Journal: Tissue Engineering. Part A

Article Title: * Skeletal Myoblast-Seeded Vascularized Tissue Scaffolds in the Treatment of a Large Volumetric Muscle Defect in the Rat Biceps Femoris Muscle

doi: 10.1089/ten.tea.2016.0523

Figure Lengend Snippet: Custom culture molds and volumetric muscle loss (VML) model in the rat biceps femoris muscle. (A) Components of the custom Teflon mold used to create vascularized constructs within individual wells of a six-well tissue culture plate, including 1—top and bottom Teflon rings between which the perforated polycaprolactone (PCL) membrane is suspended, 2—screws holding the PCL membrane stretched between the two Teflon rings, 3—the removable occluders (plastic or metal dowels) that are in place during gel formation and removed immediately to leave behind additional space to allow media circulation, 4—thin silicone membrane forming the bottom of the chamber to prevent leakage between the well base and the Teflon rings, and 5—thick silicone membrane wrapped around the Teflon mold to ensure a tight fit against the side and bottom of the well to prevent collagen solution leakage. (B) Fully assembled chamber mold with collagen gel formed around the perforated PCL mesh as shown in the schematic and photograph. (C, D) Schematic and photograph of vascularized collagen construct polymerized across the perforations in the PCL mesh (dotted line). (E) Creation of a 12-mm-diameter VML in the rat biceps femoris muscle. Plastic spatula placed underneath the biceps femoris muscle prevents injury to underlying tissues during creation of the VML. An untreated empty defect is pictured. (F) An autograft is sutured back into the defect immediately after harvest. (G) The vascularized collagen construct is sutured in place into the defect with excess PCL membrane trimmed away. This technique of suturing through the collagen gel and PCL membrane protects the collagen gel from damage during implantation. Color images available online at www.liebertpub.com/tea

Article Snippet: VML defect creation Unilateral biceps femoris muscle defects were created in 13-week-old female Sprague Dawley rats.

Techniques: Construct, Membrane

Journal: Tissue Engineering. Part A

Article Title: * Skeletal Myoblast-Seeded Vascularized Tissue Scaffolds in the Treatment of a Large Volumetric Muscle Defect in the Rat Biceps Femoris Muscle

doi: 10.1089/ten.tea.2016.0523

Figure Lengend Snippet: Vascular volume in microvessel-treated biceps femoris. (A) Representative images of the vasculature in each group show that the MVF ± Myoblast groups have comparable vascular volume to the autograft group. Vessels are colored according to their respective diameters. (B) At 2 weeks postinjury, overall vascular volumes were not significantly different between groups (*p < 0.05, mean ± SEM, n = 5–7 for injured groups). (C) Histogram of vessel diameter distribution across 21-μm-sized diameter bins, showing a peak at the smaller diameter range. (D) Subset of the histogram data of vessels ≤231 μm diameter. Notations for significant differences (p < 0.05, two-way analysis of variance [ANOVA] for simple effects within diameter bins): *MVF, MVF+My versus Autograft, Empty, $MVF, MVF+My versus Empty, #MVF+My versus Empty defects. (E) Vascular volumes reflective of the two diameter cutoffs analyzed (*p < 0.05, mean ± SEM, n = 5–7 for injured groups), and (F–I) eosin and hematoxylin-stained sections of injured muscles after 2 weeks of healing (20 × images) show large (arrow) and small (arrow head) microvessels perfused with Microfil, which appears as black pigment in the lumen. It must be noted that Microfil can be dislodged from the slide during staining. Scale bar: 150 μm. Color images available online at www.liebertpub.com/tea

Article Snippet: VML defect creation Unilateral biceps femoris muscle defects were created in 13-week-old female Sprague Dawley rats.

Techniques: Staining, Muscles

Journal: Tissue Engineering. Part A

Article Title: * Skeletal Myoblast-Seeded Vascularized Tissue Scaffolds in the Treatment of a Large Volumetric Muscle Defect in the Rat Biceps Femoris Muscle

doi: 10.1089/ten.tea.2016.0523

Figure Lengend Snippet: Qualitative histology. Transverse section of biceps femoris muscle 8 weeks postinjury, stained with Masson's trichrome, showing empty defect (A), autograft (B), MVF (C), and MVF+Myoblast (D) group 8 weeks after injury. Implanted collagen constructs appear replaced by loose fibrous tissue similar to empty defects (blue-green) and autograft tissue appeared to be maintained. Transverse sections of MVF+Myoblast-treated biceps femoris 2 weeks (E) and 8 weeks (F) after injury showing surviving myoblasts (IHC: red—GFP myoblasts, blue—nuclei) at the defect margin but not at the defect center (G). 4× Mosaic scale bar: 2 mm (A–D), 10× scale bar: 100 μm (E–G). (H–I) Masson's trichrome-stained section of an autograft-treated sample at 2 weeks (H) and 4 weeks (I) postimplantation showing fibrous tissue (arrowhead) in the defect around the autograft, and showing fibrous infiltration of muscle tissue (*). These regions appear highly vascularized as evidenced by the Microfil-perfused vessels (arrows) seen in these areas. Scale bar: 150 μm (H) and 500 μm (I). Color images available online at www.liebertpub.com/tea

Article Snippet: VML defect creation Unilateral biceps femoris muscle defects were created in 13-week-old female Sprague Dawley rats.

Techniques: Staining, Construct