J Vet Sci.  2013 Jun;14(2):175-184. 10.4142/jvs.2013.14.2.175.

Evaluation of a canine small intestinal submucosal xenograft and polypropylene mesh as bioscaffolds in an abdominal full-thickness resection model of growing rats

• Lee AJ
• Lee SH
• Chung WH
• Kim DH
• Chung DJ
• Do SH
• Kim HY

• KMID: 1705519
• DOI: http://doi.org/10.4142/jvs.2013.14.2.175

Abstract

We evaluated the biological scaffold properties of canine small intestinal submucosa (SIS) compared to a those of polypropylene mesh in growing rats with full-thickness abdominal defects. SIS is used to repair musculoskeletal tissue while promoting cell migration and supporting tissue regeneration. Polypropylene mesh is a non-resorbable synthetic material that can endure mechanical tension. Canine SIS was obtained from donor German shepherds, and its porous collagen fiber structure was identified using scanning electron microscopy (SEM). A 2.50-cm2 section of canine SIS (SIS group) or mesh (mesh group) was implanted in Sprague-Dawley rats. At 1, 2, 4, 12, and 24 weeks after surgery, the implants were histopathologically examined and tensile load was tested. One month after surgery, CD68+ macrophage numbers in the SIS group were increased, but the number of CD8+ T cells in this group declined more rapidly than that in rats treated with the mesh. In the SIS group, few adhesions and well-developed autologous abdominal muscle infiltration into the SIS collagen fibers were observed. No significant differences in the tensile load test results were found between the SIS and mesh groups at 24 weeks. Canine SIS may therefore be a suitable replacement for artificial biological scaffolds in small animals.

Keyword

biological scaffold; canine small intestinal submucosa (canine SIS); inflammatory response; polypropylene mesh; tensile load test

MeSH Terms

Abdominal Wall/*surgery
Animals
Biocompatible Materials/*therapeutic use
Dogs
Female
Intestinal Mucosa/cytology/transplantation
Intestine, Small/cytology/*transplantation
Polypropylenes/*therapeutic use
Rats
Rats, Sprague-Dawley
Tensile Strength
Tissue Adhesions
*Tissue Scaffolds
Transplantation, Heterologous/*methods
*Wound Healing
Biocompatible Materials
Polypropylenes

Figure

• 

  Fig. 1 Photograph of a naive decellularized canine SIS scaffold (insert) and an SEM image of the SIS surface. Collagen fibers formed an aporous structure.

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  Fig. 2 Results of the tensile load tests for the canine SIS and polypropylene mesh implants. Data are expressed as the mean ± SEM. Asterisks indicate significant differences (p < 0.05) in (A) implant thickness (mm), (B) maximum strain (%), and (C) maximum stress (MPa) between the two groups.

• 

  Fig. 3 Photograph of the visceral portion of the polypropylene mesh or canine SIS 24 weeks after implantation. (A) An area (2.0 × 2.0 cm) of full-thickness total resection (external abdominal oblique, transverse abdominis muscles, and peritoneum) was made in the ventral abdominal wall. (B) In the polypropylene mesh group, adhesions between the intestine or omentum and the mesh were observed along with hematomas. (C) In the canine SIS group, neither adhesion formation nor host muscle infiltration was observed.

• 

  Fig. 4 Masson's trichrome staining for monitoring host tissue morphologic changes (200× magnification). (A) Collagen connective tissue that appeared around polypropylene mesh fibers were identified 4 weeks after surgery. (B) The number of fibroblastic cells appearing around the mesh and collagen increased at 12 weeks. (C) Infiltration of skeletal muscle fibers amid the canine SIS collagen at 4 weeks. (D) The infiltrating muscle fibers developed and were enlarged (red) in the canine SIS implant (blue) by 12 weeks. Scale bars = 100 µm.

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  Fig. 5 Masson's trichrome staining for evaluating host tissue responses around the grafts 24 weeks after surgery (100× magnification). (A) Adipose tissue deposited around the mesh and necrotic changes (*) in muscle bundles were identified (B) Accumulation and development of adipose tissue along with weak development (**) of muscle and fibrous connective tissue were observed 24 weeks after surgery in the mesh group. (C) Poorly developed host muscle bundles appeared between the canine SIS collagen fibers (blue). (D) Complete host muscle bundles in the canine SIS implants were identified 24 weeks after surgery. Scale bars = 200 µm.

• 

  Fig. 6 Immunohistochemical quantification of specific mononuclear cells (CD8+ cytotoxic T lymphocytes and CD68+ macrophages) at 200× magnification. (A) CD8+ T cells in the canine SIS implant 4 weeks after surgery. (B) CD68+ M1 cells in canine SIS at 4 weeks. (C) CD8+ T cells in the canine SIS implant at 12 weeks. Significantly fewer cells were observed at this time compared to 4 weeks. (D) CD68+ M1 cells in canine SIS at 12 weeks. (E) CD8+ T cells around the mesh fibers at 4 weeks. (F) CD68+ M1 cells around the mesh fibers at 4 weeks. (G) CD8+ T cells around the mesh fibers at 12 weeks. (H) CD68+ M1 cells around the mesh fibers at 12 weeks. Scale bars = 100 µm.

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  Fig. 7 Inflammatory cell counts for the SIS and mesh groups. Asterisks (*) indicate significant differences (p < 0.05) between the canine SIS and polypropylene mesh implants. (A) The number of mast cells for the mesh group gradually increased compared to those of the canine SIS group. (B) The number of CD8+ cytotoxic T cells declined faster in the canine SIS group than in the mesh group. (C) CD 68+ macrophages were more numerous in the canine SIS group than in the mesh group 2 weeks after surgery. At 24 weeks, no significant difference between the two groups was observed.

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