English

Pickering Emulsion Templated Proteinaceous Microsphere with Bio-Stimuli Responsiveness

• Weijie Jiang ^1, ,
• Hang Jiang ^{1, ,} ,
• Wei Liu ^1, ,
• Xin Guan ^2, ,
• Yunxing Li ^{1, ,} ,
• Cheng Yang ^1, ,
• To Ngai ^2,

• 1.

  The Key Laboratory of Synthetic and Biological Colloids, Ministry of Education, School of Chemical and Material Engineering, Jiangnan University, Wuxi 214122, Jiangsu Province, China

• 2.

  Department of Chemistry, The Chinese University of Hong Kong, Shatin, N. T., Hong Kong SAR, China

Corresponding author: Hang Jiang, hangjiang@jiangnan.edu.cn Yunxing Li, yunxingli@jiangnan.edu.cn

• Received Date: 29 January 2023
  Accepted Date: 23 February 2023
  Revised Date: 23 February 2023
  Available Online: 15 December 2023
• Fund Project:

Abstract: Bio-stimuli-responsive microspheres, which can encapsulate and release actives in response to physiological triggers, have attracted increasing attention in pharmaceutical, cosmetic, food biotechnology, and agricultural industries. However, most microspheres are based on synthetic polymers and suffer from a lack of biocompatibility due to the residues of harsh organic solvents or crosslinkers used in the synthesis process. Herein, we develop a simple and sustainable method for the construction of proteinaceous microspheres templated from Pickering double emulsions. Specifically, silica nanoparticles with a diameter of 100 nm were synthesized by Stöber method and modified by reacting with dichlorodimethylsilane. The Pickering emulsions are stabilized by hydrophobic silica nanoparticles, while zein protein is dissolved in the middle phase. Subsequent ethanol removal from the emulsion template precipitated the protein skeleton. First, we stained the aqueous ethanol phase with rhodamine B and the oil phase with pyrene to demonstrate the formation of double emulsions by confocal laser scanning microscopy (CLSM). The morphology of microspheres and silica nanoparticles was characterized by scanning electron microscopy (SEM). The obtained microspheres showed high sphericity and uniformity. In addition to acting as particulate stabilizers, the silica nanoparticles could improve the mechanical strength and monodispersity of microspheres. Herein, fluorescein isothiocyanate (FITC)-labeled dextran was chosen as the model active for encapsulation into microspheres. The CLSM images showed that it was uniformly dispersed in the microspheres and had no effect on the structure of the microspheres. Next, we investigated the pH tolerance of the microspheres. Through optical microscope, it was noted that the structure was intact under pH 3–11, and thus, it has a high resistance. Finally, we investigated the bio-stimuli-responsive behavior of microspheres. Zein is rich in sulfur-containing amino acids, which can form intra- and inter-molecular disulfide bonds. Because disulfide bonds can be reduced by glutathione (GSH) and the protein itself has enzymatic hydrolysis characteristics, the proteinaceous microspheres can be triggered release in response to GSH and protease. The release profiles of FITC-dextran from microspheres at different concentrations of GSH and protease were evaluated by fluorescence spectrophotometer. The decomposition behavior of microspheres under certain concentrations of GSH and protease was further verified by CLSM and SEM. To conclude, excellent stability and tunability of emulsion templates render the resulting proteinaceous microspheres with adjustable structures. Meanwhile, the proteinaceous microspheres have high encapsulation efficiency of model actives and have shown excellent bio-stimuli-responsiveness to protease and glutathione.

Key words:
• Bio-stimuli-responsive
•  /  Proteinaceous microsphere
•  /  Pickering emulsion
•  /  Protease
•  /  Glutathione

• 1. [1]

     Kiser, P. F.; Wilson, G.; Needham, D. Nature 1998, 394, 459. doi: 10.1038/28822

  2. [2]

     Caldorera-Moore, M. E.; Liechty, W. B.; Peppas, N. A. Acc. Chem. Res. 2011, 44, 1061. doi: 10.1021/ar2001777

  3. [3]

     Kureha, T.; Aoki, D.; Hiroshige, S.; Iijima, K.; Aoki, D.; Takata, T.; Suzuki, D. Angew. Chem. Int. Ed. 2017, 56, 15393. doi: 10.1002/anie.201709633

  4. [4]

     Zhao, C.; Tian, S.; Liu, Q.; Xiu, K.; Lei, I.; Wang, Z.; Ma, P. X. Adv. Funct. Mater. 2020, 30 (21). doi: 10.1002/adfm.202000776

  5. [5]

     Ruan, L.; Su, M.; Qin, X.; Ruan, Q.; Lang, W.; Wu, M.; Chen, Y.; Lv, Q. Mater. Today Bio 2022, 16, 100394. doi: 10.1016/j.mtbio.2022.100394

