This work was supported by a Zhejiang Shuren University research project (2023R053 and 2023KJ237).

The blood used in the experiments was obtained from SPF-grade BALB/c female mice weighing 20-25 g and approximately 7 weeks old. The Animal Experimentation Ethics Committee of Zhejiang Shuren College approved all animal care and experimental procedures.
1. Solution preparation
<ol>
	<li>Weigh ALG and dissolve in ultrapure water by stirring in a water bath at 50 &#176;C to obtain 2% ALG solution.</li>
	<li>Separately prepare mass fraction with 5% (or molar mass concentration ...

Developing Sodium Alginate Hydrogel Microspheres for Biomedical Use

<ol>
	<li>Gong, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+recent+advances+of+cellulose-based+intelligent-responsive+hydrogels+as+vehicles+for+controllable+drug+delivery+system.">A review of recent advances of cellulose-based intelligent-responsive hydrogels as vehicles for controllable drug delivery system.</a> Int J BiolMacromol. 264 (2), 130525 (2024).</li><li>Zhao, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Injectable+microfluidic+hydrogel+microspheres+for+cell+and+drug+delivery.">Injectable microfluidic hydrogel microspheres for cell and drug delivery.</a> Adv Funct. 31 (31), 2103339 (2021).</li><li>Qiao, S., Chen, W., Zheng, X., Ma, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+pH-sensitive+alginate-based+hydrogel+by+microfluidic+technology+for+intestinal+targeting+drug+delivery.">Preparation of pH-sensitive alginate-based hydrogel by microfluidic technology for intestinal targeting drug delivery.</a> Int J Biol Macromol. 254 (2), 127649 (2024).</li><li>Lei, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+hydrogel+microspheres+for+protein+capture+and+wound+healing.">Antimicrobial hydrogel microspheres for protein capture and wound healing.</a> Mater Design. 215, 110478 (2022).</li><li>Ju, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zn2++incorporated+composite+polysaccharide+microspheres+for+sustained+growth+factor+release+and+wound+healing.">Zn2+ incorporated composite polysaccharide microspheres for sustained growth factor release and wound healing.</a> Mater Today Bio. 22, 100739 (2023).</li><li>El-Sayed, N. S., Hashem, A. H., Khattab, T. A., Kamel, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+antibacterial+hydrogels+based+on+sodium+alginate.">New antibacterial hydrogels based on sodium alginate.</a> Int. J Biol Macromol. 248, 125872 (2023).</li><li>Olukman &#350;ahin, M., &#350;anl&#305;, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+5-fluorouracil+release+properties+investigation+from+pH+sensitive+sodium+alginate+coated+and+uncoated+methyl+cellulose/chitosan+microspheres.">In vitro 5-fluorouracil release properties investigation from pH sensitive sodium alginate coated and uncoated methyl cellulose/chitosan microspheres.</a> Int J Biol Macromol. 258 (1), 128895 (2024).</li><li>Chi, H., Qiu, Y., Ye, X., Shi, J., Li, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+strategy+of+hydrogel+microsphere+and+its+application+in+skin+repair.">Preparation strategy of hydrogel microsphere and its application in skin repair.</a> Front Bioeng Biotechnol. 11, 1239183 (2023).