In-situ implant containing PCL-curcumin nanoparticles developed using design of experiments

Figures & data

Table 1. Full factorial design showing the effect of process variables on size.

Run order	Std order	HS (rpm) (A)	DPR (B)	PVA concn (% w/v) (C)	Mean particle size (nm)
1	8	12 000	20	2	356.4
2	6	12 000	3	2	508.1
3	3	6000	20	0.5	397.8
4	2	12 000	3	0.5	510.1
5	1	6000	3	0.5	351.6
6	5	6000	3	2	448.1
7	4	12 000	20	0.5	721.9
8	7	6000	20	2	451.5
Effect		A: 111.88	B: 27.43	C: −54.33	ABC: −80.18
		AB: 2.63	BC: −101.58	AC: −129.43
		% Contribution		A: 25.17		B: 1.51	C: 1.51	ABC: 12.93
AB: 0.014	BC: 20.75			AC: 33.69

HS, homogenization speed; DPR, drug–polymer ratio; conc, concentration.

Table 2. Central composite design to study the effect of interaction between homogenization speed and homogenization time on mean particle size.

Run order	Std order	Speed (rpm) (A)	Time (min) (B)	Size (nm)
1	11	6000	37.5	439.9
2	1	3000	15	905.9
3	6	10 242.64	37.5	361.8
4	10	6000	37.5	297.4
5	4	9000	60	448.6
6	8	6000	69.321	511.3
7	12	6000	37.5	506.9
8	13	6000	37.5	617.9
9	5	1757.36	37.5	458.4
10	9	6000	37.5	529.9
11	7	6000	5.68	1186
12	3	3000	60	461.8
13	2	9000	15	565.9
p value		0.159	0.002
		AB– 0.181

Table 3. Factorial design to study the effect of process variables on percentage drug encapsulation, size and zeta potential.

Run order	Std order	CPR	PC (% w/v)	Speed (rpm)	Size (nm)	Drug encap (%)	Zeta potential (mV)
1	3	5	1	7000	675.6	8.20	−22.5
2	8	20	1	10 000	375.9	23.05	−20.1
3	4	20	1	7000	714.2	14.98	−24.5
4	7	5	1	10 000	441.7	19.67	−28.1
5	1	5	0.1	7000	307.5	5.36	−19
6	6	20	0.1	10 000	470.9	10.84	−30.6
7	5	5	0.1	10 000	373.9	5.94	−24.8
8	2	20	0.1	7000	492.6	3.01	−18.6

CPR, curcumin-PCL ratio; PC, PVA concentration; encap, encapsulation.

Figure 1. Contour plot showing the effect of process variables analyzed using factorial design on mean particle size. Effect of various levels of homogenization speed and PVA concentration on mean particle size when the level of drug–polymer ratio is maintained at a low level (a) and high level (b).

Figure 2. Surface and overlaid contour plot showing the effect of variables during CCD. (a) Surface plot showing the effect of interaction of homogenization speed and homogenization time on mean particle size. (b) Overlaid contour plot showing level of homogenization speed and homogenization time required to achieve particle size <500 nm.

Table 4. The effect of process variables studied under factorial design on percentage drug encapsulation, size and zeta potential.

	% drug encap		Size		Zeta potential
Term	Effect	% Cont	Effect	% Cont	Effect	% Cont
A-Curcumin-PCL ratio	3.181	5.56	63.73	5.54	0.15	0.03
B-PVA concentration	10.19	56.99	140.63	26.97	−0.55	0.46
C-Speed	6.99	26.82	−131.88	23.72	−4.75	34.43
AB	1.90	1.99	−77.33	8.15	2.85	12.39
AC	0.96	0.51	−48.13	3.16	0.95	1.38
BC	2.78	4.25	−154.23	32.44	4.15	26.28
ABC	−2.66	3.90	−4.08	0.02	4.05	25.03

% drug encap – percentage drug encapsulation, % Cont, percentage contribution.

Figure 3. Effect of various levels of homogenization speed and PVA concentration on (a) percentage drug encapsulation, (b) mean particle size and (c) zeta potential when the level of curcumin-PCL ratio is maintained at highest level.

Figure 4. Mean particle size of curcumin-loaded PCL nanoparticles prepared under optimized condition.

Figure 5. Zeta potential of curcumin-loaded PCL nanoparticles prepared under optimized condition.

Figure 6. Percentage of drug released from curcumin-loaded PCL nanoparticles.

Figure 7. Effect of plain curcumin and curcumin-loaded PCL nanoparticles on (a) body weight and (b) mean survival time.

Figure 8. Effect of plain curcumin and curcumin-loaded PCL nanoparticles on hematological parameters (a) tumor volume, (b) hemoglobin level, (c) RBC level and (d) WBC level. *indicates that the treatment with plain curcumin and PCL-curcumin nanoparticles significantly (p-value < 0.05) controlled the proliferation of EAC cells.

Figure 9. Cumulative percentage of drug released from the PLGA implant containing curcumin-loaded PCL nanoparticles.