Repair of a full thickness corneoscleral defect in a German shepherd dog using porcine small intestinal submucosa

Porcine small intestinal submucosa (SIS) was used, in conjunction with a conjunctival graft, to repair a full thickness corneoscleral defect resulting from the excision of a limbal melanoma in a German shepherd dog. The SIS was found to provide adequate mechanical support and to act as a suitable physical barrier in place of the excised cornea and sclera. Corneal vascularisation was present distant to the graft by two weeks postoperatively but this was effectively controlled with topical cyclosporin. By six weeks postoperatively, the graft had become incorporated into the cornea and sclera, and the associated corneal neovascularisation had resolved. From this initial case, porcine SIS would appear to be a suitable material for the repair of corneoscleral defects in dogs.     

猪小肠黏膜下层(SIS)用于犬板层角膜移植的手术方法探索

犬的角膜由外向内分为上皮细胞层、上皮下基膜、基质层、后弹力层和内皮细胞层.板层角膜移植是指移植部分角膜,角膜的切除通常只达基质层.犬板层角膜移植对于治疗药物难以控制的角膜炎症、角膜表层肿瘤、高度睑球粘连和边缘性角膜变性等具有重要意义.     

Surgical repair of deep melting ulcers with porcine small intestinal submucosa (SIS) graft in dogs and cats

PURPOSE: To evaluate the efficacy of using a porcine small intestinal submucosa (SIS) graft for the surgical repair of deep melting ulcers in dogs and cats. METHODS: Two cats and five dogs presented with deep and large melting ulcers of the cornea. In each case, the necrotic and collagenolytic tissue of the cornea was removed by keratectomy. A SIS graft, 1 mm greater than the corneal defect, was rehydrated in sterile saline and sutured to the edges of the ulcer with a simple interrupted pattern of 9/0 polyglactin 910. A nictitating membrane flap was utilized in two cats and four dogs for 2 weeks. All cases were treated postoperatively with topical and systemic antibiotics, a systemic anti-inflammatory drug and topical atropine. All animals were re-evaluated 15 days, 4 weeks, 35-45 days, 2-3 months and 6 months postsurgery. RESULTS: At 15 days postsurgery, a superficial intense corneal neovascularization surrounded the SIS graft. No ocular discomfort was present and fluorescein staining was negative in all cases. At 4 weeks the SIS graft was thick and opaque in all cases, although in one cat the SIS graft had partially detached. Between 35 and 45 days, SIS graft integration was evident in all eyes, and corneal neovascularization had decreased progressively. All eyes healed without complications and retained corneal transparency. This occurred even in the presence of corneal perforation in two cases: one prior to and one during surgery. CONCLUSION: Results of our study suggest the SIS graft may be an effective alternative surgical treatment to the traditional conjunctival grafts commonly used to repair melting ulcers in dogs and cats. The advantages of using a SIS graft include good corneal transparency, preservation of corneal integrity and maintenance of vision.     

The use of porcine small intestinal submucosa as a biomaterial for perineal herniorrhaphy in the dog

OBJECTIVES: To develop an in vivo perineal hernia model, to develop a technique for using small intestinal submucosa (SIS) in perineal hernia repair, to further elucidate the biological behavior of SIS, and to compare SIS herniorrhaphy with the internal obturator muscle transposition (IOT) technique. STUDY DESIGN: Prospective evaluation comparing SIS herniorrhaphy with IOT. ANIMALS: Twelve adult castrated male, large-breed dogs. METHODS: All dogs had bilateral pelvic diaphragm defects created by complete excision of the levator ani muscle. Each dog had one side repaired using SIS and the other by IOT. Pain and inflammation were subjectively scored. Dogs were killed 2 weeks (n = 4), 12 weeks (n = 4), or 16 weeks (n = 4) after surgery. Each pelvic diaphragm was biomechanically tested to failure. The pelvic diaphragms from 2 normal dogs (n = 4 sides) were also biomechanically tested. Failure site, maximum pressure, displacement at failure, and initial linear stiffness values were determined. Histologic assessment was performed. Statistical analysis was performed with significance set at P <.05 RESULTS: No significant postoperative complications were noted. There were no significant differences in maximum pressure to failure, displacement, or stiffness when comparing normal, SIS, and IOT at any time point. The SIS group had significantly less displacement (P =.004) at 2 weeks than at weeks 12 or 16. For all herniorrhaphy techniques, the failure site was central (n = 22) or at the suture line (n = 2). At 2 weeks, histologic evaluation of tissues from the IOT group showed inflammation, mineralization, and necrosis, which were not present in tissues from the SIS group. Histologic examination at 12 and 16 weeks showed no microscopic differences in cell population or tissue characteristics between the IOT and SIS groups. CONCLUSIONS: SIS herniorrhaphy was successfully performed in this in vivo model of perineal hernia in the dog. CLINICAL RELEVANCE: This study suggests that SIS can be used as a primary means of repair, as augmentation when the internal obturator muscle is thin and friable, or as a salvage procedure in cases of recurrence in dogs with perineal hernia.     

