Hyaluronidase by ANT BIO PTE. LTD.: A "Molecular Scissors" in Biomedical Research

In the fields of biomedicine and cell research, effectively processing the extracellular matrix is crucial for the success of many experiments. As an efficient and specific biological tool, hyaluronidase plays an indispensable role. This article systematically introduces the definition, mechanism of action, multi-field applications of hyaluronidase, and elaborates on its application schemes in specific experiments.

1. Definition and Mechanism of Action: The Key to Precise Cleavage

Hyaluronidase is a general term for a family of enzymes capable of degrading hyaluronic acid. Hyaluronic acid is a linear macromolecular polysaccharide widely present in the extracellular matrix of animal tissues, connective tissues, and the capsules of certain bacteria. As one of the main components of the extracellular matrix, it has high hydrophilicity and viscoelasticity, which can form a hydrated gel to support, retain water and protect cells. Meanwhile, it also serves as a physical barrier for cell migration, proliferation, and signal transduction.

The mechanism of action of hyaluronidase lies in its ability to specifically hydrolyze the β-1,4-glycosidic bonds linking N-acetyl-D-glucosamine and D-glucuronic acid in hyaluronic acid molecules. Through this cleavage, hyaluronidase can decompose the huge hyaluronic acid long chains into oligosaccharide fragments, thereby achieving the following effects:

•         Reducing Tissue Viscosity: Destroying the gel-like barrier formed by hyaluronic acid.

•         Increasing Tissue Permeability: Creating channels for the diffusion and migration of other molecules, drugs, or cells.

•         Dissociating Cells from the Matrix: Helping to release cells embedded in the matrix.

It is based on this core mechanism that hyaluronidase excels in multiple research and application fields.

2. Overview of Main Applications

The applications of hyaluronidase mainly revolve around its ability to "deconstruct" hyaluronic acid, which can be divided into the following major directions:

•         Tissue Dissociation: Used to digest tissues rich in hyaluronic acid when preparing primary cells, softening them and releasing cells.

•         Assisted Drug Delivery: As a diffusing agent, it promotes the absorption and distribution of subcutaneously or intramuscularly injected drugs and reduces local swelling.

•         Cell Culture and Analysis: Used to treat certain cells that secrete a large amount of extracellular matrix during cell passage or harvesting.

•         Reproductive Biology Research: Used to treat the cell layer around oocytes in assisted reproductive technology.

•         Basic Research: Used to study the specific functions of hyaluronic acid in cell behavior, development, and diseases (such as cancer and inflammation).

3. Experimental Application Scenarios and Operational Considerations

The following elaborates on the application scenarios, key points of scheme design, and precautions of hyaluronidase in specific experiments.

3.1 Primary Cell Isolation

Application Scenario: Isolating viable and functional primary cells from animal or human tissues (such as skin, umbilical cord, cartilage, tumor tissue, etc.).

Experimental Protocol:

•         Tissue Pretreatment: Cut the tissue into small pieces (1-3 mm³).

•         Enzymatic Digestion: Soak the tissue pieces in a digestion solution containing hyaluronidase (usually with a working concentration of 0.5 - 2 mg/mL). To improve digestion efficiency, hyaluronidase is often used in combination with other hydrolases such as collagenase and trypsin to form a composite enzyme digestion solution, which synergistically decomposes different components in the extracellular matrix.

•         Incubation and Termination: Incubate with continuous shaking at 37°C for 30 minutes to several hours, depending on the type and thickness of the tissue. Regularly sample and observe the cell dissociation under a microscope. Once fully dissociated, immediately add medium containing serum (serum contains enzyme inhibitors) to terminate the reaction.

•         Cell Collection: Filter through a cell sieve to remove undigested tissue blocks, centrifuge to collect cells, resuspend for counting and culture.

Precautions:

•         Concentration and Time Optimization: Excessively high enzyme concentration or too long digestion time will damage the cell membrane, reducing cell viability and adhesion ability. Pre-experiments must be performed to determine the optimal conditions.

•         Aseptic Operation: The enzyme solution needs to be filtered and sterilized, and the entire operation process should be carried out in a ultra-clean workbench.

•         Activity Verification: The activity of enzymes from different batches or sources may vary, so attention should be paid to their specific activity units.

