Spontaneous disc degeneration in the baboon model: magnetic resonance imaging and histopathologic correlation.

Degenerative disc disease is a major source of disability in humans. The baboon model is an excellent natural disease model to study comparable human disease, because baboons are relatively large (adult males 20-26 kg, adult females 12-17 kg), long-lived (30-45 years), well defined, easy to use, and closely related to humans. Published investigations with plain radiographs of disc degeneration in baboons indicated vertebral anatomy and changes that were remarkably similar to those seen in humans, and it would be valuable to determine if magnetic resonance imaging (MRI) and histopathologic evaluation would be useful methods for studying the model, as MRI allows multi-planar visualization of tissues without the use of intravenous contrast and it is superior for evaluating disc hydration, annulus tears, and herniations. The thoracolumbar junctions from 47 randomly selected baboons, ranging in age from 2 weeks to 34 years, were evaluated with MRI and histopathology. Excellent correlation with MRI was observed for changes in disc desiccation, height, and age (P < 0.001). The pathologic analysis demonstrated P values of < 0.001 when comparing histopathology with age and MRI results. All severely degenerated discs seen by MRI were in baboons 14 years of age or older.

Evaluation of multiphase implants for repair of focal osteochondral defects in goats.

The use of biodegradable scaffolds for articular cartilage repair has been investigated by numerous researchers. The objective of this screening study was to examine how the mechanical and physical properties of four multiphase implants can affect the cartilage healing response. Multiphase implant prototypes were prepared using poly(D,L)lactide-co-glycolide as the base material. PGA fibers (FR), 45S5 Bioglass (BG) and medical grade calcium sulfate (MGCS) were used as additives to vary stiffness and chemical properties. Osteochondral defects (3 mm dia. and 4 mm in depth) were created bilaterally in the medial femoral condyle (high-weight bearing) and the distal medial portion of the patellar groove (low-weight bearing) of 16 Spanish goats. Half of the implants were loaded with autologous costochondral chondrocytes. Defect sites (total n = 64, 4 sites/treatment) were randomly treated and allowed to heal for 16 weeks, fully weight bearing. At euthanasia, gross evaluations and biomechanical testing were conducted. Histological sections of the defect sites were stained with H and E, Safranin O/Fast Green or processed to analyze collagen architecture. Sections were semi-quantitatively scored for repair tissue structure. Qualitative evaluations showed that all groups had a high percentage of hyaline cartilage and good bony restoration, with new tissue integrating well with the native cartilage. Gross and histology scoring indicated a significantly higher score for defect healing in the condyle than in the patellar groove, but no difference in healing for implant types or addition/omission of cells was found. This investigation demonstrates that focal, osteochondral defects in caprine distal femurs treated with various implant constructs were repaired with hyaline-like cartilage and good underlying bone. The multiphase implants show potential for treatment of osteochondral defects and long-term studies need to be undertaken to confirm the longevity of the regenerated tissue.

Bone morphogenetic protein but not transforming growth factor-beta enhances bone formation in canine diaphyseal nonunions implanted with a biodegradable composite polymer.

BACKGROUND: The purpose of the present study was to create an effective bone-graft substitute for the treatment of a diaphyseal nonunion. METHODS: A standardized nonunion was established in the midportion of the radial diaphysis in thirty mongrel dogs by creating a three-millimeter segmental bone defect (at least 2 percent of the total length of the bone). The nonunion was treated with implantation of a carrier comprised of poly(DL-lactic acid) and polyglycolic acid copolymer (50:50 polylactic acid-polyglycolic acid [PLG50]) containing canine purified bone morphogenetic protein (BMP) or recombinant human transforming growth factor-beta (TGF-beta1), or both, or the carrier without BMP or TGF-beta1. Five groups, consisting of six dogs each, were treated with implantation of the carrier alone, implantation of the carrier with fifteen milligrams of BMP, implantation of the carrier with 1.5 milligrams of BMP, implantation of the carrier with fifteen milligrams of BMP and ten nanograms of TGF-beta1, or implantation of the carrier with ten nanograms of TGF-beta1. At twelve weeks after implantation, the radii were examined radiographically and the sites of nonunion were examined histomorphometrically. RESULTS: We found that implantation of the polylactic acid-polyglycolic acid carrier alone or in combination with ten nanograms of TGF-beta1 failed to induce significant radiographic or histomorphometric evidence of healing at the site of the nonunion. The radii treated with the carrier enriched with either 1.5 or fifteen milligrams of BMP showed significantly increased periosteal and endosteal bone formation on histomorphometric (p < 0.05) and radiographic (p < 0.02) analysis. CONCLUSIONS: Bone formation in a persistent osseous defect that is similar to an ununited diaphyseal fracture is increased when species-specific BMP incorporated into a polylactic acid-polyglycolic acid carrier is implanted at the site of the nonunion. TGF-beta1 at a dose of ten nanograms per implant did not induce a similar degree of bone formation or potentiate the effect of BMP in this model. CLINICAL RELEVANCE: The biodegradable implant containing BMP that was used in the present study to treat diaphyseal nonunion is an effective bone-graft substitute.

