Bacterial Synthesis of Nanoparticles: Current Trends in Biotechnology and Biomedical Fields

On estimation scales ranging from 0.1 nm to 100 nm, the nanoscale is part of the capacitance components of the physical-synthetic and natural environment. Dimensionality, morphology, structure, uniformity, and agglomeration are all used to classify nanoparticles. Its functionality and effect on the environment and species are influenced by its shape and morphology. The priority research is to determine the effects of nanoparticles on any biological entity that is necessary when designing nanotechnology-based biotechnological and biomedical products. Bacteria have a remarkable ability to reduce metal ions, making them one of the most promising candidates for nanoparticle biosynthesis. Nanoparticles have been researched in the biomedical field for antimicrobial, biosensor, diagnostic imaging, and drug delivery applications. These natural technologies appear to be capable of producing stable nanoparticles with well-defined dimensions, morphologies, and compositions by optimizing reaction conditions and selecting the best bacteria. This work includes a list of the most commonly used microorganisms and associated Nanoparticles, as well as a discussion of current biotechnology and biomedical developments

Microfluidic Live-Imaging Technology to Perform Research Activities in 3D Models

Morphological dissimilarity and its evolution over time are one of the most unexpected variations found when comparing cell cultures in 2D and 3D. Monolayer cells appear to flatten in the lower part of the plate, adhering to and spreading in the horizontal plane while not extending vertically. Consequently, cells developed in two dimensions have a forced apex-basal polarity. Co-cultivation and crosstalking between multiple cell types, which control development and formation in the in vivo counterpart, are possible in 3D cultures. With or without a scaffold matrix, 3D model culture may exhibit more in vivo-like morphology and physiology. 3D cultures mimic relevant physiological cellular processes, transforming them into one-of-a- kind drug screening platforms. The structures and dynamics of regulatory networks, which are increasingly studied with live-imaging microscopy, must be considered to help and guarantee the functional maintenance of a 3D structure. However, commercially available technologies that can be used for current laboratory needs are minimal, despite the need to make it easier to acquire cellular kinetics with high spatial and temporal resolution, in order to improve visual efficiency and, as a result, experimentation performance. The CELLviewer is a newly developed multi-technology instrument that integrates and synchronizes the work of various scientific disciplines. The aim of this study is to test the device using two different models: a single Jurkat cell and an MCF-7 1 spheroid. The two models are loaded into the microfluidic cartridge for each experiment after they have been grown and captured in time-lapse for a total of 4 hours. The samples used are tracked under the operation of the optics after adaptive autofocus, while slipping inside the cartridge chamber and the 3D rotation was successfully obtained experimentally. The MitoGreen dye, a fluorescence marker selectively permeable to live cells, was then used to determine cell viability. To measure the model diameter, construct fluorescence intensity graphs along a straight line passing through the cell, and visualize the spatial fluorescence intensity distribution in 3D, Image J software was used

UPDATED MODIFICATIONS ABOUT SPIN BIOREACTOR

This paper carries out the implementation of a Raspberry-controlled bioreactor. The previously proposed model was analyzed and assembled to determine its effectiveness and solutions to the problems that emerged during construction were highlighted. The realization of the project is possible, as now some elements can be replaced. Overall, these modifications on this miniature rotary bioreactor make the apparatus exceptionally easy to build following designated guidelines. This updated device is susceptible to various experiments of varying lengths without having to worry about possible malfunctions such as unexpected events or hardware and software problems

Germplasm Conservation

With the increase in risk of extinction of various plants, the trend has been shifted to employment of many biotechnological techniques for preservation of genetic resources of plant and is the area of research which needs to be revolutionized after a specific time period because it allows the production and selection of crop varieties with desirable characteristics during breeding process such as improved fuel, food and health facilities. Having an immense research in conservation of non-threatened species, there is a small collection of knowledge available for conservation of endangered ones. This chapter aims to highlight the various techniques in germplasm conservation of endangered or the species which are at extent of extinction and also the future directions in this field. In developing countries where most of agriculture depends upon food crops, the maintenance of genetic variation is of immense importance. On farm conservation provides the best example of preservation and evolution based on genetic variability which can occur ex-situ and in- situ environment in farms or gene bank. So, it presents the best option for conservation or maintenance of ecosystem and biodiversity which ensures survival of endangered species via germplasm. The most point to consider is that germplasm or genes have to be conserved instead of genotype. In situ conservation involves preservation of plant crops in the field condition in ecosystem where plant is adopted to grow in order to maintain self –sustaining process in natural ecosystem. Similarly ex-situ involve the collections of seed banks of genes collected from plant under natural conditions to produce desirable varieties or from tissue culture in laboratory also referred as in-vitro methodology. In –vitro techniques include cryopreservation which include freezing at much lower temperature than that of freezing point i.e. -196 °C in liquid nitrogen for preserving species which are near to extent of endangerment. Cold storage and storing at lower temperature provides best opportunity for protection against damage caused by rapid freezing. Germplasm exchange has become now a usual practice ensuring exchange of varieties between cultivated and wild types as for example in potatoes specie etc. DNA as well as gene or seed banks provide molecular sources for conservation at biotechnological level. The techniques of introgression and incorporation are basic approaches for germplasm conservation. So there is need to revolutionize and practice germplasm conservation for fulfilling future needs being aimed at conserving endangered or threatened species from conservation hotspots

