In Vitro Synergistic Photodynamic, Photothermal, Chemodynamic, and Starvation Therapy Performance of Chlorin e6 Immobilized, Polydopamine-Coated Hollow, Porous Ceria-Based, Hypoxia-Tolerant Nanozymes Carrying a Cascade System.

A synergistic therapy agent (STA) with photothermal, photodynamic, chemodynamic, and starvation therapy (PTT, PDT, CDT, and ST) functions was developed. Hollow, mesoporous, and nearly uniform CeO2 nanoparticles (H-CeO2 NPs) were synthesized using a staged shape templating sol-gel protocol. Chlorin e6 (Ce6) was adsorbed onto H-CeO2 NPs, and a thin polydopamine (PDA) layer was formed on Ce6-adsorbed H-CeO2 NPs. Glucose oxidase (GOx) was bound onto PDA-coated Ce6-adsorbed H-CeO2 NPs to obtain the targeted STA (H-CeO2@Ce6@PDA@GOx NPs). A reversible photothermal conversion behavior with the temperature elevations up to 34 °C was observed by NIR laser irradiation at 808 nm. A cascade enzyme system based on immobilized GOx and intrinsic catalase-like activity of H-CeO2 NPs was rendered on STA for enhancing the effectiveness of PDT by elevation of ROS generation and alleviation of hypoxia in a tumor microenvironment. Glucose-mediated generation of highly toxic hydroxyl radicals (·OH) was evaluated for CDT. The effectiveness of PDT on glioblastoma T98G cells was markedly enhanced by O2 generation started by the decomposition of glucose. A similar increase in cell death was also observed when ST and CDT functions were enhanced by photothermal action. The viability of T98G cells decreased to 10.6% by in vitro synergistic action including ST, CDT, PDT, and PTT without using any antitumor agent.

Investigation of UV-treated mesenchymal stem cells in an in vitro wound model.

This study examines the effects of ultraviolet-induced adipose tissue-derived mesenchymal stem cells and their supernatants on wound healing regarding cell viability, percentage of wound healing, released cytokine, and growth factors. It has been reported in previous studies that mesenchymal stem cells are resistant to ultraviolet light and have a protective effect on skin cells against ultraviolet-induced damage. At the same time, there are many studies in the literature about the positive effects of cytokines and growth factors secreted by mesenchymal stem cells. Based on this information, the effects of ultraviolet-induced adipose-derived stem cells and supernatants containing their secreted cytokines and growth factors on an in vitro two-dimensional wound model created with two different cell lines were investigated in this study. It was determined from the results that the highest cell viability and the least apoptotic staining were 100 mJ in mesenchymal stem cells (**p < 0.01). Furthermore, analysis of cytokines and growth factors collected from supernatants also supported 100 mJ as the optimal ultraviolet dose. It was observed that cells treated with ultraviolet and their supernatants significantly increased cell viability and wound-healing rate over time compared to other groups. In conclusion, with this study, it has been shown that adipose-derived stem cells exposed to ultraviolet light can have an important use in wound healing, both with their potential and with the more cytokines and growth factors they secrete. However, further analysis and animal experiments should be performed before clinical use.

DMSA-coated cubic iron oxide nanoparticles as potential therapeutic agents.

Aim: Superparamagnetic cubic iron oxide nanoparticles (IONPs) were synthesized and functionalized with meso-2,3-dimercaptosuccinic acid (DMSA) as a potential agent for cancer treatment. Methods: Monodisperse cubic IONPs with a high value of saturation magnetization were synthesized by thermal decomposition method and functionalized with DMSA via ligand exchange reaction, and their cytotoxic effects on HeLa cells were investigated. Results: DMSA functionalized cubic IONPs with an edge length of 24.5 ± 1.9 nm had a specific absorption rate value of 197.4 W/gFe (15.95 kA/m and 488 kHz) and showed slight cytotoxicity on HeLa cells when incubated with 3.3 × 1010, 6.6 × 1010 and 9.9 × 1010 NP/mL for 24, 48 and 72 h. Conclusion: To the best of our knowledge, this is the first study to investigate both the cytotoxic effects of DMSA-coated cubic IONPs on HeLa cells and hyperthermia performance of these nanoparticles.

Comparative Effects of Single-Dose Cardioplegic Solutions Especially in Repeated Doses During Minimally Invasive Aortic Valve Surgery.

OBJECTIVE: This study aims to compare del Nido cardioplegia (DNC) and histidine-tryptophan-ketoglutarate (HTK) cardioplegic solutions in minimally invasive aortic valve replacement (mini-AVR) surgery to discuss the safety level of myocardial protection and rationale for redosing intervals. METHODS: During the period from January 2017 to June 2019, 200 patients undergoing mini-AVR (solely or with concomitant procedures) were prospectively randomized to DNC (n = 100) andHTK (n = 100), both up to 90 minutes ischemic time. Patients with ischemic time over 90 minutes, needing a redosing, were further analyzed in 2 subgroups with DNC-R (n = 30) and HTK-R (n = 36). Sensitive biomarkers, in addition to routine biochemistry, were also documented at baseline (T1), after cessation of cardiopulmonary bypass (T2), and on the first postoperative day (T3). Transmural myocardial biopsies were sampled for staining. RESULTS: No statistical differences could be demonstrated in DNC and HTK groups with up to 90 minutes cross-clamp times in routine biochemical measurements and basic perioperative clinical outcomes. DNC-R showed significantly more arrhythmia/AV block incidence resulting in more extended intensive care unit (ICU) stay. Interleukin-6 and syndecan-1 in DNC and DNC-R groups were substantially higher at T2. Aquaporin-4 levels were significantly lower in the DNC-R group, demonstrating unsatisfactory response of cells to an excessive volume at T2. CONCLUSIONS: DNC and HTK provided acceptable myocardial protection as single-dose applications. DNC-R had significantly unbalanced levels of biomarkers, and more arrhythmia/AV block incidence resulting in more extended ICU stay. For patients who may need redosing HTK may be preferable to DNC.

Effects of N-acetyl cysteine, vitamin E and vitamin C on liver glutathione levels following amiodarone treatment in rats.

INTRODUCTION: Amiodarone, a pharmaceutical extensively used to suppress atrial and ventricular tachyarrhythmias, is also known to cause many side effects on many tissues. N-acetyl-cysteine (NAC), vitamin E and vitamin C are known as antioxidants for their ability to minimize oxidative stress. In the peer-reviewed literature, there is no study reporting on the protective effects of these antioxidant agents against its hepatotoxicity. AIM: We investigated the oxidative effects of NAC, vitamins E and C on liver tissue after amiodarone treatment. MATERIAL AND METHODS: Rats were randomly assigned to: control; amiodarone group; amiodarone + NAC treated group; amiodarone + Vit. E group and amiodarone + Vit. C group. Liver tissues were isolated from animals and total glutathione levels were measured. RESULTS: In all time intervals, the level of glutathione increased. When all time intervals were compared, the amiodarone group revealed the lowest levels. The antioxidant co-administered group was studied; the glutathione levels were statistically significantly higher than the sole amiodarone group. When vitamins E, C or N-acetyl cysteine were examined, there was no statistically significant difference among them. CONCLUSIONS: In this study we found that hepatotoxicity capacity of amiodarone may be reduced by taking up antioxidants. In addition, the effect documented here may be reproducible and may be applied to clinical settings.

Investigation of survival and migration potential of differentiated cardiomyocytes transplanted with decellularized heart scaffold.

Mesenchymal stem cell-derived cardiomyocytes are employed as a source for myocardial cell transplantation as well as for tissue engineering in decellularized tissue scaffolds. The present study aimed at investigating the survival and migration potential of differentiated cardiomyocytes integrated to decellularized scaffolds after implantation into retroperitoneum of rats, and to assess the feasibility of their ectopic use for future cardiovascular tissue engineering. For this purpose, adipose tissue-derived mesenchymal stem cells (AdMSCs) were first isolated. Cells were labeled by bromodeoxyuridine (BrdU). Decellularized cardiac tissue scaffolds were acquired by application of ionic and non-ionic detergents and the labeled differentiated cells were seeded onto these tested decellularized scaffolds. After 1, 2, and 4 weeks of implantation, either cell free scaffold (CFS) or cell scaffold (CS) composites were examined by various techniques for ectopic migration potential of the implanted cells and interaction between the seeded cells on scaffolds. Throughout the first and second weeks of implantation, positively stained cells were observed in renal tissue samples. Observations, for cardiomyocytes-specific gene expression during weeks 1, 2, and 4, showed potential increased over each time period. A reverse transcription polymerase chain reaction (RT-PCR) results revealed an increased interaction between cells seeded on scaffolds, however CFS test groups showed no significant difference in gene expression. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 561-570, 2019.