Osteochondral progenitor cells in acute and chronic canine nonunions.

This study examined the ability of cells isolated from early healing segmental defects and from tissue from chronic nonunions to support bone and cartilage formation in vivo and their response to transforming growth factor-beta1 in vitro. Ostectomies (3 mm) were created in the radial diaphysis of four dogs. The dogs were splinted 3-5 days postoperatively and then allowed to bear full weight. At 7 days, tissue in the defect was removed and any periosteum was discarded; cells in the defect tissue were released by enzymatic digestion. The dogs were splinted again and allowed to bear full weight for 12 weeks. Radiographs confirmed a persistent nonunion in each dog. Defect tissue was again removed, any periosteum was discarded, and cells were isolated. Cells were also obtained from the defect tissue by nonenzymatic means with use of explant cultures. One-half of the tissue and one-half of any preconfluent, first-passage cultures were shipped to Cleveland by overnight carrier. At second passage, cells were loaded into ceramic cubes and implanted into immunocompromised mice for 3 or 6 weeks. Harvested cubes were examined histologically for cartilage and bone with use of a semiquantitative scoring system. Confluent fourth-passage cultures of 7 and 84-day defect tissue cells were cultured with 0.03-0.88 ng/ml transforming growth factor-beta1 for 24 hours, and [3H]thymidine incorporation and alkaline phosphatase specific activity were determined. Donor-dependent differences were noted in the rate at which defect cells achieved confluence; in general, cells from 7-day tissue divided most rapidly. Seven-day defect cells formed less bone and at a slower rate than was seen in the ceramic cubes containing samples from day 84. Cells derived enzymatically behaved similarly to those from explant cultures. Ceramic cubes contained fibrous connective tissue, cartilage, bone, and fat, indicating that multipotent cells were present. Stimulation of [3H]thymidine incorporation in response to transforming growth factor-beta1 was donor dependent and variable; only two of six separate isolates of cells exposed to it had measurable alkaline phosphatase activity (which was relatively low), and none of the cultures exhibited an increase in response to transforming growth factor-beta1 for 24 hours. This indicates that mesenchymal progenitor cells are present in the healing defect tissue at 7 and 84 days and that the relative proportion of osteochondroprogenitor cells is greater at the later time. The response to transforming growth factor-beta1 is typical of multipotent mesenchymal cells but not of committed chondrocytes or osteoblasts, indicating that these committed and differentiated cells are not present in early stages of healing and suggesting that their differentiation is inhibited in chronic nonunion.

TP53 overexpression in radiation-induced osteosarcoma of the rabbit mandible.

The objective of this study was to investigate the incidence of overexpression of TP53 (formerly known as p53) in osteosarcomas occurring after treatment of rabbit mandibles with high-dose external-beam radiation. As part of a protocol investigating hyperbaric oxygen treatment for osteoradionecrosis, 102 female New Zealand-White rabbits underwent mandibular radiation treatments with a total dose of 64 Gy in 20 treatment fractions. Twelve animals died during irradiation, leaving 90 animals at risk for tumor development. These animals were divided into one control group and 12 other groups each treated with different schedules of postirradiation hyperbaric oxygen. All animals were sacrificed after the hyperbaric oxygen treatment, approximately 8 months after completion of irradiation. Seventeen of the 90 animals that survived after irradiation developed high-grade osteosarcomas, for a 19% incidence of malignancy. Tumor sizes ranged from 1-4 cm. Immunohistochemistry staining of the 17 tumors detected a 59% overall incidence of TP53 overexpression. There was no correlation between the intensity of hyperbaric oxygen treatment and development of osteosarcoma. The high incidence and short interval of development of osteosarcoma suggest that the study animals may have had a genetic predisposition to radiation-induced osteosarcoma. Additionally, our data provide further evidence that TP53 mutations may play an important role in radiation-induced osteosarcoma.

"Mandibular and facial reconstruction" rehabilitation of the head and neck cancer patient.

Cancer surgery and its reconstruction today can result in functional and cosmetically supportive results for most patients. Today's better understanding of malignant tumor kinetics has evolved preservative and precision extirpative surgery which has on one hand enhanced cure rates and on the other hand has preserved function and appearance in such patients. Today's reconstructive techniques have a sound basis in wound healing and bone regeneration science so as to be predictable and long lasting. We now consistently reconstruct cancer patients with minimal complications and without adding undue deformity and disability. Within just the past ten years the additional advancement of osseointegrated implants has taken functional results related to chewing force and efficiency to a higher level. These implants provide for an optimal return of eating ability to closely match those with natural dentitions. Such implants have also been taken a step further with their application to facial unit restorations. Today facial units with the excellent tissue color and consistency matches made possible by available elastic materials are worn with confidence and comfort by those who require them. The messy and ineffectual adhesives of the past have been discarded. At the time of this writing the hope for the immediate future is the availability of recombinant human BMP. As the next anticipated advance in complete cancer reconstruction it has the potential to regenerate physiologically normal bone without bone grafting. The reduction in morbidity and operating room time would be an enormous step forward, as would the applicability of reconstruction to more people at a reduced cost.

