Clodronate Liposomes

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Clodronate liposomes represent a pivotal advancement in the field of cellular biology, particularly in the study of the immune system. This sophisticated tool is instrumental for the selective depletion of macrophages and other immune cells, facilitating a deeper understanding of their roles in various biological processes and disease models. BOC Sciences provides various types of clodronate liposomes and control liposomes to our customers. All liposomes are prepared under sterile conditions. In addition, we offer a range of custom liposome services for you to choose from. At BOC Sciences, you're sure to make a difference in liposomes.

What are Clodronate Liposomes?

Clodronate liposomes are lipid-based nanoparticles that encapsulate clodronate, a bisphosphonate compound traditionally used to treat bone-related disorders. These liposomes are composed of phospholipid bilayers that serve as a delivery vehicle for clodronate, allowing for targeted and efficient cellular uptake. The unique structure of clodronate liposomes facilitates the selective elimination of phagocytic cells, primarily macrophages, by exploiting the natural propensity of these cells to engulf lipid-based particles. The typical structure of clodronate liposomes includes phospholipids, cholesterol, and clodronate. The phospholipids form a bilayer, while cholesterol stabilizes the liposome structure, providing fluidity and integrity. This configuration ensures the effective encapsulation of clodronate, which can then be released intracellularly upon liposome uptake by target cells, primarily macrophages.

Clodronate

Clodronate, also known as clodronic acid, is a synthetic bisphosphonate that plays a significant role in inhibiting bone resorption. While it is widely used in the treatment of conditions such as osteoporosis and metastatic bone disease, its pharmacological potential extends beyond skeletal health. Clodronate exhibits immunomodulatory properties by inducing apoptosis in macrophages when delivered intracellularly. This characteristic is particularly advantageous in research settings, as it allows scientists to manipulate macrophage populations and explore their contributions to different biological processes, including inflammation, tissue repair, and immune response regulation.

Clodronate Liposomes from BOC Sicences

Clodronate molecules are encapsulated in small liposomes, which have excellent activity, physical and chemical stability, and are easy to use and handle. After a single intravenous or intraperitoneal administration of clodronate liposomes, 90% of macrophages in the spleen can be depleted.

The Fluorescent-Control-for-Clodronate product line from BOC Sciences includes fluorescently labeled liposomes that serve as negative control markers for imaging in microfluidic structures. Utilizing five fluorescent dyes (DiA, DiD, DiI, DiO, and DiR), these liposomes enable specific fluorescence and are ideal for studying macrophage uptake mechanisms.

BOC Sciences offers lyophilized clodronate liposomes, including neutral and anionic formulations, effectively addressing the stability issues faced by traditional liposomes during long-term storage. Utilizing advanced freeze-drying technology, these products extend shelf life and can be easily reconstituted by adding sterile pure water, ensuring optimal performance in various applications.

The Mannosylated Clodronate Liposomes product category from BOC Sciences includes multilamellar liposome suspensions that achieve efficient drug delivery targeting macrophages through mannose modification. Products in this category demonstrate excellent cellular uptake characteristics and biodistribution, aiding in immune research and related drug development.

The Standard Clodronate Liposomes from BOC Sciences are effective tools for specific macrophage depletion, featuring both neutral and anionic liposomes with control options. This series is highly active, long-lasting, and non-toxic, suitable for various administration routes to enhance understanding of macrophage functions in pathological conditions.

Clodronate Liposomes Preparation

The preparation of clodronate liposomes requires a series of meticulous steps to ensure high encapsulation efficiency, stability, and reproducibility. The process typically begins with the formation of a lipid film, wherein phospholipids—commonly phosphatidylcholine and cholesterol—are dissolved in an organic solvent. After the solvent is evaporated, the resulting lipid film is hydrated with an aqueous phase containing clodronate. This hydration process leads to the formation of multilamellar vesicles (MLVs), which can be further processed to obtain unilamellar vesicles with a consistent size distribution. Size reduction techniques such as sonication or extrusion are employed to achieve the desired particle size, typically in the range of 100-200 nm. The final product is characterized for encapsulation efficiency, particle size, and stability to ensure it meets the rigorous standards expected in biological applications. During the preparation process, the liposomes are filtered through a 2 μm polycarbonate membrane to ensure that large particles that may be toxic to animals are removed from the suspension. All products are prepared and packaged under aseptic conditions. Quality control measures are essential to guarantee that the clodronate liposomes are effective and reliable for research purposes.

The preparation of liposomal clodronate.Schematic representation of steps involved in the preparation of liposomal clodronate. (Ravichandran, S.; et al, 2022)

Clodronate Liposome Function

The primary function of clodronate liposomes is to facilitate the targeted depletion of macrophages and other phagocytic cells. Following administration, these liposomes are preferentially taken up by macrophages via phagocytosis. Once inside the macrophages, clodronate is released, leading to the accumulation of toxic metabolites that induce apoptosis. This selective action enables researchers to study the effects of macrophage depletion on various biological processes, including inflammation, tissue repair, and tumorigenesis. By employing clodronate liposomes, scientists can elucidate the roles of macrophages in immune responses and their interactions with other cell types, thereby advancing our understanding of complex biological systems.

