Frontiers of stem cell engineering for nanotechnology-mediated drug delivery systems.

Background and purpose: Cell biology approaches have gained a successful integration, development and application of nanotechnology with stem cell engineering and have led to the emergence of a new interdisciplinary field known as stem cell nanotechnology (SCN). Recent studies showed the potential and the advancement of developments for SCN applications in drug delivery systems. Cancer, neurodegenerative, muscle and blood diseases, cell and gene therapies, and tissue engineering and regenerative medicine applications are the important targets of SCN. Experimental approach: In this overview, we searched the literature using the common online websites for research and read the open access, full-text available articles since 2013. Key results: The studies vary according to the type of disease they targeted and the strategies they proposed, whether diagnostic or therapeutic. In addition to the use of stem cells, the utilisation of their membranes, secretomes, exosomes and extracellular vesicles with an appropriate nanotechnology strategy is also an aspect of the research. Conclusion: This brief overview of stem cell nanotechnology over the last ten years aims to provide insight into the frontiers of stem cell engineering for nanotechnology-mediated drug delivery systems.

Optimization of a rapid and sensitive nucleic acid lateral flow biosensor for hepatitis B virus detection.

BACKGROUND AND OBJECTIVE: The utilization of direct amplification of nucleic acid from lysate has attracted interest in the advancement of straightforward and economical point-of-care assays. Consequently, this study primarily focuses on the development of a rapid, precise, and cost-effective lateral flow biosensor for the convenient detection of HBV nucleic acid at the point-of-care. Furthermore, the study evaluates the effectiveness of the direct amplification method in comparison to purified nucleic acid samples within the context of LAMP-LF biosensing approaches. METHODS: The experiments conducted in this study utilized clinical serum samples that were confirmed as HBV-positive through real-time PCR assays. Sample preparation involved employing spin column nucleic acid purification and serum heat treatment. To amplify a 250 bp fragment of the HBV polymerase gene, three pairs of specific LAMP primers were utilized, which were biotin-labeled and FITC-labeled for detection purposes. Various incubation temperatures (ranging from 64 to 68 °C) and durations (30 min, 45 min, and 1 h) were investigated to determine the optimal conditions for the LAMP assay. The results were subsequently assessed through fluorometric analysis, white turbidity measurements, and lateral flow assay. Milenia HybriDetect1 strips, designed for immediate use, were employed to visualize the LAMP amplicons. Furthermore, the performance of the lateral flow biosensor was evaluated using 10-fold serial dilutions of a secondary standard containing a viral load of 108 IU/ml. RESULTS: The optimization of the LAMP reaction was achieved at a temperature of 67 °C, resulting in significant turbidity after a 30-minute incubation period. When the spin column purification method was employed, varying test bands were observed for templates ranging from 108 IU/ml to 101 IU/ml viral load. However, when serum samples underwent heat treatment and the resulting supernatant was directly used for LAMP, the lateral flow assay was capable of detecting a minimum viral load of 103 IU/ml. CONCLUSION: In resource-limited settings, the LAMP-LF assay presents a promising solution for HBV testing. However, it is important to note that direct amplification without DNA purification may diminish the performance of the approach.

A new approach for development of vaccine against visceral leishmaniasis: Lipophosphoglycan and polyacrylic acid conjugates.

OBJECTIVE: To determine the antileishmanial vaccine effectiveness of lipophosphoglycan (LPG) and polyacrylic acids (PAA) conjugates on in vivo mice models. METHODS: LPG molecule was isolated and purified from large-scale Leishmania donovani parasite culture. Protection efficacies of LPG alone, in combination with Freund's adjuvant, in a physical mixture and in conjugate (consisting of various LPG concentrations) with PAA, were comparatively determined by various techniques, such as cultivation with the micro-culture method, assessment of in vitro infection rates of peritoneal macrophages, determination of parasite load in liver with Leishman-Donovan Units, and detection of cytokine responses. RESULTS: Obtained results demonstrated that the highest vaccine-mediated immune protection was provided by LPG-PAA conjugate due to all parameters investigated. According to the Leishman-Donovan Units results, the sharpest decline in parasite load was seen with a ratio of 81.17% when 35 µg LPG containing conjugate was applied. This value was 44.93% for the control group immunized only with LPG. Moreover, decreases in parasite load were 53.37%, 55.2% and 65.8% for the groups immunized with 10 µg LPG containing LPG-PAA conjugate, a physical mixture of the LPG-PAA, and a mixture of LPG + Freund's adjuvant, respectively. Furthermore, cytokine results supported that Th1 mediated protection occurred when mice were immunized with LPG-PAA conjugate. CONCLUSIONS: It has been demonstrated in this study that conjugate of LPG and PAA has an antileishmanial vaccine effect against visceral leishmaniasis. In this respect, the present study may lead to new vaccine approaches based on high immunogenic LPG molecule and adjuvant polymers in fighting against Leishmania infection.

