induced experimental autoimmune myocarditis (EAM) in ANKRD1-deficient mice, and evaluated post-MC consequences at the DCM stage mice hearts. We demonstrated that ANKRD1 does not significantly modulate heart failure; nevertheless, the genetic ablation of Ankrd1 blunted the cardiac damage/remodeling and preserved heart function during post-MC DCM.

The Effect of UV Treatment on Surface Contact Angle, Fibroblast Cytotoxicity, and Proliferation with Two Types of Zirconia-Based Ceramics.

UV photofunctionalization of Zirconia-based materials for abutment fabrication is a promising approach that might influence the formation of a sound peri-implant seal, thus promoting long-term soft and hard tissue implant integration. This study aimed to evaluate the effect of UV treatment of test specimens made by two different ZnO2-based ceramic materials on the hydrophilicity, cell cytotoxicity, and proliferation of human gingival fibroblasts (HGFs). Two Zirconia-based materials, high-translucent and ultra-translucent multi-layered Zirconia (Katana, Kuraray Noritake, Japan), were used to prepare a total of 40 specimens distributed in two equally sized groups based on the material (n = 20). The same surface finishing protocol was applied for all specimens, as suggested by the manufacturer. Half the specimens from each group were treated with UV-C light for 48 h. Water contact angle (WCA), fibroblast cytotoxicity, and proliferation were investigated. The WCA values for the high-translucent Zirconia ranged from 69.9° ± 6.4° to 73.7° ± 13.9° for the treated/non-treated specimens and from 79.5° ± 12.8° to 83.4° ± 11.4° for the ultra-translucent multi-layered Zirconia, respectively. However, the difference was insignificant (F(16) = 3.50, p = 0.292). No significant difference was observed for the fibroblast cytotoxicity test. The results for proliferation revealed a significant difference, which was material-dependent (F(8) = 9.58, p = 0.005). We found that UV surface photofunctionalization of ZrO2-based materials alters the human gingival fibroblast cell viability, which might produce favourable results for cell proliferation.

Cytotoxicity and Efficacy in Debris and Smear Layer Removal of HOCl-Based Irrigating Solution: An In Vitro Study.

In the present study we evaluated the cytotoxicity of super-oxidized water on human gingival fibroblasts and its efficacy in debris and smear layer removal from root canal walls. Cultured gingival fibroblasts were exposed to super-oxidized water (Sterilox), which was diluted in Iscove's modified Dulbecco's medium (IMDM) at 30%, 40%, 50%, 60% and 70% concentrations. The control group was maintained in IMDM. The cell viability was evaluated by means of an MTT assay after incubation periods of 1 h, 2 h, 24 h and 48 h. Pathological cellular changes were also observed under fluorescence and phase contrast microscopes. The efficacy in debris and smear layer removal was evaluated in comparison to the conventional application of sodium hypochlorite (NaOCl) and ethylenediaminetetraacetic acid (EDTA). Forty maxillary premolars were randomly divided into two equal groups (n = 20) and shaped with ProTaper NEXT rotary instruments using Sterilox or NaOCl/EDTA for irrigation. Afterwards, roots were split longitudinally and examined under a scanning electron microscope. The results revealed that super-oxidized water and sterile distilled water have acceptable biological properties for endodontic applications at concentrations up to 50% (p > 0.05). Moreover, super-oxidized water is equally effective in debris and smear layer removal as compared to NaOCl/EDTA (p > 0.05).

Dental pulp stem cell-derived extracellular matrix: autologous tool boosting bone regeneration.

BACKGROUND AIMS: To facilitate artificial bone construct integration into a patient's body, scaffolds are enriched with different biologically active molecules. Among various scaffold decoration techniques, coating surfaces with cell-derived extracellular matrix (ECM) is a rapidly growing field of research. In this study, for the first time, this technology was applied using primary dental pulp stem cells (DPSCs) and tested for use in artificial bone tissue construction. METHODS: Rat DPSCs were grown on three-dimensional-printed porous polylactic acid scaffolds for 7 days. After the predetermined time, samples were decellularized, and the remaining ECM detailed proteomic analysis was performed. Further, DPSC-secreated ECM impact to mesenchymal stromal cells (MSC) behaviour as well as its role in osteoregeneration induction were analysed. RESULTS: It was identified that DPSC-specific ECM protein network ornamenting surface-enhanced MSC attachment, migration and proliferation and even promoted spontaneous stem cell osteogenesis. This protein network also demonstrated angiogenic properties and did not stimulate MSCs to secrete molecules associated with scaffold rejection. With regard to bone defects, DPSC-derived ECM recruited endogenous stem cells, initiating the bone self-healing process. Thus, the DPSC-secreted ECM network was able to significantly enhance artificial bone construct integration and induce successful tissue regeneration. CONCLUSIONS: DPSC-derived ECM can be a perfect tool for decoration of various biomaterials in the context of bone tissue engineering.

