Ionically Cross-Linked Chitosan Nanoparticles for Sustained Delivery of Docetaxel: Fabrication, Post-Formulation and Acute Oral Toxicity Evaluation

Figures & data

Table 1 Composition of Chitosan Loaded Nanoparticles Containing Docetaxel

Formulation Code	Chitosan Concentration^a	DTXL Concentration^b	STPP Concentration^c
CNP1	75 mg/25 mL, i.e. 3 mg/mL	10 mg/3 mL	10 mg/10 mL
CNP2	75 mg/25 mL, i.e. 3 mg/mL	10 mg/3 mL	15 mg/10 mL
CNP3	75 mg/25 mL, i.e. 3 mg/mL	10 mg/3 mL	20 mg/10 mL
CNP4	75 mg/25 mL, i.e. 3 mg/mL	10 mg/3 mL	25 mg/10 mL
CNP5	75 mg/25 mL, i.e. 3 mg/mL	10 mg/3 mL	30 mg/10 mL

Notes: ^aIn 25 mL of 1.5% acetic acid solution in water to produce the concentration of 3mg/mL. ^bIn 3 mL of ethanol. ^cIn 10 mL of distilled water.

Table 2 Evaluation of Different Formulation Parameters of Docetaxel Loaded Nanoparticles

Formulation Code	Percentage Yield (%)±SD	Entrapment Efficiency (%)±SD	Loading Capacity (%)±SD	Particle Size ±SD	PDI ±SD	Zeta Potential ±SD
CNP1	66.79±0.008	60.124 ± 0.93	9.002 ± 0.14	172.6 ± 4.08	0.319 ± 0.013	35.95 ± 0.69
CNP2	73.79±0.012	66.758 ± 1.31	9.025 ± 0.18	195.3 ± 1.92	0.448 ± 0.012	33.65 ± 0.37
CNP3	77.81±0.022	72.728 ± 1.12	9.347 ± 0.14	270.9 ± 5.31	0.219 ± 0.004	33.0 ± 0.82
CNP4	69.20±0.016	64.022 ± 0.82	9.252 ± 0.12	337.4 ± 2.45	0.398 ± 0.016	33.5 ± 0.57
CNP5	71.57±0.005	56.393 ± 0.61	7.880 ± 0.09	479.65 ± 11.47	0.487 ± 0.011	30.45 ± 0.37

Figure 1 Scanning electron micrographs of docetaxel loaded chitosan nanoparticles.

Figure 2 DSC and TGA (Drug, physical mixture, formulation CNP3).

Figure 3 XRD of DTXL (A), chitosan (B), STPP (C), physical mixture (D) and formulation CNP3 (E).

Figure 4 Comparative FTIR spectra of DTXL (A), chitosan (B), STPP (C), physical mixture (D) and formulation CNP3 (E).

Figure 5 Cumulative drug release of polymeric nanoparticles.

Table 3 Kinetic Modeling of Drug Release Profile of Chitosan Nanoparticles

Formulation Code	Zero Order	First Order	Higuchi Model	Korsmeyer-Peppas Model
	R^2	R^2	R^2	R^2	n
CNP1	0.1796	0.6131	0.8573	0.9734	0.338
CNP2	0.1639	0.5962	0.8616	0.9734	0.340
CNP3	0.1598	0.6739	0.8624	0.9744	0.340
CNP4	0.7360	0.2047	0.7018	0.9769	0.279
CNP5	0.9048	0.0139	0.6478	0.9786	0.264

Table 4 Clinical Observations of an Acute Oral Toxicity Study for Prepared Chitosan Loaded Nanoparticles

Observation	Group I (Treated with CNPs) Mean ± SD	Group II (Control) Mean ± SD
Signs of illness	Nil	Nil
Body Weight (in grams)
Day 1	172.42 ± 0.42	167.25 ± 0.35
Day 7	193.08 ± 0.31	197.17 ± 0.31
Day 14	206.83 ± 0.47	207.83 ± 0.66
Any visible skin irritation/ toxicity	Nil	Nil
Ocular toxicity	Nil	Nil
Mortality	Nil	Nil

Table 5 Biochemical Blood Analysis

Hematology	Group I (CNPs) Mean ± SD	Group II (Control) Mean ± SD
Hemoglobin (g/dL)	13.3±0.01	13.20±0.14
Red blood cells ×10^6 (µL)	6.99±0.07	6.94±0.05
Platelets × 10^3 (µL)	924.75±3.96	928.50±4.72
White blood cells× 10^3 (µL)	7.58±0.38	7.93±0.33
Lymphocytes (%)	89.08±1.06	88.38±0.41
Neutrophils (%)	8.23±0.59	8.75±0.38
Mean corpuscular volume (fL)	67.38±0.71	67.83±0.48
Mean corpuscular hemoglobin concentration (g/dL)	28.34±0.03	28.41±0.06
Mean corpuscular hemoglobin (pg)	18.95±0.11	19.20±0.22

Note: All values are given as mean plus or minus the standard deviation (n=3).

Table 6 Liver and Kidney Function Tests

Liver and Kidney Tests	Group I (CNPs) Mean ± SD	Group II (Control) Mean ± SD
ALT (U/L)	73.88±0.74	73.13±1.88
Alkaline phosphatase (g/dL)	115.75±3.77	113.25±4.66
Bilirubin (mg/dL)	0.31±0.01	0.37±0.02
AST (U/L)	122±2.24	123±2.45
Urea (mg/dL)	46.75±2.28	45.75±1.92
Creatinine (mg/dL)	0.38±0.06	0.41±0.04

Note: All values are given as mean plus or minus the standard deviation (n=3).

Figure 6 Histopathological microscopic examination of different tissues of wistar rats of group-I (test), and group-II (control).