Human olfactory stem cells for injured facial nerve reconstruction in a rat model.

BACKGROUND: The purpose of this study was to show the efficacy of olfactory stem cells for injured facial nerve reconstruction in a rat model. METHODS: Olfactory stem cells were isolated from the olfactory mucosa of human participants. A 2-mm excision was performed on the right facial nerve of all rats. Reconstruction was performed with a conduit in group 1 (n = 9); a conduit and phosphate-buffered saline in group 2 (n = 9); and a conduit and labeled olfactory stem cell in group 3 (n = 9). Rats were followed for whisker movements and electroneuronography (ENoG) analyses. RESULTS: The whisker-movement scores for group 3 were significantly different from other groups (p < .001). ENoG showed that the amplitude values for group 3 were significantly different from group 1 and group 2 (p = .030; p < .001). Group 3 showed marked olfactory stem cell under a fluorescence microscope. CONCLUSION: This study suggests that olfactory stem cells may be used as a potent cellular therapy for accelerating the regeneration of peripheral nerve injuries. © 2016 Wiley Periodicals, Inc. Head Neck 38: E2011-E2020, 2016.

Investigation of antileishmanial activities of Tio2@Ag nanoparticles on biological properties of L. tropica and L. infantum parasites, in vitro.

Leishmaniasis is a public health problem which is caused by protozoon parasites belonging to Leishmania species. The disease threatens approximately 350 million people in 98 countries all over the world. Cutaneous Leishmaniasis (CL) and Visceral Leishmaniasis (VL) are the mostly commonly seen forms of the disease. Treatment of the disease has remained insufficient since current antileishmanial drugs have several disadvantages such as toxicity, costliness and drug-resistance. Therefore, there is an immediate need to search for new antileishmanial compounds. TiO2@Ag nanoparticles (TiAg-Nps) have been demonstrated as promising antimicrobial agents since they provide inhibition of several types of bacteria. The basic antimicrobial mechanism of TiAg-Nps is the generation of reactive oxygen species (ROS). Even though Leishmania parasites are sensitive to ROS, there is no study in literature indicating antileishmanial activities of TiAg-Nps. Herein, in this study, TiAg-Nps are shown to possess antileishmanial effects on Leishmania tropica and Leishmania infantum parasites by inhibiting their biological properties such as viability, metabolic activity, and survival within host cells both in the dark and under visible light. The results indicate that TiAg-Nps decreased viability values of L. tropica, and L. infantum promastigotes 3- and 10-fold, respectively, in the dark, while these rates diminished approximately 20-fold for each species in the presence of visible light, in contrast to control. On the other hand, non-visible light-exposed TiAg-Nps inhibited survival of amastigotes nearly 2- and 2.5-fold; while visible light-exposed TiAg-Nps inhibited 4- and 4.5-fold for L. tropica and L. infantum parasites, respectively. Consequently, it was determined that non-visible light-exposed TiAg-Nps were more effective against L. infantum parasites while visible light-exposed TiAg-Nps exhibited nearly the same antileishmanial effect against both species. Therefore, we think that a combination of TiAg-Nps and visible light can be further used for treatment of CL, while application of TiAg-Nps alone can be a promising alternative in VL treatment.

Microcapillary culture method: a novel tool for in vitro expansion of stem cells from scarce sources.