</li><li>Yang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bio-inspired+dual-adhesive+particles+from+microfluidic+electrospray+for+bone+regeneration.">Bio-inspired dual-adhesive particles from microfluidic electrospray for bone regeneration.</a> Nano Res. 16 (4), 5292-5299 (2022).</li><li>Zheng, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Celastrol-encapsulated+microspheres+prepared+by+microfluidic+electrospray+for+alleviating+inflammatory+pain.">Celastrol-encapsulated microspheres prepared by microfluidic electrospray for alleviating inflammatory pain.</a> Biomater Adv. 149, 213398 (2023).</li><li>Yang, L., Yang, W., Xu, W., Zhao, Y., Shang, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bio-inspired+Janus+microcarriers+with+sequential+actives+release+for+bone+regeneration.">Bio-inspired Janus microcarriers with sequential actives release for bone regeneration.</a> Chem Eng J. 476, 146797 (2023).</li><li>Yang, L., Sun, L., Zhang, H., Bian, F., Zhao, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ice-inspired+lubricated+drug+delivery+particles+from+microfluidic+electrospray+for+osteoarthritis+treatment.">Ice-inspired lubricated drug delivery particles from microfluidic electrospray for osteoarthritis treatment.</a> ACS Nano. 15 (12), 20600-20606 (2021).</li><li>Li, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultra-flexible+stretchable+liquid+metal+circuits+with+antimicrobial+properties+through+selective+laser+activation+for+health+monitoring.">Ultra-flexible stretchable liquid metal circuits with antimicrobial properties through selective laser activation for health monitoring.</a> Chem Eng J. 482, 149173 (2024).</li><li>Sukhodub, L., Kumeda, M., Sukhodub, L., Bielai, V., Lyndin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metal+ions+doping+effect+on+the+physicochemical,+antimicrobial,+and+wound+healing+profiles+of+alginate-based+composite.">Metal ions doping effect on the physicochemical, antimicrobial, and wound healing profiles of alginate-based composite.</a> Carbohydr Polym. 304, 120486 (2023).</li><li>Daly, A. C., Riley, L., Segura, T., Burdick, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrogel+microparticles+for+biomedical+applications.">Hydrogel microparticles for biomedical applications.</a> Nat Rev Mater. 5 (1), 20-43 (2020).</li><li>Bertsch, P., Diba, M., Mooney, D. J., Leeuwenburgh, S. C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Self-healing+injectable+hydrogels+for+tissue+regeneration.">Self-healing injectable hydrogels for tissue regeneration.</a> Chem Rev. 123 (2), 834-873 (2023).</li><li>Yang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomass+microcapsules+with+stem+cell+encapsulation+for+bone+repair.">Biomass microcapsules with stem cell encapsulation for bone repair.</a> Nanomicro Lett. 14 (1), (2021).</li><li>Wu, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NK-Cell-Encapsulated+porous+microspheres+via+microfluidic+electrospray+for+tumor+immunotherapy.">NK-Cell-Encapsulated porous microspheres via microfluidic electrospray for tumor immunotherapy.</a> ACS Appl Mater. 11 (37), 33716-33724 (2019).</li><li>Nan, L., Zhang, H., Weitz, D. A., Shum, H. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+future+of+droplet+microfluidics.">Development and future of droplet microfluidics.</a> LOC. 24 (5), 1135-1153 (2024).</li></ol>