The Effect of a Unique Region of Parvovirus B19 Capsid Protein VP1 on Endothelial Cells.

Parvovirus B19 (B19V) is a widespread human pathogen possessing a high tropism for erythroid precursor cells. However, the persistence or active replication of B19V in endothelial cells (EC) has been detected in diverse human pathologies. The VP1 unique region (VP1u) of the viral capsid has been reported to act as a major determinant of viral tropism for erythroid precursor cells. Nevertheless, the interaction of VP1u with EC has not been studied. We demonstrate that recombinant VP1u is efficiently internalized by rats' pulmonary trunk blood vessel-derived EC in vitro compared to the human umbilical vein EC line. The exposure to VP1u was not acutely cytotoxic to either human- or rat-derived ECs, but led to the upregulation of cellular stress signaling-related pathways. Our data suggest that high levels of circulating B19V during acute infection can cause endothelial damage, even without active replication or direct internalization into the cells.

Anti-inflammatory effect of different curcumin preparations on adjuvant-induced arthritis in rats.

BACKGROUND: Curcumin, a natural polyphenolic substance, has been known for more than two millennia as having strong anti-inflammatory activity towards multiple ailments, including arthritis. The main drawback of curcumin is its poor solubility in water, which leads to low intestinal absorption and minimal bioavailability. In this study, we aimed to compare the anti-arthritic in vivo effect of different curcumin preparations - basic curcumin extract, micellar curcumin, curcumin mixture with piperine, and microencapsulated curcumin. METHODS: Arthritis was induced in Wistar rats by complete Freund's adjuvant, and the severity of arthritis was evaluated daily using the arthritis score system. Curcumin preparations were given to animals per os daily for 20 consecutive days, starting at 6th day after arthritis induction. To determine the inflammatory background, pro-inflammatory cytokines were determined using the ELISA test. In addition, hematologic test, weight change, and limb swelling were tracked. RESULTS: Our results indicate that curcumin had a rather weak effect on arthritis progression in the Wistar rat model, microencapsulated curcumin effectively prevented the progression of arthritis - the disease stabilized after 10 days of supplementation. It also reduced the levels of immune cells (neutrophils and leukocytes), as well as pro-inflammatory cytokines - TNFα, IL-1, and IL-6, which levels were close to arthritis-free control. Other formulations of curcumin had lower or no effect on arthritis progression. CONCLUSION: Our study shows that the same concentrations of curcumin had a distinctly expressed positive anti-inflammatory effect depending on the form of its delivery. Specifically, we found that microencapsulated curcumin had the most promising effect for treatment.

The Microbiota-Gut-Brain Axis and Alzheimer's Disease: Neuroinflammation Is to Blame?

For years, it has been reported that Alzheimer's disease (AD) is the most common cause of dementia. Various external and internal factors may contribute to the early onset of AD. This review highlights a contribution of the disturbances in the microbiota-gut-brain (MGB) axis to the development of AD. Alteration in the gut microbiota composition is determined by increase in the permeability of the gut barrier and immune cell activation, leading to impairment in the blood-brain barrier function that promotes neuroinflammation, neuronal loss, neural injury, and ultimately AD. Numerous studies have shown that the gut microbiota plays a crucial role in brain function and changes in the behavior of individuals and the formation of bacterial amyloids. Lipopolysaccharides and bacterial amyloids synthesized by the gut microbiota can trigger the immune cells residing in the brain and can activate the immune response leading to neuroinflammation. Growing experimental and clinical data indicate the prominent role of gut dysbiosis and microbiota-host interactions in AD. Modulation of the gut microbiota with antibiotics or probiotic supplementation may create new preventive and therapeutic options in AD. Accumulating evidences affirm that research on MGB involvement in AD is necessary for new treatment targets and therapies for AD.

