Name: formulation of zinc-based nanomaterials using the eucommia ulmoides bark extract and their wound healing potential
Uploaded: 2024-12-27T14:20:00.000+00:00
Duration: 6 min 54 s
Description: The aqueous extract from the bark of Eucommia ulmoides serves as a rich source of bioactive compounds with numerous health benefits. The protocol here ...

The authors would like to sincerely thank the Department of Cell Biology, Central South University, Changsha, China, for providing the instrumentation facilities.

NOTE: Before preparing the extract, the obtained bark material was washed twice using deionized water and dried in a shaded place. The shade-dried barks were stored in an airtight container.
1. Preparation of Eucommia ulmoides bark extract
<ol>
	<li>Chop the bark collected from the Eucommia ulmoides tree into small pieces using scissors.</li>
	<li>Wash the chopped bark materials twice with double distilled water.</li>
	<li>Dry the bark pieces under 37 &#176;C for 24 h at shaded conditions.
	<ol>
		<li>Adjust the duration of the drying process based on the quantity of bark utilized for the study. Ensure the barks are completely dried before chopping the barks into small pieces. Avoid direct sunlight.</li>
	</ol>
	</li>
	<li>Transfer 20 g of shade-dried barks into a conical flask containing 220 mL of sterile double distilled water and heat at 130 &#176;C for 20 min.
	<ol>
		<li>The color of the reaction solution changes to light yellow. The changes in color of the solution occur after 10 min. Allow the solution to heat for another 10 min. Adjust the volume of double distilled water based on the sample quantity.</li>
	</ol>
	</li>
	<li>Store the crude extract containing polyisoprene at 4 &#176;C for further use. The formation of a thread-like structure indicates the presence of polyisoprene in the extracts.</li>
</ol>
2. Biosynthesis of E. ulmoides bark-mediated ZnO nanoparticles
<ol>
	<li>Add 1 M Zinc nitrate dihydrate Zn (NO3)2 to 50 mL of deionized water in a 500 mL conical flask. Stir continuously using magnetic stirring (60 RPM). The Zn (NO3)2 takes 30 min to dissolve completely.</li>
	<li>Add 15 mL of E. ulmoides bark extract dropwise to 20 mL of 1 M Zinc nitrate dihydrate (Zn (NO3)2) solution.</li>
	<li>Place the covered reaction mixture onto a magnetic stirrer, switch on the stirrer, and spin at (60 RPM) for 3 h.</li>
	<li>Add 1 N sodium hydroxide NaOH (3 mL) solution dropwise to the reaction mixture to adjust the pH to 9. Add NaOH until the solution mixture turns milky white and the pH is not more than 9. The solution becomes a milky white color when ZnO nanoparticles are formed.
	<ol>
		<li>The preparation of 1 M of Zinc nitrate dihydrate Zn (NO3)2 solution volume may vary based on the experimental needs. Make sure to use the freshly prepared bark extracts. If the pH exceeds 10, it will result in nanoparticle aggregation.</li>
	</ol>
	</li>
	<li>Transfer the synthesized Eu-ZnO-NPs to a 50 mL centrifuge tube and centrifuge it at 100 x g for 5 min at 4 &#176;C. 
	NOTE: The impurities can be avoided by immediate washing.</li>
	<li>Collect the washed Eu-ZnO-NPs in a glass plate and dry them at 45-50 &#176;C for 1 h in a hot air oven. 
	NOTE: If the Eu-ZnO-NPs are kept for more than 30 min can affect the physicochemical nature of the nanoparticles</li>
</ol>
3. Size confirmation using TEM
<ol>
