Polymers for induction of revascularization in the rat fascial flap: application of vascular endothelial growth factor and pancreatic islet cells.
One of the major obstacles in transplanting avascular tissue or metabolically active cells for ischemic diseases is the loss of transplanted cells due to lack of oxygen and nutrients in the early posttransplantation period. Biodegradable polymeric tissue engineering scaffolds and hydrogels have a potential to incorporate cells or cellular organoids such as islets of Langerhans and growth factors. In this study, we tested the efficiency of two types of polymeric materials to carry recombinant human vascular endothelial growth factor (rhVEGF) or pancreatic tumor cell lines, namely Ins-1 and AR42J, for the induction of new vessels. Chitosan hydrogel fibers with micropores were prepared and molded into a cylinder construct (5 mm phi; 8 mm height). Macroporous PLGA scaffolds with a pore size of 250-400 microm were prepared and cut into cylinders (6 mm phi; 3 mm height). Both chitosan and PLGA constructs were loaded with rhVEGF (3 microg) or seeded with the cell lines (5 x 10(5) cells and 3 x 10(5) cells/construct, respectively, for AR42J and INS-1 cells), and transplanted into the fascial flaps of Wistar rats. At distinct time points up to 4 weeks postimplantation, polymers were explanted, fixed, and vessel density was counted on sections stained with anti-Factor-VIII antibody. Additionally, the kinetics of rhVEGF release from PLGA microspheres (phi of 50-80 microm) was determined using VEGF Elisa. Endogenous VEGF release from pancreatic rat cell lines was also determined. Light microscopy study was performed on H&E-stained paraffin sections of the islet-polymer samples. The vascular density of rhVEGF-loaded chitosan constructs was increased fourfold 2 weeks after subcutaneous transplantation compared with rhVEGF-unloaded controls (465 +/- 144 vs. 104 +/- 80 vessels per mm2, p < 0.05). Protein leakage occurred, but was not observed after 2 weeks. Higher insulin content was detected in rat islet grafts transplanted following VEGF application. More than 50% of total rhVEGF was released on the first day of in vitro culture of PLGA microspheres. rhVEGF secretion had another, but smaller, peak on the third day followed by a constant release. By comparison, endogeneous VEGF secretion of pancreatic tumor cells was measured within a 3-day culture period. Biodegradable polymer scaffolds and hydrogels may have potential use as solid supports to induce angiogenesis for pancreatic cell transplantation.
Linn
Thomas
T
Justus Liebig University, Medical Clinic and Policlinic 3, 35392 Giessen, Germany. thomas.linn@innere.med.uni-giessen.de
Erb
Doris
D
Schneider
Darius
D
Kidszun
Andre
A
Elçin
A Eser
AE
Bretzel
Reinhard G
RG
Elçin
Y Murat
YM
D000818:Animals; D002460:Cell Line; D002686:Chitin / Q000031:analogs & derivatives* / Q000737:chemistry*; D048271:Chitosan; D005205:Fascia / Q000098:blood supply; D006801:Humans; D000860:Hypoxia; D007150:Immunohistochemistry; D007328:Insulin / Q000032:analysis; D016381:Islets of Langerhans Transplantation; D019344:Lactic Acid / Q000737:chemistry*; D008863:Microspheres; D018919:Neovascularization, Physiologic; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers / Q000737:chemistry*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics; D023822:Tissue Engineering; D042461:Vascular Endothelial Growth Factor A / Q000008:administration & dosage* / Q000235:genetics / Q000494:pharmacology
D007328:Insulin; D011108:Polymers; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D002686:Chitin; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D048271:Chitosan