TY - JOUR
T1 - Chitosan Nanoparticles
T2 - shedding Light on Immunotoxicity and Hemocompatibility
AU - Jesus, Sandra
AU - Marques, Ana Patrícia
AU - Duarte, Alana
AU - Soares, Edna
AU - Costa, João Panão
AU - Colaço, Mariana
AU - Schmutz, Mélanie
AU - Som, Claudia
AU - Borchard, Gerrit
AU - Wick, Peter
AU - Borges, Olga
N1 - Funding Information:
This work was financed by the European Regional Development Fund (ERDF), through the Centro 2020 Regional Operational Programme and through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalization and Portuguese national funds via FCT – Fundação para a Ciência e Tecnologia, under project PROSAFE/0001/2016 and POCI-01-0145-FEDER-030331, and the strategic project UIDB/04539/2020. This work is part of the GoNanoBioMat project and has received funding from the Horizon 2020 framework program of the European Union, ProSafe Joint Transnational Call 2016 and from the CTI (1.1.2018 Innosuisse), under grant agreement Number 19267.1 PFNM-NM.
Funding Information:
The authors would like to thank Dra Ana Donato from the Faculty of Pharmacy Clinical Laboratory Analysis (University of Coimbra) for the hematological studies support, Dra Mónica Zuzarte from iLAB - Bioimaging Laboratory of the Faculty of Medicine of the University of Coimbra for TEM analysis, and Primex for supplying Chitoclear©R chitosan polymers. NMR data was collected at the UC-NMR facility which is supported in part by FEDER – European Regional Development Fund through the COMPETE Programme (Operational Programme for Competitiveness) and by National Funds through FCT – Fundação para a Ciência e a Tecnologia (Portuguese Foundation for Science and Technology) through grants RECI/QEQ-QFI/0168/2012, CENTRO-07-CT62-FEDER-002012, and also through support to Rede Nacional de Ressonância Magnética Nuclear (RNRMN) and to Coimbra Chemistry Centre through grant UID/QUI/00313/2019.
Funding Information:
The authors would like to thank Dra Ana Donato from the Faculty of Pharmacy Clinical Laboratory Analysis (University of Coimbra) for the hematological studies support, Dra M?nica Zuzarte from iLAB - Bioimaging Laboratory of the Faculty of Medicine of the University of Coimbra for TEM analysis, and Primex for supplying Chitoclear? chitosan polymers. NMR data was collected at the UC-NMR facility which is supported in part by FEDER ? European Regional Development Fund through the COMPETE Programme (Operational Programme for Competitiveness) and by National Funds through FCT ? Funda??o para a Ci?ncia e a Tecnologia (Portuguese Foundation for Science and Technology) through grants RECI/QEQ-QFI/0168/2012, CENTRO-07-CT62-FEDER-002012, and also through support to Rede Nacional de Resson?ncia Magn?tica Nuclear (RNRMN) and to Coimbra Chemistry Centre through grant UID/QUI/00313/2019. Funding. This work was financed by the European Regional Development Fund (ERDF), through the Centro 2020 Regional Operational Programme and through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalization and Portuguese national funds via FCT ? Funda??o para a Ci?ncia e Tecnologia, under project PROSAFE/0001/2016 and POCI-01-0145-FEDER-030331, and the strategic project UIDB/04539/2020. This work is part of the GoNanoBioMat project and has received funding from the Horizon 2020 framework program of the European Union, ProSafe Joint Transnational Call 2016 and from the CTI (1.1.2018 Innosuisse), under grant agreement Number 19267.1 PFNM-NM.
Publisher Copyright:
© Copyright © 2020 Jesus, Marques, Duarte, Soares, Costa, Colaço, Schmutz, Som, Borchard, Wick and Borges.