	<li>Prepare 1 mg/mL of nanoparticles in DDH2O and load 5 &#181;L of Eu-ZnO-NP sample in the copper grid and wait until it completely dries. Vortex the sample prior to loading on the copper grid.</li>
	<li>Load the copper grid containing Eu-ZnO-NPs on the TEM sample holder and acquire images at 50x and 100x magnifications. 
	NOTE: Grids must be picked up using tweezers during this step.</li>
</ol>
4. Cytotoxicity assessment
<ol>
	<li>Seed 1 &#215; 104 HUVECs in each well of a 96 well plate and place it in a 5% CO2 37 &#176;C incubator.</li>
	<li>Add 10 &#181;L of various concentrations 0, 10, 20, 30, 40, and 50 &#181;g/mL of Eu-ZnO-NPs into the 90% confluent cells and incubate it for 24 h.</li>
	<li>After incubation, remove the old medium without disturbing the cells, add 10 &#181;L of CCK-8 solution to each well containing 90 &#181;L of fresh DMEM medium, and incubate in a 5% CO2, 37 &#176;C incubator.</li>
	<li>Measure the absorbance of cells treated with CCK8 solution at 450 nm using a spectrophotometer. 
	NOTE: The absorbance was measured immediately within 15 min to avoid the changes in absorbance.</li>
</ol>
5. Preparation of HUVEC cells for scratch assay
<ol>
	<li>Seed an appropriate (1 &#215; 105) quantity of HUVECs in 12 well culture plates containing Dulbecco&#39;s modified Eagle medium with 10% fetal bovine serum (FBS) and 1% pen-strep and incubate it in a 5% CO2 37 &#176;C incubator.
	<ol>
		<li>Before performing the scratch assay, check the confluency using the inverted microscope. 
		NOTE: For performing the scratch assay 6 well plates or 12 plates will be utilized based on the requirements, and the cell density will vary for different culture plates. The usage of 70%-80% confluent cells is ideal and recommended for scratch assay. The volume of DMEM medium may vary based on the culture plates utilized in the study. For instance, 6 well plates require 1-1.5 mL of culture medium, and 12 well plates require 0.5-1.0 mL of growth medium.</li>
	</ol>
	</li>
	<li>Gently make a scratch using a sterile 200 &#181;L pipette tip in the representative wound with a wound width of 200 &#181;m.</li>
	<li>Ensure that the tip used to make scratches on the cell monolayer makes contact with the surface of the cells. 
	NOTE: Every time a scratch is made, use the sterile tip.</li>
	<li>Remove the complete medium and wash the HUVEC monolayer using 1 mL of 1x PBS to remove the detached cells. 
	NOTE: Ensure the detached monolayer cells are completely removed from the respective wells. Confirm that no damage has occurred in the wound-created area.</li>
	<li>To evaluate the wound healing potential of synthesized Eu-ZnO-NPs, add 0 (Control), 10, and 20 &#181;g/mL concentrations of Eu-ZnO nanoparticles combined with a complete medium with 10% FBS into the wells. Maintain the experimental plates at 37 &#176;C in a 5% CO2 incubator. 
	NOTE: Do not disturb the cell monolayer while adding the fresh, complete medium. Ensure that the monolayer cells remain undisturbed during the incubation.</li>
	<li>Acquire microphotograph at 0 h and 24&#160;h using an inverted microscope. Use the annotation tool to measure the wound closure in different time intervals.
	<ol>
		<li>Calculate the percentage of wound closure using the following formula: Wound closure (%) = [(cell migration (in &#181;m) at 0 h - cell migration (in &#181;m) at 24 h)/ cell migration (in &#181;m) at 0 h] x 100.</li>
	</ol>
	</li>
</ol>