Bone marrow derived mesenchymal stem cells ameliorate inflammatory response in an in vitro model of familial hemophagocytic lymphohistiocytosis 2.

BACKGROUND: Familial hemophagocytic lymphohistiocytosis 2 (FHL2) is the most common familial type of hemophagocytic lymphohistiocytosis with immune dysregulation. FHL2 patients have mutations in the perforin gene which cause overactivation and proliferation of cytotoxic T lymphocytes and natural killer cells. Perforin is the key component of the cytolytic granule response function of cytotoxic T lymphocytes and natural killer cells. Perforin dysfunction causes a cytotoxic immune deficiency with a clinical outcome of uncontrolled and continuous immune stimulation response. This excessive stimulation leads to continuous systemic inflammation and, ultimately, multiorgan failure. Radical therapy is hematopoietic stem cell transplantation which is limited by the availability of a donor. Exacerbations of inflammatory attacks require a palliative immunosuppressive regimen. There is a need for an alternative or adjuvant therapy to maintain these patients when immunosuppression is ineffective or a donor is not available. Beneficial actions of mesenchymal stem cells (MSCs) have been shown in autoimmune diseases in clinical trials and are attributed to their immune-modulatory properties. This study aimed to assess the immune-modulatory effect of MSCs in an in-vitro model of FHL2. METHODS: We generated a targeted mutation in the perforin gene of NK92 cells to create an in-vitro FLH2 model using Crispr/Cas technology. A coculture setup was employed to assess the immunomodulatory efficacy of MSCs. RESULTS: Engineered NK92 clones did not show PRF1 mRNA expression and failed to secrete perforin upon phorbol myristate acetate-ionomycin stimulation, providing evidence for a valid FHL2 model. Coculture media of the engineered cells were investigated for the abundance of several cytokines. Coculture with MSCs revealed a reduction in major proinflammatory cytokines and an induction in anti-inflammatory and immunomodulatory cytokines compared to the parental NK92 cells. CONCLUSIONS: This study shows the ameliorating effect of MSCs as an adjuvant immune modulator toward the therapy of FHL2 patients. MSCs are supportive therapy candidates for FHL2 patients under circumstances where prolonged immunosuppression is required to gain time before allogeneic hematopoietic stem cell transplantation.

Cellular localization and biological effects of 20nm-gold nanoparticles.

Gold nanoparticles (AuNPs) have recently emerged as prominent vehicles for many biomedical applications from sensing to delivery. The relevant literature contains conflicting data about the effects of AuNPs on living cells. The aim of present study is the synthesis and characterization of AuNPs at nanoscale, tracking their cellular localization and determining their effects on cell viability, migration and angiogenesis. Within this scope, 20 nm AuNPs were synthesized and characterized using various spectrometric techniques to determine their size, shape and surface properties such as charge and texture. Two main cell types including mouse fibroblast (L929) and human cervix adenocarcinoma (HeLa) were used in the study to compare the biological effects of colloidal gold on both non-cancer and cancer cells. AuNPs were allowed to interact with HeLa cells to determine their intracellular localization. AuNPs were mainly attached to the cell membrane/membranous compartments and to be captured in small amounts in cytoplasmic vacuoles or to be distributed freely in the cytosol. Scratch assay results showed that AuNPs reduced cancer cell migration especially at increasing concentrations. According to the chick chorioallantoic membrane assay, AuNPs exhibited strong anti-angiogenetic properties and can inhibit vascularization during angiogenesis. In addition, the MTT assay confirmed that AuNP-treated cells caused concentration dependent cytotoxic effects on both cell types. As a result, AuNPs not only have inhibitory effects on cancer cells, but also possess antiangiogenic activity, thus making them a multipotent agent for cancer therapy. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1708-1721, 2018.

Effects of propolis on warfarin efficacy.

