Synthesis and in vivo evaluation of three fluid spray dried hybrid ciprofloxacin microparticles in Sprague Dawley rats

Figures & data

Figure 1. Scanning electron microscopy (SEM) images of silica nanoparticles prepared by 2-fluid nozzle spray drying (A), silica coated silver nanoparticles prepared by 2-fluid nozzle spray drying (B), silica coated silver nanoparticles and entrapped with a layer of CFX prepared by 3-fluid nozzle spray drying (C) and silica coated silver nanoparticles inside CFX/chitosan prepared by 3-fluid nozzle spray drying (D).

Figure 2. X-ray dispersive spectrometry (EDX) results showing surface abundance of (A) silicon (Si) and (B) silver (Ag) of samples prepared by spray drying.

Figure 3. FTIR spectra of prepared solid dispersions prepared using two and 3-fluid nozzle spray drying along with corresponding spectra for ciprofloxacin (CFX) and chitosan. The dotted lines show peak positions in CFX where a shift was seen compared to S-AgNPs-CSCFX microparticles signifying structural changes.

Figure 4. X-ray powder diffraction diffractograms of spray dried microparticles of silica, silica coated silver particles, silica coated silver nanoparticles loaded with CFX, and silica coated silver nanoparticles loaded with CFX and chitosan.

Figure 5. Transmission electron microscopy (TEM) images showing dissolution of silica coated nanoparticles inside matrix of CFX/chitosan (A) and CFX (B) prepared by 3-fluid nozzle spray drying.

Table 1. Body weight of rats treated with blank and loaded silica-coated silver nanoparticles with ciprofloxacin.

Time Treatments	Day-0	Day-7	Day-14	Day-21
Control Negative	^A 136.2 ± 1.83^A	^A162.7 ± 28.80^A	^A188.50 ± 40.27^A	^A205.7 ± 54.71^A
blank LD	^A125.5 ± 9.13^AB	^AB143.2 ± 10.99^A	^B167.2 ± 15.77^A	^BC183.7 ± 10.40^A
blank MD	^A138.2 ± 7.72^AB	^B174.5 ± 11.73^A	^BC196.2 ± 9.98^A	^C210.0 ± 12.66^A
blank HD	^A138.0 ± 8.32^AB	^AB157.2 ± 16.74^A	^B179.2 ± 27.97^A	^BC197.5 ± 11.96^A
Loaded LD	^A140.7 ± 4.39^AB	^B168.2 ± 16.78^A	^BC189.7 ± 10.71^A	^C209.2 ± 14.08^A
loaded MD	^A150.2 ± 10.61^B	^B174.7 ± 13.63^A	^BC198.7 ± 12.92^A	^C216.7 ± 9.25^A
loaded HD	^A139.5 ± 9.13^AB	^AB167.3 ± 10.96^A	^BC186.2 ± 8.54^A	^C197.5 ± 7.02^A

*CN: Control negative; LD: Low dose (30 mg/Kg body weight); MD: Median Dose (60 mg/Kg body weight); and HD: High Dose (90 mg/Kg body weight); S-AgNPs: Silica coated silver nanoparticles; S-AgNPs-CSCFX: Silica coated silver nanoparticles loaded with ciprofloxacin.

The data are mean ± Std. Deviation. Bonferroni Post Hoc test was used. The superscript letters on the right of each data represent the statistical differences between groups. While the superscripts letters on the left represent the statistical differences within the groups. P-value less than 0.05 was considered statistical differences.

Table 2. Serum liver function tests of rats treated with blank and loaded silica-coated silver nanoparticles with ciprofloxacin.