Bioactive ZnO Nanoparticles from Eucommia ulmoides for Wound Healing Enhancement

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The authors have nothing to disclose

The E. ulmoides bark, seed, and leaves are believed to exhibit numerous health benefits. Our results have shown that the synthesis of EU-ZnO nanoparticles was achieved using a simple and cost-effective approach. The aqueous extract was utilized to synthesize the nanoparticles. Heating the bark material at high temperatures may cause degradation of some phytoconstituents and lower the extraction efficiency. The phytoconstituents present in the aqueous herbal extract can reduce the metal ions to form nanoparticles...

Medicinal plants and plant-derived compounds have been shown to exhibit numerous health benefits1. The World Health Organization (WHO) reported that 80% of the global population depends upon traditional medicinal plants for primary health care. China is well-recognized and popular for its Traditional Chinese Medicine (TCM) practices. Chinese medicinal herbs have been reported to treat various diseases and utilized for their biological potential. Medicinal plants serve as reservoirs for bioactive compounds and multiple therapeutic roles. Medicinal plants have also been utilized to treat the wounds. There are several types of approaches applied to treat chronic wounds2. A recent investigation revealed that medicinal plants were involved in the wound healing process by providing favorable conditions for healing, free from infections, and fastening tissue regeneration3. Meanwhile, the antibacterial and antifungal properties of bioactive compounds present in medicinal plants can help to cure wounds and fasten wound healing efficiency4.
Metal-based nanomaterials are gaining attention due to their biocompatible and biodegradable properties. Eucommia ulmoides, commonly called the Chinese rubber tree, is a native species of China. The leaves and bark of the tree are used in medicinal practices. Most importantly, the plant species was cultivated in the central and western provinces of China5. Peng et al.6reported that leaves, barks, and staminate flowers were edible with therapeutic potential. In addition, E. ulmoides serve as the best source of lignans, phenylpropanoids, iridoids, flavonoids, amino acids, and trace elements. Furthermore, the bark has been utilized for various biomedical applications such as controlling blood pressure, lowering fat, and promoting antiosteroporosis and hypoglycemic activity7. Hence, undoubtedly, it has been proven that E. ulmoides bark extract has a long history in traditional Chinese medicine. Earlier reports suggested that natural polymer polyisoprene is rich in the barks of Eucommia ulmoides8. Based on the information above, the present work aims to fabricate nanomaterials using Eucommia ulmoides bark extracts. The combination of zinc with bark extract is an attractive choice for preparing nanomaterials. Overall, the ultimate aim of the present investigation was to fabricate a novel hybrid nanomaterial for wound healing applications.

<table><tbody><tr><td>12 well plate</td><td>NEST</td><td>703011</td><td>Used for cell culture and assay</td></tr><tr><td>Centrifuge</td><td>SCILOGEX</td><td>SC1406</td><td>Used to separate the nanoparticles from the colloidal mixture</td></tr><tr><td>Centrifuge Tube &amp;ndash; 15 mL</td><td>BIOFIL</td><td>CFT-312150</td><td>To centrifuge the synthesized solution&amp;nbsp;</td></tr><tr><td>Centrifuge tube &amp;ndash; 5 mL</td><td>Biosharp&amp;nbsp;</td><td>BS-50-CM-S</td><td>To store the nanoparticles</td></tr><tr><td>CO&lt;sub&gt;2&lt;/sub&gt; incubator</td><td>Thermo scientific</td><td>3010</td><td>To culture the HUVEC cells</td></tr><tr><td>Denoised water</td><td>Millipore</td><td>Not applicable&amp;nbsp;</td><td>For preparation of the extract&amp;nbsp;</td></tr><tr><td>DMEM medium</td><td>Cytiva</td><td>SH30243.01</td><td>Used for cell culture work</td></tr><tr><td>Funnel</td><td>Thermo scientific</td><td>42600060</td><td>To hold the filter paper during the filtration</td></tr><tr><td>Glass beakers</td><td>Borosilicate&amp;nbsp;</td><td>1102-50</td><td>Used to prepare the aqueous extract</td></tr><tr><td>Hot air oven</td><td>Genetimes</td><td>Not Applicable&amp;nbsp;</td><td>Used to dry the nanoparticles and collect in the powder form</td></tr><tr><td>Magnetic stirrer</td><td>KYLIN-BELL</td><td>GL-5250-A</td><td>Used for nanoparticles synthesis</td></tr><tr><td>Microscope</td><td>Nikon Eclipse</td><td>Ts2</td><td>Used to take microphotographs&amp;nbsp;</td></tr><tr><td>Petri dish</td><td>NEST</td><td>753001</td><td>Used to collect the nanoparticles&amp;nbsp;</td></tr><tr><td>Pipette 1 mL</td><td>Lab Science&amp;nbsp;</td><td>YEA17AD0055580</td><td>To take/add the specific volume of solution/extract</td></tr><tr><td>Pipette tips 1 mL</td><td>SAINING&amp;nbsp;</td><td>3014200-T</td><td>To take/add the specific volume of solution/extract</td></tr><tr><td>PTFE Magnetic Mixer Stir Bars</td><td>LAN RAN</td><td>Not applicable&amp;nbsp;</td><td>Used for nanomaterial synthesis process</td></tr><tr><td>Sodium hydroxide</td><td>Sigma Alrich</td><td>71690</td><td>Used to adjust pH during the synthesis</td></tr><tr><td>Stainless Scissor</td><td>Deli</td><td>6034</td><td>Used for chopping the bark materials</td></tr><tr><td>T25 tissue culture flask</td><td>NEST</td><td>707001</td><td>Used to maintain the cells&amp;nbsp;</td></tr><tr><td>Weighing Balance&amp;nbsp;</td><td>Radwag&amp;nbsp;</td><td>AS220R2</td><td>Used to weigh the chemicals&amp;nbsp;</td></tr><tr><td>Whatman filter paper No.1</td><td>Newstar</td><td>GB/T1914-2017</td><td>Used to filter the extract for synthesis</td></tr><tr><td>Zinc nitrate&amp;nbsp;</td><td>Sigma Alrich</td><td>13778-30-8</td><td>Used as precursor for the nanoparticle&amp;rsquo;s synthesis</td></tr></tbody></table>

