Cholesterol-linoleic acid liposomes induced extracellular vesicles secretion from immortalized adipose-derived mesenchymal stem cells for in vitro cell migration

Figures & data

Figure 1. Stability of different types of liposomes. Data are represented as mean ± SD (n = 3).

Table 1. Characterization of the size and zeta potential for different types of liposomes.

Types of liposomes	Size (nm) (mean ± SD)	Zeta potential (mV) (mean ± SD)
DOTAP liposomes	242.3 ± 0.8	22.3 ± 0.8
Chitosan–coated DOTAP liposomes	694 ± 20	24.9 ± 0.7
Cholesterol–DOTAP liposomes	358 ± 1	26 ± 1
LA liposomes	202.2 ± 0.1	−31 ± 2
Chitosan–coated LA liposomes	275 ± 5	−40.0 ± 0.8
Cholesterol–LA liposomes	161.8 ± 0.6	−57.8 ± 0.7

The size and zeta potential for different types of liposomes were determined between days 14 to 21 after preparation. Data are represented as mean ± SD (n = 3).

Figure 2. Transmission electron microscopy of (a) DOTAP liposomes, (b) chitosan–coated DOTAP liposomes, (c) cholesterol–DOTAP liposomes, (d) LA liposomes, (e) chitosan–coated LA liposomes and (f) cholesterol–LA liposomes with a magnification of ×25,000. The chitosan coating layer is indicated by arrows.

Figure 3. The secretion of isolated EVs from immortalized adipose-derived mesenchymal stem cells after being treated with different dosages and types of liposomes: (a) DOTAP liposomes, (b) chitosan–coated DOTAP liposomes, (c) cholesterol–DOTAP liposomes, (d) LA liposomes, (e) chitosan–coated LA liposomes and (f) cholesterol–LA liposomes. Data are represented as mean ± SD (n = 3), **p < .01, ***p < .001.

Figure 4. The percentage of cell viability after being treated with different dosages and types of liposomes. Data are represented as mean ± SD (n = 3), *p < .05, **p < .01, ***p < .001.

Figure 5. Fluorescence microscopy of (a) LA liposomes before and after being separated from the conditioned medium and (b) cholesterol–LA liposomes before and after being separated from the conditioned medium. The green spherical shape represents the presence of liposomes in conditioned medium before separation.

Table 2. Characterization of the size and zeta potential for different types of liposomal-EVs with wound healing properties.

Types of liposomal-EVs	Size (nm) (mean ± SD)	Zeta potential (mV) (mean ± SD)
Non-induced EVs	280 ± 10	−2.5 ± 0.4
LA liposomal-EVs (LA-EVs)	557 ± 20	−4.8 ± 0.8
Cholesterol–LA liposomal-EVs (cholesterol–LA-EVs)	320 ± 9	−4.6 ± 0.7

Data are represented as mean ± SD (n = 3).

Figure 6. Transmission electron microscopy of (a) non-induced EVs, (b) LA liposomal-EVs and (c) cholesterol–LA liposomal-EVs. All EVs with a magnification of ×10,000 are indicated by arrows, respectively.

Figure 7. The percentage of wound closure in HaCaT after treatment with different dosages and types of liposomal-induced EVs. Data are represented as mean ± SEM (n = 3), *p < .05, **p < .01 .

Data availability statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.