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Tissue Engineering of Ureteral Grafts: Preparation of Biocompatible Crosslinked Ureteral Scaffolds of Porcine Origin.

Koch H, Hammer N, Ossmann S, Schierle K, Sack U, Hofmann J, Wecks M, Boldt A - Front Bioeng Biotechnol (2015)

Bottom Line: After decellularization, scaffold morphology and composition of ECM were maintained, all cellular components were removed, DNA destroyed and strongly reduced.In vitro: GP and CDI scaffolds revealed a higher number of ingrown 3T3 and SMC cells as compared to untreated scaffolds.TIMP1 was below the detection limit.

View Article: PubMed Central - PubMed

Affiliation: Translational Centre for Regenerative Medicine (TRM), University of Leipzig , Leipzig , Germany.

ABSTRACT
The surgical reconstruction of ureteric defects is often associated with post-operative complications and requires additional medical care. Decellularized ureters originating from porcine donors could represent an alternative therapy. Our aim was to investigate the possibility of manufacturing decellularized ureters, the characteristics of the extracellular matrix (ECM) and the biocompatibility of these grafts in vitro/in vivo after treatment with different crosslinking agents. To achieve these goals, native ureters were obtained from pigs and were decellularized. The success of decellularization and the ECM composition were characterized by (immuno)histological staining methods and a DNA-assay. In vitro: scaffolds were crosslinked either with carbodiimide (CDI), genipin (GP), glutaraldehyde, left chemically untreated or were lyophilized. Scaffolds in each group were reseeded with Caco2, LS48, 3T3 cells, or native rat smooth muscle cells (SMC). After 2 weeks, the number of ingrown cells was quantified. In vivo: crosslinked scaffolds were implanted subcutaneously into rats and the type of infiltrating cells were determined after 1, 9, and 30 days. After decellularization, scaffold morphology and composition of ECM were maintained, all cellular components were removed, DNA destroyed and strongly reduced. In vitro: GP and CDI scaffolds revealed a higher number of ingrown 3T3 and SMC cells as compared to untreated scaffolds. In vivo: at day 30, implants were predominantly infiltrated by fibroblasts and M2 anti-inflammatory macrophages. A maximum of MMP3 was observed in the CDI group at day 30. TIMP1 was below the detection limit. In this study, we demonstrated the potential of decellularization to create biocompatible porcine ureteric grafts, whereas a CDI-crosslink may facilitate the remodeling process. The use of decellularized ureteric grafts may represent a novel therapeutic method in reconstruction of ureteric defects.

No MeSH data available.


Related in: MedlinePlus

Immunohistochemical DAB-staining of decellular porcine ureteral scaffolds for collagen I (A), collagen III (B), fibronectin (C), collagen IV (D), and elastin (E) in comparison to natural tissue (small pictures). Decellular ureteral scaffolds were shown to maintain native extracellular matrix composition. The arrows mark vessels. Bar = 100 μm.
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Figure 2: Immunohistochemical DAB-staining of decellular porcine ureteral scaffolds for collagen I (A), collagen III (B), fibronectin (C), collagen IV (D), and elastin (E) in comparison to natural tissue (small pictures). Decellular ureteral scaffolds were shown to maintain native extracellular matrix composition. The arrows mark vessels. Bar = 100 μm.

Mentions: After decellularization, the ECM composition is similar to native ureteral tissue. Azan staining of natural (Figure 1A) and decellular ureteral scaffolds (Figure 1B) revealed morphologically intact structures, ideal matrix geometry, and no remaining cellular structures. Histological analysis after lyophilization also showed preserved morphological structures and matrix geometry (Figure 1C) similar to the native and decellular conditions. After decellularization, collagen I (Figure 2A), collagen III (Figure 2B), and fibronectin (Figure 2C) could be observed in large amounts in all tissue areas similar to native conditions (small pictures, Figures 2A–C). The vessels expressed collagen IV (Figure 2D) and elastin, which are also located in tunica muscularis (Figure 2E).