Test Group	Total protein (TP) (g/L)	Albumin (ALB) (g/L)	Globulin (GLB) (g/L)	Alanine Aminotransferase (ALT) (U/L)	Aspartate Aminotransferase (AST) (U/L)	Alkaline Phosphatase (ALP) (U/L)	Total Serum Bilirubin (TSB) (𝜇mol/L)
CN	6.03 ± 0.24^a	3.4 ± 0.07^a	3.55 ± 75.15^a	85.25 ± 28.77^a	142.5 ± 12.84^a	595.25 ± 86.04^b	0.0 ± 0.0^a
LD-S-AgNPs	6.07 ± 0.15^a	3.4 ± 0.1^a	3.93 ± 1.25^a	82.0 ± 3.51^a	169.0 ± 15.72^a	563.67 ± 67.77^b	0.0 ± 0.0^a
MD-S-AgNPs	7.03 ± 0.11^c	3.7 ± 0.11^ab	3.2 ± 0.11^a	73.75 ± 5.66^a	159.25 ± 24.77^a	480.75 ± 41.58^ab	0.0 ± 0.0^a
HD-S-AgNPs	6.43 ± 0.17^ab	3.63 ± 0.14^ab	2.8 ± 0.11^a	89.5 ± 23.6^a	192.0 ± 23.23^a	500.75 ± 84.17^b	0.0 ± 0.0^a
LD-S-AgNPs-CSCFX	7.08 ± 0.11^c	4.08 ± 0.08^c	3.0 ± 0.08^a	95.25 ± 6.49^a	172.25 ± 12.17^a	507.5 ± 36.36^b	0.023 ± 0.023^a
MD-S-AgNPs-CSCFX	6.75 ± 0.12^bc	3.8 ± 0.0^bc	2.95 ± 0.12^a	88.50 ± 7.9^a	161.25 ± 5.34^a	549.75 ± 55.65^b	0.0 ± 0.0^a
HD-S-AgNPs-CSCFX	6.83 ± 0.26^bc	3.95 ± 0.16^bc	3.08 ± 0.27^a	63.0 ± 6.15^a	144.25 ± 15.61^a	406.5 ± 26.08^a	0.0 ± 0.0^a

CN: Control negative; LD: Low dose (30 mg/Kg body weight); MD: Median Dose (60 mg/Kg body weight); and HD: High Dose (90 mg/Kg body weight); S-AgNPs: Silica coated silver nanoparticles; S-AgNPs-CFX: Silica coated silver nanoparticles loaded with ciprofloxacin.

Superscripts on the right of the means indicate statistical differences (p < 0.05) between the groups. Values are means ± standard error. Values are means ± standard errors for N = 4. Statistical analysis was by one way ANOVA.

Table 3. Serum kidney function tests of rats treated with blank and loaded silica-coated silver nanoparticles with ciprofloxacin.

Test Group	Urea (mmol/L)	Creatinine (μmol/L)	Total Calcium (mmol/L)	Phosphate (mmol/L)
CN	34.0 ± 2.12^a	0.37 ± 0.22^ab	11.53 ± 0.48^a	7.99 ± 0.91^a
LD-S-AgNPs	30.33 ± 4.37^a	0.45 ± 0.05^ab	13.4 ± 0.75^b	9.7 ± 0.69^b
MD-S-AgNPs	33.5 ± 1.32^a	0.58 ± 0.24^b	12.78 ± 0.37^b	8.33 ± 0.37^a
HD-S-AgNPs	30.0 ± 3.92^a	0.55 ± 0.01^a	13.08 ± 0.39^b	8.23 ± 0.49^a
LD-S-AgNPs-CSCFX	34.6 ± 2.91^a	0.33 ± 0.05^ab	13.23 ± 0.25^b	8.13 ± 0.23^a
MD-S-AgNPs-CSCFX	36.5 ± 1.55^a	0.39 ± 0.06 ^ab	13.20 ± 0.35^b	8.2 ± 0.32^a
HD-S-AgNPs-CSCFX	36.25 ± 2.17^a	0.34 ± 0.05^ab	12.95 ± 0.21^b	8.1 ± 0.30^a

CN: Control negative; LD: Low dose (30 mg/Kg body weight); MD: Median Dose (60 mg/Kg body weight); and HD: High Dose (90 mg/Kg body weight); S-AgNPs: Silica coated silver nanoparticles; S-AgNPs-CFX: Silica coated silver nanoparticles loaded with ciprofloxacin.

Superscripts on the right of the means indicate statistical differences (p < 0.05) between the groups.Values are means ± standard error. Values are means ± standard errors for N = 4. Statistical analysis was by one way ANOVA.

Table 4. Lipid profile and hormone values of rats treated with blank and loaded silica-coated silver nanoparticles with ciprofloxacin.