  6. [6]

     Rosenblum, D.; Joshi, N.; Tao, W.; Karp, J. M.; Peer, D. Nat. Commun. 2018, 9, 1410. doi: 10.1038/s41467-018-03705-y

  7. [7]

     Lu, Y.; Aimetti, A. A.; Langer, R.; Gu, Z. Nat. Rev. Mater. 2016, 2, 16075. doi: 10.1038/natrevmats.2016.75

  8. [8]

     Wu, C.; Schwibbert, K.; Achazi, K.; Landsberger, P.; Gorbushina, A.; Haag, R. Biomacromolecules 2017, 18, 210. doi: 10.1021/acs.biomac.6b01527

  9. [9]

     Huang, X.; Patil, A. J.; Li, M.; Mann, S. J. Am. Chem. Soc. 2014, 136, 9225. doi: 10.1021/ja504213m

  10. [10]

      Li, J.; Zhang, L.; Liu, Y.; Wen, J.; Wu, D.; Xu, D.; Segura, T.; Jin, J.; Lu, Y.; Wang, H. Chem. Commun. 2016, 52, 13608. doi: 10.1039/c6cc05099a

  11. [11]

      Sun, Z.; Zhao, Q.; Haag, R.; Wu, C. Angew. Chem. Int. Ed. 2021, 60, 8410. doi: 10.1002/anie.202013737

  12. [12]

      Ju, S.; Shin, G.; Lee, M.; Koo, J. M.; Jeon, H.; Ok, Y. S.; Hwang, D. S.; Hwang, S. Y.; Oh, D. X.; Park, J. Green Chem. 2021, 23, 6953. doi: 10.1039/d1gc01588e

  13. [13]

      Jo, Y. K.; Lee, D. Small 2020, 16 (Suppl. 9), 1903736. doi: 10.1002/smll.201903736

  14. [14]

      王义洲, 刘晔宏, 徐首红, 刘洪来. 物理化学学报, 2019, 35, 876. doi: 10.3866/PKU.WHXB201901019Wang, Y.; Liu, Y.; Xu, S.; Liu, H. Acta Phys. -Chim. Sin. 2019, 35, 876. doi: 10.3866/PKU.WHXB201901019

  15. [15]

      廖雨瑶, 范震, 杜建忠. 物理化学学报, 2021, 37, 1912053. doi: 10.3866/PKU.WHXB201912053Liao, Y.; Fan, Z.; Du, J. Acta Phys. -Chim. Sin. 2021, 37, 1912053. doi: 10.3866/PKU.WHXB201912053

  16. [16]

      Stoppel, W. L.; Ghezzi, C. E.; McNamara, S. L.; Black, L. D., Ⅲ; Kaplan, D. L. Ann. Biomed. Eng. 2015, 43, 657. doi: 10.1007/s10439-014-1206-2

  17. [17]

      Li, Y.; Maciel, D.; Rodrigues, J.; Shi, X.; Tomás, H. Chem. Rev. 2015, 115, 8564. doi: 10.1021/cr500131f

  18. [18]

      Sun, M.; Yin, W.; Chen, J.; Wang, W.; Guo, T.; Meng, T. Green Chem. 2021, 23, 740. doi: 10.1039/d0gc02999h

  19. [19]

      Yang, X.; Wang, Y.; Bai, R.; Ma, H.; Wang, W.; Sun, H.; Dong, Y.; Qu, F.; Tang, Q.; Guo, T.; et al. Green Chem. 2019, 21, 2229. doi: 10.1039/c8gc03573c

  20. [20]

      Patel, A. R.; Remijn, C.; Cabero, A. -i. M.; Heussen, P. C. M.; ten Hoorn, J. W. M. S.; Velikov, K. P. Adv. Funct. Mater. 2013, 23, 4710. doi: 10.1002/adfm.201300320

  21. [21]

      Reddy, N.; Yang, Y. Trends Biotechnol. 2011, 29, 490. doi: 10.1016/j.tibtech.2011.05.003

  22. [22]

      Lawton, J. W. Cereal Chem. 2002, 79, 1. doi: 10.1094/CCHEM.2002.79.1.1

  23. [23]

      Argos, P.; Pedersen, K.; Marks, M. D.; Larkins, B. A. J. Biol. Chem. 1982, 257, 9984. doi: 10.1016/s0021-9258(18)33974-7

  24. [24]

      Giteru, S. G.; Ali, M. A.; Oey, I. Food Hydrocolloids 2021, 120, 106948. doi: 10.1016/j.foodhyd.2021.106948

  25. [25]

      Dong, S.; Wang, J. -M.; Cheng, L. -M.; Lu, Y. -L.; Li, S. -H.; Chen, Y. J. Agric. Food Chem. 2017, 65, 7352. doi: 10.1021/acs.jafc.7b02205

