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Cellular Internalization and Exiting Behavior of Zwitterionic 4-Armed Star-Shaped Polymers.

Yuta Yoshizaki1, Tomohiro Konno1

  • 1Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai 980-8578, Japan.

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Star-shaped phospholipid polymers (4armPMB) demonstrate rapid cellular uptake and release, similar to linear polymers, with no observed cytotoxicity. This architecture offers distinct cellular interaction dynamics for biomaterial applications.

Keywords:
amphiphilic polymeratom transfer radical polymerizationcell shuttlecellular internalizationphospholipid polymerstar polymer

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Area of Science:

  • Biomaterials Science
  • Polymer Chemistry
  • Cell Biology

Background:

  • Zwitterionic phospholipid polymers (PMB) are amphiphilic and biocompatible, known for cell membrane penetration.
  • Conventional linear PMBs are synthesized via free radical polymerization.
  • Branched polymer architectures, like star-shaped polymers, exhibit unique properties such as altered viscosity due to excluded volume effects.

Purpose of the Study:

  • To synthesize a 4-armed star-shaped PMB (4armPMB) using atom transfer radical polymerization (ATRP).
  • To compare the effects of branched (4armPMB) versus linear PMB architecture on cytotoxicity and cellular uptake.
  • To investigate the cellular internalization and exiting behaviors influenced by polymer architecture.

Main Methods:

  • Synthesis of 4-armed star-shaped PMB (4armPMB) and linear PMB using ATRP (a living radical polymerization technique).
  • Verification of water solubility for both polymer types.
  • Assessment of polymer aggregate behavior using pyrene fluorescence.
  • Evaluation of cytotoxicity and cell membrane damage.
  • Quantification of cellular uptake and back-diffusion rates.

Main Results:

  • Both 4armPMB and linear PMB were successfully synthesized and confirmed to be water-soluble.
  • Polymer architecture did not influence polymer aggregate behavior in solution.
  • Neither 4armPMB nor linear PMB exhibited cytotoxicity or caused cell membrane damage.
  • Both polymer architectures showed similar rates of cellular penetration after short incubation.
  • 4armPMB demonstrated significantly faster back-diffusion from cells compared to linear PMB, indicating rapid cellular exiting.

Conclusions:

  • The study successfully synthesized water-soluble 4-armed star-shaped and linear PMBs using ATRP.
  • Both polymer architectures are cytocompatible and do not damage cell membranes.
  • While cellular uptake rates are similar, the star-shaped architecture facilitates faster cellular exiting (back-diffusion) compared to the linear counterpart.
  • The distinct cellular internalization and exiting dynamics of 4armPMB suggest potential for tailored biomaterial design.