Test Group	Cholesterol (mg/dL)	Triglyceride (TG) (mg/dL)	High-Density Lipoprotein (HDL) (mg/dL)	Low-Density Lipoprotein (LDL) (mg/dL)	Very low-Density Lipoprotein (VLDL)(mg/dL)	Procalcitonin% (PCT)
CN	78.25 ± 11.65^a	62.75 ± 18.74^a	18.13 ± 1.95^a	27.45 ± 5.29^a	12.55 ± 3.75^a	0.38 ± 0.03^b
LD-S-AgNPs	78.0 ± 2.31^a	99.33 ± 15.93^b	17.23 ± 1.04^a	22.77 ± 4.48^a	19.87 ± 3.19^b	0.29 ± 0.01^a
MD-S-AgNPs	80.23 ± 2.46^a	102.0 ± 7.67^b	18.9 ± 0.20^a	24.70 ± 1.59^a	20.4 ± 1.53^b	0.36 ± 0.02^ab
HD-S-AgNPs	74.75 ± 10.55^a	99.0 ± 5.82^b	18.05 ± 1.06^a	23.18 ± 2.189^a	19.8 ± 1.16^b	0.32 ± 0.04^ab
LD-S-AgNPs-CSCFX	78.5 ± 1.94^a	88.50 ± 9.95^ab	20.38 ± 0.96^b	25.08 ± 0.21^a	17.65 ± 1.97^ab	0.38 ± 0.03^b
MD-S-AgNPs-CSCFX	80.36 ± 9.11^a	93.75 ± 8.29^ab	18.55 ± 1.18^a	23.90 ± 2.25^a	18.75 ± 1.66^ab	0.37 ± 0.02 ^ab
HD-S-AgNPs-CSCFX	74.5 ± 4.17^a	72.0 ± 1.63^ab	17.4 ± 0.426^a	22.55 ± 1.07^a	14.3 ± 0.33^ab	0.34 ± 0.03^ab

CN: Control negative; LD: Low dose (30 mg/Kg body weight); MD: Median Dose (60 mg/Kg body weight); and HD: High Dose (90 mg/Kg body weight); S-AgNPs: Silica coated silver nanoparticles; S-AgNPs-CFX: Silica coated silver nanoparticles loaded with ciprofloxacin.

Superscripts on the right of the means indicate statistical differences (p < 0.05) between the groups.Values are means ± standard error. Values are means ± standard errors for N = 4. Statistical analysis was by one way ANOVA.

Table 5A. Hematological values of rats treated with blank and loaded silica-coated silver nanoparticles with ciprofloxacin.

T Test Group	Red Blood Cell (RBC) (10^6/μL)	Hemoglobin (Hgb) (g/dL)	Hematocrit% (HCT)	Mean Corpuscular Volume (MCV) (μmol)	Mean Corpuscular Hemoglobin (MCH) (Pg)	Mean Corpuscular Hemoglobin Concentration (MCHC) (g/dL)	RDWC%	RDWS (μm)
CN	5.93 ± 0.40^a	13.98 ± 0.84^a	45.78 ± 3.20^a	71.55 ± 1.7^a	21.95 ± 1.13^a	30.65 ± 0.96^a	18.18 ± 0.74^a	49.5 ± 42.88^a
LD-S-AgNPs	6.02 ± 0.17^a	14.13 ± 0.31^a	43.7±.97^a	72.65 ± 1.34^a	23.48 ± 45^ab	32.35 ± 0.94^a	17.8 ± 0.51^a	49.4 ± 40.77^a
MD-S-AgNPs	6.16 ± 0.16^a	14.4 ± 0.27^a	44.8 ± 1.69^a	72.7 ± 1.81^a	23.4 ± 0.63^ab	32.23 ± 0.85^a	16.3 ± 0.69^a	43.73 ± 35.89^a
HD-S-AgNPs	6.02 ± 0.21^a	14.28 ± 0.47^a	45.2 ± 1.53^a	75.43 ± 3.98^a	23.75 ± 0.15^ab	31.73 ± 1.58^a	17.88 ± 1.47^a	52.6 ± 21.52^a
LD-S-AgNPs-CSCFX	6.39 ± 0.199^a	15.1 ± 0.17^a	47.73 ± 1.28^a	75.08 ± 3.97^a	23.7 ± 0.70^ab	31.73 ± 0.71^a	17.35 ± 1.55^a	48.65 ± 27.61^a
MD-S-AgNPs-CSCFX	5.97 ± 0.166^a	14.63 ± 0.13^a	44.85 ± 0.66^a	75.18 ± 1.29^a	24.55 ± 0.86^b	32.65 ± 0.66^a	15.9 ± 0.22^a	44.05 ± 41.93^a
HD-S-AgNPs-CSCFX	6.14 ± 0.34^a	14.4 ± 0.46^a	44.33 ± 1.28^a	72.68 ± 2.93^a	23.55 ± 0.59^ab	32.48 ± 0.69^a	17.73 ± 1.18^a	49.85 ± 32.35^a

Table 5B. Hematological values of rats treated with blank and loaded silica-coated silver nanoparticles with ciprofloxacin.