formulation of zinc-based nanomaterials using the eucommia ulmoides bark extract and their wound healing potential

The aqueous extract from the bark of Eucommia ulmoides serves as a rich source of bioactive compounds with numerous health benefits. The protocol here aims to explore the preparation of zinc oxide (ZnO) nanoparticles using the Eucommia ulmoides bark-mediated polyisoprene-rich aqueous extract. Meanwhile, the proposed protocol is associated with the preparation of wound healing material by easing the process. In addition, the wound-healing potential of the synthesized nanoparticles (Eu-ZnO-NPs) was evaluated using a simple scratch assay on a human umbilical vein endothelial cell (HUVEC) monolayer. After 24 h of treatment with Eu-ZnO-NPs, the cell proliferation and migration of HUVEC cells were assessed. At the end of the study, cell proliferation and migration were observed in scratched monolayer treated with different concentrations of Eu-ZnO-NPs, whereas poor cell migration and proliferation rates were observed in control cells. Of the chosen concentrations, 20 &#181;g/mL Eu-ZnO nanomaterials showed better cell migration and enhanced wound healing potential.

The present investigation aims to synthesize nanoparticles using the barks of Eucommia ulmoides tree. The bark material was dried completely in a shaded environment (Figure 1). The bark materials were utilized to prepare the hot water aqueous crude extract by heating the samples at 130 &#176;C for 20 min. A slight alteration in the temperature and duration may disrupt the phytocompounds and make them unsuitable for preparing the aqueous extract. The aqueous crude bark extracts of <e...

Formulation of Zinc-Based Nanomaterials using the Eucommia ulmoides Bark Extract and their Wound Healing Potential

Here, we present a protocol to synthesize the zinc oxide (ZnO) nanoparticles using the polyisoprene-rich aqueous extract obtained from Eucommia ulmoides tree bark. The wound-healing potential exhibited by the synthesized ZnO nanoparticles on human umbilical vein endothelial cells (HUVECs) was evaluated using scratch assay, a simple, cost-effective, and efficient method.

The aqueous extract from the bark of Eucommia ulmoides serves as a rich source of bioactive compounds with numerous health benefits. The protocol here ...

formulation-zinc-based-nanomaterials-using-eucommia-ulmoides-bark

Research

JoVE Journal

Biology

272 Views.  Central South University. Ons onderzoek richt zich op het blootleggen van het wondgenezingspotentieel van bio-engineered metalen nanodeeltjes. We willen begrijpen hoe deze nanodeeltjes het genezingsproces en hun werkingsmechanisme beïnvloeden om de belangrijkste vragen over hun therapeutische toepassing bij weefselherstel te beantwoorden. We hebben met succes verschillende op nano's gebaseerde wondverbanden ontwikkeld met behulp van natuurlijke polymeren, waarmee we hun potentieel vo...

