Dear Beheshteh Babazadeh

To prepare nanoliposomes and some other vesicles you do not need high temperature. As you see in the attached figure and as explained in many papers such as the one below, depending on your drug, a mild temperature is enough.

Let me know if you have more questions.

Khorasani, S., Danaei, M., & Mozafari, M. R. (2018). Nanoliposome technology for the food and nutraceutical industries. Trends in Food Science & Technology, 79, 106-115.

Dear Beheshteh,

As explained by Patcharida, all the necessary information is summarized in the illustration as well as the attached review paper. Thus, only a mild heating process is sufficient. What is more, this procedure is renowned as it is green, economically feasible, free of toxic-solvents as well as its potential in the industry and mass production which is not seen in other strategies like thin-film hydration, etc.

Best

Dear Beheshteh,

Incresing temperature in Mozafari method helps to solving insolouble materials in aqueous solution (also glycerol plays a pivotal role)

I strongly recommend using Mozafari method. I compared this method with thin lipid film hydration method in my master thesis and Mozafari method was much more effective (due to non-toxicity and other advantages which their explanation is beyond the scope of this discussion).

I hope you found this useful.

Hi Beheshteh Babazadeh ,

Thanks for your question. As explained above by Dr Patcharida Chaosri ,

Dr Iman Katouzian , and Dr Mehrnoush Mehrarya , the method is safe, robust and easily scalable. The procedure (Mozafari method) is proven to be reproducible by scientists in many different countries. You just need to follow the following SOP:

1. Add the liposomal ingredients to a preheated (60°C, 5 min) mixture of your water-soluble drug and a polyol such as glycerol (final concentration 3%, v/v) in a heat-resistant flask such as a pyrex beaker. Alternatively, a heat-resistant bottle with six baffles can be used as explained and pictured in reference [1] below.

2. Heat the mixture at 60°C while stirring (approx. 1,000 rpm) on a hotplate stirrer (e.g. RET basic IKAMAG1 Safety Control, IKA, Malaysia) for a period of 45–60 min under nitrogen atmosphere.

3. For the preparation of cholesterol-containing formulations, first dissolve the cholesterol in the aqueous phase at elevated temperatures (c. 120°C) while stirring (approx. 1,000 rpm) for a period of 15–30 min under nitrogen atmosphere before adding the other phospholipid components.

4. Depending on the formulation ingredients, sample volume, type of flask used and its number of baffles, as well as type, speed and duration of mixing, nanometric vesicles can be produced in one step, without the need to perform filtration or sonication.

5. Leave the nanoliposome suspension at temperatures above Tc under an inert atmosphere such as nitrogen or argon for 1 h to allow the sample to anneal and stabilize.

Reference [1]:

Colas, J. C., Shi, W., Rao, V. M., Omri, A., Mozafari, M. R., & Singh, H. (2007). Microscopical investigations of nisin-loaded nanoliposomes prepared by Mozafari method and their bacterial targeting. Micron, 38(8), 841-847.

Let us know if you have any further questions. Our team at ANNI will be happy to help you.

Hi, Dear professor

Many thanks for your answering.

We have used methods of Prof. Mozafari in different projects successfully. In the first stage of Mozafari method, we introduce phospholipids (and lipid soluble drug) into a mixture of water (or buffer) at 60 degree C.

Let me know if you have more questions.

Hello Beheshteh Babazadeh

Please note:

Currently, Mozafari method is widely used to encapsulate VO and polyunsaturated fatty acids (Table 2). When compared to thin hydration process, Mozafari method produce liposomes with higher stability. Rasti et al. (2012) investigated the difference between the oxidative stability of liposomes (before sonication) and nanoliposomes (after sonication) produced by Mozafari and conventional thin hydration techniques. For both methods, polyunsaturated fatty acids entrapment efficiency values of nanoliposomes were higher than those of liposomes. Authors related this result to the fact that size reduction increases the surface area of the liposomes resulting in increasing the contact and bond between the fatty acids and phospholipids in the bilayers. Moreover, liposomes prepared by Mozafari process showed higher physical stability than those prepared by conventional method (Rasti et al., 2012).

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From:

Lammari, N., Louaer, O., Meniai, A. H., Fessi, H., & Elaissari, A. (2021). Plant oils: From chemical composition to encapsulated form use. International Journal of Pharmaceutics, 120538.