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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 CDI crosslinked implants for MMP3 and TIMP1 in comparison to chemical untreated scaffolds after 1, 9, and 30 days in vivo (n = 6/day/group). One day post implantation, MMP3 were detectable in both untreated (A) and CDI crosslinked (B) scaffolds, whereas CDI crosslinked scaffolds showed a weaker intensity. However, scaffolds of both groups were negative for TIMP1 [untreated: (C), CDI: (D)].
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Figure 9: Immunohistochemical DAB-staining of decellular CDI crosslinked implants for MMP3 and TIMP1 in comparison to chemical untreated scaffolds after 1, 9, and 30 days in vivo (n = 6/day/group). One day post implantation, MMP3 were detectable in both untreated (A) and CDI crosslinked (B) scaffolds, whereas CDI crosslinked scaffolds showed a weaker intensity. However, scaffolds of both groups were negative for TIMP1 [untreated: (C), CDI: (D)].

Mentions: At day 1 after implantation, MMP3 was activated in all groups (untreated: Figure 9A, crosslinked: Figure 9B) and increased to day 9 (untreated: Figure 10A, crosslinked: Figure 10B), whereas the reactivity was weaker in crosslinked than in untreated scaffolds. At day 30, MMP3 was detectable in both untreated (Figure 11A) and crosslinked (Figure 11B) scaffolds with same intensity. Scaffolds of all groups were negative for TIMP1 at all time-points (Figures 9C,D; Figures 10C,D; Figures 11C,D).


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 CDI crosslinked implants for MMP3 and TIMP1 in comparison to chemical untreated scaffolds after 1, 9, and 30 days in vivo (n = 6/day/group). One day post implantation, MMP3 were detectable in both untreated (A) and CDI crosslinked (B) scaffolds, whereas CDI crosslinked scaffolds showed a weaker intensity. However, scaffolds of both groups were negative for TIMP1 [untreated: (C), CDI: (D)].
© Copyright Policy
Related In: Results  -  Collection

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

Figure 9: Immunohistochemical DAB-staining of decellular CDI crosslinked implants for MMP3 and TIMP1 in comparison to chemical untreated scaffolds after 1, 9, and 30 days in vivo (n = 6/day/group). One day post implantation, MMP3 were detectable in both untreated (A) and CDI crosslinked (B) scaffolds, whereas CDI crosslinked scaffolds showed a weaker intensity. However, scaffolds of both groups were negative for TIMP1 [untreated: (C), CDI: (D)].
Mentions: At day 1 after implantation, MMP3 was activated in all groups (untreated: Figure 9A, crosslinked: Figure 9B) and increased to day 9 (untreated: Figure 10A, crosslinked: Figure 10B), whereas the reactivity was weaker in crosslinked than in untreated scaffolds. At day 30, MMP3 was detectable in both untreated (Figure 11A) and crosslinked (Figure 11B) scaffolds with same intensity. Scaffolds of all groups were negative for TIMP1 at all time-points (Figures 9C,D; Figures 10C,D; Figures 11C,D).

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