In this protocol, we present a method for preparing ALGMS based on microfluidic electrospray technology. The method is simple to operate and yields a large number of microspheres with uniform roundness and controllable diameter. This approach offers convenience to researchers and can promote the research and application of hydrogel microspheres. In addition, by cross-linking with different metal ions, the stability and bioactivity of the ALGMS were improved. In the antimicrobial experiments, Cu-ALGMS and Zn-ALGMS exhibit...

Drug delivery systems are a research hotspot in the field of bio-tissue engineering, aiming to improve drug delivery efficiency and efficacy and reduce adverse reactions and side effects1. Among these systems, hydrogel microspheres, characterized by good biocompatibility, tunable mechanical properties, and functional plasticity, are one of the most commonly used vehicles for drug loading and delivery2. They can be used for both slow and controlled release of drugs, provide good protective effects for drugs, avoid or minimize non-specific effects of drugs in other tissues, and target drug delivery to specific tissue structures3. Therefore, hydrogel microspheres have become a new and efficient drug delivery system, with research in this field gradually emerging4.
Hydrogel microspheres are typically synthesized from biodegradable materials, including polysaccharides, proteins, and natural polymers5. Among them, ALG is a biocompatible, biodegradable polysaccharide extracted from marine brown algae6. Its molecular chain contains free hydroxyl and carboxyl groups that can crosslink with most divalent or multivalent cations to form a water-insoluble hydrogel structure with a three-dimensional network5. The hydrogel microspheres formed by ALG can be converted into negatively charged polyelectrolytes in neutral and alkaline solutions. This repulsion between negative charges causes the microspheres to swell, allowing the release of the encapsulated active ingredient or drug. These properties have led to the consideration of ALG microspheres as promising drug carriers widely used for drug loading and controlled release7.
Various methods exist for the preparation of hydrogel microspheres. Traditional ALGMS preparation methods usually include the sol-gel method or the emulsion-sol method. These methods involve steps such as precipitation, co-precipitation, and gelation reactions to obtain the target microspheres8. In recent years, with the continuous development of microfluidic technology, the microfluidic electrospray method has gradually become an efficient and precise microsphere preparation method9. This method utilizes microfluidic technology to electrospray a polymer solution through a microfine nozzle to form micrometer-sized droplets and microspheres during the subsequent curing or cross-linking process10. Compared to the traditional method, microfluidic electrospray offers precise control of microsphere particle size and morphology by adjusting parameters such as solution flow rate, voltage, and fine nozzle size11. It also enables high-speed continuous preparation of microspheres, improving preparation efficiency and maintaining mild reaction conditions. In addition, ALGMS can be prepared to possess various functions, such as controlled-release drugs and loaded catalysts, enabling their easy application in various fields.
Here, we present a protocol for the preparation of ALG microspheres using the microfluidic electrospray method. The process involves passing an ALG solution through a microfluidic device and subjecting it to electrospray. The resulting droplets were collected in the solution containing different metal ions (Ca2+, Cu2+, Zn2+, and Fe3+) to initiate the cross-linking reaction. This reaction improves the stability and adhesion of the microspheres and endows them with different functionalities. This method is easy to perform, and the synthesized microspheres exhibit good size uniformity in their morphology. In addition, we investigated their antimicrobial properties, slow drug-release ability, and biocompatibility. This protocol will be useful for further drug development and production.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>120 mesh screen</td>
			<td>Solarbio,China</td>
			<td>YA0946</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol burner</td>
			<td>Solarbio,China</td>
			<td>YA2320</td>
			<td></td>
		</tr>
		<tr>
			<td>BALB/c mice</td>
			<td>Wukong Biotechnology,China</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Bicinchoninic Acid Assay reagent</td>
			<td>Meilunbio,China</td>
			<td>MA0082</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Lablead,China</td>
			<td>9048-46-8</td>
			<td></td>
		</tr>
		<tr>
			<td>CaCl2&#160; powder</td>
			<td>Aladdin,China</td>
			<td>10043-52-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcein-AM/PI</td>
			<td>Biosharp,China</td>
			<td>BL130A</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge tubes</td>
			<td>Corning,America</td>
			<td>430290</td>
			<td></td>
		</tr>
		<tr>
			<td>CuSO4&#160; powder</td>
			<td>Jnxinyuehuagong,China</td>
			<td>7758-99-8</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM</td>
			<td>Gibicol,China</td>
			<td>C11995500BT</td>
			<td></td>
		</tr>
		<tr>
			<td>FeCl3&#160; powder</td>
			<td>Aladdin,China</td>
			<td>7705-08-0</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>HAKATA,China</td>
			<td>HN-FBS</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass tubes</td>
			<td>Sartorius,Germany</td>
			<td>CC0028</td>
			<td></td>
		</tr>
		<tr>
			<td>Light microscopy</td>
			<td>Evidentscientific,Japan</td>
			<td>BX53(LED)</td>
			<td></td>
		</tr>
		<tr>
			<td>Microfluidic syringe pump</td>
			<td>Longerpump,England</td>
			<td>LSP01-3A</td>
			<td></td>
		</tr>
		<tr>
			<td>NIH3T3</td>
			<td>HyGyte,China</td>
			<td>TCM-C752</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dish</td>
			<td>Thermofisher,America</td>
			<td>150464</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate buffer saline</td>
			<td>Thermofisher,America</td>
			<td>3002</td>
			<td></td>
		</tr>
		<tr>
			<td>Scanning electron microscope</td>
			<td>Thermofisher,America</td>
			<td>Axia ChemiSEM</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium alginate powder&#160;</td>
			<td>Bjbalb,China</td>
			<td>Y13095</td>
			<td></td>
		</tr>
		<tr>
			<td>ZnSO4 powder</td>
			<td>Jnxinyuehuagong,China</td>
			<td>7733-02-0</td>
			<td></td>
		</tr>
	</tbody>
</table>