Video: Formulering van nanomaterialen op basis van zink met behulp van het Eucommia ulmoides-schorsextract  en hun wondgenezingspotentieel

Rapid Genotyping of Mouse Tissue Using Sigma's Extract-N-Amp Tissue PCR Kit

Genomic detection of DNA via PCR amplification and detection on an electrophoretic gel is a standard way that the genotype of a tissue sample is determined. Conventional preparation of tissues for PCR-ready DNA often take several hours to days, depending on the tissue sample. The genotype of the sample may thus be delayed for several days, which is not an option for many different types of experiments. Here we demonstrate the complete genotyping of a mouse tail sample, including tissue digestion and PCR readout, in one and a half hours using Sigma's SYBR Green Extract-N-Amp Tissue PCR Kit. First, we demonstrate the fifteen-minute extraction of DNA from the tissue sample. Then, we demonstrate the real time read-out of the PCR amplification of the sample, which allows for the identification of a positive sample as it is being amplified. Together, the rapid extraction and real-time readout allow for a prompt identification of genotype of a variety different types of tissues through the reliable method of PCR.

The complete genotyping of a mouse tail sample, including tissue digestion and PCR readout, is done in one and a half hours using Sigma's SYBR Green Extract-N-Amp Tissue PCR kit.

Genomic detection of DNA via PCR amplification and detection on an electrophoretic gel is a standard way that the genotype of a tissue sample is ...

Dispersion of Nanomaterials in Aqueous Media: Towards Protocol Optimization

The sonication process is commonly used for de-agglomerating and dispersing nanomaterials in aqueous based media, necessary to improve homogeneity and stability of the suspension. In this study, a systematic step-wise approach is carried out to identify optimal sonication conditions in order to achieve a stable dispersion. This approach has been adopted and shown to be suitable for several nanomaterials (cerium oxide, zinc oxide, and carbon nanotubes) dispersed in deionized (DI) water. However, with any change in either the nanomaterial type or dispersing medium, there needs to be optimization of the basic protocol by adjusting various factors such as sonication time, power, and sonicator type as well as temperature rise during the process. The approach records the dispersion process in detail. This is necessary to identify the time points as well as other above-mentioned conditions during the sonication process in which there may be undesirable changes, such as damage to the particle surface thus affecting surface properties. Our goal is to offer a harmonized approach that can control the quality of the final, produced dispersion. Such a guideline is instrumental in ensuring dispersion quality repeatability in the nanoscience community, particularly in the field of nanotoxicology.

Here, we present a step-wise protocol for the dispersion of nanomaterials in aqueous media with real-time characterization to identify the optimal sonication conditions, intensity, and duration for improved stability and uniformity of nanoparticle dispersions without impacting the sample integrity.

The sonication process is commonly used for de-agglomerating and dispersing nanomaterials in aqueous based media, necessary to improve homogeneity and ...

Environment

Preparation of Expanded Chitin Foams and their Use in the Removal of Aqueous Copper

Chitin is an underexploited, naturally abundant, mechanically robust, and chemically resistant biopolymer. These qualities are desirable in an adsorbent, but chitin lacks the necessary specific surface area, and its modification involves specialized techniques and equipment. Herein is described a novel chemical procedure for expanding chitin flakes, derived from shrimp shell waste, into foams with higher surface area. The process relies on the evolution of H2 gas from the reaction of water with NaH trapped in a chitin gel. The preparation method requires no specialized equipment. Powder X-ray diffraction and N2-physisorption indicate that the crystallite size decreases from 6.6 nm to 4.4 nm and the specific surface area increases from 12.6 &#177; 2.1 m2/g to 73.9 &#177; 0.2 m2/g. However, infrared spectroscopy and thermogravimetric analysis indicate that the process does not change the chemical identity of the chitin. The specific Cu adsorption capacity of the expanded chitin increases in proportion to specific surface area from 13.8 &#177; 2.9 mg/g to 73.1 &#177; 2.0 mg/g. However, the Cu adsorption capacity as a surface density remains relatively constant at an average of 10.1 &#177; 0.8 atom/nm2, which again suggests no change in the chemical identity of the chitin. This method offers the means to transform chitin into a higher surface area material without sacrificing its desirable properties. Although the chitin foam is described here as an adsorbent, it can be envisioned as a catalyst support, thermal insulator, and structural material.

This study describes a method to expand chitin into a foam by chemical techniques that require no specialized equipment.

Chitin is an underexploited, naturally abundant, mechanically robust, and chemically resistant biopolymer. These qualities are desirable in an adsorbent, ...

