Optimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on Macrophages

Yee Huang,1 Li Nan,1,2 Chenwen Xiao,1 Quanan Ji,1 Ke Li,1 Qiang Wei,1 Yan Liu,1 Guolian Bao1 1Institute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Science, Hangzhou 310021, People’s Republic of China; 2Zhejiang Normal University, Jinhua 321000, People&...

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Autores principales:	Huang Y, Nan L, Xiao C, Ji Q, Li K, Wei Q, Liu Y, Bao G
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2019
Materias:	rehmannia glutinosa polysaccharide pegylation preparation technology lyophilization method macrophages Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/58f43544a9f34b46a71111480f7e1484
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spelling	oai:doaj.org-article:58f43544a9f34b46a71111480f7e14842021-12-02T06:20:52ZOptimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on Macrophages1178-2013https://doaj.org/article/58f43544a9f34b46a71111480f7e14842019-11-01T00:00:00Zhttps://www.dovepress.com/optimum-preparation-method-for-self-assembled-pegylation-nano-adjuvant-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Yee Huang,1 Li Nan,1,2 Chenwen Xiao,1 Quanan Ji,1 Ke Li,1 Qiang Wei,1 Yan Liu,1 Guolian Bao1 1Institute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Science, Hangzhou 310021, People’s Republic of China; 2Zhejiang Normal University, Jinhua 321000, People’s Republic of ChinaCorrespondence: Yan Liu; Guolian BaoInstitute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Science, Hangzhou 310021, People’s Republic of ChinaEmail 35792191@qq.com; baoguolian@163.comBackground: Rehmannia glutinosa polysaccharide is the main reason that contributes to the immunological function of R. glutinosa. Due to its disadvantages in clinical use, here we designed the PEGylation nano-RGP (pRL) to improve the drug-targeting effect and the immunological function. Our present work aims to establish the optimum condition of preparing the pRL and to investigate its immunological function on macrophages.Methods: pRL was prepared by thin film hydration method combined with ultra-sonication technique. And its preparation conditions were optimized with response surface methodology. Also, the lyophilization method was optimized. The characteristics of the pRL were evaluated, including particle size, drug loading, encapsulation efficiency and morphology. The immunological function of pRL on macrophage was investigated through CCK-8 test, ELISA and flow cytometry.Results: The lipid-to-cholesterol molar ratio of 8:1, the addition of DSPE-PEG2000 of 9% and the lipid-to-drug ratio of 5.4:1 were the optimum preparation technology for pRL. The encapsulation efficiency (EE) of pRL under this preparation technology was 95.81±1.58%, with a particle size of 31.98 ± 2.6 nm. The lactose-to-lipid ratio (2:1) was the optimal lyophilization method. pRL promoted macrophage proliferation, which is significantly better than that of nano-RGP without PEGylation (RL). pRL-stimulated RAW264.7 cells showed a high secretion of pro-inflammatory cytokines, which is the characteristic indicator of M1 polarization. Enhanced cellular uptake through macropinocytosis-dependent and caveolae-mediated endocytosis was observed in pRL-treated RAW264.7 cells.Conclusion: Our study concluded that PEGylation effectively overcame the poor targeting effect of Rehmannia glutinosa polysaccharide (RGP) and significantly improved the immunological profile of its nano-formulation, which suggested that pRL could serve as an immune adjuvant in clinical application.Keywords: Rehmannia glutinosa polysaccharide, PEGylation, preparation technology, lyophilization method, macrophagesHuang YNan LXiao CJi QLi KWei QLiu YBao GDove Medical Pressarticlerehmannia glutinosa polysaccharidepegylationpreparation technologylyophilization methodmacrophagesMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 14, Pp 9361-9375 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	rehmannia glutinosa polysaccharide pegylation preparation technology lyophilization method macrophages Medicine (General) R5-920
spellingShingle	rehmannia glutinosa polysaccharide pegylation preparation technology lyophilization method macrophages Medicine (General) R5-920 Huang Y Nan L Xiao C Ji Q Li K Wei Q Liu Y Bao G Optimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on Macrophages
description	Yee Huang,1 Li Nan,1,2 Chenwen Xiao,1 Quanan Ji,1 Ke Li,1 Qiang Wei,1 Yan Liu,1 Guolian Bao1 1Institute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Science, Hangzhou 310021, People’s Republic of China; 2Zhejiang Normal University, Jinhua 321000, People’s Republic of ChinaCorrespondence: Yan Liu; Guolian BaoInstitute of Animal Husbandry and Veterinary Medicine, Zhejiang Academy of Agricultural Science, Hangzhou 310021, People’s Republic of ChinaEmail 35792191@qq.com; baoguolian@163.comBackground: Rehmannia glutinosa polysaccharide is the main reason that contributes to the immunological function of R. glutinosa. Due to its disadvantages in clinical use, here we designed the PEGylation nano-RGP (pRL) to improve the drug-targeting effect and the immunological function. Our present work aims to establish the optimum condition of preparing the pRL and to investigate its immunological function on macrophages.Methods: pRL was prepared by thin film hydration method combined with ultra-sonication technique. And its preparation conditions were optimized with response surface methodology. Also, the lyophilization method was optimized. The characteristics of the pRL were evaluated, including particle size, drug loading, encapsulation efficiency and morphology. The immunological function of pRL on macrophage was investigated through CCK-8 test, ELISA and flow cytometry.Results: The lipid-to-cholesterol molar ratio of 8:1, the addition of DSPE-PEG2000 of 9% and the lipid-to-drug ratio of 5.4:1 were the optimum preparation technology for pRL. The encapsulation efficiency (EE) of pRL under this preparation technology was 95.81±1.58%, with a particle size of 31.98 ± 2.6 nm. The lactose-to-lipid ratio (2:1) was the optimal lyophilization method. pRL promoted macrophage proliferation, which is significantly better than that of nano-RGP without PEGylation (RL). pRL-stimulated RAW264.7 cells showed a high secretion of pro-inflammatory cytokines, which is the characteristic indicator of M1 polarization. Enhanced cellular uptake through macropinocytosis-dependent and caveolae-mediated endocytosis was observed in pRL-treated RAW264.7 cells.Conclusion: Our study concluded that PEGylation effectively overcame the poor targeting effect of Rehmannia glutinosa polysaccharide (RGP) and significantly improved the immunological profile of its nano-formulation, which suggested that pRL could serve as an immune adjuvant in clinical application.Keywords: Rehmannia glutinosa polysaccharide, PEGylation, preparation technology, lyophilization method, macrophages
format	article
author	Huang Y Nan L Xiao C Ji Q Li K Wei Q Liu Y Bao G
author_facet	Huang Y Nan L Xiao C Ji Q Li K Wei Q Liu Y Bao G
author_sort	Huang Y
title	Optimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on Macrophages
title_short	Optimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on Macrophages
title_full	Optimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on Macrophages
title_fullStr	Optimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on Macrophages
title_full_unstemmed	Optimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on Macrophages
title_sort	optimum preparation method for self-assembled pegylation nano-adjuvant based on rehmannia glutinosa polysaccharide and its immunological effect on macrophages
publisher	Dove Medical Press
publishDate	2019
url	https://doaj.org/article/58f43544a9f34b46a71111480f7e1484
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Optimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on Macrophages

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