Slow-Freezing Cryopreservation Ensures High Ovarian Tissue Quality Followed by In Vivo and In Vitro Methods and Is Safe for Fertility Preservation.

Background and objectives: Cancer incidence is growing with younger patients diagnosed with this disease every year. Improved cancer diagnostics and treatment lead to better survival of cancer patients. However, after aggressive chemo- or radiotherapy, cancer survivors suffer from various degrees of subfertility or infertility. Several fertility preservation technologies have been developed for young cancer patients: cryopreservation of germ cells, embryos, or reproductive tissues. The best results have been shown by cryopreservation of sperm and embryos. Yet the success of using cryopreserved oocytes or reproductive tissues (ovarian and testicular) is still insufficient. Therefore, this study was designed to assess the vitality, viability, general quality, and safety of frozen-thawed human ovarian tissue for retransplantation using modern molecular tests. Materials and Methods: The new miRNA array test was used to evaluate miRNA expression in thawed ovarian tissue in combination with standard xenotransplantation and pathological examination of microslides. Results: Our results demonstrated that slow freezing is an efficient way (80%) to cryopreserve ovarian tissue with no structural damage afterwards. We have shown that xenotransplantation into immunodeficient mice, histology, and immunohistochemistry could be potentially replaced by more recent molecular methods. Conclusions: The latter method has shown that altered expression of miRNAs might be used as identifiers of normal/damaged tissue after further analysis. Newer, safer, and more specific approaches need to be developed in order to eliminate the risk of disease reoccurrence.

The Effect of Different Cleaning Protocols of Polymer-Based Prosthetic Materials on the Behavior of Human Gingival Fibroblasts.

Dental implant abutment and prosthetic materials, their surface treatment, and cleaning modalities are important factors for the formation of a peri-implant soft tissue seal and long-term stability of bone around the implant. This study aimed to investigate the influence of a polymeric material surface cleaning method on the surface roughness, water contact angle, and human gingival fibroblasts (HGF) proliferation. Polymeric materials tested: two types of milled polymethylmethacrylate (PMMA-Ker and PMMA-Bre), three-dimensionally (3D) printed polymethylmethacrylate (PMMA-3D), polyetheretherketone (PEEK), and polyetherketoneketone (PEKK). Titanium (Ti) and zirconia oxide ceramics (ZrO-HT) were used as positive controls. A conventional surface cleaning protocol (CCP) was compared to a multi-step research cleaning method (RCP). Application of the RCP method allowed to reduce Sa values in all groups from 0.14-0.28 µm to 0.08-0.17 µm (p < 0.05 in PMMA-Ker and PEEK groups). Moreover, the water contact angle increased in all groups from 74-91° to 83-101° (p < 0.05 in the PEKK group), except ZrO-HT-it was reduced from 98.7 ± 4.5° to 69.9 ± 6.4° (p < 0.05). CCP resulted in higher variability of HGF viability after 48 and 72 h. RCP application led to higher HGF viability in PMMA-3D and PEKK groups after 48 h, but lower for the PMMA-Ker group (p < 0.05). After 72 h, no significant differences in HGF viability between both cleaning methods were observed. It can be concluded that the cleaning method of the polymeric materials affected surface roughness, contact angle, and HGF viability at 48 h.

In vitro comparison of 3D printed polylactic acid/hydroxyapatite and polylactic acid/bioglass composite scaffolds: Insights into materials for bone regeneration.

3D printing of polylactic acid (PLA) and hydroxyapatite (HA) or bioglass (BG) bioceramics composites is the most promising technique for artificial bone construction. However, HA and BG have different chemical composition as well as different bone regeneration inducing mechanisms. Thus, it is important to compare differentiation processes induced by 3D printed PLA + HA and PLA + BG scaffolds in order to evaluate the strongest osteoconductive and osteoinductive properties possessing bioceramics. In this study, we analysed porous PLA + HA (10%) and PLA + BG (10%) composites' effect on rat's dental pulp stem cells fate in vitro. Obtained results indicated, that PLA + BG scaffolds lead to weaker cell adhesion and proliferation than PLA + HA. Nevertheless, osteoinductive and other biofriendly properties were more pronounced by PLA + BG composites. Overall, the results showed a strong advantage of bioceramic BG against HA, thus, 3D printed PLA + BG composite scaffolds could be a perspective component for patient-specific, cheaper and faster artificial bone tissue production.