3.2 Cell Culture and Passage

Application Scenario: Certain cell lines (such as some cancer cells or mesenchymal stem cells) secrete abundant extracellular matrix during culture, forming a dense cell layer. When using conventional trypsin digestion, the cells fall off in sheets and are not completely dissociated.

Experimental Protocol:

•         Before trypsin digestion, the cells can be pretreated with a buffer containing hyaluronidase (for example, 0.1 - 0.5 mg/mL) for 5-15 minutes.

•         Hyaluronidase first degrades hyaluronic acid in the matrix, weakening the connection between cells and the matrix, and then adding trypsin can more gently and efficiently dissociate the cells into a single cell suspension, which is conducive to subsequent counting, passage, or seeding.

3.3 Assisted Reproductive Technology Research

Application Scenario: In in vitro fertilization experiments, it is necessary to remove the cumulus cells and corona radiata around the oocytes to observe and evaluate oocyte maturity or perform micromanipulation (such as ICSI).

Experimental Protocol:

•         Place the collected cumulus-oocyte complexes in a culture medium containing hyaluronidase (usually a lower concentration is used, such as 80-100 IU/mL).

•         Act at 37°C for 30 seconds to several minutes, and observe closely. Once the cumulus cells spread out, immediately wash the oocytes with culture medium multiple times to completely remove the enzyme solution and shed granulosa cells.

•         This method can quickly and relatively gently expose the oocytes, avoiding physical damage that may be caused by mechanical methods.

3.4 Disease Model and Mechanism Research

Application Scenario: Studying the role of hyaluronic acid in tumor invasion, metastasis, angiogenesis, and inflammatory response.

Experimental Protocol:

•         Loss-of-Function Research: Add hyaluronidase locally in the cell culture system or animal models (such as chick chorioallantoic membrane model) to degrade local hyaluronic acid.

•         Observation Indicators: By comparing the treatment group with the control group, the following can be observed:
       

￮       Cell invasion/migration ability: Use Transwell or scratch test to see if the invasion ability of cancer cells is weakened after degrading hyaluronic acid.

￮       Angiogenesis: Observe the promotion or inhibition of neovascularization after degrading hyaluronic acid.

￮       Drug permeability: Study whether degrading hyaluronic acid in the tumor matrix can enhance the penetration efficiency of chemotherapeutic drugs.

Such research provides direct evidence for clarifying the function of hyaluronic acid in physiological and pathological processes.

4. Summary

As an efficient and specific biocatalyst, hyaluronidase is a powerful tool for researchers to break through the extracellular matrix barrier. From basic primary cell preparation to cutting-edge disease mechanism exploration, its application runs through multiple levels of biomedical research. The key to successful application lies in a deep understanding of its mechanism of action, and carefully optimizing the enzyme concentration, action time, and compatibility scheme according to the specific experimental system (such as tissue type, cell type, research purpose). Mastering this "molecular scissors" will open up more possibilities for your scientific research.

5. Recommended Hyaluronidase Products by ANT BIO PTE. LTD.

ANT BIO PTE. LTD. provides high-quality hyaluronidase products that have undergone strict quality control and verification, with stable activity and reliable performance, which can fully meet the needs of various scientific research experiments. The detailed product information is shown in the following table:

6. Brand Mission

ANT BIO PTE. LTD. is committed to advancing life science research by providing high-quality, reliable reagents and comprehensive solutions. Our hyaluronidase products, as an important part of the molecular interaction research series, are carefully developed and optimized to meet the diverse needs of researchers in extracellular matrix-related experiments and emerging research fields.

Guided by the principles of innovation, quality, and customer-centricity, our three specialized sub-brands (Absin, Starter, and UA) cover a full spectrum of research needs, from general reagents to specialized experimental kits. We strive to establish long-term and trusted partnerships with researchers worldwide, supporting them in achieving breakthroughs in life science research and contributing to the development of fields such as cell biology, reproductive biology, and oncology.

7. Disclaimer

This article is compiled and interpreted based on relevant core content. All intellectual property rights related to the original content belong to the corresponding author. For any infringement, please contact us promptly and we will take immediate action.

8. Brand Promotion Copy

ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs

At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our hyaluronidase products, with stable activity and strict quality control, provide strong support for various research fields such as primary cell isolation, cell culture, and disease mechanism exploration. Backed by professional technical support, we strive to be your trusted partner in unlocking scientific mysteries. Explore our product portfolio today and elevate your research to new heights.