  26. [26]

      Manickam, D. S.; Li, J.; Putt, D. A.; Zhou, Q. H.; Wu, C.; Lash, L. H.; Oupicky, D. J. Control. Release 2010, 141, 77. doi: 10.1016/j.jconrel.2009.08.022

  27. [27]

      Zhang, M.; Liu, J.; Kuang, Y.; Li, Q.; Chen, H.; Ye, H.; Guo, L.; Xu, Y.; Chen, X.; Li, C.; et al. J. Mater. Chem. B 2016, 4, 3387. doi: 10.1039/c5tb02548f

  28. [28]

      Zhang, P.; Wu, J.; Xiao, F.; Zhao, D.; Luan, Y. Med. Res. Rev. 2018, 38, 1485. doi: 10.1002/med.21485

  29. [29]

      Su, L.; Li, R.; Khan, S.; Clanton, R.; Zhang, F.; Li, Y. N.; Song, Y.; Wang, H.; Fan, J.; Hernandez, S.; et al. J. Am. Chem. Soc. 2018, 140, 1438. doi: 10.1021/jacs.7b11462

  30. [30]

      Ding, Y.; Kang, Y.; Zhang, X. Chem. Commun. 2015, 51, 996. doi: 10.1039/c4cc05878j

  31. [31]

      de la Rica, R.; Aili, D.; Stevens, M. M. Adv. Drug Delivery Rev. 2012, 64, 967. doi: 10.1016/j.addr.2012.01.002

  32. [32]

      Luo, Y.; Wang, Q. J Appl. Polym. Sci. 2014, 131, 40696. doi: 10.1002/app.40696

  33. [33]

      Zhang, Y.; Cui, L.; Li, F.; Shi, N.; Li, C.; Yu, X.; Chen, Y.; Kong, W. Int. J. Pharm. 2016, 513, 191. doi: 10.1016/j.ijpharm.2016.09.023

  34. [34]

      Jiang, H.; Hu, X.; Li, Y.; Ngai, T. Mater. Chem. Front. 2021, 5, 3897. doi: 10.1039/d0qm01010c

  35. [35]

      Jiang, H.; Hu, X.; Li, Y.; Yang, C.; Ngai, T. Chem. Sci. 2021, 12, 12463. doi: 10.1039/d1sc03693a

  36. [36]

      Jiang, H.; Hu, X.; Jiang, W.; Guan, X.; Li, Y.; Ngai, T. Langmuir 2022, 38, 12273. doi: 10.1021/acs.langmuir.2c01904

  37. [37]

      Stöber, W.; Fink, A.; Bohn, E. J. Colloid Interface Sci. 1968, 26, 62. doi: 10.1016/0021-9797(68)90272-5

  38. [38]

      Binks, B. P.; Fletcher, P. D. I.; Thompson, M. A.; Elliott, R. P. Langmuir 2013, 29, 5723. doi: 10.1021/la4008697

  39. [39]

      孙冠卿, 易宗霖, 魏涛. 物理化学学报, 2020, 36, 1910005. doi: 10.3866/PKU.WHXB201910005Sun, G.; Yi, Z.; Ngai, T. Acta Phys. -Chim. Sin. 2020, 36, 1910005. doi: 10.3866/PKU.WHXB201910005

  40. [40]

      邹后兵, Ettelaie, R., 闫帅, 薛楠, 杨恒权. 物理化学学报, 2020, 36, 1910006. doi: 10.3866/PKU.WHXB201910006Zou, H.; Ettelaie, R.; Yan, S.; Xue, N.; Yang, H. Acta Phys. -Chim. Sin. 2020, 36, 1910006. doi: 10.3866/PKU.WHXB201910006

  41. [41]

      Jiang, H.; Sheng, Y.; Ngai, T. Curr. Opin. Colloid Interface Sci. 2020, 49, 1. doi: 10.1016/j.cocis.2020.04.010

  42. [42]

      Huang, D.; Sun, M.; Bu, Y.; Luo, F.; Lin, C.; Lin, Z.; Weng, Z.; Yang, F.; Wu, D. J. Mater. Chem. B 2019, 7, 2330. doi: 10.1039/c8tb02928h

  43. [43]

      Vian, A.; Amstad, E. Soft Matter 2019, 15, 1290. doi: 10.1039/c8sm02047g

  44. [44]

      Webber, M. J.; Appel, E. A.; Meijer, E. W.; Langer, R. Nat. Mater. 2016, 15, 13. doi: 10.1038/nmat4474

  45. [45]

      Zhou, Y.; Song, J.; Wang, L.; Xue, X.; Liu, X.; Xie, H.; Huang, X. Biomacromolecules 2017, 18, 2446. doi: 10.1021/acs.biomac.7b00598

•