The Role of Cardiac T-Cadherin in the Indicating Heart Failure Severity of Patients with Non-Ischemic Dilated Cardiomyopathy.

Background and objectives: T-cadherin (T-cad) is one of the adiponectin receptors abundantly expressed in the heart and blood vessels. Experimental studies show that T-cad sequesters adiponectin in cardiovascular tissues and is critical for adiponectin-mediated cardio-protection. However, there are no data connecting cardiac T-cad levels with human chronic heart failure (HF). The aim of this study was to assess whether myocardial T-cad concentration is associated with chronic HF severity and whether the T-cad levels in human heart tissue might predict outcomes in patients with non-ischemic dilated cardiomyopathy (NI-DCM). Materials and Methods: 29 patients with chronic NI-DCM and advanced HF were enrolled. Patients underwent regular laboratory investigations, echocardiography, coronary angiography, and right heart catheterization. TNF-α and IL6 in serum were detected by enzyme-linked immunosorbent assay (ELISA). Additionally, endomyocardial biopsies were obtained, and the levels of T-cad were assessed by ELISA and CD3, CD45Ro, CD68, and CD4- immunohistochemically. Mean pulmonary capillary wedge pressure (PCWP) was used as a marker of HF severity, subdividing patients into two groups: mean PCWP > 19 mmHg vs. mean PCWP < 19 mmHg. Patients were followed-up for 5 years. The study outcome was composite: left ventricular assist device implantation, heart transplantation, or death from cardiovascular causes. Results: T-cad shows an inverse correlation with the mean PCWP (rho = -0.397, p = 0.037). There is a tendency towards a lower T-cad concentration in patients with more severe HF, as indicated by the mean PCWP > 19 mmHg compared to those with mean PCWP ≤ 19 mmHg (p = 0.058). Cardiac T-cad levels correlate negatively with myocardial CD3 cell count (rho = -0.423, p = 0.028). Conclusions: Univariate Cox regression analysis did not prove T-cad to be an outcome predictor (HR = 1, p = 0.349). However, decreased T-cad levels in human myocardium can be an additional indicator of HF severity. T-cad in human myocardium has an anti-inflammatory role. More studies are needed to extend the role of T-cad in the outcome prediction of patients with NI-DCM.

Assessment of the morphology and dimensional accuracy of 3D printed PLA and PLA/HAp scaffolds.

INTRODUCTION: In complex clinical conditions when physiological bone regeneration is insufficient, there is a need to develop synthetic material-based scaffolds. The morphologic properties of porous scaffolds are of crucial importance. The dimensional accuracy of 3D printed scaffolds can be affected by a variety of factors. MATERIALS AND METHODS: Three groups of 3D printed scaffolds were investigated: PLA1 (pure polylactic acid) printed with an FDM Ultimaker Original printer, PLA2 and composite PLA/hydroxyapatite (PLA/HAp) scaffolds printed with a Pharaoh XD 20. PLA/HAp filament was created with hot-melt extrusion (HME) equipment. The morphology of the prepared scaffolds was investigated with SEM, micro-CT and superimposition techniques, gravimetric and liquid displacement methods. RESULTS: Layer heights of PLA1 scaffolds varied the most. PLA1 scaffold volume statistically significantly differed from PLA2 (p < 0.001) and PLA/HAp (p < 0.01) groups. Filament composition had no effect on the volumes of the scaffolds printed with the Pharaoh XD 20 printer (p > 0.05). The total porosity of printed PLA/HAp scaffolds deviated the least from the original STL model. CONCLUSIONS: This study showed that PLA/10% HAp filament fabricated with HME and printed with FFF 3D printer produced equal or even better accuracy of printed scaffolds than scaffolds printed with pure PLA filament. Further research is needed to analyze the effect of HAp on 3D scaffold morphology, accuracy, mechanical and biologic properties.

