TABLE 1.
Summary of Chit-Ins NPs production protocols by ionotropic-gelation method.
| Preparation method | NP characterization | ||||||||||||
| System | Insulin, source | Chitosan, source | Chitosan solution | Insulin solution | TPP solution | Final pH | Size | Zeta Potential | Insulin AE% | Toxicity assay | Anti-insulin IgG | References | |
| Chit NPs | Porcine pancreas insulin, Sigma | 186 kDa; 85% DDA Aldrich Chemicals | 8 mL chitosan 2 mg/mL | Insulin 31.65 μg/mL to 235.25 | 1 mL premixed with TPP or | 4 mL TPP 1 mg/mL | pH 2.8 to 6.1 | 237 nm ±53 nm to 325 nm ±45 nm | – | 2–85% | – | – | Ma et al., 2002 |
| Chit NPs | Insulin 27.6 I.U/mg, Xuzhou biochemical plant | ? kDa 88.9% DDA; viscosity 45 mPa.s Shenyang | 4 mL chitosan 2.6 mg/mL | Concen- tration? mins/mChit = 0.1 | Solution Premixed with TPP solution | ? mL TPP 0.45 mg/mL | – | 265.3 nm ±34.1 nm | +40.71 mV ±0.69 mV | 88.6% ± 2.4% | – | – | Pan et al., 2002 |
| Chit NPs | Porcine pancreas insulin 27.8 USP/mg, Sigma Chemicals | 186 kDa; 85% DDA Aldrich Chemical, Milwaukee | 8 mL chitosan 2 mg/mL | Insulin 2 mg/mL in 0.01 M HCl | 1 mL premixed with TPP solution | 4 mL TPP 1 mg/mL in 0.05 M NaOH | pH 5.3 | 269 nm ± 7 nm | +34.9 mV ± 0.9 mV | 38.5% ± 1.5% | – | – | Ma et al., 2005 |
| 4 mL TPP 1 mg/mL in 0.075 M NaOH | pH 6.1 | 339 nm ± 8 nm | +21.8 mV ± 0.6 mV | 78.5% ± 2.3% | – | – | |||||||
| Chit NPs | Human insulin Novolin R®, 100 IU/mL | Low viscosity chitosan ? kDa; DDA ? | 5 mL chitosan 4 mg/mL | Insulin solution 4.6 mg/Ml | Premixed with TPP solution | 2 ml TPP 1 mg/mL | pH 6.1 | 312.8 nm PDI 0.48 | +23 mV ± 2 mV | 69.37% ± 4.71% | – | – | Azevedo et al., 2011 |
| Chit NPs | Bovine pancreas insulin (27 USP/mg) Sigma-Aldrich, United States | 200 kDa DDA ? Sigma-Aldrich, United States | ? mL chitosan 2 mg/mL | Insulin solution 0.5 mg/ml | Premixed with TPP solution | ? ml TPP 0.5 mg/mL | pH 5.5 | 215 nm PDI 0.16 | +20.7 mV ± 0.7 mV | 49.43% ± 0.44% | – | – | Makhlof et al., 2011 |
| Chit NPs | Crystalline recombinant human insulin Novo Nordisk, Denmark | LMWC; 98% DDA; viscosity 22 cP | 10 mL chitosan 1 mg/mL or 3 mg/mL | Insulin 0.5 mg/mL and 1 mg/mL (concentration in TPP) | Premixe d with TPP solution | ? ml TPP solution 1 mg/mL and 3 mg/mL | – | 261 nm PDI 0.4 or 419 nm PDI 0.45 | +27.2 mV or +48.4 mV | 61.61% ± 4.52% or 61.88% ± 5.59% | – | – | Kouchak et al., 2012 |
| MMWC; 92% DDA; viscosity 715 cP Primex, Iceland | 10 mL; chitosan 0.5 mg/mL or 1 mg/mL | 132 nm PDI 0.28 or 343 nm PDI 0.49 | +25.1 mV or +39.3 mV | 70.89% ± 3.32% or 70.59% ± 1.70% | – | – | |||||||
| HMWC; 96% DDA; viscosity 1234 cP Primex, Iceland | 10 mL; chitosan 0.5 mg/mL or 1 mg/mL | 112 nm PDI 0.27 or 160 nm PDI 0.28 | +27.5 mV or +29.0 mV | 53.50% ± 2.61% or 53.73% ± 2.29% | – | – | |||||||
| Chit NPs | Zinc-free human insulin | 150 kDa; 87% DDA; viscosity 2.37 dL/g Sigma-Aldrich, Missouri | ? mL chitosan 2.5 mg/mL (in acetic acid) | 4 mg/mL insulin solution | Premixed with TPP solution | ? mL TPP 0.25 mg/mL | pH 5.5 | 330 nm ± 36 nm | +30 mV ± 4 mV | 55% ± 8% | No death or inflammatory response (CAM assay in fertilized chicken eggs) | – | Rampino et al., 2013 |
| Chit NPs | Insulin 27.5 IU/mg Jiangsu Wangbang Bio- Technology | 400 kDa; DDA? Haixin Biological Product | ? mL chitosan 50 mg (in acetic acid) | 4 mg insulin solution in NaOH | Premixed with chitosan solution | 3 mL TPP solution 0.5 mg/mL | pH 3 | 91.28 nm ± 7.9 nm to 220.2 nm ± 9.5 nm | +14.4 mV ± 2.9 mV | 93.1% | – | – | Zhao et al., 2014 |
| Chit NPs into transdermal patch | Pure insulin powder Sigma- Aldrich | LMWC; DDA? Sigma-Aldrich | ? mL chitosan 1.5 mg/mL or 2 mg/mL (in acetic acid) | 1 mL of insulin 20 mg/mL | Premixed with chitosan solution | ? mL TPP 2.5 mg/mL | pH 5 | 465 nm or 661 nm | – | 77.3% ± 0.5% to 78.9% ± 0.25% | – | – | Sadhasivam et al., 2015 |
| Chit-TPP- micro emulsion | Recombinant human insulin (Humulin R 100 IU/mL) Eli Lilly and Company, | MMWC; 75% to 85% DDA Sigma-Aldrich, United States | ? mL chitosan 3 mg/mL (in acetic acid) | – | Insulin added to solution after NPs formation | ? mL TPP solution 1 mg/mL | – | 80.8 nm ± 7.0 nm to 401.8 nm ± 41.7 nm | +38.1 mV to +47.0 mV | – | Viability depend on concentration (XTT assay) | – | Erel et al., 2016 |
Cs NPs, chitosan nanoparticles; TPP, tripolyphosphate; AE, association efficiency; CAM assay, Chick Chorioallantoic Membrane assay; XTT assay, Cell Viability Assay. Units were converted to standardization, so they can differ from the ones at the original paper. ? refers to data that could not be confirmed in the respective publication and thus remain unknown.