Lee Andrus

Evaluation of 45S5 bioactive glass combined as a bone substitute in the reconstruction of critical size calvarial defects in rabbits.

Biomaterial research and tissue engineering have guided new developments in bone replacement. In this study, the osteoconductive and osteoinductive properties of 45S5 Bioglass® (Novabone-C/M®, Porex Surg., Newnan, GA), granules as a bone replacement material for large calvarial defects were evaluated. Rabbit periosteal cells were expanded in culture and used in vivo. Alkaline-phosphatase assay, collagen type I, and calcium expression were applied to confirm osteoblast phenotype. In the in vivo phase, a 15-mm diameter critical size calvarial defect was created in rabbits (n = 14). The defect was reconstructed according to four treatment groups: autogenous bone (n = 2), Bioglass alone (n = 2), Bioglass + bone (n = 5), Bioglass + periosteal cells (n = 5). The animals were killed 12 weeks after surgery, and the samples were analyzed. Periosteal cells grew successfully in vitro. Because of their fast proliferation and potential to differentiate into osteoblasts, they were an excellent source of cells for bone tissue engineering. The best ossification was seen when autogenous bone was used (79.4% ossified), whereas only 8.2% of the defect in the Bioglass group showed ossification. Addition of bone or cells to the Bioglass increased the area of ossification to 42.7% and 30.2%, respectively. Defects replaced with Bioglass showed varying degrees of inflammatory reaction because of the intense cell-mediated biodegradation process. Based on these findings, the use of Bioglass granules to repair large craniofacial defects cannot be advised.

Topical gentamicin does not provide any additional anastomotic strength when combined with fibrin glue

BACKGROUND We evaluated the effect of a combination of fibrin sealant and topical gentamicin on a colonic anastomosis in a rat model. METHODS Partial anastomosis in the transverse colon was performed in 70 male Sprague-Dawley rats aged 6 to 10 weeks using 5 interrupted sutures. The rats were divided into 4 groups (control, gentamicin, fibrin glue, and combination). On postoperative days 3 and 5, the rats in each group were killed, anastomotic bursting pressures scores and bowel loop adhesions were determined, and histologic examination was performed. RESULTS No significant difference was noted in the bursting pressures, adhesions, inflammatory infiltrates, fibroblasts, or neoangiogenesis between the fibrin-glue only and the combination groups for both the day 3 and day 5 subgroups. CONCLUSIONS The combination of topical gentamicin and fibrin glue had little effect because the combination did not provide additional anastomotic strength or decrease the number of adhesions when compared with fibrin glue alone.

Transphyseal bioabsorbable screws cause temporary growth retardation in rabbit femur.

A self-reinforced bioabsorbable poly-L-lactide/polyglycolide (SR-PLGA) 80/20 screw 2.0 mm in diameter was implanted transphyseally across the distal growth plate of the right femur in 24 immature rabbits. Radiologic evaluation revealed a mean shortening of 3.1 mm at 3 weeks, 11.1 mm at 6 weeks, 9.3 mm at 24 weeks, 9.0 mm at 48 weeks, and 12.6 mm at 72 weeks compared with the intact contralateral femur. In 13 control rabbits, drilling without screw placement did not cause any statistically significant femoral shortening. Therefore, the transphyseal SR-PLGA 80/20 screw caused growth retardation for 6 weeks postoperatively, after which the normal growth tendency was recovered until the growth plate was closed. The duration of temporary growth retardation correlated with that of strength retention of the SR-PLGA 80/20 copolymer. These findings suggest that SR-PLGA 80/20 screws can be applied in transphyseal bone fixation. The use of bioabsorbable screws for temporary epiphysiodesis seems attractive but requires further study.

The effect of low-molecular-weight heparin in the survival of a rabbit congested skin flap.