Chemistry

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation

Stable integration of cloned gene products into the Xenopus genome is necessary to control the time and place of expression, to express genes at later stages of embryonic development, and to define how enhancers and promoters regulate gene expression within the embryo. The protocol demonstrated here can be used to efficiently produce transgenic Xenopus laevis embryos. This transgenesis approach involves three parts: 1. Sperm nuclei are isolated from adult X. laevis testis by treatment with lysolecithin, which permeabilizes the sperm plasma membrane. 2. Egg extract is prepared by low speed centrifugation, addition of calcium to cause the extract to progress to interphase of the cell cycle, and a high-speed centrifugation to isolate interphase cytosol. 3. Nuclear transplantation: the nuclei and extract are combined with the linearized plasmid DNA to be introduced as the transgene and a small amount of restriction enzyme. During a short reaction, egg extract partially decondenses the sperm chromatin and the restriction enzyme generates chromosomal breaks that promote recombination of the transgene into the genome. The treated sperm nuclei are then transplanted into unfertilized eggs. Integration of the transgene usually occurs prior to the first embryonic cleavage such that the resulting embryos are not chimeric. These embryos can be analyzed without any need to breed to the next generation, allowing for efficient and rapid generation of transgenic embryos for analyses of promoter and gene function. Adult X. laevis resulting from this procedure also propagate the transgene through the germline and can be used to generate lines of transgenic animals for multiple purposes.

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation

This video protocol demonstrates a method for generating transgenic Xenopus laevis by introduction of transgenes into sperm nuclei followed by nuclear transplantation into unfertilized eggs.

Stable integration of cloned gene products into the Xenopus genome is necessary to control the time and place of expression, to express genes at later ...

Formulation and Characterization of Bioactive Agent Containing Nanodisks

The term nanodisk refers to a discrete type of nanoparticle comprised of a bilayer forming lipid, a scaffold protein, and an integrated bioactive agent. Nanodisks are organized as a disk-shaped lipid bilayer whose perimeter is circumscribed by the scaffold protein, usually a member of the exchangeable apolipoprotein family. Numerous hydrophobic bioactive agents have been efficiently solubilized in nanodisks by their integration into the hydrophobic milieu of the particle&#39;s lipid bilayer, yielding a largely homogenous population of particles in the range of 10-20 nm in diameter. The formulation of nanodisks requires a precise ratio of individual components, an appropriate sequential addition of each component, followed by bath sonication of the formulation mixture. The amphipathic scaffold protein spontaneously contacts and reorganizes the dispersed bilayer forming lipid/bioactive agent mixture to form a discrete, homogeneous population of nanodisk&#160;particles. During this process, the reaction mixture transitions from an opaque, turbid appearance to a clarified sample that, when fully optimized, yields no precipitate upon centrifugation. Characterization studies involve the determination of bioactive agent solubilization efficiency, electron microscopy, gel filtration chromatography, ultraviolet visible (UV/Vis) absorbance spectroscopy, and/or fluorescence spectroscopy. This is normally followed by an investigation of biological activity using cultured cells or mice. In the case of nanodisks harboring an antibiotic (i.e., the macrolide polyene antibiotic amphotericin B), their ability to inhibit the growth of yeast or fungi as a function of concentration or time can be measured. The relative ease of formulation, versatility with respect to component parts, nanoscale particle size, inherent stability, and aqueous solubility permits myriad in vitro and in vivo applications of nanodisk technology. In the present article, we describe a general methodology to formulate and characterize nanodisks containing amphotericin&#160;B as the hydrophobic bioactive agent.

Here, we describe the production and characterization of bioactive agents containing nanodisks. Amphotericin B nanodisks are taken as an example to describe the protocol in a stepwise manner.

The term nanodisk refers to a discrete type of nanoparticle comprised of a bilayer forming lipid, a scaffold protein, and an integrated bioactive agent. ...

