An efficient calcium phosphate nanoparticle-based nonviral vector for gene delivery

Figures & data

Figure 1 Representative atomic force microscopic images of the classical calcium phosphate particle (0PS-CaP) and protamine sulfate-coated calcium phosphate particle (1PS-CaP) after standing for two hours and seven days, respectively.

Abbreviations: PS-CaP, protamine sulfate coated-calcium phosphate; 0PS-CaP, PS-CAP with 0% protamine sulfate concentration; 1PS-CaP, PS-CAP with 1% protamine sulfate concentration.

Table 1 Results of all transfection experiments

Cell type	Storage time	Transfection efficiency (%)*
		0PS-CaP	1PS-CaP	2PS-CaP	4PS-CaP
HEK 293	2 hours	2.64	3.57	6.15	8.33
	7 days	1.08	1.94	3.64	4.94
293 FT	20 minutes	3.98	15.56	22.81	49.43
NIH 3T3	20 minutes	0.75	1.97	3.81	4.71

Note:

* Transfection efficiency was computed by calibration of blank value from the result of flow cytometry analysis.

Figure 2 Fluorescence confocal microscopy (20× magnification) of NIH 3T3 and 293 FT cells transfected with the classical calcium phosphate particle (0PS-CaP) and 1PS-CaP, 2PS-CaP, and 4PS-CaP.

Abbreviations: PS-CaP, protamine sulfate coated-calcium phosphate; 0PS-CaP, PS-CAP with 0% protamine sulfate concentration; 1PS-CaP, PS-CAP with 1% protamine sulfate concentration; 2PS-Cap, PS-CaP with 2% protamine sulfate concentration; 4PS-CaP, PS-CAP with 4% protamine sulfate concentration.

Figure 3 Fluorescence confocal microscopy (magnification 20× in all cases) of HEK 293 cells that were transfected with the classical calcium phosphate particle (0PSCaP) and 1PS-CaP, 2PS-CaP, and 4PS-CaP, after standing for two hours and for seven days.

Abbreviations: PS-CaP, protamine sulfate-coated calcium phosphate; 0PS-CaP, PS-CAP with 0% protamine sulfate concentration; 1PS-CaP, PS-CAP with 1% protamine sulfate concentration; 2PS-Cap, PS-CaP with 2% protamine sulfate concentration; 4PS-CaP, PS-CAP with 4% protamine sulfate concentration.

Figure 4 Effect of PS-CaP and classical calcium phosphate particle (0PS-CaP) dosage on cell viability. A series of different doses (0–2 times that of transfection dosage) of the 0PS-CaP and 1PS-CaP, 2PS-CaP, and 4PS-CaP were used to treat HEK 293 cells, and their inhibitory effect on cell proliferation was evaluated by measuring MTT absorption.

Note: *Indicates that 1PS-CaP is significantly different from 0PS-CaP (P < 0.05).

Abbreviations: PS-CaP, protamine sulfate-coated calcium phosphate; 0PS-CaP, PSCAP with 0% protamine sulfate concentration; 1PS-CaP, PS-CAP with 1% protamine sulfate concentration; 2PS-Cap, PS-CaP with 2% protamine sulfate concentration; 4PS-CaP, PS-CAP with 4% protamine sulfate concentration.