Cellsome® made from Cardiolipin Lipids
Cardiolipin (CL) is a unique phospholipid with a very interesting chemical and specific ultrastructural characteristics. It is a highly acid dimer of phosphatidylglycerol (PG) and phosphatidic acid (PA), containing four acyl chains; three glycerols and two phosphate headgroups. Due to deprotonation of one of one of these phosphate groups, cardiolipin is negatively charged in physiological pH [1,2].
Cardiolipin (CL) is known as mitochondria-specific phospholipid since it is almost exclusively biosynthesized and located in the inner mitochondrial membranes. The name “cardiolipin” is derived from fact that it was first found and isolated from animal heart. Cardiolipin is considered to be be intimately linked to mitochondrial bioenergetic process. It plays a functional role in mitochondrial membrane stability and dynamics, interacts with a number of inner mitochondrial membrane metabolite carriers, enzymes and proteins. During apoptosis in presence of H2O2, CL-bound Cyt c catalyzes the peroxidation of cardiolipin, releasing Cyt c, which is a death-inducing protein. CL peroxidation and depletion have important implications to age-related mitochondrial dysfunction, resulting in a number of pathophysiological conditions, such as hypo/hyperthyroid states [3-7], heart ischemia–reperfusion [8-12], nonalcoholic fatty liver disease , diabetes [14,15], Barth syndrome [16,17] and aging [18-21]. According Birk et al. , the main functions of cardiolipin are: “(i) to support spatial organization of mitochondrial cristae; (ii) to create the proton trap necessary for sustaining the proton gradient and ATP synthesis by the F0F1 ATP synthase; (iii) to act as a scaffold for assembly of respiratory complexes and super-complexes to facilitate optimal electron transfer among the redox partners.”
Extensive studies [23-29] of pharmacological, toxicological, and therapeutic effects have shown that the incorporation of doxorubicin in cardiolipin liposomes improved the antitumor activity of doxorubicin, while the histopathologic lesions in cardiac tissue of mice significantly decreased. It has been reported that cardiolipin-containing liposomes have lower (at least 2-fold lower than that observed with conventional doxorubicin) cardiotoxicity associated with doxorubicin by altering the pharmacokinetics and tissue distribution of the drug in mice . Also, it has been indicated that cardiolipin provides two types of binding possibility for drugs; one mostly exposed at the surface, and the other deeply buried in the membrane [30,31]. Hence, the cardiolipin-liposomes has been suggested as a convenient carrier for doxorubicin delivery to increase the therapeutic index of the drug .
Cardiolipin is a negatively charged lipid. Cellsome® made from cardiolipin lipid catalog containing many different type of saturated and unsaturated cardiolipin based liposomes made from 0.5 up to 100 percent of cardiolipin.
Cellsome® made from Cardiolipin Lipids
For more information on the lipid composition of the liposomes mentioned above click here.
|Buffer and Liposome Size||Specification|
|Buffer||Phosphate Buffered Saline*|
|Liposome size||100 nm|
|* If you prefer different buffers, please specify in your order.|
Liposome Particle Calculator
Cellsome®-Cardiolipin liposomes are unilamellar and sized to 100 nm. The molar concentration of liposome is 10 mM. By having the liposome diameter (nm) and lipid concentration (µM), you can calculate the total number of the lipids in one liposome and number of the liposomes in one milliliter of the liposome solution. To use the calculator click here.
Each 100-nm liposome is composed of almost 80,000 phospholipid molecules. 10 mM liposome solution (phospholipid concentration of 10 mM) has 75.2 trillion (75.2×1012) liposome particles per milliliter of the solution. When using the liposomes calculator, you need to keep two things in mind:
- Enter the concentration in µM. As an example, 10 mM solution is 10,000 µM.
- Only include the concentration of phospholipids in your calculation. Cholesterol does not count since it inserts itself into the hydrophobic fatty acid chains of the lipid.
This calculation only works for the unilamellar liposomes which are below 200 nm in size. It does not work for larger multilamellar liposomes. For more information about the calculation see the video below:
- Due to very rapid hydrolysis of adenosine triphosphate (ATP), ATP liposome (ATPsome®) formulations are all freeze dried.
