1. A Novel Tissue-Engineered Approach to Problems of the Postpneumonectomy Space 
Nobuo Tsunooka, MD, PhD, Shin Hirayama, MD, PhD, Jeffrey A. Medin, PhD, W. Conrad Liles, MD, PhD, Shaf Keshavjee, MD, MS, Thomas K. Waddell, MD, PhD 
The Annals of Thoracic Surgery 
Volume 91, Issue 3, Pages (March 2011)
DOI: /j.athoracsur
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

2. Fig 1
Experimental time course and bone marrow cell (BMC) proliferation in collagen matrices ex vivo. (A) Cultured BMCs were seeded into collagen matrices at 3 × 106/mL and cultured for another week. Cellularized matrices were implanted into the thoracic cavity immediately after left pneumonectomy. The mice were sacrificed on days 1, 4, 7, 14, and 21. (B) Hematoxylin and eosin staining for collagen matrices after co-culture for 7 days with BMCs. Cells can be seen on the wall of matrices (magnification ×20). (C) Cell count in collagen matrices ex vivo. The BMCs proliferated in collagen matrices ex vivo over time.
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

3. Fig 2
Angiogenesis in collagen matrices in vivo. Angiogenesis was measured by immunohistochemistry for von Willebrand factor (vWF). Hematoxylin and eosin staining for (A) no-cell group and (B) bone marrow cell (BMC) group on day 21 (magnification ×20). The BMC group showed more vWF-positive vessels (arrowheads) than the no-cell group. (C) Count of vWF-positive vessels. Vessels within the collagen matrices were much more common in the BMC group (shaded bars; no-cell group indicated by open bars). *p = in two-way analysis of variance; **p < 0.05; ***p < 0.01 in t test with Bonferroni correction.
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

4. Fig 3
Bioluminescence imaging (BLI) activity analysis. (A) Change of bioluminescence activity after luciferin injection. Mesenchymal stromal cells with luciferase expression (MSC-Luc) were used as donor cells. (a) Slope of bioluminescence activity at early time point after luciferin injection depends on the distribution speed of luciferin which reflects angiogenesis into collagen matrices. (b) Peak bioluminescence activity, which appeared 30 minutes after luciferin injection, mainly depends on MSC-Luc survival. (B) Bioluminescence activity in vivo. These showed time differences of bioluminescence activity 10 minutes after injection of luciferin. (C) Early slope difference of bioluminescence activity. Slope calculated with bioluminescence difference between 2.5 and 10 minutes after luciferin injection was used as Δbioluminescence. Day 1 showed high bioluminescence values, presumably because of extravasation after surgery. Although it decreased on days 2 to 4, it increased thereafter up to day 6. *p = one-way analysis of variance; **p < 0.05 versus day 9.
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

5. Fig 4
Bioluminescence activity (BLI) changes of Staphylococcus aureus with luciferase expression (SA-Luc). (A) Bioluminescence activity of SA-Luc with or without bone marrow cells (BMC) on day 1. (B) Effect of BMC against SA-Luc in vitro. The BMC group showed less bioluminescence activity than the control group on day 1. (C) Effect of cellularized collagen matrices in a model of thoracic empyema over time. On day minus 4, pneumonectomy and collagen matrices implantation were performed. After 4 days, 0.5 mL of SA-Luc at 1.0 × 107 CFU/mL was injected into the thoracic cavity, and bioluminescence activity was measured at each time point. (Control group: without collagen matrices). Both the BMC group and the no-cell group (n = 5 in each group) showed less bioluminescence activity than the control group (n = 3) on days 4 and 7. *p < two-way analysis of variance for time and between groups. **p < control group versus no-cells group, ***p < between control group and BMC group.
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions