Generation-dependent effect of PAMAM dendrimers on human insulin fibrillation and thermal stability

Nowacka, Olga; Miłowska, Katarzyna; Belica-Pacha, Sylwia; Pałecz, Bartłomiej; Šipošová, Katarína; Gažová, Zuzana; Bryszewska, Maria

doi:10.1016/j.ijbiomac.2015.10.029

Cited by 15 publications

(7 citation statements)

References 35 publications

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“…All the endothermic and exothermic insulin peaks were indicated with arrows. Figure 03 revealed a minor endothermic peak centered at 77°C (∆H = 0.04 kJ mol -1 ) followed by a major exothermic peak at 85.30 °C (∆H = 0.4 kJ mol -1 ) for insulin, which has been already reported for dendrimer-loaded insulin (Nowacka et al, 2016). The sequence of endothermic followed by exothermic peak could be correlated with protein denaturation followed by aggregation (Sarmento, Veiga, Ferreira, 2006).…”

Section: Resultssupporting

confidence: 72%

Insulin-loaded polymeric mucoadhesive nanoparticles: development, characterization and cytotoxicity evaluation

Gatti

Eloy

Ferreira

et al. 2018

Braz. J. Pharm. Sci.

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Mucoadhesive nanoparticles are particularly interesting for delivery through nasal or pulmonary routes, as an approach to overcome the mucociliary clearance. Moreover, these nanoparticles are attractive for peptide and protein delivery, particularly for insulin to treat diabetes, as an alternative to conventional parenteral administration. Thus, chitosan, a cationic mucoadhesive polysaccharide found in shells of crustaceans, and the negatively-charged dextran sulfate are able to form nanoparticles through ionic condensation, representing a potential insulin carrier. Herein, chitosan/dextran sulfate nanoparticles at various ratios were prepared for insulin loading. Formulations were characterized for particle size, zeta potential, encapsulation efficiency, scanning electron microscopy, differential scanning calorimetry, and in vitro drug release. Moreover, the interaction with mucin and the cytotoxicity against a lung cell line were studied, which altogether have not been addressed before. Results evidenced that a proper selection of polyelectrolytes is necessary for smaller particle size formation and also the composition and zeta potential impact encapsulation efficiency, which is benefited by the positive charge of chitosan. Insulin remained stable after encapsulation as evidenced by calorimetric assays, and was released in a sustained manner in the first 10 h. Positively-charged nanoparticles based on chitosan/dextran-sulfate at the ratio of 6:4 successfully interacted with mucin, which is a prerequisite for delivery to mucus-containing tissues. Finally, insulin-loaded nanoparticles displayed no cytotoxicity effect against lung cells at tested concentrations, suggesting the potential for further in vivo studies.

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Section: Resultssupporting

confidence: 72%

Insulin-loaded polymeric mucoadhesive nanoparticles: development, characterization and cytotoxicity evaluation

Gatti

Eloy

Ferreira

et al. 2018

Braz. J. Pharm. Sci.

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“…The activity of the enzyme without incubation was defined as 100%. T m values [the temperature at which half protein molecule denature] were determined by differential scanning calorimetry (DSC) as previously described [ 36 , 37 ]. The samples were diluted to 0.2 mg mL −1 in 20 mM Tris/HCl buffer (pH 7.0) and heated from 40°C to 105°C at a rate of 2°C min −1 .…”

Section: Methodsmentioning

confidence: 99%

Improving the Thermostability of Acidic Pullulanase from Bacillus naganoensis by Rational Design

et al. 2016

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Pullulanase (EC 3.2.1.41) plays an important role in the specific hydrolysis of branch points in amylopectin. Enhancing its thermostability is required for its industrial application. In this study, rational protein design was used to improve the thermostability of PulB from Bacillus naganoensis (AB231790.1), which has strong enzymatic properties. Three positive single-site mutants (PulB-D328H, PulB-N387D, and PulB-A414P) were selected from six mutants. After incubation at 65°C for 5 min, the residual activities of PulB-D328H, PulB-N387D, and PulB-A414P were 4.5-, 1.7-, and 1.47-fold higher than PulB-WT, and their Tm values (the temperature at which half protein molecule denature) were 1.8°C, 0.4°C, and 0.9°C higher than PulB-WT, respectively. Then the final combined mutant PulB-328/387/414 was constructed. The t1/2 of it was 12.9-fold longer than that of PulB-WT at 65°C and the total increase in Tm of it (5.0°C) was almost 60% greater than the sum of individual increases (3.1°C). In addition, kinetic studies revealed that the kcat and the kcat/Km of PulB-328/387/414 increased by 38.8% and 12.9%. The remarkable improvement in thermostability and the high catalytic efficiency of PulB-328/387/414 make it suitable for industrial applications.

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“…For example, Agarwal et al 21 reported that PAMAM-functionalized filter papers extracted eight and three times more BSA and lysozyme contents from solutions, respectively, compared to regular filter papers. Klajnert et al 22 also showed that PAMAM has a high affinity for BSA, and Nowacka et al 40 demonstrated that insulin interacts strongly with various generations of PAMAM dendrimers.…”

Section: Analyticalmentioning

confidence: 99%

PAMAM-Functionalized Paper as a New Substrate for the Paper Spray Mass Spectrometry Measurement of Proteins

et al. 2023

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Paper surface functionalization with polyamidoamine (PAMAM) dendrimers has been developed for increased sensitivity analysis of proteins by paper spray mass spectrometry (PS-MS). PAMAM is a branched polymeric compound with an ethylenediamine core linked to repeating PAMAM units that generates an outer surface rich in primary amines. These positively charged amine groups can interact electrostatically with negatively charged residues (e.g., aspartate, glutamate) on the protein surface. PAMAM inner amide moieties can also promote hydrogen bonding with protein surface oxygens, making PAMAM a useful material for protein extraction. PAMAM-functionalized PS-MS paper strips were used to extract proteins from biofluids, dipped in acetonitrile to remove unbound constituents, dried, and then measured with PS-MS. The use of this strategy was optimized and compared with unmodified paper strips. PAMAM-functionalized paper substrates provided sixfold greater sensitivity for albumin, 11-fold for hemoglobin, sevenfold for insulin, and twofold for lysozyme. The analytical performance of the functionalized paper substrate was evaluated through the analysis of albumin in urine, achieving linearity with R 2 > 0.99, LOD of 1.1 μg mL −1 , LOQ of 3.8 μg mL −1 , precision better than 10%, and relative recovery 70− 83%. The method was applied to quantify urinary albumin from nine anonymous patient samples (concentrations ranged from 6.5 to 77.4 μg mL −1 ), illustrating its potential for the diagnosis of microalbuminuria. These data demonstrate the utility of paper modification with the PAMAM dendrimer for sensitive PS-MS analysis of proteins, opening a path for further applications in clinical diagnosis through the analysis of disease-related proteins.

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Generation-dependent effect of PAMAM dendrimers on human insulin fibrillation and thermal stability

Cited by 15 publications

References 35 publications

Insulin-loaded polymeric mucoadhesive nanoparticles: development, characterization and cytotoxicity evaluation

Insulin-loaded polymeric mucoadhesive nanoparticles: development, characterization and cytotoxicity evaluation

Improving the Thermostability of Acidic Pullulanase from Bacillus naganoensis by Rational Design

PAMAM-Functionalized Paper as a New Substrate for the Paper Spray Mass Spectrometry Measurement of Proteins

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