The Use of a Green Fluorescent Protein Porcine Model to Evaluate Host Tissue Integration into Extracellular Matrix Derived Bionanocomposite Scaffolds

When using heterogeneous extracellular matrix (ECM) derived scaffolds for soft tissue repair, current methods of in vivo evaluation can fail to provide a clear distinction between host collagen and implanted scaffolds making it difficult to assess host tissue integration and remodeling. The purpose of this study is both to evaluate novel scaffolds conjugated with nanoparticles for host tissue integration and biocompatibility and to assess green fluorescent protein (GFP) expressing swine as a new animal model to evaluate soft tissue repair materials. Human-derived graft materials conjugated with nanoparticles were subcutaneously implanted into GFP expressing swine to be evaluated for biocompatibility and tissue integration through histological scoring and confocal imaging. Histological scoring indicates biocompatibility and remodeling of the scaffolds with and without nanoparticles at 1, 3, and 6 months. Confocal microscope images display host tissue integration into scaffolds although nonspecificity of GFP does not allow for quantification of integration. However, the confocal images do allow for spatial observation of host tissue migration into the scaffolds at different depths of penetration. The study concludes that the nanoparticle scaffolds are biocompatible and promote integration and that the use of GFP expressing swine can aid in visualizing the scaffold/host interface and host cell/tissue migration.

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Further structural characterization of ovine forestomach matrix and multi-layered extracellular matrix composites for soft tissue repair

Journal of Biomaterials Applications ◽

10.1177/08853282211045770 ◽

2021 ◽

pp. 088532822110457

Author(s):

Matthew J Smith ◽

Sandi G Dempsey ◽

Robert W Veale ◽

Claudia G Duston-Fursman ◽

Chloe A F Rayner ◽

...

Keyword(s):

Extracellular Matrix ◽

Hyaluronic Acid ◽

Soft Tissue ◽

Tissue Repair ◽

Matrix Composites ◽

Soft Tissue Repair ◽

Decellularized Extracellular Matrix ◽

Biological Functionality ◽

Traditional Approaches

Decellularized extracellular matrix (dECM)–based biomaterials are of great clinical utility in soft tissue repair applications due to their regenerative properties. Multi-layered dECM devices have been developed for clinical indications where additional thickness and biomechanical performance are required. However, traditional approaches to the fabrication of multi-layered dECM devices introduce additional laminating materials or chemical modifications of the dECM that may impair the biological functionality of the material. Using an established dECM biomaterial, ovine forestomach matrix, a novel method for the fabrication of multi-layered dECM constructs has been developed, where layers are bonded via a physical interlocking process without the need for additional bonding materials or detrimental chemical modification of the dECM. The versatility of the interlocking process has been demonstrated by incorporating a layer of hyaluronic acid to create a composite material with additional biological functionality. Interlocked composite devices including hyaluronic acid showed improved in vitro bioactivity and moisture retention properties.

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Sexual Pheromone Induces Diffusion of the Pheromone-Homologous Polypeptide in the Extracellular Matrix of Volvox carteri

Eukaryotic Cell ◽

10.1128/ec.00304-07 ◽

2007 ◽

Vol 6 (11) ◽

pp. 2157-2162 ◽

Cited By ~ 5

Author(s):

Koichi Ishida

Keyword(s):

Extracellular Matrix ◽

Green Fluorescent Protein ◽

Fluorescent Protein ◽

Sexual Pheromone ◽

Volvox Carteri ◽

Terminal Domain ◽

Induction Signal ◽

C Terminus ◽

Green Fluorescent

ABSTRACT The C-terminal domain of pherophorin II is homologous to the sexual pheromone of Volvox carteri and is released from other domains during sexual induction. Green fluorescent protein fused to the C terminus of pherophorin II was located at the extracellular matrix directly surrounding the gonidium, the final target of the sexual-induction signal.

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Host protection against deliberate bacterial contamination of an extracellular matrix bioscaffold versus Dacron? mesh in a dog model of orthopedic soft tissue repair

Journal of Biomedical Materials Research ◽

10.1002/jbm.b.10062 ◽

2003 ◽

Vol 67B (1) ◽

pp. 648-654 ◽

Cited By ~ 41

Author(s):

Stephen F. Badylak ◽

Ching Ching Wu ◽

Melissa Bible ◽

Edward McPherson

Keyword(s):

Extracellular Matrix ◽

Soft Tissue ◽

Tissue Repair ◽

Bacterial Contamination ◽

Soft Tissue Repair ◽

Host Protection ◽

Dog Model ◽

Dacron Mesh

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Intravital Confocal and Two-Photon Imaging of Dual-Color Cells and Extracellular Matrix Mimics

Microscopy and Microanalysis ◽

10.1017/s1431927612014080 ◽

2013 ◽

Vol 19 (1) ◽

pp. 201-212 ◽

Cited By ~ 6

Author(s):

Ufuk Bal ◽

Volker Andresen ◽

Brenda Baggett ◽

Urs Utzinger

Keyword(s):

Extracellular Matrix ◽

Green Fluorescent Protein ◽

Fluorescent Protein ◽

Second Harmonic ◽

Three Dimensional ◽

Confocal Imaging ◽

Collagen I ◽

Two Photon ◽

Significant Difference ◽

Green Fluorescent

AbstractWe report our efforts in identifying optimal scanning laser microscope parameters to study cells in three-dimensional culture. For this purpose we studied contrast of extracellular matrix (ECM) mimics, as well as signal attenuation, and bleaching of red and green fluorescent protein labeled cells. Confocal backscattering, second harmonic generation (SHG), and autofluorescence were sources of contrast in ECM mimics. All common ECM mimics exhibit contrast observable with confocal reflectance microscopy. SHG imaging on collagen I based hydrogels provides high contrast and good optical penetration depth. Agarose is a useful embedding medium because it allows for large optical penetration and exhibits minimal autofluorescence. We labeled breast cancer cells' outline with DsRed2 and nucleus with enhanced green fluorescent protein (eGFP). We observed significant difference both for the bleaching rates of eGFP and DsRed2 where bleaching is strongest during two-photon excitation (TPE) and smallest during confocal imaging. But for eGFP the bleaching rate difference is smaller than for DsRed2. After a few hundred microns depth in a collagen I hydrogel, TPE fluorescence of DsRed2 becomes twice as strong compared to confocal imaging. In fibrin and agarose gels, the imaging depth will need to be beyond 1 mm to notice a TPE advantage.

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