Test Group	Platelets (PLT) (103/μL)	Mean Platelet Volume (MPV) (μm3)	Platelet Distribution Width% (PDW)	Platelet-Large Cell Ratio% (PLCR)	White Blood Cells (WBC) (103 /μL)	Lymphocyte%	Monocyte%	Granulocyte%
CN	456.0 ± 59.17^b	8.58 ± 0.73^b	17.7 ± 0.73^a	17.35 ± 5.35^ab	7.8±.69^a	74.90 ± 4.84^a	6.98 ± 0.73^b	18.1 ± 4.74^b
LD-S-AgNPs	303.75 ± 16.40^a	9.65 ± 0.29^a	16.7 ± 0.88^a	17.3 ± 2.15 ^cd	10.23±.84^a	83.45 ± 1.67^b	5.43 ± 0.67^ab	11.13 ± 1.01^a
MD-S-AgNPs	443.25 ± 26.73^b	8.25 ± 0.20^ab	17.25 ± 1.04^a	14.6 ± 1.81^a	14.58 ± 2.14^b	87.73±.54^b	4.98 ± 0.23^a	7.3 ± 0.53^a
HD-S-AgNPs	406.75 ± 57.79^ab	7.88 ± 0.31^ab	17.05 ± 0.75^a	12.5 ± 1.87^a	15.23 ± 1.37^b	86.88 ± 2.48^b	4.98±.71^a	8.15 ± 1.80^a
LD-S-AgNPs-CSCFX	373.5 ± 33.93^ab	10.25 ± 0.45^b	17.25 ± 0.76^a	19.45 ± 3.73^d	15.95±.96^b	83.18 ± 2.72^b	5.15 ± 0.46^ab	10.68 ± 2.23^a
MD-S-AgNPs-CSCFX	419.5 ± 15.44^ab	8.85 ± 0.12^ab	18.3 ± 0.39^a	18.35 ± 1.1^abc	15.55 ± 1.95^b	87.23±.74^b	5.3 ± 0.51^ab	7.48 ± 0.51^a
HD-S-AgNPs-CSCFX	359.5 ± 21.75^ab	9.53 ± 0.23^ab	17.50 ± 1.12^a	24.95 ± 3.07^bcd	18.73 ± 1.63^b	89.38±.87^b	5.05 ± 0.44^a	5.58 ± 0.50^a

Figure 6. Photomicrograph of liver from groups; G1: Control negative group received (D.W); G2: Low dose group (G2) treated with 30 mg/kg of S-AgNPs-CSCFX; G3: Medium dose group (G3), treated with 60 mg/kg of S-AgNPs-CSCFX; G4: High dose group (G4) treated with 90 mg/kg of S-AgNPs-CSCFX; G5: Low dose group (G5) treated with 30 mg/kg of blank S-AgNPs; G6: Medium dose group (G6) treated with 60 mg/kg of blank S-AgNPs; G7: High dose group (G7) treated with 90 mg/kg of blank S-AgNPs. H&E. Scale bar: 4 mm.

Figure 7. Photomicrograph of kidney from groups; G1: Control negative group received (D.W); G2: Low dose group (G2) treated with 30 mg/kg of S-AgNPs-CSCFX; G3: Medium dose group (G3), treated 60 mg/kg of S-AgNPs-CSCFX; G4: High dose group (G4) treated with 90 mg/kg of S-AgNPs-CSCFX; G5: Low dose group (G5), treated with 30 mg/kg of blank S-AgNPs; G6: Medium dose group (G6), treated with 60 mg/kg of blank S-AgNPs; G7: high dose group (G7), treated with 90 mg/kg of blank S-AgNPs; H&E. Scale bar: 4 mm.

Data availability Statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.