Salinomycin and dichloroacetate synergistically inhibit Lewis lung carcinoma cell proliferation, tumor growth and metastasis.

Drug combination is considered to be the cornerstone of cancer treatment. Simultaneous administration of two or more drugs but at lower doses not only increases cytotoxic effects on tumor cells, but also reduces side effects and possibly overcomes drug resistance. Salinomycin is a well-known cancer stem cell killer, and dichloroacetate is a pyruvate dehydrogenase kinase inhibitor that exclusively targets cells with altered mitochondrial activity, a characteristic being common to most of the cancer cells. In our recent study, we have demonstrated that salinomycin exerted a cytotoxic effect on colorectal carcinoma cells in the 2D and 3D cultures and provided evidence that the mechanism of their synergy was mediated by dichloroacetate-dependent inhibition of the activity of multidrug resistance proteins. In the current work, we confirmed the synergistic cytotoxic properties of salinomycin and dichloroacetate in the 2D and 3D cultures of Lewis lung carcinoma (LLC1) cells. To verify if a synergistic effect of these compounds persisted in vivo, we performed series of experiments using a syngeneic LLC1-C57BL/6 mouse model and demonstrated that combination therapy with salinomycin and DCA increased the survival rate of allografted mice, inhibited metastatic site formation and reduced the populations of cancer stem cells as well as cells that underwent the epithelial-to-mesenchymal transition. Our results demonstrate that a synergistic effect of salinomycin and dichloroacetate exists not only in vitro but also in vivo and suggest their benefits in the treatment of metastatic cancers.

Biocompatibility Investigation of Hybrid Organometallic Polymers for Sub-Micron 3D Printing via Laser Two-Photon Polymerisation.

Hybrid organometallic polymers are a class of functional materials which can be used to produce structures with sub-micron features via laser two-photon polymerisation. Previous studies demonstrated the relative biocompatibility of Al and Zr containing hybrid organometallic polymers in vitro. However, a deeper understanding of their effects on intracellular processes is needed if a tissue engineering strategy based on these materials is to be envisioned. Herein, primary rat myogenic cells were cultured on spin-coated Al and Zr containing polymer surfaces to investigate how each material affects the viability, adhesion strength, adhesion-associated protein expression, rate of cellular metabolism and collagen secretion. We found that the investigated surfaces supported cellular growth to full confluency. A subsequent MTT assay showed that glass and Zr surfaces led to higher rates of metabolism than did the Al surfaces. A viability assay revealed that all surfaces supported comparable levels of cell viability. Cellular adhesion strength assessment showed an insignificantly stronger relative adhesion after 4 h of culture than after 24 h. The largest amount of collagen was secreted by cells grown on the Al-containing surface. In conclusion, the materials were found to be biocompatible in vitro and have potential for bioengineering applications.

Bioavailability of Different Vitamin D Oral Supplements in Laboratory Animal Model.

Background and Objectives: The major cause of vitamin D deficiency is inadequate exposure to sunlight. It is difficult to supplement it with food because sufficient concentrations of vitamin D naturally occur only in a handful of food products. Thereby, deficiency of this vitamin is commonly corrected with oral supplements. Different supplement delivery systems for improved vitamin D stability and bioavailability are proposed. In this study, we compared efficiency of three vitamin D delivery systems: microencapsulated, micellized, and oil-based. Materials and Methods: As a model in this medical testing, laboratory rats were used for the evaluation of bioavailability of different vitamin D vehicles. Animals were divided into three groups: the first one was given microencapsulated vitamin D3, the second-oil-based vitamin D3, and the third-micellized vitamin D3. Test substances were given per os to each animal for 7 days, and vitamin D concentration in a form of 25-hydroxyvitamin D (25(OH)D) in the blood was checked both during the vitamin delivery period and later, up to the 24th day. Results: Comparison of all three tested products showed that the microencapsulated and oil-based vitamin D3 vehicles were the most bioavailable in comparison to micellized vitamin D3. Even more, the effect of the microencapsulated form of vitamin D3 remained constant for the longest period (up to 14 days). Conclusions: The results of this study suggest that the oral vitamin D supplement vehicle has an impact on its bioavailability, thus it is important to take into account how much of the suppled vitamin D will be absorbed. To maximize the full exploit of supplement, the best delivery strategy should be employed. In our study, the microencapsulated form of vitamin D was the most bioavailable.