preparation of cross-linked sodium alginate microspheres with different metal ions using the microfluidic electrospray technology

Microspheres are micrometer-sized particles that can load and gradually release drugs via physical encapsulation or adsorption onto the surface and within polymers. In the field of biomedicine, hydrogel microspheres have been extensively studied for their application as drug carriers owing to their ability to reduce the frequency of drug administration, minimize side effects, and improve patient compliance. Sodium alginate (ALG) is a naturally occurring linear polysaccharide with three backbone glycosidic linkages. There are two auxiliary hydroxyl groups present in each of the moieties of the polymer, which have the characteristics of an alcohol hydroxyl moiety. The synthetic ALG units can undergo chemical cross-linking reactions with metal ions, forming a cross-linked network structure of polymer stacks, ultimately forming a hydrogel. Hydrogel microspheres can be prepared using a simple process involving the ionic cross-linking properties of ALG. In this study, we prepared ALG-based hydrogel microspheres (ALGMS) using a microfluidic electrodeposition strategy. The prepared hydrogel microspheres were uniformly sized and well-dispersed, owing to accurate control of the microfluidic electrospray flow. ALGMS cross-linked with different metal ions were prepared using a microfluidic electrospray technique combining microfluidic and high electric field, and its antimicrobial properties, slow drug release ability, and biocompatibility were investigated. This technology holds promise for application in advanced drug development and production.

Author Spotlight: Advancing Therapeutics with Biocompatible Sodium Alginate Hydrogel Microspheres

Preparation of Cross-Linked Sodium Alginate Microspheres with Different Metal Ions Using the Microfluidic Electrospray Technology

Hydrogen microspheres are characterized by good compatibility, design flexibility, and high degree of customization. They have a broad application protector in biomedical fields such as therapeutic drug delivery and the construction of TC repair discoveries. In this study, we prepared hydrogen microspheres using a simple microfluidic device.Recently we have been trying to use hydrogen microspheres loaded with some small molecules from traditional Chinese medicine to promote all the healing. In this study, we found that zinc and copper ion cross-link the sodium alginate microspheres have good anti-microbial properties and thus copper ARGMS, and zinc ARGMS are suitable for the treatment of bacterial infectious diseases.

Characterization of ALGMS cross-linked with different metal ions 
The optical morphology of Ca-ALGMS, Cu-ALGMS, Zn-ALGMS, and Fe-ALGMS is shown in Figure 2, exhibiting good sphericity, smooth surface, uniform particle size distribution (Supplementary Figure 2) and excellent monodispersity. We further performed microscopic characterization using scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) analysis. As shown in <strong clas...

The protocol describes the preparation of sodium alginate microspheres cross-linked with different metal ions using a microfluidic device for drug carrier design. The antimicrobial properties and slow drug release of these microspheres were also investigated.

Microspheres are micrometer-sized particles that can load and gradually release drugs via physical encapsulation or adsorption onto the surface and within ...

preparation-cross-linked-sodium-alginate-microspheres-with-different

Research

JoVE Journal

Bioengineering

690 Views.  Zhejiang Shuren University. Hydrogen microspheres are characterized by good compatibility, design flexibility, and high degree of customization. They have a broad application protector in biomedical fields such as therapeutic drug delivery and the construction of TC repair discoveries. In this study, we prepared hydrogen microspheres using a simple microfluidic device.Recently we have been trying to use hydrogen microspheres loaded with some small molecules from traditional Chinese medicine to promote all the hea...