Clodronate Liposomes Monocyte Depletion

Monocytes, the precursors to macrophages, can also be targeted for depletion using clodronate liposomes. The selective depletion of monocytes is critical for studying their differentiation into macrophages and the subsequent roles they play in immune responses. By employing clodronate liposomes, researchers can effectively investigate the impact of monocyte depletion on inflammation, tissue homeostasis, and immune regulation. The ability to deplete monocytes provides a clearer understanding of the dynamics within the mononuclear phagocyte system and allows for the exploration of compensatory mechanisms that may arise in response to cellular depletion.

Clodronate Liposomes Mechanism

Once clodronate is encapsulated in liposomes, it can specifically target phagocytes. Liposomes can be quickly recognized and swallowed by macrophages, allowing an effective clodronate concentration threshold to trigger target cell apoptosis. Clodronate encapsulated in liposomes is recognized as a foreign body by phagocytes and is taken into internal vesicles called phagosomes. After phagocytosis, the phagosome fuses with the lysosome containing phospholipase, which destroys the phospholipid bilayer of the liposome, thereby allowing clodronate to be released in the cell. Once clodronate is released in the cytoplasm, aminoacyl-tRNA synthetase will mistakenly identify it as pyrophosphate and will use clodronate to generate the non-hydrolyzable ATP analogue (AppCCl2p). Then, this ATP analog translocates to the mitochondria, where it irreversibly binds to the ATP/ADP translocase, causing blockage of the respiratory chain, leading to changes in mitochondrial function and integrity. These altered mitochondria release molecular signals that will initiate cell apoptosis. Unencapsulated clodronate cannot pass through the cell membrane to trigger cell death. Since the control liposome does not contain clodronate, the phagocytes will not be killed.

Clodronate Liposomes Monocyte Depletion Protocol

A typical protocol for monocyte depletion utilizing clodronate liposomes includes the following detailed steps:

This protocol provides a robust framework for utilizing clodronate liposomes in monocyte depletion studies, facilitating insights into their roles within the immune system.

Advantages of Clodronate Liposomes

Clodronate liposomes offer several advantages for researchers, making them a preferred choice for studies involving macrophage and monocyte depletion.

Applications of Clodronate Liposomes

Immunology

Clodronate liposomes are used to study macrophages and phagocytic cells in immunology. By selectively depleting macrophages, researchers can investigate their impact on inflammation, antigen presentation, and autoimmune diseases like rheumatoid arthritis and inflammatory bowel disease, as well as T cell activation and differentiation, which provide insights into immune tolerance and vaccine efficacy.

Neuroscience

In neuroscience, clodronate liposomes are valuable for studying microglia, the CNS's immune cells. Their depletion aids in understanding their role in neuroinflammatory diseases such as Alzheimer's and Parkinson's, as well as their impact on synaptic function and memory.

Cancer

In cancer research, clodronate liposomes help explore tumor-associated macrophages (TAMs), which promote tumor growth and metastasis. Depleting TAMs allows researchers to assess their effects on tumor progression and responses to therapies, including immunotherapy and chemotherapy.

Tissue Engineering

In tissue engineering and regenerative medicine, clodronate liposomes are used to modulate macrophage activity during wound healing. By depleting macrophages, researchers can identify strategies to enhance tissue repair and understand how macrophages influence stem cell activity and differentiation in regenerative therapies.

Hot Spot Research on Clodronate Liposomes

Clodronate Liposomes Microglia

Microglia, the immune cells of the central nervous system (CNS), are essential for neuronal health and play a role in neuroinflammation and neurodegenerative diseases. Clodronate liposomes selectively deplete microglia, enabling researchers to study the effects on neuronal health and brain homeostasis. This approach is particularly useful for understanding Alzheimer's disease, multiple sclerosis, and traumatic brain injury, where microglial activity impacts disease progression.

Clodronate Liposomes Neutrophils

Clodronate liposomes primarily target macrophages but also affect neutrophil dynamics. Neutrophils are crucial for acute inflammation, and their interactions with macrophages shape the immune response. By depleting macrophages, researchers can investigate compensatory mechanisms in neutrophils and their roles in inflammation, offering insights into immune coordination and therapeutic targets.

Clodronate Liposomes Dendritic Cells

Dendritic cells (DCs) are key for initiating adaptive immune responses as antigen-presenting cells. Clodronate liposomes help explore the interplay between DCs and macrophages in antigen presentation and T cell activation. Depleting macrophages allows researchers to understand how DCs function without them, informing studies on immune tolerance, vaccine efficacy, and immunotherapy development.

Advantages of Clodronate Liposomal Products from BOC Sciences

These advantages position BOC Sciences as a leader in providing clodronate liposomal products, especially suitable for effective macrophage research and manipulation.

Reference

  1. Ravichandran, S.; et al. Liposome encapsulated clodronate mediated elimination of pathogenic macrophages and microglia: A promising pharmacological regime to defuse cytokine storm in COVID-19. Medicine in Drug Discovery. 2022, 15: 100136.

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