Swelling and congestion of flaps are frequently seen postoperatively and can cause unexpected necrosis. According to previous reports, venous thrombosis seems to be a more frequent problem than arterial occlusion in both experimental and clinical surgery. Few satisfactory venous trauma models exist, and reports on experimental venous thrombosis are rare. The object of this study was to create a rabbit venous occlusion flap model and to evaluate the effect of low-molecular-weight heparin on this flap. Eight New Zealand rabbits were used in the pilot study, in which the ideal congested flap was investigated using a flap pedicle based on the central auricular artery with a skin pedicle 0, 1, 2, or 3 cm wide. The flap (3 x 6 cm) was designed on the central part of the left ear, and the central auricular vein and nerve, the former for venous return, were cut out at the base of the flap. The flaps with skin pedicles 0, 1, 2, or 3 cm wide showed mean necrosis length of 60.0, 9.3, 4.2, and 0.0 mm, respectively. The flaps with skin pedicles 0, 1, 2, or 3 cm wide showed mean necrosis of 100, 15.5, 7, and 0 percent, respectively. Therefore, the flap, based on a 1-cm-wide skin pedicle and the central auricular artery, was selected as an optimal congested flap model showing 15.5 percent necrosis. The congested flap was then elevated on the left ear of another 10 rabbits. Subcutaneous low-molecular-weight heparin (320 IU/kg) was administered immediately after surgery to five of the rabbits (the low-molecular-weight heparin group), and the remaining five were used as a control group. Fluorescein was injected 15 minutes after surgery to evaluate the circulatory territory of the flap, and the circulatory territory was measured 5 minutes after injection. The flaps were assessed 7 days after surgery by angiography, histology, and clinical findings. The circulatory territory was significantly greater in the low-molecular-weight heparin group (mean +/- SD, 39.2 +/- 3.0 mm) than the control group (mean +/- SD, 48.0 +/- 1.0 mm) (p < 0.001) assessed 7 days after surgery. The longest flap survival length in group A and group B ranged from 40 to 55 mm (mean +/- SD. 49.4 +/- 5.6 mm) and complete survival (mean +/- SD, 60.0 +/- 0.0 mm). The improvement in survival was statistically significant for group B compared with group A (p < 0.015). Histologic evaluation revealed moderate to severe venous congestion and inflammation in the control group, whereas there were minimal changes in the low-molecular-weight heparin group. Angiography of the flap revealed obvious venous occlusion in the periphery in the control group compared with the low-molecular-weight heparin group. The authors conclude that subcutaneous administration of low-molecular-weight heparin has a great potential to improve the survival length of a congested flap without major complications.

Easy tissue expansion of prelaminated mucosa-lined flaps for cheek reconstruction in a canine model

In head and neck reconstruction, there is sometimes the need for a skin flap lined with mucosa. The object of this study was to determine whether small pieces of mucosa grafted onto the undersurface of a skin flap can be expanded in a reasonable time to provide the material required to reconstruct a full-thickness cheek defect as a free flap. The study consisted of two phases: prelamination and expansion of the flap, and vascularized free-tissue transfer of the flap. Six adult mongrel dogs were used. First, a 5 x 10-cm flap based on the saphenous vessels was elevated on the lower leg, and then four 1 x 2-cm pieces of mucosa harvested from the tongue were grafted onto the undersurface of the flap. A tissue expander (5 x 10 cm) was then placed under the flap, and the incision was closed primarily. The expanders were initially filled with just enough normal saline to obliterate dead space immediately after surgery. The expansion was continued twice weekly for 3 weeks until sufficient expansion was obtained. Two of six flaps were followed for an additional 6 weeks after the 3-week expansion period to observe whether additional mucosa could be obtained. After measurement of the mucosal area, each flap was transferred as free flap to reconstruct an iatrogenic cheek defect. The increase of mucosal surface area was compared with the original graft, and differences were analyzed using the paired t test. All flaps were successfully expanded without any complications. Histologic evaluation revealed that grafted mucosa took well without evidence of graft necrosis, and the intergraft area was covered with histiocytes. Angiography revealed well-defined vascular structures covering the entire area of the flap. The new mucosal area (23.5 +/- 2.4 cm2) was significantly larger than the original mucosal graft (8.7 +/- 0.9 cm2) (p < 0.001). The net increase of the mucosal area was 172.9 +/- 32.4 percent. The increase of mucosal area in two flaps, following a 6-week consolidation period after 3 weeks of expansion, was only slightly greater (25.9 +/- 1.3 cm2) than those without the consolidation period (22.3 +/- 1.8 cm2). This increase of the mucosal area appears to be related to the amount of expansion, and not to the length of the consolidation period. The flaps were successfully transferred as free flaps to reconstruct the full-thickness cheek defects without major complications. Although a staged operation to allow flaps to mature is needed, the present procedure has the advantages of providing a mucosa-lined flap and allowing primary closure of the donor site. The authors conclude that expansion of this flap has great potential in reconstructive surgery.