Video: Author Spotlight: Advancing Therapeutics with Biocompatible Sodium Alginate Hydrogel Microspheres

The protocol describes the preparation of sodium alginate microspheres cross-linked with different metal ions using a microfluidic device for drug carrier design. The antimicrobial properties and slow drug release of these microspheres were also...

https://cloudfront.jove.com/CDNSource/teasers/66871_b.jpg

biocompatibility tests to assess the use of sodium alginate hydrogel microspheres for drug delivery

Biocompatibility Tests to Assess the Use of Sodium Alginate Hydrogel Microspheres for Drug Delivery

creating alginate hydrogel microspheres using microfluidic electrospray for controlled drug release

Creating Alginate Hydrogel Microspheres Using Microfluidic Electrospray for Controlled Drug Release

generation of alginate microspheres for biomedical applications

Generation of Alginate Microspheres for Biomedical Applications

In the following sections, we outline procedures for the preparation of alginate microspheres for use in biomedical applications. We specifically illustrate a technique for creating multilayered alginate microspheres for the dual purpose of cell and protein encapsulation as a potential treatment for type 1...

development and characterization of fusidic acid-loaded alginate-aloe vera based hydrogel film

Author Spotlight: Optimizing &lt;i&gt;Aloe vera&lt;/i&gt;-Alginate Hydrogel Films for Enhanced Fusidic Acid Delivery and Wound Healing

A formulation of fusidic acid-loaded hydrogel film was developed with different Aloe vera ratios and characterized in this...

Fusidic Acid-Loaded Hydrogel Films with &lt;i&gt;Aloe vera&lt;/i&gt; for Enhanced Drug Delivery

https://cloudfront.jove.com/CDNSource/teasers/67068_b.jpg

constructing a collagen hydrogel for the delivery of stem cell-loaded chitosan microspheres

Constructing a Collagen Hydrogel for the Delivery of Stem Cell-loaded Chitosan Microspheres

A major hurdle in current stem cell therapies is determining the most effective method to deliver these cells to host tissues. Here, we describe a chitosan-based delivery method that is efficient and simple in approach, while allowing adipose-derived stem cells to maintain their...

synthesis and assay of vibrio quorum sensing inhibitors

Author Spotlight: Advancing Therapeutics to Treat Vibriosis in Humans and Aquatic Organisms

Thiophenesulfonamide compounds are potent and specific inhibitors of Vibrio quorum sensing regulators LuxR/HapR that block their activity in vivo, thus preventing transcription of genes for virulence, motility, and biofilms. This protocol details how these compounds are synthesized, modeled in silico, and assayed in vivo for activity against...

Biological Assessment of Thiophenesulfonamides in Quorum Sensing Inhibition

https://cloudfront.jove.com/CDNSource/teasers/66582_b.jpg

bioprintable alginate/gelatin hydrogel 3d in vitro model systems induce cell spheroid formation

Bioprintable Alginate/Gelatin Hydrogel 3D In Vitro Model Systems Induce Cell Spheroid Formation

We developed a heterogeneous breast cancer model consisting of immortalized tumor and fibroblast cells embedded in a bioprintable alginate/gelatin bioink. The model recapitulates the in vivo tumor microenvironment and facilitates the formation of multicellular tumor spheroids, yielding insight into the mechanisms driving...

Bioprintable Alginate/Gelatin Hydrogel 3D In Vitro Model Systems Induce Cell Spheroid Formation

Development and Characterization of Fusidic Acid-Loaded Alginate-Aloe vera Based Hydrogel Film

a hydrogel construct and fibrin-based glue approach to deliver therapeutics in a murine myocardial infarction model.

A Hydrogel Construct and Fibrin-based Glue Approach to Deliver Therapeutics in a Murine Myocardial Infarction Model.

