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&...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Dove Medical Press
2019-11-01
|
| Series: | International Journal of Nanomedicine |
| Subjects: | |
| Online Access: | https://www.dovepress.com/optimum-preparation-method-for-self-assembled-pegylation-nano-adjuvant-peer-reviewed-article-IJN |
| _version_ | 1818992366343684096 |
|---|---|
| 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 |
| collection | DOAJ |
| 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 |
| first_indexed | 2024-12-20T20:25:01Z |
| format | Article |
| id | doaj.art-58f43544a9f34b46a71111480f7e1484 |
| institution | Directory Open Access Journal |
| issn | 1178-2013 |
| language | English |
| last_indexed | 2024-12-20T20:25:01Z |
| publishDate | 2019-11-01 |
| publisher | Dove Medical Press |
| record_format | Article |
| series | International Journal of Nanomedicine |
| spelling | doaj.art-58f43544a9f34b46a71111480f7e14842022-12-21T19:27:29ZengDove Medical PressInternational Journal of Nanomedicine1178-20132019-11-01Volume 149361937550091Optimum Preparation Method for Self-Assembled PEGylation Nano-Adjuvant Based on Rehmannia glutinosa Polysaccharide and Its Immunological Effect on MacrophagesHuang YNan LXiao CJi QLi KWei QLiu YBao GYee 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, macrophageshttps://www.dovepress.com/optimum-preparation-method-for-self-assembled-pegylation-nano-adjuvant-peer-reviewed-article-IJNrehmannia glutinosa polysaccharidepegylationpreparation technologylyophilization methodmacrophages |
| spellingShingle | 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 International Journal of Nanomedicine rehmannia glutinosa polysaccharide pegylation preparation technology lyophilization method macrophages |
| title | 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_short | 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 |
| topic | rehmannia glutinosa polysaccharide pegylation preparation technology lyophilization method macrophages |
| url | https://www.dovepress.com/optimum-preparation-method-for-self-assembled-pegylation-nano-adjuvant-peer-reviewed-article-IJN |
| work_keys_str_mv | AT huangy optimumpreparationmethodforselfassembledpegylationnanoadjuvantbasedonrehmanniaglutinosapolysaccharideanditsimmunologicaleffectonmacrophages AT nanl optimumpreparationmethodforselfassembledpegylationnanoadjuvantbasedonrehmanniaglutinosapolysaccharideanditsimmunologicaleffectonmacrophages AT xiaoc optimumpreparationmethodforselfassembledpegylationnanoadjuvantbasedonrehmanniaglutinosapolysaccharideanditsimmunologicaleffectonmacrophages AT jiq optimumpreparationmethodforselfassembledpegylationnanoadjuvantbasedonrehmanniaglutinosapolysaccharideanditsimmunologicaleffectonmacrophages AT lik optimumpreparationmethodforselfassembledpegylationnanoadjuvantbasedonrehmanniaglutinosapolysaccharideanditsimmunologicaleffectonmacrophages AT weiq optimumpreparationmethodforselfassembledpegylationnanoadjuvantbasedonrehmanniaglutinosapolysaccharideanditsimmunologicaleffectonmacrophages AT liuy optimumpreparationmethodforselfassembledpegylationnanoadjuvantbasedonrehmanniaglutinosapolysaccharideanditsimmunologicaleffectonmacrophages AT baog optimumpreparationmethodforselfassembledpegylationnanoadjuvantbasedonrehmanniaglutinosapolysaccharideanditsimmunologicaleffectonmacrophages |