- Leftover liposomes in liquid form that are made after hydration of the lyophilized proliposome formulation can not be used or kept in refrigerator for future use due to rapid hydrolysis on ATP.
- Liposomes are formed upon hydration of the lyophilized formulation. Very large percentage of ATP molecules will remain encapsulated inside the liposomes but some percentage will be free and in non-encapsulated form after hydration of the freeze dried formulation. Free ATP molecules usually does not have any negative impact upon injection of the formulation to the animal or upon adding the formulation to the cell cultures.
- Trehalose is used as a lyoprotectant in all freeze dried liposome formulation. The size distribution after hydration of the freeze dried formulation will be around 100 nm.
- Freeze dried liposomes should be kept at -40 °C or -80 °C.
Cellsome®-Cardiolipin is a white translucent liquid made of nano size unilamellar liposomes. Usually due to the small size of liposomes no settling will occur in the bottom of the vial. The liposomes are packaged in an amber vial.
- All liposome based formulations are shipped on blue ice at 4 °C in insulated packages using overnight shipping or international express shipping.
- Liposomes should NEVER be frozen. Crystal ice that is formed in the lipid membrane can rupture the membrane and change the size of the liposomes. Liposomes in liquid form should always be kept in the refrigerator.
- Clients who order from outside of the United States of America are responsible for their government import taxes and customs paperwork. Encapsula NanoSciences is NOT responsible for importation fees to countries outside of the United States of America.
- We strongly encourage the clients in Japan, Korea, Taiwan and China to order via a distributor. Tough customs clearance regulations in these countries will cause delay in custom clearance of these perishable formulations if ordered directly through us. Distributors can easily clear the packages from customs. To see the list of the distributors click here.
- Clients ordering from universities and research institutes in Australia should keep in mind that the liposome formulations are made from synthetic material and the formulations do not require a “permit to import quarantine material”. Liposomes are NOT biological products.
- If you would like your institute’s FedEx or DHL account to be charged for shipping then please provide the account number at the time of ordering.
- Encapsula NanoSciences has no control over delays due to inclement weather or customs clearance delays. You will receive a FedEx or DHL tracking number once your order is confirmed. Contact FedEx or DHL in advance and make sure that the paperwork for customs is done on time. All subsequent shipping inquiries should be directed to Federal Express or DHL.
Storage and Shelf Life
Cellsome® products should always be stored at in the dark at 4 °C, except when brought to room temperature for brief periods prior to animal dosing. DO NOT FREEZE. If the suspension is frozen, the encapsulated drug can be released from the liposomes thus limiting its effectiveness. In addition, the size of the liposomes will also change upon freezing and thawing.
Cellsome®-Cardiolipin is made on daily basis. The batch that is shipped is manufactured on the same day. It is advised to use the products within 4 months of the manufacturing date.
References and background reading
2. Paradies G, Paradies V, De Benedictis V, Ruggiero FM, Petrosillo G. Functional role of cardiolipin in mitochondrial bioenergetics. Biochimica et Biophysica Acta (BBA)-Bioenergetics. 2014 Apr 1;1837(4):408-17.
3. Paradies G, Ruggiero FM. Enhanced activity of the tricarboxylate carrier and modification of lipids in hepatic mitochondria from hyperthyroid rats. Archives of biochemistry and biophysics. 1990 May 1;278(2):425-30.
5. Paradies G, Ruggiero FM, Dinoi P. The influence of hypothyroidism on the transport of phosphate and on the lipid composition in rat-liver mitochondria. Biochimica et Biophysica Acta (BBA)-Biomembranes. 1991 Nov 18;1070(1):180-6.
6. Paradies G, Ruggiero FM, Petrosillo G, Quagliariello E. Stimulation of carnitine acylcarnitine translocase activity in heart mitochondria from hyperthyroid rats. FEBS letters. 1996 Nov 18;397(2-3):260-2.
8. Paradies G, Petrosillo G, Pistolese M, Di Venosa N, Federici A, Ruggiero FM. Decrease in mitochondrial complex I activity in ischemic/reperfused rat heart: involvement of reactive oxygen species and cardiolipin. Circulation research. 2004 Jan 9;94(1):53-9.