From isolation of adult adipose tissue derived stem cells ADAS to labelling with superparamagnetic iron oxide nanoparticles: first approaches to unleash the potential

The use of adult adipose-derived stem cells (ADAS) as a treatment for neurological diseases is in promising development. Extracellular vesicles such as exosomes (EXO), which impact surrounding cells, are the main biological agents of ADAS. Exosome localization and tracking techniques need to be effective and non-invasive in the current development of exosome therapies. Exosomes must be labeled with contrast agents, such as ferrous superparamagnetic nanoparticles (NPs). The current research project aims to validate the therapeutic efficacy of ADAS-derived EXOs labelled with different NPs in models of neurodegenerative diseases, capable of providing an imaging and cell therapy approach

Automation in 3D cellular system in Live-Imaging with Microfluidic Technology CELLviewer®

Differences observed when comparing cell cultures in 2D and 3D is morphological dissimilarity and their evolution over time. Cells grown in a monolayer tend to flatten on the bottom of the plate by adhering and spreading on the horizontal plane without expanding into the vertical dimension; § Mitochondria are involved in crucial cellular tasks controlling the cell cycle and growth such as cell signaling, differentiation, and death. Damage to and subsequent dysfunction of mitochondria play a role in various diseases like diabetes, myopathy and other systemic disorders; § CELLviewer® enables the simultaneous 3D cell culture and live cell imaging as well, featuring microfluidics and time-lapse multicolour epifluorescence microscopy; § Single cell tracking in 3D space is now possible and is combined with subsequent biochemical analyses of individually tracked cells, keeping their identity traceable with CELLviewer® system; q Jurkat (ATCC) Cells grown at 37°C and 5% CO2; q Medium RPMI 1640 soil (Gibco, Life Technologies, Thermo Fisher Scientific), with 2 mM of L-glutamine, 10% FBS, 100 units/mL of penicillin and 100 mg/mL of streptomycin; q MitoGreen (PromoKine, PromoCell) incubated for 20 minutes in the dark at 37°C with MitoGreen 200 mM; q The sample is then piped inside a 50ml Falcon tube closed with a 50ml CELLviewer® DOCK and flowed inside the cartridge chamber; q CELLviewer® automatically captures sample images in Brightfield channel and GFP channel; q ImageJ software was used for image analysis using the Measure function to calculate the diameter of a single cell; q 3D surface plot plug-in to display in 3D the distribution of the intensity of spatial fluorescence; Staining of mitochondria with fluorescent dyes, antibodies or fluorescent molecules can greatly facilitate studies of their function and distribution and the viability of cells in healthy and diseased individuals. The preliminary experience conducted with CELLviewer indicates that this equipment responds to the needs of individual operators as it consists of a synthesis of different integrated tools, which works both with manual and automated control. A microfluidic system has been developed and demonstrated that the 3D model can locate the 3D model spatially, it's possible to carry out experiments in direct time in terms of physiology, toxicology and clinical pharmacology. The entire automated system allows full autonomy and protocol management thanks to the software making the operator free to conduct other work, thus increasing the productivity of his project. In summary, the proposed microfluidic technology can serve as a new platform approach, which has the potential to advance studies at the cellular level. Single-cell Jurkat cells was isolated and imaged for 4 and 7 hours respectively and intensified labelling of the mitochondria and fluidic transport were observed over time. CELLviewer® can obtain detailed images of current cellular morphology with resolution and high-quality data; employing time-lapse imaging can be achieved, the evolution of cells and their 3D morphology

