Ex vivo multi-modal multi-photon imaging of peripheral nerve regeneration induced by collagen scaffolds

Tzeranis, Dimitrios S.; Soller, Eric C.; Lin, Rebecca; Yannas, Ioannis V.; So, Peter T. C.

doi:10.1364/biomed.2012.bw1b.5

Cited by 2 publications

(9 citation statements)

References 3 publications

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“…Degradation rate in vivo depends primarily on the conditions of the crosslinking reaction that lead to specific levels of the crosslink density (Yannas et al 1975b, Huang andYannas 1977), corresponding to corresponding values of the average molecular weight between crosslinks (Yannas 1981(Yannas , 1990. Finally, the identity and density of ligands for specific integrins depend on the presence of particular hexapeptides in the collagen molecule and is, therefore, a property of the basic collagen protein (Tzeranis 2013).…”

Section: Preparation Of Active Collagen Scaffoldsmentioning

confidence: 99%

“…Binding of an integrin on a ligand appears to be mediated entirely by the I domain of the integrin (Hynes 2002, Luo et al 2007. I domains, isolated using protocols that have been published (Tzeranis et al 2010), have been used as markers for adhesion ligands of the parent integrin, providing means for assay of ligands on DRT; see below (Tzeranis 2013).…”

Section: Binding Of Cells On Surface Of Collagenmentioning

confidence: 99%

“…A novel methodology has been described for quantifying the density of ligands of a particular adhesion receptor on the surface of a 3D matrix in situ (Tzeranis et al 2010, 2014, Tzeranis 2013. It consists of: (i) developing soluble fluorescently-labeled markers whose binding properties mimic those of particular adhesion receptors, (ii) using a binding assay of the markers, based on 3D microscopy to detect the fluorescent markers bound on the matrix, and (iii) assaying the density of adhesion ligands using a novel model that describes binding of soluble receptors on ligands present on an insoluble surface.…”

Section: Quantitative Description Of the Scaffold Surfacementioning

confidence: 99%

“…The methodology has been used to measure the adhesion ligand density for the two major collagenbinding integrins (α1β1, α2β1) in two kinds of porous collagen scaffolds, named here 'active' and 'inactive' (Tzeranis et al 2010, Tzeranis 2013. The active scaffold had previously shown strong regenerative activity in a peripheral nerve study while the inactive control was minimally active (Soller et al 2012).…”

Section: Quantitative Description Of the Scaffold Surfacementioning

confidence: 99%

See 3 more Smart Citations

Surface biology of collagen scaffold explains blocking of wound contraction and regeneration of skin and peripheral nerves

Yannas¹,

Tzeranis²,

So³

2015

Biomed. Mater.

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We review the details of preparation and of the recently elucidated mechanism of biological (regenerative) activity of a collagen scaffold (dermis regeneration template, DRT) that has induced regeneration of skin and peripheral nerves (PN) in a variety of animal models and in the clinic. DRT is a 3D protein network with optimized pore size in the range 20–125 μm, degradation half-life 14 ± 7 d and ligand densities that exceed 200 μ α1β1 or α2β1 ligands. The pore has been optimized to allow migration of contractile cells (myofibroblasts, MFB) into the scaffold and to provide sufficient specific surface for cell–scaffold interaction; the degradation half-life provides the required time window for satisfactory binding interaction of MFB with the scaffold surface; and the ligand density supplies the appropriate ligands for specific binding of MFB on the scaffold surface. A dramatic change in MFB phenotype takes place following MFB-scaffold binding which has been shown to result in blocking of wound contraction. In both skin wounds and PN wounds the evidence has shown clearly that contraction blocking by DRT is followed by induction of regeneration of nearly perfect organs. The biologically active structure of DRT is required for contraction blocking; well-matched collagen scaffold controls of DRT, with structures that varied from that of DRT, have failed to induce regeneration. Careful processing of collagen scaffolds is required for adequate biological activity of the scaffold surface. The newly understood mechanism provides a relatively complete paradigm of regenerative medicine that can be used to prepare scaffolds that may induce regeneration of other organs in future studies.

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Section: Preparation Of Active Collagen Scaffoldsmentioning

confidence: 99%

Section: Binding Of Cells On Surface Of Collagenmentioning

confidence: 99%

Section: Quantitative Description Of the Scaffold Surfacementioning

confidence: 99%

Section: Quantitative Description Of the Scaffold Surfacementioning

confidence: 99%

See 2 more Smart Citations

Surface biology of collagen scaffold explains blocking of wound contraction and regeneration of skin and peripheral nerves

Yannas¹,

Tzeranis²,

So³

2015

Biomed. Mater.

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show abstract

“…Since cells in tissue interact with and are regulated by their extracellular matrix, cells cultured in 3D matrix can better mimic the behavior of cells in tissue than cells cultured in 2D well plate or slide glass. For example, cell–matrix interaction has proven to be an important factor in determining the outcome of the nerve regeneration . In these 3D systems, image cytometer with 3D sectioning can better quantify cellular behavior in their native state.…”

mentioning

confidence: 99%

Three‐dimensional image cytometer based on widefield structured light microscopy and high‐speed remote depth scanning

Choi

Wadduwage

et al. 2014

Cytometry Pt A

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A high throughput 3D image cytometer have been developed that improves imaging speed by an order of magnitude over current technologies. This imaging speed improvement was realized by combining several key components. First, a depth-resolved image can be rapidly generated using a structured light reconstruction algorithm that requires only two wide field images, one with uniform illumination and the other with structured illumination. Second, depth scanning is implemented using the high speed remote depth scanning. Finally, the large field of view, high NA objective lens and the high pixelation, high frame rate sCMOS camera enable high resolution, high sensitivity imaging of a large cell population. This system can image at 800 cell/sec in 3D at submicron resolution corresponding to imaging 1 million cells in 20 min. The statistical accuracy of this instrument is verified by quantitatively measuring rare cell populations with ratio ranging from 1:1 to 1:105.

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Ex vivo multi-modal multi-photon imaging of peripheral nerve regeneration induced by collagen scaffolds

Cited by 2 publications

References 3 publications

Surface biology of collagen scaffold explains blocking of wound contraction and regeneration of skin and peripheral nerves

Surface biology of collagen scaffold explains blocking of wound contraction and regeneration of skin and peripheral nerves

Three‐dimensional image cytometer based on widefield structured light microscopy and high‐speed remote depth scanning

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