Effects of porcine small intestinal submucosa on elution characteristics of gentamicin-impregnated plaster of Paris

OBJECTIVE: To evaluate effects of small intestinal submucosa (SIS) on elution properties of plaster of Paris (POP). SAMPLE POPULATION: 27 POP cylinders, 27 POP spheres, and 9 polymethylmethacrylate (PMMA) spheres. PROCEDURES: Pellets were loaded with gentamicin (50 mg/g) and divided into 7 groups of 9 beads each: PMMA spheres; POP cylinders coated with 0, 4, or 8 layers of SIS; and POP spheres coated with 0, 4, or 8 layers of SIS. Gentamicin concentration was measured 6, 12, 18, 24, 32, 40, and 48 hours and 3, 4, 5, 7, 14, 21, 28, 35, and 42 days after wrapping. Porosity was evaluated via scanning electron microscopy. Curvature factor of elution curves, total amount of drug released (TDR), time required to reach 50% of total release (TDR(t50)), and number of days with concentrations > or = 1 microg/mL were compared among groups. RESULTS: SIS decreased the curvature factor and increased the TDR(t50) and TDR of POP spheres and cylinders. Curvature factor of the PMMA-release curve remained lower than that for any POP group, but all POP groups wrapped in SIS released more gentamicin than PMMA spheres. Gentamicin concentrations remained > or = 1 microg/mL in SIS-wrapped POP and PMMA groups throughout the study. Wrapping POP in SIS minimized the increase in porosity of pellets. CONCLUSIONS AND CLINICAL RELEVANCE: Wrapping POP with SIS slows the release and increases the amount of gentamicin leaching from spheres and cylinders. All groups wrapped in SIS maintained antimicrobial concentrations greater than the minimum inhibitory concentration of most pathogens.     

The use of porcine small intestinal submucosa in ten cases of feline corneal disease

Porcine small intestinal submucosa (SIS) was used as a novel graft material in the management of 10 cases of feline corneal disease. Five cases had stromal ulceration associated with trauma, ocular foreign body and/or suspected infection and required a grafting procedure. Five cases had feline sequestra that were managed by a keratectomy prior to placement of SIS as a graft material. Eight eyes healed with minimal corneal scarring with a very good cosmetic and visual result. One eye with continued aqueous leakage in the immediate postoperative period required a conjunctival pedicle graft to reinforce the SIS graft site. One eye required enucleation 48 h following grafting due to progressive keratomalacia but the SIS material remained intact.     

Invasion of enterocytes in cultured porcine small intestinal mucosal explants by Salmonella choleraesuis.

Porcine small intestinal explants maintained in vitro were inoculated with Salmonella choleraesuis to study the characteristics of its invasion of enterocytes. The explants were fixed at selected intervals for up to 12 hours after inoculation and examined by conventional light microscopy, immunoperoxidase staining, and transmission electron microscopy. Although there was diffuse loss of villous enterocytes during the first hour of incubation, the villi were reepithelialized by the end of 2 hours of culture, and the mucosal epithelium remained intact and appeared to be viable through 12 hours of culture. Intraepithelial S choleraesuis were not detected before 6 hours after inoculation, but after 12 hours of incubation, bacteria were numerous within enterocytes. Ultrastructurally, penetration of the brush border by S choleraesuis resulted in focal loss of microvilli. Bacteria were endocytosed into membrane-bound vacuoles where most remained, but a few were free within the cytoplasm of enterocytes. Invasion of the explants closely resembled that described for live animal and cell culture models of Salmonella spp invasion.     

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

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.     