INTRODUCTION: Warfarin is commonly used to avoid thromboembolism, predominantly for cardiovascular pathologies. However, the consumption of several herbal products is not permitted during its use due to the associated interactions. Propolis is a popular phytotherapy product made by honey bees. The use of propolis has been dramatically increasing in recent times. AIM: To evaluate the possible interactions between propolis and warfarin in a mouse model with determination of the international normalized ratio (INR) values. MATERIAL AND METHODS: CD-1 mice were employed in the experimental model. The mice were warfarinized, and propolis was administered simultaneously. The INR values were obtained. All animals were sacrificed at the end of the study. RESULTS: The baseline INR value was 0.8 ±0.1. After 72 h, the INR value increased as expected. The INR value was 7.28 ±1.08 in the control group and 5.8 ±2.88 in the propolis group. At the end of the study, the INR value was 1.3 ±0.37. Propolis interacted with warfarin and caused a decrease in the INR value. CONCLUSIONS: Propolis interactions, especially with warfarin, should be kept in mind and further studied. Healthcare specialists should be aware of this possible interaction between warfarin and propolis and inform patients about it.

Effects of VEGF and MSCs on vascular regeneration in a trauma model in rats.

In the human body, vascular injuries that are caused by trauma, vessel lumen stenosis, and occlusions are often irreversible and can lead to sequelae formation as the vessels cannot reproduce fast enough. To solve this problem, the blood flow must be returned to the region as fast as possible. The adipose tissue contains progenitor cells with angiogenic potential and can be used to resolve the issue. In the present study, mesenchymal stem cells (MSCs) derived from rat adipose tissue, vascular endothelial growth factor (VEGF), and their mixture were applied on the dorsum of a rat, which was traumatized and its contribution to vascular regeneration was reviewed. No application was made to the control group. The results showed that the percentage of necrotic area was significantly lower in the MSC group than that of all the other groups. When the VEGF group was compared to the VEGF + MSCs, the percentage of necrotic area was observed to be similiar. However, VEGF showed effects only when a large quantites of VEGF was applied to the flap area. VEGF could not fully respond to the needs, whereas MSCs can produce VEGF according to the needs of tissue. This makes them superior to stem cells.

Clinical performance and biocompatibility of hyaluronan-based heparin-bonded extracorporeal circuits in different risk cohorts.

This prospective randomized study compares novel hyaluronan-based heparin-bonded circuits vs. uncoated controls across EuroSCORE patient risk strata including biomaterial evaluation. Over a two-year period, 90 patients undergoing coronary artery bypass grafting were prospectively randomized to one of the two perfusion protocols: Group 1 was treated with hyaluronan-based heparin-bonded preconnected circuits (Vision HFO-GBS, Gish, CA, USA) and Group 2 with identical uncoated controls. Each group was composed of three subgroups (n=15) with respect to preoperative evaluation of low (EuroSCORE 0-2), medium (3-5) and high (6+) risk patients. Blood samples were collected after induction (T1) and heparinization (T2), 15 min after cardiopulmonary bypass start (T3), before cessation of CPB (T4), 15 min after reversal (T5), and the first postoperative day (T6). In high-risk patients, platelet counts demonstrated significant preservation at T4, T5 and leukocyte counts were lower at T5 in hyaluronan group (P

Vascular smooth muscle contraction/relaxation of rat carotid artery is not altered by bone grafting substitutes in vitro.

PURPOSE: The aim of this study was to explore the effects of various bone grafting substitutes (Osteosponge, Perioglas, Tutoplast, and Surgibone) on vascular smooth muscle tonus. METHODS: Bilateral carotid arteries were removed from rats and contraction/relaxation of isolated vessel rings were measured before and after contact with the biomaterials and then, for dose-dependent epinephrine and papaverin administrations, by a force displacement transducer. The data of each biomaterial group were collected by a computerized system and corresponding software at a sample rate of 1,000 kHz and were converted to contraction force. RESULTS: Vascular contraction forces were influenced in response to biomaterials tested except for Osteosponge (P < 0.05), although the differences between groups were insignificant (P > 0.05). There was a dose-dependent vascular response to epinephrine and papaverine administration upon biomaterial contact (P < 0.05). The dose-dependent vascular responses to epinephrine and papaverine administration were almost similar for all biomaterials tested (P < 0.05), suggesting that the biomaterials led to reversible effects on vascular contraction/relaxation behavior, which resulted in recovery. CONCLUSIONS: Osteosponge, Perioglas, Tutoplast, and Surgibone do not alter vascular smooth muscle tonus and vitality and therefore would, presumably, not jeopardize the angiogenesis of fresh blood vessels and full vascularization during tissue healing.