PY - 2020/2/21
Y1 - 2020/2/21
N2 - Nanoparticles (NPs) assumed an important role in the area of drug delivery. Despite the number of studies including NPs are growing over the last years, their side effects on the immune system are often ignored or omitted. One of the most studied polymers in the nano based drug delivery system field is chitosan (Chit). In the scientific literature, although the physicochemical properties [molecular weight (MW) or deacetylation degree (DDA)] of the chitosan, endotoxin contamination and appropriate testing controls are rarely reported, they can strongly influence immunotoxicity results. The present work aimed to study the immunotoxicity of NPs produced with different DDA and MW Chit polymers and to benchmark it against the polymer itself. Chit NPs were prepared based on the ionic gelation of Chit with sodium tripolyphosphate (TPP). This method allowed the production of two different NPs: Chit 80% NPs (80% DDA) and Chit 93% NPs (93% DDA). In general, we found greater reduction in cell viability induced by Chit NPs than the respective Chit polymers when tested in vitro using human peripheral blood monocytes (PBMCs) or RAW 264.7 cell line. In addition, Chit 80% NPs were more cytotoxic for PBMCs, increased reactive oxygen species (ROS) production (above 156 μg/mL) in the RAW 264.7 cell line and interfered with the intrinsic pathway of coagulation (at 1 mg/mL) when compared to Chit 93% NPs. On the other hand, only Chit 93% NPs induced platelet aggregation (at 2 mg/mL). Although Chit NPs and Chit polymers did not stimulate the nitric oxide (NO) production in RAW 264.7 cells, they induced a decrease in lipopolysaccharide (LPS)-induced NO production at all tested concentrations. None of Chit NPs and polymers caused hemolysis, nor induced PBMCs to secrete TNF-α and IL-6 cytokines. From the obtained results we concluded that the DDA of the Chit polymer and the size of Chit NPs influence the in vitro immunotoxicity results. As the NPs are more cytotoxic than the corresponding polymers, one should be careful in the extrapolation of trends from the polymer to the NPs, and in the comparisons among delivery systems prepared with different DDA chitosans.
AB - Nanoparticles (NPs) assumed an important role in the area of drug delivery. Despite the number of studies including NPs are growing over the last years, their side effects on the immune system are often ignored or omitted. One of the most studied polymers in the nano based drug delivery system field is chitosan (Chit). In the scientific literature, although the physicochemical properties [molecular weight (MW) or deacetylation degree (DDA)] of the chitosan, endotoxin contamination and appropriate testing controls are rarely reported, they can strongly influence immunotoxicity results. The present work aimed to study the immunotoxicity of NPs produced with different DDA and MW Chit polymers and to benchmark it against the polymer itself. Chit NPs were prepared based on the ionic gelation of Chit with sodium tripolyphosphate (TPP). This method allowed the production of two different NPs: Chit 80% NPs (80% DDA) and Chit 93% NPs (93% DDA). In general, we found greater reduction in cell viability induced by Chit NPs than the respective Chit polymers when tested in vitro using human peripheral blood monocytes (PBMCs) or RAW 264.7 cell line. In addition, Chit 80% NPs were more cytotoxic for PBMCs, increased reactive oxygen species (ROS) production (above 156 μg/mL) in the RAW 264.7 cell line and interfered with the intrinsic pathway of coagulation (at 1 mg/mL) when compared to Chit 93% NPs. On the other hand, only Chit 93% NPs induced platelet aggregation (at 2 mg/mL). Although Chit NPs and Chit polymers did not stimulate the nitric oxide (NO) production in RAW 264.7 cells, they induced a decrease in lipopolysaccharide (LPS)-induced NO production at all tested concentrations. None of Chit NPs and polymers caused hemolysis, nor induced PBMCs to secrete TNF-α and IL-6 cytokines. From the obtained results we concluded that the DDA of the Chit polymer and the size of Chit NPs influence the in vitro immunotoxicity results. As the NPs are more cytotoxic than the corresponding polymers, one should be careful in the extrapolation of trends from the polymer to the NPs, and in the comparisons among delivery systems prepared with different DDA chitosans.
KW - Chitosan nanoparticle
KW - Deacetylation degree
KW - Endotoxin-free
KW - Hemocompatibility
KW - Immunotoxicity
KW - Inflammation
KW - PBMCs
KW - Reactive oxygen species
UR - http://www.scopus.com/inward/record.url?scp=85081661818&partnerID=8YFLogxK
U2 - 10.3389/fbioe.2020.00100
DO - 10.3389/fbioe.2020.00100
M3 - Article
C2 - 32154232
AN - SCOPUS:85081661818
SN - 2296-4185
VL - 8
JO - Frontiers in Bioengineering and Biotechnology
JF - Frontiers in Bioengineering and Biotechnology
M1 - 100
ER -