Rat neonatal intestinal wall-free graft

An experimental study was carried out in order to evaluate the regeneration capacity of the neonatal intestinal wall in ischaemia and its repercussion over organs of the immune system such as the spleen. We isolated a reservoir of small intestine in adult Sprague-Dawley rats that, after having been everted and placed at a subcutaneous level, was grafted with a free small intestine segment of neonatal Sprague-Dawley rats and analysed 3, 15 and 30 days after grafting. Samples obtained were stained with Martin's trichromic stain and studied at the light microscopic level. A total regeneration of the crypt architecture, formed by absorptive enterocytes, was observed in the interior of the reservoirs. A quantitative immunohistochemical study with monoclonal antibodies against CD4, CD8, MHC I and MHC II was carried out in the spleen of these animals. An additional immunohistochemical study was also performed in the small intestine reservoirs and spleen of transplanted animals using nitric oxide synthase (NOS) antibodies. Three days after transplantation NOS immunoreactive cells were observed in the reservoirs with transplanted neonatal small intestine. Antigenic stimulation produced, in the spleen red pulp of transplanted animals, a significant increase (P < 0.001) in the percentage of NOS, CD8 and MHC I immunoreactive cells in relation with values observed in control animals. These morphometric changes could be related with the stimulation that nitric oxide produces in the proliferation of the cytotoxic T cells.     

Use of porcine small intestinal submucosa for corneal reconstruction in dogs and cats: 106 cases

Objectives : To describe the efficacy of porcine small intestinal submucosa in corneal reconstructive surgery in dogs and cats through a large retrospective study. Methods : A retrospective evaluation of 106 cases of surgical reconstruction of the cornea with small intestinal submucosa seen between May 2005 and January 2010 was carried out. The corneal defect was filled by microsurgical grafting of porcine small intestinal submucosa. The biomaterial implant was deposited in one or several layers depending on the depth of the defect. The animals were examined 3, 6 and 12 weeks after surgery. Results : Vision was preserved in all eyes at three months post‐surgery. In 74 cases (69.8%) the corneal scar was either transparent or discrete, whilst in 32 cases (30.2%) a mild or marked scar was observed. Minor complications occurred in 9 cases (8.5%) with partial integration of the small intestinal submucosa and in 24 cases (22.6%) with faint or mild corneal pigmentation, without impairing vision. In cases followed over a period longer than three months, major complications occurred in five dogs resulting in vision impairment because of pronounced pigmentation. Clinical Significance : Corneal grafting of porcine small intestinal submucosa is an effective method for corneal reconstruction resulting in corneal transparency in most cases. It is an excellent alternative to conventional conjunctival grafts.     

Rabbit small intestinal beta-galactosidases

The beta-galactosidase activities of the rabbit small intestinal mucosa were studied over the pH range of 2.6 to 8.4, using different substrates, and in the presence or absence of the enzyme inhibitor p-chloromercuribenzoic acid. The results indicated the presence of 4 beta-galactosidases: (i) a neutral beta-galactosidase (lactase) with optimum pH of 5.8, (ii) an acid beta-galactosidase I with optimum pH of 3.4, (iii) an acid beta-galactosidase II with optimum pH of 4.6, and (iv) a hetero beta-galactosidase with pH optimum of 7.6. Seemingly, 3 of these beta-galactosidases were comparable with those in other mammals, and the 4th has not been reported in other species.     

Grafts of porcine small intestinal submucosa seeded with cultured homologous smooth muscle cells for bladder repair in dogs

Background

Due to numerous complications associated to gastrointestinal augmented cystoplasty, this study aimed to analyze the anatomic repair of the bladder of 10 female dogs using grafts of porcine small intestinal submucosa (SIS) seeded with cultured homologous smooth muscle cells, and compare them with the acellular SIS grafts.

Results

We assessed the possible side effects and complications of each type of graft by clinical examination, abdominal ultrasound and laboratory findings. Anatomic repair of neoformed bladder was assessed by histological staining for H/E and Masson''s Trichrome, analyzed with a Nikon Photomicroscope connected to the system of image analysis Image J.

Conclusions

We propose that SIS associated to homologous smooth cells can improve the quality of tissue repair, and consequently decrease the potential complications inherent to acellular SIS.     

Ultrasonography of small intestinal obstructions: a contemporary approach

Objectives : To assess the accuracy of intestinal ultrasound for diagnosis of intestinal obstruction in dogs and cats. Methods : A prospective clinical study was performed. Inclusion criteria were dogs and cats with clinical signs suggestive of gastrointestinal obstruction. Animals with no obstruction detected on ultrasound were excluded if they could not be monitored for 48 hours to confirm absence of obstruction. Sonographic diagnosis of small intestinal obstruction was based on identification of at least two findings suggestive of intestinal obstruction. Results : Ninety‐two patients suspected of having intestinal obstruction were included. Correct diagnosis of intestinal obstruction was made in 21 cases (23%), and in 68 (74%) this diagnosis was excluded. Interpretation of the images on prospective analysis had sensitivity, positive predictive, specificity and negative predictive values of 100%, 87.5%, 95.8% and 100%, respectively. Clinical Significance : Ultrasonography is an excellent method for investigation of animals with gastrointestinal disorders, and is particularly useful for excluding obstructive processes.     

