Controlled release of the fibronectin central cell binding domain from polymeric microspheres

C. Bouissou, U. Potter, H. Altroff, H.J. Mardon, C.F. Van Der Walle

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Non-ionic surfactants have been employed as alternatives to PVA for the emulsification–encapsulation of a conformationally labile protein (FIII9′-10) into PLGA microspheres. FIII9′-10 was encapsulated using a w/o/w double emulsification–evaporation technique and the microspheres fabricated were characterized by SEM and CLSM. The peptide backbone integrity of FIII9′-10 was assayed by SDS-PAGE and the degree of unfolding of FIII9′-10 following emulsification–encapsulation was assessed using a fibroblast cell-attachment assay. The encapsulation efficiency for FIII9′-10 was 25% when using PVA, compared to 50–60% when using Igepal CA-630 or Triton-X100, with values below for the other surfactants. FIII9′-10 released from microspheres promoted cell attachment in a concentration-dependent manner, only Igepal CA-630 and Triton X-100 maintaining near-maximal cell attachment, indicating that the conformation of the relatively unstable FIII9′ domain was preserved. All non-ionic surfactants reduced microsphere surface porosity, compared to PVA, and an increasing surface rugosity (leading to minor ‘ridges’) could be correlated with decreasing surfactant HLB. Low surface porosities did not effect the diffusion of FIII9′-10 from the microspheres' internal pores in a ‘burst release’, as may have been imagined. In summary, non-ionic surfactants should be considered over PVA for the maintenance of biological activity of conformationally labile proteins during encapsulation.
LanguageEnglish
Pages557-566
Number of pages10
JournalJournal of Controlled Release
Volume95
DOIs
Publication statusPublished - 2004

Fingerprint

Microspheres
Fibronectins
Surface-Active Agents
Porosity
Octoxynol
Polyacrylamide Gel Electrophoresis
Proteins
Fibroblasts
Peptides
Nonidet P-40

Keywords

  • non-ionic surfactants
  • poly(lactic-co-glycolic acid)
  • microsphere
  • fibronectin
  • surface morphology
  • cell attachment

Cite this

Bouissou, C. ; Potter, U. ; Altroff, H. ; Mardon, H.J. ; Van Der Walle, C.F. / Controlled release of the fibronectin central cell binding domain from polymeric microspheres. In: Journal of Controlled Release. 2004 ; Vol. 95. pp. 557-566.
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Controlled release of the fibronectin central cell binding domain from polymeric microspheres. / Bouissou, C.; Potter, U.; Altroff, H.; Mardon, H.J.; Van Der Walle, C.F.

In: Journal of Controlled Release, Vol. 95, 2004, p. 557-566.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Controlled release of the fibronectin central cell binding domain from polymeric microspheres

AU - Bouissou, C.

AU - Potter, U.

AU - Altroff, H.

AU - Mardon, H.J.

AU - Van Der Walle, C.F.

PY - 2004

Y1 - 2004

N2 - Non-ionic surfactants have been employed as alternatives to PVA for the emulsification–encapsulation of a conformationally labile protein (FIII9′-10) into PLGA microspheres. FIII9′-10 was encapsulated using a w/o/w double emulsification–evaporation technique and the microspheres fabricated were characterized by SEM and CLSM. The peptide backbone integrity of FIII9′-10 was assayed by SDS-PAGE and the degree of unfolding of FIII9′-10 following emulsification–encapsulation was assessed using a fibroblast cell-attachment assay. The encapsulation efficiency for FIII9′-10 was 25% when using PVA, compared to 50–60% when using Igepal CA-630 or Triton-X100, with values below for the other surfactants. FIII9′-10 released from microspheres promoted cell attachment in a concentration-dependent manner, only Igepal CA-630 and Triton X-100 maintaining near-maximal cell attachment, indicating that the conformation of the relatively unstable FIII9′ domain was preserved. All non-ionic surfactants reduced microsphere surface porosity, compared to PVA, and an increasing surface rugosity (leading to minor ‘ridges’) could be correlated with decreasing surfactant HLB. Low surface porosities did not effect the diffusion of FIII9′-10 from the microspheres' internal pores in a ‘burst release’, as may have been imagined. In summary, non-ionic surfactants should be considered over PVA for the maintenance of biological activity of conformationally labile proteins during encapsulation.

AB - Non-ionic surfactants have been employed as alternatives to PVA for the emulsification–encapsulation of a conformationally labile protein (FIII9′-10) into PLGA microspheres. FIII9′-10 was encapsulated using a w/o/w double emulsification–evaporation technique and the microspheres fabricated were characterized by SEM and CLSM. The peptide backbone integrity of FIII9′-10 was assayed by SDS-PAGE and the degree of unfolding of FIII9′-10 following emulsification–encapsulation was assessed using a fibroblast cell-attachment assay. The encapsulation efficiency for FIII9′-10 was 25% when using PVA, compared to 50–60% when using Igepal CA-630 or Triton-X100, with values below for the other surfactants. FIII9′-10 released from microspheres promoted cell attachment in a concentration-dependent manner, only Igepal CA-630 and Triton X-100 maintaining near-maximal cell attachment, indicating that the conformation of the relatively unstable FIII9′ domain was preserved. All non-ionic surfactants reduced microsphere surface porosity, compared to PVA, and an increasing surface rugosity (leading to minor ‘ridges’) could be correlated with decreasing surfactant HLB. Low surface porosities did not effect the diffusion of FIII9′-10 from the microspheres' internal pores in a ‘burst release’, as may have been imagined. In summary, non-ionic surfactants should be considered over PVA for the maintenance of biological activity of conformationally labile proteins during encapsulation.

KW - non-ionic surfactants

KW - poly(lactic-co-glycolic acid)

KW - microsphere

KW - fibronectin

KW - surface morphology

KW - cell attachment

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JO - Journal of Controlled Release

T2 - Journal of Controlled Release

JF - Journal of Controlled Release

SN - 0168-3659

ER -