Modulation of inflammation and angiogenesis and changes in ECM GAG-activity via dual delivery of nucleic acids.
Tissue-engineered organs and implants hold promise for the replacement of damaged and diseased organs. However, the foreign body response (FBR) is a major obstacle that compromises the function of tissue-engineered constructs, typically causing them to fail. Two components of FBR are an inflammatory response and a lack of vascularization. To overcome these limitations, a collagen system was developed to release interleukin-6 (IL-6) siRNA and endothelial nitric oxide synthase (eNOS) pDNA in a staggered manner. Hollow collagen microspheres were assembled into a collagen sphere-in-hydrogel system that displayed a staggered release profile in vitro. This system was assessed in vivo in a subcutaneous rat model. The doses of IL-6 siRNA and eNOS pDNA were first individually optimized for their ability to reduce the volume fraction of inflammatory cells (7 days) and increase the length density of blood vessels (14 days), respectively. The identified optimal doses were combined, and the ability of the system to decrease the volume fraction of inflammatory cells and increase the length density of blood vessels was confirmed at both 7 and 14 days. Analysis of the tissue using Raman microspectroscopy revealed that in addition to changes in inflammation and angiogenesis, there were also changes in the extracellular matrix (ECM) at seven days. While changes in sulfated glycosaminoglycan (sGAG) content of the ECM were not detected, changes in the binding of sGAG of the ECM to growth factors were observed. Two growth factors tested, VEGF165 and bFGF showed increased binding to sGAG extracted from eNOS pDNA-treated samples at seven days, increasing the angiogenic potential of the ECM. Thus, we observe that changes in the tissue in terms of the balance of inflammation and angiogenesis as well changes in the activity of sGAG of the ECM occurs following dual delivery of nucleic acids from the collagen sphere-in-hydrogel system.
Centre for Research in Medical Devices (CÚRAM), National University of Ireland Galway, Ireland.
Shane
S
Browne
Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology, Stuttgart, Germany.
Michael G
MG
Monaghan
Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology, Stuttgart, Germany.
Eva
E
Brauchle
Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology, Stuttgart, Germany.
Daniel Carvajal
DC
Berrio
Laboratoire Croissance Cellulaire, Réparation et Régénération Tissulaires, CRRET/EA-4397/CNRS ERL-9215, Université Paris Est Créteil, Université Paris Est, F-94000, Créteil, France.
Sandrine
S
Chantepie
Laboratoire Croissance Cellulaire, Réparation et Régénération Tissulaires, CRRET/EA-4397/CNRS ERL-9215, Université Paris Est Créteil, Université Paris Est, F-94000, Créteil, France.
Dulce
D
Papy-Garcia
Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology, Stuttgart, Germany; Dept. of Women's Health, Research Institute for Women's Health, Eberhard Karls University, Tübingen, Germany; Department of Medicine/Cardiology, Cardiovascular Research Laboratories, University of California, Los Angeles, CA, USA.
Katja
K
Schenke-Layland
Centre for Research in Medical Devices (CÚRAM), National University of Ireland Galway, Ireland. Electronic address: abhay.pandit@nuigalway.ie.
Abhay
A
Pandit
D000818:Animals; D002417:Cattle; D003094:Collagen / Q000737:chemistry; D004270:DNA, Circular / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use; D005109:Extracellular Matrix / Q000276:immunology*; D005260:Female; D015316:Genetic Therapy; D006025:Glycosaminoglycans / Q000276:immunology*; D007249:Inflammation / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D015850:Interleukin-6 / Q000235:genetics*; D018919:Neovascularization, Physiologic; D052250:Nitric Oxide Synthase Type III / Q000235:genetics*; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use; D011917:Rats, Inbred Lew; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry
D004270:DNA, Circular; D006025:Glycosaminoglycans; D015850:Interleukin-6; D034741:RNA, Small Interfering; C013786:A73025; D003094:Collagen; D052250:Nitric Oxide Synthase Type III
Angiogenesis; Collagen; Extracellular matrix (ECM); Gene therapy; Inflammation