BACKGROUND AND AIMS: Although increasing numbers of studies report the derivation of stem cells from a variety of different tissues, derivation efficiencies greatly vary among different studies even for the same tissue source. Hence, a consistent and efficient isolation protocol has not yet been established to date. Several factors have so far been documented that influence and limit mesenchymal stem cell (MSC) isolation and cultivation, including the age and gender of the tissue donor, origin of the tissue, amount of sampled tissue material and cell culture characteristics including the choice of basal media, serum, gas composition, etc. The aim of the study was to investigate the microcapillary culture method (MCM) to establish an efficient and consistent isolation as well as cultivation protocol by comparing the results with other classic culture systems (flasks, center wells). METHODS: MSCs isolated from adipose tissue of different donors were observed comparatively under different culture systems (flasks, center wells, microcapillary tubes) and their proliferation and differentiation were investigated. Flow cytometry was used for immunophenotypic characterization of derived cells and histochemical staining (Oil Red O and Alizarin Red S) was applied for determining their differentiation capacity. RESULTS: It has been shown for the first time that AD-MSCs can consistently and efficiently be derived from a scarce amount of adipose tissue by MCM. CONCLUSIONS: Further and similar studies should be performed to determine whether this methodology can also be applicable for other MSC sources.

Detection of antileishmanial antibodies in blood sampled from blood bank donors in Istanbul.

AIMS: According to the WHO, only 5-20% of the total cases of leishmaniasis are symptomatic leishmaniasis; the other cases are identified as asymptomatic leishmaniasis. In recent studies, it has been demonstrated that donor blood plays an important role in the epidemiology of asymptomatic leishmaniasis. However, the number of the studies on this subject is still insufficient. Additionally, donor blood samples obtained from Istanbul, which is the biggest metropolitan area in Turkey, have not been investigated with regard to Leishmania. Moreover, there is no information about the sensitivity of noninvasive serological methods that are used in the detection of leishmaniasis donor blood samples. Accordingly, this study aimed to investigate the presence of antileishmanial antibodies in blood samples obtained from blood bank donors in Istanbul, by using different serologic methods, and to determine the most sensitive detection method. MATERIALS & METHODS: Blood samples were taken from 188 healthy blood bank donors to the Capa Turkish Red Crescent Blood Bank (Istanbul, Turkey), and the presence of antileishmanial antibodies was measured by indirect immunofluorescent antibody test (IFAT), ELISA, immunochromatographic dipstick rapid test, and western blot (WB). RESULTS: Antileishmanial antibodies were determined in 12 out of 188 samples by IFAT (6.4%), and six out of these 12 donors were found to be positive at diagnostic titer 1:128 (3.2%). One hundred and eighty eight samples were investigated by ELISA and one (0.5%) of them gave a positive result. None of 188 samples provided a positive result by immunochromatographic test. WB applied to the 12 seroreactive donors showed that three out of 12 donors were positive. CONCLUSION: In this study, the presence of antileishmanial antibodies in blood samples of blood bank donors from Istanbul has been demonstrated by using feasible and low-cost serological methods. Additionally, in comparison with other simple and low-cost detection methods, WB was used for confirmation. IFAT has a higher sensitivity and therefore may be preferred as a prescreening method in endemic or nonendemic areas.

Adipose tissue-derived mesenchymal stem cells as a new host cell in latent leishmaniasis.

Some protozoan infections such as Toxoplasma, Cryptosporidium, and Plasmodium can be transmitted through stem cell transplantations. To our knowledge, so far, there is no study about transmission of Leishmania parasites in stem cell transplantation and interactions between parasites and stem cells in vitro. Therefore, the aim of this study was to investigate the interaction between different species of Leishmania parasites and adipose tissue-derived mesenchymal stem cells (ADMSCs). ADMSCs have been isolated, cultured, characterized, and infected with different species of Leishmania parasites (L. donovani, L. major, L. tropica, and L. infantum). Infectivity was examined by Giemsa staining, microculture, and polymerase chain reaction methods. As a result, infectivity of ADMSCs by Leishmania parasites has been determined for the first time in this study. According to our findings, it is very important that donors are screened for Leishmania parasites before stem cell transplantations in regions where leishmaniasis is endemic.