Germplasm Conservation

With the increase in risk of extinction of various plants, the trend has been shifted to employment of many biotechnological techniques for preservation of genetic resources of plant and is the area of research which needs to be revolutionized after a specific time period because it allows the production and selection of crop varieties with desirable characteristics during breeding process such as improved fuel, food and health facilities. Having an immense research in conservation of non-threatened species, there is a small collection of knowledge available for conservation of endangered ones. This chapter aims to highlight the various techniques in germplasm conservation of endangered or the species which are at extent of extinction and also the future directions in this field. In developing countries where most of agriculture depends upon food crops, the maintenance of genetic variation is of immense importance. On farm conservation provides the best example of preservation and evolution based on genetic variability which can occur ex-situ and in- situ environment in farms or gene bank. So, it presents the best option for conservation or maintenance of ecosystem and biodiversity which ensures survival of endangered species via germplasm. The most point to consider is that germplasm or genes have to be conserved instead of genotype. In situ conservation involves preservation of plant crops in the field condition in ecosystem where plant is adopted to grow in order to maintain self –sustaining process in natural ecosystem. Similarly ex-situ involve the collections of seed banks of genes collected from plant under natural conditions to produce desirable varieties or from tissue culture in laboratory also referred as in-vitro methodology. In –vitro techniques include cryopreservation which include freezing at much lower temperature than that of freezing point i.e. -196 °C in liquid nitrogen for preserving species which are near to extent of endangerment. Cold storage and storing at lower temperature provides best opportunity for protection against damage caused by rapid freezing. Germplasm exchange has become now a usual practice ensuring exchange of varieties between cultivated and wild types as for example in potatoes specie etc. DNA as well as gene or seed banks provide molecular sources for conservation at biotechnological level. The techniques of introgression and incorporation are basic approaches for germplasm conservation. So there is need to revolutionize and practice germplasm conservation for fulfilling future needs being aimed at conserving endangered or threatened species from conservation hotspots

MICROFLUIDIC LIVE-IMAGING WITH CELLVIEWER TECHNOLOGY TO PERFORM BIOTECHNOLOGICAL TASKS

Cells grown in a monolayer tend to flatten in the lower part of the plate adhering to and spreading in the horizontal plane without expanding in the vertical dimension. The result is that cells grown in 2D have a forced apex-basal polarity. Microfluidic Live-Imaging with CellViewer technology is an ideal solution to observe the maintenance of a cell in excellent health, trying to bridge the gap between the 2D and 3D model. In this work we propose to test the system on a single isolated Jurkat cell in the microfluidic cartridge and record the timelapse for 4 hours. After adaptive autofocus, when sliding inside the cartridge chamber, the single cell is tracked under the action of the optics and the 3D rotation was experimentally successfully achieved. Then a single cell viability assessment was used using MitoGreen-dye a fluorescence marker selectively permeable to live cells. ImageJ software was used to: calculate the diameter of a single cell, create fluorescence intensity graphs along a straight line passing through the cell, visualize spatial fluorescence intensity distribution in 3D

Magnetic resonance imaging of adipose-derived adult stem cells labelled with superparamagnetic iron oxide nanoparticles

The application of mesenchymal stem cells (MSCs) represents a new promising approach for treating neurodegenerative diseases. Recently, considerable attention has been paid to adipose-derived adult MSC (ADAS), thanks to the easy availability of adipose tissue and to the possibility of autologous cells transplantation. Any possible application of therapies based on ADAS in the clinics cannot occur without elucidation of their homing. Superparamagnetic iron-oxide nanoparticles (NPs) can be used to label and track cells in vivo via Magnetic Resonance Imaging (MRI

In vitro 3D Model of Female Reproductive Tract: An Overview and Future Aspect

Hormones must be balanced and dynamically controlled for the Female Repro- ductive Tract (FRT) to function correctly during the menstrual cycle, pregnancy, and delivery. Gamete selection and successful transfer to the uterus, where it implants and pregnancy occurs, is supported by the mucosal epithelial lining of the FRT ovaries, uterus, cervix, fallopian tubes, and vagina. Successful implantation and placentation in humans and other animals rely on complex interactions between the embryo and a re- ceptive female reproductive system. The FRT’s recent breakthroughs in three-dimen- sional (3D) organoid systems now provide critical experimental models that match the organ’s physiological, functional, and anatomical characteristics in vitro. (Haycock, [1]). This article summarizes the current state of the art on organoids generated from various parts of the FRT. The current analysis examines recent developments in the creation of organoid models of reproductive organs, as well as their future directions