This protocol aims to alleviate the limitation of poor cell engraftment for stem cell treatment of myocardial infarctions through the use of a hydrogel system and a fibrin-based glue. With this approach, cell-to-tissue contact post-infarction can be maintained, increasing the therapeutic potential of beneficial agents at the site of...

an open-source framework for mass calculation of antibody-based therapeutic molecules

Author Spotlight: Advancing Biotherapeutic Mass Calculation by Introducing mAbScale, a Python-Based Desktop Application

This article describes the use of a software application, mAbScale, for the calculation of masses for monoclonal antibody-based protein...

A Python-Based Tool for Accurate Mass Calculation of Biotherapeutic Entities

3d culture of porcine cocs: a procedure to encapsulate cumulus oocyte complexes in fibrin-alginate polymer hydrogel beads

This video describes the protocols for encapsulating porcine cumulus-oocyte complexes (COCs) in fibrin-alginate beads to establish a 3D culture.&#160; This in-vitro maturation system maintains normal cell-to-cell communication, ensuring oocyte maturation and retaining...

3D Culture of Porcine COCs: A Procedure to Encapsulate Cumulus Oocyte Complexes in Fibrin-alginate Polymer Hydrogel Beads

3d printing bacteria to study motility and growth in complex 3d porous media

Author Spotlight: Studying Bacterial Growth in 3D Hydrogel Matrices

This protocol describes a procedure for three-dimensional (3D) printing of bacterial colonies to study their motility and growth in complex 3D porous hydrogel matrices that are more akin to their natural habitats than conventional liquid cultures or Petri dishes.

Bioprinting Bacteria in Hydrogel Matrices for the Study of Motility and Growth

https://cloudfront.jove.com/CDNSource/teasers/66166_b.jpg

3d bioprinting phototunable hydrogels to study fibroblast activation

Author Spotlight: Understanding Chronic Lung Diseases Using 3D Printed Phototunable Hydrogels 

This article describes how to 3D bioprint phototunable hydrogels to study extracellular matrix stiffening and fibroblast activation.

Phototunable Bioprinted Hydrogels to Probe Cellular Responses to ECM Stiffening

https://cloudfront.jove.com/CDNSource/teasers/65639_b.jpg

hydrogel arrays enable increased throughput for screening effects of matrix components and therapeutics in 3d tumor models

Hydrogel Arrays Enable Increased Throughput for Screening Effects of Matrix Components and Therapeutics in 3D Tumor Models

The present protocol describes an experimental platform to assess the effects of mechanical and biochemical cues on chemotherapeutic responses of patient-derived glioblastoma cells in 3D matrix-mimetic cultures using a custom-made UV illumination device facilitating high-throughput photocrosslinking of hydrogels with tunable mechanical...

in vitro culturing technique for studying cellular dynamics in zebrafish scales

Author Spotlight: Advancing Bone Cell Activity Research with Zebrafish Scales 

This protocol offers a method to study cellular dynamics using a simple in vitro culture technique. It provides an opportunity for zebrafish researchers and educators to study cellular processes, such as those related to bone homeostasis and basic cell biology, by visualizing fluorescent nuclei and apoptotic cells within the...

Removing Zebrafish Scales for Culturing and Bone Biology Studies

https://cloudfront.jove.com/CDNSource/teasers/66619_b.jpg

operating and biocontainment procedures of a facility for laboratory mice with a natural microbiome: immunophenotyping procedure

Author Spotlight: Advancing Murine Research with a Facility for Handling Wildling Mice

Here, we describe the structure and operating procedures, including microbial containment measures of a facility for "Wilding mice" using blood sampling for immunophenotyping as an...

Operational Protocols for Handling Wildling Mice in Research Facilities

Prepare incubation chambers by applying thin strips of paraffin film to glass microscope slides. Pipette 7.5-microliter drops of the FA mixture onto the paraffin-film-coated glass slide with separating spacers, placing 8 to 10 drops arranged in two...

design of a biocompatible drug-eluting tracheal stent in mice with laryngotracheal stenosis

Design of a Biocompatible Drug-Eluting Tracheal Stent in Mice with Laryngotracheal Stenosis

Laryngotracheal stenosis results from pathologic scar deposition that critically narrows the tracheal airway and lacks effective medical therapies. Using a PLLA-PCL (70% poly-L-lactide and 30% polycaprolactone) stent as a local drug delivery system, potential therapies aimed at decreasing scar proliferation in the trachea can be...