9. Petrosillo G, Di Venosa N, Pistolese M, Casanova G, Tiravanti E, Colantuono G, Federici A, Paradies G, Ruggiero FM. Protective effect of melatonin against mitochondrial dysfunction associated with cardiac ischemia-reperfusion: role of cardiolipin. The FASEB journal. 2006 Feb 1;20(2):269-76.
10. Paradies G, Petrosillo G, Pistolese M, Di Venosa N, Serena D, Ruggiero FM. Lipid peroxidation and alterations to oxidative metabolism in mitochondria isolated from rat heart subjected to ischemia and reperfusion. Free Radical Biology and Medicine. 1999 Jul 1;27(1-2):42-50.
11. Petrosillo G, Colantuono G, Moro N, Ruggiero FM, Tiravanti E, Di Venosa N, Fiore T, Paradies G. Melatonin protects against heart ischemia-reperfusion injury by inhibiting mitochondrial permeability transition pore opening. American Journal of Physiology-Heart and Circulatory Physiology. 2009 Oct 1;297(4):H1487-93.
12. Petrosillo G, Ruggiero FM, Di Venosa N, Paradies G. Decreased complex III activity in mitochondria isolated from rat heart subjected to ischemia and reperfusion: role of reactive oxygen species and cardiolipin. The FASEB Journal. 2003 Apr 1;17(6):714-6.
13. Petrosillo G, Portincasa P, Grattagliano I, Casanova G, Matera M, Ruggiero FM, Ferri D, Paradies G. Mitochondrial dysfunction in rat with nonalcoholic fatty liver: involvement of complex I, reactive oxygen species and cardiolipin. Biochimica et Biophysica Acta (BBA)-Bioenergetics. 2007 Oct 1;1767(10):1260-7.
14. Han X, Yang J, Cheng H, Yang K, Abendschein DR, Gross RW. Shotgun lipidomics identifies cardiolipin depletion in diabetic myocardium linking altered substrate utilization with mitochondrial dysfunction. Biochemistry. 2005 Dec 20;44(50):16684-94.
15. Han X, Yang J, Yang K, Zhao Z, Abendschein DR, Gross RW. Alterations in myocardial cardiolipin content and composition occur at the very earliest stages of diabetes: a shotgun lipidomics study. Biochemistry. 2007 May 29;46(21):6417-28.
17. Vreken P, Valianpour F, Nijtmans LG, Grivell LA, Plecko B, Wanders RJ, Barth PG. Defective remodeling of cardiolipin and phosphatidylglycerol in Barth syndrome. Biochemical and biophysical research communications. 2000 Dec 20;279(2):378-82.
20. Paradies G, Paradies V, Ruggiero FM, Petrosillo G. Changes in the mitochondrial permeability transition pore in aging and age-associated diseases. Mechanisms of ageing and development. 2013 Jan 1;134(1-2):1-9.
21. Petrosillo G, Matera M, Moro N, Ruggiero FM, Paradies G. Mitochondrial complex I dysfunction in rat heart with aging: critical role of reactive oxygen species and cardiolipin. Free Radical Biology and Medicine. 2009 Jan 1;46(1):88-94.
22. Birk AV, Chao WM, Bracken C, Warren JD, Szeto HH. Targeting mitochondrial cardiolipin and the cytochrome c/cardiolipin complex to promote electron transport and optimize mitochondrial ATP synthesis. British journal of pharmacology. 2014 Apr 1;171(8):2017-28.
24. Rahman A, Fumagalli A, Barbieri B, Schein PS, Casazza AM. Antitumor and toxicity evaluation of free doxorubicin and doxorubicin entrapped in cardiolipin liposomes. Cancer chemotherapy and pharmacology. 1986 Jan 1;16(1):22-7.
28. Rahman A, White G, More N, Schein PS. Pharmacological, toxicological, and therapeutic evaluation in mice of doxorubicin entrapped in cardiolipin liposomes. Cancer research. 1985 Feb;45(2):796-803.
31. Henry N, Fantine EO, Bolard J, Garnier-Suillerot A. Interaction of adriamycin with negatively charged model membranes: evidence of two types of binding sites. Biochemistry. 1985 Dec 1;24(25):7085-92.