Biochemistry

Ultrathin Porated Elastic Hydrogels As a Biomimetic Basement Membrane for Dual Cell Culture

The basement membrane is a critical component of cellular bilayers that can vary in stiffness, composition, architecture, and porosity. In vitro studies of endothelial-epithelial bilayers have traditionally relied on permeable support models that enable bilayer culture, but permeable supports are limited in their ability to replicate the diversity of human basement membranes. In contrast, hydrogel models that require chemical synthesis are highly tunable and allow for modifications of both the material stiffness and the biochemical composition via incorporation of biomimetic peptides or proteins. However, traditional hydrogel models are limited in functionality because they lack pores for cell-cell contacts and functional in vitro migration studies. Additionally, due to the thickness of traditional hydrogels, incorporation of pores that span the entire thickness of hydrogels has been challenging. In the present study, we use poly-(ethylene-glycol) (PEG) hydrogels and a novel zinc oxide templating method to address the previous shortcomings of biomimetic hydrogels. As a result, we present an ultrathin, basement membrane-like hydrogel that permits the culture of confluent cellular bilayers on a customizable scaffold with variable pore architectures, mechanical properties, and biochemical composition.

Current bilayer culture models do not allow for functional in vitro studies that mimic in vivo microenvironments. Using polyethylene glycol and a zinc oxide templating method, this protocol describes the development of an ultrathin biomimetic basement membrane with tunable stiffness, porosity, and biochemical composition that closely mimics in vivo extracellular matrices.

The basement membrane is a critical component of cellular bilayers that can vary in stiffness, composition, architecture, and porosity. In vitro studies ...

Bioengineering

Lignin Micro-/Submicron Particle Synthesis for Bioflavonoid Release

Green Synthesis, Characterization, Encapsulation, and Measurement of the Release Potential of Novel Alkali Lignin Micro-/Submicron Particles

The applicability of biopolymer micro-/nano- technology in human, veterinary medicine, pharmaceutical, and food technology is rapidly growing due to the great potential of biopolymer-based particles as effective carrier systems. The use of lignin as a basic heteropolymer biomatrix for the design of innovative micro-/submicron formulations allows the achievement of increased biocompatibility and offers various active functional groups presenting opportunities for customization of the physicochemical properties and bioactivities of the formulations for diverse applications. The aim of the present study was to develop a simple and ecofriendly methodology for the synthesis of lignin particles with micro- and submicron size; to evaluate their physicochemical, spectral, and structural characteristics; and to examine their capacity for encapsulation of biologically active molecules and potential for in vitro release of bioflavonoids in simulated gastrointestinal media. The presented methodologies apply cheap and green solvents; easy, straightforward, quick, and sensitive processes requiring little equipment, non-toxic substances, and simple methods for their characterization, the determination of encapsulation capacity towards the poorly water-soluble bioactive compounds morin and quercetin, and the in vitro release potential of the lignin matrices.

Author Spotlight: Development and Characterization of Eco-Friendly Lignin-Based Microparticles for Enhanced Delivery of Bioflavonoids

We describe novel, simple methodologies of synthesis and characterization of biocompatible lignin micro- and submicron particles. These formulations provide a facile approach for the utilization of the heteropolymer, as well as an alternative for the rational design of multifunctional carrier matrices with potential applicability in biomedicine, pharmaceutical technology, and the food industry.

The applicability of biopolymer micro-/nano- technology in human, veterinary medicine, pharmaceutical, and food technology is rapidly growing due to the ...

Long-term Culture of Human Breast Cancer Specimens and Their Analysis Using Optical Projection Tomography

Breast cancer is a leading cause of mortality in the Western world. It is well established that the spread of breast cancer, first locally and later distally, is a major factor in patient prognosis. Experimental systems of breast cancer rely on cell lines usually derived from primary tumours or pleural effusions. Two major obstacles hinder this research: (i) some known sub-types of breast cancers (notably poor prognosis luminal B tumours) are not represented within current line collections; (ii) the influence of the tumour microenvironment is not usually taken into account.
We demonstrate a technique to culture primary breast cancer specimens of all sub-types. This is achieved by using three-dimensional (3D) culture system in which small pieces of tumour are embedded in soft rat collagen I cushions. Within 2-3 weeks, the tumour cells spread into the collagen and form various structures similar to those observed in human tumours1. Viable adipocytes, epithelial cells and fibroblasts within the original core were evident on histology. Malignant epithelial cells with squamoid morphology were demonstrated invading into the surrounding collagen. Nuclear pleomorphism was evident within these cells, along with mitotic figures and apoptotic bodies.
We have employed Optical Projection Tomography (OPT), a 3D imaging technology, in order to quantify the extent of tumour spread in culture. We have used OPT to measure the bulk volume of the tumour culture, a parameter routinely measured during the neo-adjuvant treatment of breast cancer patients to assess response to drug therapy. 
Here, we present an opportunity to culture human breast tumours without sub-type bias and quantify the spread of those ex vivo. This method could be used in the future to quantify drug sensitivity in original tumour. This may provide a more predictive model than currently used cell lines.