Laparoscopic repair of a small intestinal mesenteric rent in a broodmare

OBJECTIVES : To describe standing laparoscopic repair of mesoduodenal rent inaccessible by ventral median celiotomy. STUDY DESIGN : Clinical case report. SAMPLE POPULATION : A 6-year-old Thoroughbred broodmare with a right displacement of the dorsal colon and small intestinal incarceration in a mesoduodenal rent. METHODS : Two days after correction of a right dorsal displacement of the ascending colon, small intestinal incarceration, and partial closure of mesoduodenal rent, standing left and right flank laparoscopy was used to explore the abdomen to identify the mesoduodenal defect. The mesenteric rent was repaired from right flank portals by approximation with hemostatic clips. RESULTS : Approximation of the mesoduodenal rent was achieved with laparoscopic clips. No further complications or signs of abdominal pain occurred over the following year. CONCLUSIONS : Mesenteric rents in the mesoduodenum can be repaired by laparoscopic techniques in the standing horse. CLINICAL RELEVANCE : Use of laparoscopic techniques in the standing horse should be considered for mesoduodenal rents that are not accessible form a ventral median celiotomy.     

Granular mucosal lymphocytes in porcine small intestine

A subpopulation of purified, interepithelial lymphocytes from porcine small intestinal mucosa contained cytoplasmic granules. Toluidine blue staining revealed metachromatic granules in 13.64% (606/4,450) cells. The cells had scant organelles, a single large nucleus with obvious invagination of the nuclear membrane, and prominent chromatin. Each cell contained 1 to 10 cytoplasmic membrane-bound granules, 0.6 to 1.5 micron in diameter. These findings indicated that the granular mucosal lymphocytes are related morphologically to mucosal mast cells. The presence of serotonin in the granules, confirmed by the serotonin releasing test, provided functional evidence that granular mucosal lymphocytes are related to mucosal mast cells.     

Lamellar keratoplasty with a graft of lyophilized acellular porcine corneal stroma in the rabbit

Objective To evaluate the efficacy of lamellar keratoplasty in the rabbit using a graft of lyophilized acellular porcine corneal stroma (APCS). Animal studied Twelve adult 2–2.5 kg Zealand white rabbits were studied. Procedure The cell components of the porcine cornea were removed by the means of enzymatic digestion, freezing, and thawing and then APCS was lyophilized. The 6.5 mm diameter APCS was implanted on a 6.0‐mm diameter keratectomy wound each of 12 rabbits. The postoperative clinical and histological evaluations were performed in the early, intermediate, and late periods. Results All corneal wounds healed. Ten of the 12 grafts of APCS were integrated completely with the receptive cornea except two grafts scraped partially off by the eyelid. The blepharospasm, ocular discharge, and edema of the cornea were marked 1 week after transplantation. New vessels invaded the graft after week 2 and regressed after week 8. The cornea became transparent gradually. The histological evaluation showed that the epithelium on the graft stratified normally post surgery. The keratocytes of the recipient grew into the graft and were proliferative at week 4. The inflammatory cells and new vessels were observed before week 8. The fibrosis in the graft was revealed at week 4 and lessened at week 8. The histological structure of the cornea after surgery was similar to the normal cornea at week 32. Conclusions APCS can recover the integrity of the rabbit's cornea and become transparent in vivo. APCS is an effective graft for lamellar keratoplasty in the rabbit.     

Small intestinal morphology in weaned piglets fed a diet containing spray-dried porcine plasma

The hypothesis tested in this study was that the reported beneficial effects of spray-dried porcine plasma (SDPP) on piglet post-weaning performance and health are associated with a trophic effect on small intestinal mucosa. At 24 days of age, the piglets of seven sows were assigned to one of three treatment groups. One group continued to be suckled. The other two groups were weaned and offered a diet containing either 15 per cent SDPP or casein. From each treatment group, one piglet was anaesthetised and samples were taken from the small intestinal wall at 26, 28 and 31 days of age. There were no significant effects of SDPP versus casein on villus length. On average, there was less mitotic activity in the SDPP -fed piglets than in those fed casein on days 4 and 7 after weaning. As less mitotic activity leads to less immature enterocytes, this may provide a mechanism for the reported beneficial effects of SDPP on performance and health.