Protective effect of methylprednisolone on warm ischemia-reperfusion injury in a cholestatic rat liver.

BACKGROUND Cholestasis has been identified as a risk factor for oxidative stress, and it potentially enhances after ischemic-reperfusion injury. The aim of this study was to evaluate the role of methylprednisolone on warm ischemia-reperfusion injury in the presence of cholestasis. METHODS A reversible cholestatic rat model was created. After 7 days, rats received 30 mg/kg of intravenous methylprednisolone 2 hours before ischemia, followed by 30 minutes of ischemia. Rats were euthanized 24 hours after ischemia. Serum aspartate aminotransferase and interleukin-6 were measured, and the liver was harvested for histology and myeloperoxidase estimation. RESULTS Methylprednisolone had a protective effect, with a statistically significant decrease in aspartate aminotransferase (P=.01) and a trend toward decreased levels of interleukin-6 (P=.07). Histology showed a significant difference in architectural distortion (P=.01), cytoplasmic vacuolation (P=.01), and nodular hepatocellular necrosis (P=.04). CONCLUSIONS Methylprednisolone attenuated the ischemic-reperfusion injury in the presence of cholestasis and can be considered for clinical use in the presence of cholestasis.

Prefabricated buccal mucosa-lined flap in an animal model that could be used for vaginal reconstruction.

Congenital vaginal aplasia, gynecological tumor excision, and male-to-female sex surgery are three clinical conditions in which the plastic surgeon is involved in vaginal reconstruction. Skin-lined or skin-grafted local flaps are currently used, but for many reasons, keratinized skin is not the ideal lining for such a moist cavity because it leads to dryness, desiccation, maceration of the skin, and even hair growth in the cavity. The purpose of this study was to create a subcutaneous cavity lined with mucosa in an area with a predictable blood supply. The abdominal area supplied by the deep circumflex iliac vessels was chosen. Six minipigs were used. Strips of tongue buccal mucosa formed the lining; if additional tissue was required, it was taken from the mucosal aspect of the cheek. The mucosa was expanded by using multiple stab incisions. The mucosa was sutured onto the fascia supplied by the deep circumflex iliac vessels, and the skin incision was closed over a silicone sheet to prevent adhesion to the underlying mucosa. This was left for 1 week to allow the mucosa to take. The prefabricated fascial flap was rolled over a silicone stent and was closed longitudinally to form a cylindrical shape. The flap was placed in a subcutaneous pocket in the right inguinal area. The caudal end was left open and was sutured to the surrounding skin. The silicone stent was used to keep the cavity patent and to prevent adhesions in the early stage of the healing process. Regular digital examination was performed to assess patency and contour; endoscopy allowed assessment of mucosa viability. This method of producing a mucosa-lined flap may provide a solution to the difficult problem of vaginal reconstruction.

Rabbit as a distraction model—pitfalls

The principles of distraction osteogenesis have been successfully applied to the craniofacial skeleton of different animals. The rabbit, in particular, has been evaluated as a model by enumerable authors. To our knowledge, however, none of the studies either report the causes of premature euthanization or the pitfalls leading to the untimely death of the animal. We here describe our experience with 30 rabbits used as a model for mandibular distraction osteogenesis and suggest precautions to take in order to avoid unforeseen problems. Thirty skeletally mature New Zealand white rabbits were used. Fifteen animals had bilateral distraction devices placed on the anterior mandible, and another 15 underwent unilateral distraction osteogenesis. In both groups, 12 animals were euthanized prematurely due to complications that included excessive weight loss (malnutrition), anesthesia/animal-related problems, and distraction device failure. The remaining 18 animals tolerated the operative procedure well. Indisputably, rabbit is an excellent choice for craniofacial experiments, but because of its complex anatomy and physiology, an unexpected outcome frequently occurs. We believe that the following suggestions in relation to the pre-operative selection of a suitable animal model, operative technique, and management of eating problems may help the researcher to choose an appropriate animal and avoid complications leading to early death.