We have developed a collagen-based in vitro assay which promotes proliferation and invasion from samples of all breast cancer subtypes. Optical Projection Tomography, a three dimensional microscopy technique was utilised to visualise and quantify tumour expansion. This assay may be used to quantify drug response of individual tumour samples.

Breast cancer is a leading cause of mortality in the Western world. It is well established that the spread of breast cancer, first locally and later ...

 Zinc Oxide Nanoparticles Synthesis and Analysis of its Antibacterial Effects

Preparation of Zinc Oxide Nanoparticles and the Evaluation of their Antibacterial Effects

Nosocomial bacterial infections have become increasingly challenging due to their inherent resistance to antibiotics. The emergence of multidrug-resistant bacterial strains in hospitals has been attributed to the extensive and varied use of antibiotics, further exacerbating the problem of antibiotic resistance. Metal nanomaterials have been widely studied as an alternative solution for eradicating antibiotic-resistant bacterial cells. Metallic nanoparticles attack bacterial cells through various mechanisms, such as the release of antibacterial ions, generation of reactive oxygen species, or physical disruption, against which bacteria cannot develop resistance. Among the actively researched antimicrobial metal nanoparticles, zinc oxide nanoparticles, which are FDA-approved, are known for their biocompatibility and antibacterial properties. In this study, we focused on successfully developing a precipitation method for synthesizing zinc oxide nanoparticles, analyzing the properties of these nanoparticles, and conducting antimicrobial tests. Zinc oxide nanoparticles were characterized using transmission electron microscopy (TEM), dynamic light scattering (DLS), ultraviolet/visible spectroscopy, and X-ray diffraction (XRD). Antibacterial tests were conducted using the broth microdilution test with the multidrug-resistant strains of methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. This study demonstrated the potential of zinc oxide nanoparticles in inhibiting the proliferation of antibiotic-resistant bacteria.

Author Spotlight: Exploring the Antibacterial Effects of Zinc Oxide Nanoparticles in Overcoming Antibiotic Resistance

In this study, zinc oxide nanoparticles were synthesized using a precipitation method. The antibacterial effect of the synthesized particles was tested against multidrug-resistant methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa bacterial strains.

Nosocomial bacterial infections have become increasingly challenging due to their inherent resistance to antibiotics. The emergence of multidrug-resistant ...

A Standardized Procedure of Dressing Management for Toxic Epidermal Necrolysis

Toxic epidermal necrolysis (TEN) is a severe cutaneous adverse drug reaction with high mortality rates, demanding specialized wound care to address epidermal detachment and bullae formation. This study summarizes a standardized dressing management protocol to enhance the healing process, reduce complications, and improve patient comfort during dressing changes for TEN patients. The protocol emphasizes a systematic approach to patient preparation, environmental control, and the utilization of silver-ion-based dressing materials. Specifically, it integrates gentle cleansing techniques with a warmed povidone-iodine saline solution, precise necrotic tissue removal, and silver sulfadiazine lipid hydrocolloid dressings to maintain an optimal healing environment. The effectiveness of this protocol was further validated through a retrospective analysis, which showed a significant reduction in the onset of re-epithelization, wound healing time, and hospital stays, resulting in diminished pain during dressing changes. Furthermore, this protocol also provides tailored strategies for dressing changes in sensitive areas, ensuring comprehensive care. The standardized protocol streamlines the dressing process and contributes to more efficient allocation of healthcare resources, establishing a robust foundation for TEN treatment that can be adopted in clinical practice and inform future research.

This is a standardized dressing protocol for patients with toxic epidermal necrolysis. This robust protocol utilizes silver ion dressings designed to accelerate healing and alleviate pain, minimizing hospital stays.

Toxic epidermal necrolysis (TEN) is a severe cutaneous adverse drug reaction with high mortality rates, demanding specialized wound care to address ...

Formulation of Zinc-Based Nanomaterials using the Eucommia ulmoides Bark Extract and their Wound Healing Potential

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