The effect of larger than cell diameter polylactic acid surface patterns on osteogenic differentiation of rat dental pulp stem cells.

Topography of the scaffold is one of the most important factors defining the quality of artificial bone. However, the production of precise micro- and nano-structured scaffolds, which is known to enhance osteogenic differentiation, is expensive and time-consuming. Meanwhile, little is known about macro-patterns (larger than cell diameter) effect on cell fate, while this kind of structures would significantly facilitate the manufacturing of artificial skeleton. Therefore, this research is focused on polylactic acid scaffold's macro-pattern impact on rat's dental pulp stem cells (DPSCs) morphology, proliferation, and osteogenic differentiation. For this study, two types of scaffolds were 3D printed: wavy and porous. Wavy scaffolds consisted of 188 µm wide joined threads, meaning that cells might have been curved on the filament as well as compressed in the groove. Porous scaffolds were designed to avoid groove formation and consisted of 500 µm threads, arranged in the woodpile manner, forming 300 µm diameter pores. We found that both macro-surfaces influenced DPSC morphology compared to control. As a consequence, enhanced DPSC proliferation and increased osteogenic differentiation potential was registered in cells grown on these scaffolds. Finally, our results showed that the construction of an artificial bone did not necessarily require the precise structuring of the scaffold, because both types of macro-topographic PLA scaffolds were sufficient enough to induce spontaneous DPSC osteogenic differentiation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 174-186, 2019.

Modelling of silk-reinforced PDMS properties for soft tissue engineering applications.

BACKGROUND: Polydimethylsiloxane (PDMS) is widely used in biomedical research and technology, but its mechanical properties should be tuned according to the desired product specifications. Mixing ratio of base polymer to curing agent or additives enables its mechanical properties to be manipulated and fit to mechanical properties of biological tissues. OBJECTIVE: In this paper, we analysed the effect of mechanical load on silk-reinforced PDMS depending on silk concentration. METHODS: We prepared cylinder-type PDMS samples with different silk concentrations and performed cyclic uniaxial compression tests with a fixed magnitude of applied strain. Next, we analysed the mechanical charascteristics of PDMS using computational modelling. RESULTS: The stress-strain data within the large-strain region of different PDMS cylinders without silk and with 1%, 5% and 10% silk concentrations was fitted to non-linear second order Mooney-Rivlin, and third-order Ogden models. The results show the equivalence of both models for investigated strain region of PDMS. On the other hand, PDMS cylinders with 10% silk concentration allowed the successful fitting of experimental data just for the second-order Mooney-Rivlin model, while all numerical probes to find an appropriate fitting parameters for third-order Ogden models were unsuccessful. CONCLUSIONS: The second-order Mooney-Rivlin model is preferable for analysing the properties of silk-reinforced PDMS over the entire measurement range.

Study of the cytotoxic effects of 2,5-diaziridinyl-3,6-dimethyl-1,4-benzoquinone (MeDZQ) in mouse hepatoma cells.

A number of quinones have been shown to be efficient anticancer agents. However, some mechanisms of their action, in particular cell signaling are not well understood. The aim of this study was to partly fill this gap by characterizing the mode of cytotoxicity of 2,5-diaziridinyl-3,6-dimethyl-1,4-benzoquinone (MeDZQ) in malignant mouse hepatoma cells (MH-22A) with regard to the expression and activation of main molecules in MAPK cell signaling pathway. The study revealed unequal roles of MAP kinases in MeDZQ-induced cell death: the compound did not induce significant changes in ERK expression or its phosphorylation; JNK appeared to be responsible for cell survival, however, p38 kinase was shown to be involved in cell death. In order to assess the enzymatic activation mechanisms responsible for the action of MeDZQ, we have also found that the antioxidant N,N'-diphenyl-p-phenylene diamine, the iron-chelating agent desferrioxamine, and DT-diaphorase inhibitor, dicoumarol, partly protected the cells from MeDZQ cytotoxicity. It points to parallel oxidative stress and bioreductive alkylation modes of the cytotoxicity of MeDZQ.