To begin, mix ALG with ultrapure water in a water bath, tempered at 50 degrees Celsius. Separately, prepare mass fraction with 5%solutions of calcium chloride, ferric chloride, zinc sulfate, and copper sulfate in ultrapure water. Pour each solution separately into a collecting dish.Place a glass capillary tube over a blowtorch flame to soften it and stretch it to various diameters. With a glass cutter, cut off the fine parts and gently polish the tip port with silicon carbide sandpaper. Connect the thick end of the capillary tube to a long tubing and the other end to a dispensing needle in a five-milliliter syringe.Attach the syringe to the microfluidic syringe pump and the capillary tubing at one end to the holder. Connect the red high-voltage end clip of the high-voltage power supply to the dispensing needle of the syringe and place the silver clip in the collection fluid. This configuration creates a simple microfluidic electrospray device.Place the collection dishes containing different metal ions directly under the capillary tube. Turn on the microfluidic syringe pump. Select fast forward to prime the tube with the ALG and set the flow rate to two milliliters per hour.Switch on the high-voltage power switch and turn the knob to set the voltage to five to seven kilovolts. Collect ALG microspheres cross-linked with different metal ions. After the formation of ALG hydrogel microspheres, transfer them to individual labeled 1.5-milliliter centrifuge tubes.The micrographs of ALG microspheres showed good sphericity, smooth surface, and uniform particle size distribution. Scanning electron microscopy showed that the microspheres were generally spherical with well-defined roundness.

To begin, prepare zinc, calcium, copper, and ferric alginate hydrogel micro-spheres. For the antimicrobial performance test, add 10 milliliters of each of the two bacterial solutions into a 15 milliliter sterile centrifuge tube. Then incubate the cultures with 0.5 milliliters of hydrogel microspheres in a constant temperature shaker.Dilute each bacterial solution, 10 to the power of five times, with sterile water. Using sterile glass beads, evenly spread 200 microliters of bacterial solution on an LB solid medium. Place the plates in an incubator at 37 degrees Celsius for 12 hours to assess colony formation.Add one milligram per milliliter of bovine serum albumin or BSA suspension, to 500 microliters of each microsphere and incubate for 24 hours. Add each saturated sample to one milliliter PBS, and agitate for 15 minutes at 37 degrees Celsius with continuous shaking at 80 revolutions per minute. After aliquoting, replace the medium with a fresh one at one, three, six, 12, 24, 36, and 48 hours.Quantify the supernatant protein concentrations at specific times. Add the prepared ALG microspheres to PBS, and incubate them at 37 degrees Celsius for 24 hours. Obtain whole blood from healthy mice in anticoagulation tubes containing sodium citrate.After vortexing centrifuge the tube for 15 minutes at 1, 500 revolutions per minute to obtain red blood cells. After flushing the cells, prepare a 10%red blood cell solution with PBS, and add 20 microliters of blood cell solution to the microsphere experimental and control groups. Incubate the samples at 37 degrees Celsius for four hours, and centrifuge them at 1, 500 RPM for 15 minutes.After keeping the supernatant aside, photograph the red blood cells in each group following hemolysis. Finally, measure the absorbance values of the supernatants in each group and compute the hemolysis rate using the formula. Different microspheres exhibited antibacterial activity against staphylococcus aureus and escherichia coli with copper and zinc, ALG microspheres, showing the strongest antibacterial properties.The drug release assay revealed that the release rate of the iron microsphere was relatively faster than that of the other three ions. Biocompatibility assessment showed that the red blood cells in the suspension remained intact upon contact with different microspheres, indicating minimal hemolysis by the microspheres.