Tissue Engineering of Ureteral Grafts: Preparation of Biocompatible Crosslinked Ureteral Scaffolds of Porcine Origin.

Koch H, Hammer N, Ossmann S, Schierle K, Sack U, Hofmann J, Wecks M, Boldt A - Front Bioeng Biotechnol (2015)

Immunohistochemical DAB-staining of decellular porcine ureteral scaffolds for collagen I (A), collagen III (B), fibronectin (C), collagen IV (D), and elastin (E) in comparison to natural tissue (small pictures). Decellular ureteral scaffolds were shown to maintain native extracellular matrix composition. The arrows mark vessels. Bar = 100 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4477215&req=5

Figure 2: Immunohistochemical DAB-staining of decellular porcine ureteral scaffolds for collagen I (A), collagen III (B), fibronectin (C), collagen IV (D), and elastin (E) in comparison to natural tissue (small pictures). Decellular ureteral scaffolds were shown to maintain native extracellular matrix composition. The arrows mark vessels. Bar = 100 μm.
Mentions: After decellularization, the ECM composition is similar to native ureteral tissue. Azan staining of natural (Figure 1A) and decellular ureteral scaffolds (Figure 1B) revealed morphologically intact structures, ideal matrix geometry, and no remaining cellular structures. Histological analysis after lyophilization also showed preserved morphological structures and matrix geometry (Figure 1C) similar to the native and decellular conditions. After decellularization, collagen I (Figure 2A), collagen III (Figure 2B), and fibronectin (Figure 2C) could be observed in large amounts in all tissue areas similar to native conditions (small pictures, Figures 2A–C). The vessels expressed collagen IV (Figure 2D) and elastin, which are also located in tunica muscularis (Figure 2E).

Bottom Line: After decellularization, scaffold morphology and composition of ECM were maintained, all cellular components were removed, DNA destroyed and strongly reduced.In vitro: GP and CDI scaffolds revealed a higher number of ingrown 3T3 and SMC cells as compared to untreated scaffolds.TIMP1 was below the detection limit.

View Article: PubMed Central - PubMed

Affiliation: Translational Centre for Regenerative Medicine (TRM), University of Leipzig , Leipzig , Germany.

ABSTRACT
The surgical reconstruction of ureteric defects is often associated with post-operative complications and requires additional medical care. Decellularized ureters originating from porcine donors could represent an alternative therapy. Our aim was to investigate the possibility of manufacturing decellularized ureters, the characteristics of the extracellular matrix (ECM) and the biocompatibility of these grafts in vitro/in vivo after treatment with different crosslinking agents. To achieve these goals, native ureters were obtained from pigs and were decellularized. The success of decellularization and the ECM composition were characterized by (immuno)histological staining methods and a DNA-assay. In vitro: scaffolds were crosslinked either with carbodiimide (CDI), genipin (GP), glutaraldehyde, left chemically untreated or were lyophilized. Scaffolds in each group were reseeded with Caco2, LS48, 3T3 cells, or native rat smooth muscle cells (SMC). After 2 weeks, the number of ingrown cells was quantified. In vivo: crosslinked scaffolds were implanted subcutaneously into rats and the type of infiltrating cells were determined after 1, 9, and 30 days. After decellularization, scaffold morphology and composition of ECM were maintained, all cellular components were removed, DNA destroyed and strongly reduced. In vitro: GP and CDI scaffolds revealed a higher number of ingrown 3T3 and SMC cells as compared to untreated scaffolds. In vivo: at day 30, implants were predominantly infiltrated by fibroblasts and M2 anti-inflammatory macrophages. A maximum of MMP3 was observed in the CDI group at day 30. TIMP1 was below the detection limit. In this study, we demonstrated the potential of decellularization to create biocompatible porcine ureteric grafts, whereas a CDI-crosslink may facilitate the remodeling process. The use of decellularized ureteric grafts may represent a novel therapeutic method in reconstruction of ureteric defects.

No MeSH data available.


Related in: MedlinePlus