Comparison of polymethoxyethylacrylate-coated circuits with leukocyte filtration and reduced heparinization protocol on heparin-bonded circuits in different risk cohorts.

OBJECTIVES: The relative benefits of strategic leukofiltration on polymer-coated and low-dose heparin protocol on heparin-coated circuits were studied across EuroSCORE patient risk strata for three different cohorts. METHODS: In a prospective, randomized study, 270 patients undergoing coronary artery bypass grafting were allocated into three groups (n = 90): Group 1 - polymethoxyethylacrylate-coated circuits + leukocyte filters; Group 2 - polypeptide-based heparin-bonded circuits with reduced heparinization; and Group 3--CONTROL: uncoated circuits. Each group was further divided into three subgroups (n = 30), with respect to low- (EuroSCORE 0-2), medium- (3-5), and high- (6+) risk patients. Blood samples were collected at T1: following induction of anesthesia; T2: following heparin administration; T3: 15 min after CPB; T4: before cessation of CPB; T5: 15 min after protamine reversal; and T6: ICU. RESULTS: In high-risk cohorts, leukocyte counts demonstrated significant differences at T4 and T5 in Group 1, and at T4 in Group 2. Platelet counts were preserved significantly better at T4 and T5 in both groups (p < 0.05 versus control). Serum IL-2 and C3a levels were significantly lower at T3, T4 and T5 in Group 1, and T4 and T5 in Group 2 (p < 0.05). Postoperative bleeding, respiratory support time and incidence of atrial fibrillation were lower in the study groups versus control. Cell counts on filter mesh and heparin-coated fibers/ circuits were significantly higher in the high-risk cohorts versus uncoated fibers. Phagocytic capacity increased on filter mesh, especially in high-risk specimens. SEM evaluation demonstrated better preserved coated circuits. CONCLUSION: Leukofiltration and coating reduced platelet adhesion, protein adsorption, atrial fibrillation and reduced heparinization acted via modulation of systemic inflammatory response in high-risk groups.

In vivo study to assess the viability and mass effect of fascia injection in comparison with block fascia graft.

Recently, fat injection has gained favor among plastic surgeons for soft tissue augmentation. However, fat injections lose 50% of their volume after 1 year. The profession is in need of an injectable bulking material that gives a long-lasting improvement. Using 30 male rats, this study investigated the stability of the size and structure of the injected fascia autograft and compared it with surgically transplanted fascia. The fascia graft was harvested from the back of the rat, big, and divided into two equal pieces. The first piece was minced into a paste and injected subcutaneously on the anterior surface of the right ear. The other piece was transplanted subcutaneously on the anterior surface of the left ear. The grafts were observed for any sign of resorption over 1 to 6 months.Grossly, injection and transplantation sites were palpable at the end of the observation periods. Microscopic examination showed that injected fascia maintains its histomorphologic structure. These findings indicate that the injected fascia graft is well tolerated, and the size of the graft remained stable. According to this study, fascia injection can result in bulking material that gives a long-lasting improvement, and can be a viable alternative to other methods.

Morphologic, functional and behavioral effects of titanium dioxide exposure on nerves.

OBJECTIVES: The purpose of this study was to explore morphologic, functional, and behavioral effects of titanium dioxide (TiO(2)) on nerves. MATERIAL AND METHODS: A total of 17 albino rats were used for nerve conduction experiments, hot-plate tests, and histological evaluation. TiO(2) was implanted unilaterally on the sciatic nerves of five rats. Ten days after surgery, test and control nerves were dissected and their signal transduction speeds were quantified by suction electrodes in a bath containing a Tyrode solution. Twelve rats were divided into three equal groups resulting in equal number of nerves (n=8) for TiO(2) implantation, surgical exposure of the nerves, and for use as controls. One week after surgery, hot-plate tests were undertaken for 10 consecutive days to determine response latencies of the nerves. At the termination of the experiments, the nerves were harvested, processed, and examined under a microscope. RESULTS: The signal transduction speeds of TiO(2)-implanted nerves was similar to control specimens (P>0.05). The avoidance responses of TiO(2)-implanted, surgically exposed, and control nerves were comparable (P>0.05). At the cellular level, TiO(2) did not lead to any signs of adverse reactions on nerves. CONCLUSIONS: TiO(2), the main oxide surrounding endosseous titanium implants, does not alter the structure and the function of myelinated nerves.