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Enhanced PLGA Microparticle Spheronisation with PEG Coating for Improved Biocompatibility

Explore the effects of PEG coating on PLGA microparticles through a series of experiments involving spheronisation process, scaffold strength, porosity analysis, and biocompatibility testing. The addition of PEG-400 enhances particle properties, such as reduced swelling, better spheronisation, and minimized agglomeration, leading to improved biocompatibility results during cytotoxicity testing and injectability assessment. Additionally, the scaffolds exhibit enhanced strength and porosity, making them promising candidates for various biomedical applications.

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Enhanced PLGA Microparticle Spheronisation with PEG Coating for Improved Biocompatibility

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  1. Spheronisation – PVA suspension and heating • PLGA (85:15) 100-200 micron • Stirred 0.5 g in 50ml 0.2 % PVA in PBS at 80⁰C for 4 h • Top • Untreated • Middle • PEG 400 added (final 28% v/v) after 4 h • Bottom • No PEG added

  2. Spheronisation – Rheology of spheronised PLGA microparticles +/- PEG 400 • PLGA (85:15) 100-200 micron • Stirred 0.5 g in 50ml 0.2 % PVA in PBSat 80⁰C for 4 h • Addition of PEG400 after 4h to “coat” particles

  3. Spheronisation – PVA suspension and heating100-200 µm 6.5% PEG-PLGA 70 ⁰C in 0.2 % PVA in ddH2O 70 ⁰C C in 0.2 % PVA /6.5% PEG400 in ddH2O PEG 400 • Addition of PEG 400 leads to • Less swelling • Better spheronisation • Less agglomeration

  4. Scaffold strength and porosity100-200 µm 6.5% PEG-PLGA - untreated or Spheronised for 4 h in 0.2 % PVA in ddH2O at 70 ⁰C

  5. Biocompatability testing of carrier formulations • Assay setup • Linearity of plate reader response • linear to ODs ~ 3.0 • Linearity of cell response • linear to OD 1.1 • This response was from 4 x 105 cells/well 48 h after seeding (assay length) • OD ≥ 0.2 required by ISO10993-5. • OD > 0.4 for 1 x 105 cells/well 48 h after seeding.

  6. Biocompatability testing of carrier formulations • Cytotoxicity testing • Addition of carriers directly to monolayers • 15 min, 1 h, 4 h and 24 h exposure. • Carrier aspirated and replaced with complete media after 15 min, 1 h and 4 h. • MTT assay carried out after 24h after initial exposure to carrier

  7. Biocompatability testing of carrier formulations • Cytotoxicity testing • 2 % Pluronics F127 now used to improve injectability • No increased cytotoxic effects

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