Surgical correction of severe vaginal introital stenosis in female baboons (Papio sp.) infected with simian agent 8.

Simian agent 8 (SA8) is a neurotropic, endemic alphaherpesvirus in Papio sp. Lesions associated with simian agent 8 infections usually involve the genitalia of sexually mature baboons. In females, secondary bacterial infections may lead to scarring of the vulva and perineum, with resulting vaginal obstruction. Affected baboons are poor breeders and often develop urinary tract infections due to retention of urine in the vagina. Reconstructive vaginal surgery was performed on seven baboons with vaginal obstruction. Four weeks before surgery, 50 mg of Depo-Provera was administered intramuscularly to each animal to arrest the estrous cycle, reduce swelling, and allow adequate surgical exposure of the vagina. Ventral and circular incisions were made around the constricted vagina. The peripheral skin was excised to break down the underlying connective tissue, allowing access to the vaginal mucosa. Healthy vaginal tissue was then opposed to the incised skin edges with 3-0 Vicryl in an interrupted pattern. Once healed, the vaginas remained patent, allowing normal mating and urination. Nine months after surgery, five baboons had healthy infants, and pregnancy was confirmed in the other two animals. The described procedure may allow female baboons previously infertile and at risk of urinary tract infections to be returned to a healthy reproductive state.

Total reconstruction of the temporomandibular joint with cryogenically preserved allograft mandibular condyle, meniscus, and fossa in the adult goat.

OBJECTIVE: The purpose of this study was to determine the feasibility of total temporomandibular joint reconstruction with the use of cryogenically preserved allograft mandibular condyle, disk, and fossa in the adult goat. STUDY DESIGN: This study consisted of an experimental group of 10 adult goats that underwent total resection of their temporomandibular joints. These animals were immediately reconstructed with cryogenically preserved mandibular condyles, disks, and fossa. These allograft were harvested from 10 donor animals matched to the recipient animal with respect to age, sex, and head dimensions. These grafts were harvested and stored at -185 degrees C for several months before implantation. At 2 years after surgery, the animals were killed. They were evaluated, clinically, radiographically, and histologically. RESULTS: Fifty percent of the eight surviving animals had a normal clinically functional joint with an intact superior joint space and viable articular surfaces. CONCLUSION: The authors conclude that this temporomandibular joint reconstruction technique is feasible in the adult goat model and is deserving of further research.

Balloon angioplasty with stent implantation in experimental coarctation of the aorta.

BACKGROUND: Balloon angioplasty of coarctation of the aorta is an effective method of treatment but is complicated by tearing of the aortic intima, formation of aneurysms, and restenosis. Stent placement at the time of balloon dilation could prevent restenosis and could also prevent progression of intimal tears to aneurysms. The purpose of this study was to evaluate the feasibility of balloon dilation and implantation of balloon-expandable stents in an experimental model of coarctation and to examine the effect of stent placement at the site of surgically created stenosis. METHODS AND RESULTS: Coarctation of the aorta was surgically produced in 11 juvenile swine. Simultaneous coarctation angioplasty and stent implantation was performed in 10 animals 34 +/- 7.8 days after surgery. Repeat catheterization was performed 59 +/- 6 days after stent implantation. Five animals underwent reexpansion of stents with subsequent follow-up catheterization. Aortic specimens were examined by light microscopy and scanning electron microscopy. Coarctation angioplasty with stent implantation was successful in all, with an increase in coarctation diameter from 46 +/- 8.5% to 90 +/- 12.2% of proximal aortic diameter (P = .0001). Systolic pressure gradient decreased from 32 +/- 19.8 to 0.5 +/- 2.8 mm Hg (P < .001). All stents were patent at follow-up catheterization, with no evidence of intraluminal thrombosis. Reexpansion in five animals increased the stent diameter from a mean of 77.4 +/- 12.1% to 93 +/- 11.0% of proximal aortic diameter (P = .02). Gross examination of aortic specimens demonstrated formation of neointima over the stent wherever the stent struts were in contact with the aortic wall. The stent occupied a subintimal position and produced minimal compression of the underlying media. Medial compression was noted immediately beneath stent struts, but there was no evidence of intimal or medial dissection. CONCLUSIONS: Balloon angioplasty with simultaneous implantation of balloon-expandable stents is effective in relieving aortic obstruction in experimental coarctation. Reexpansion of the rigid stent can be performed in an area of surgical aortotomy and coarctation without significant intimal or medial injury. Stent implantation may be useful in preventing restenosis and aneurysm formation after angioplasty of coarctation.

A comparison of the form and function of the human, monkey, and goat temporomandibular joint.

The anatomic form and function of the human temporomandibular joint is compared with the monkey (Macaca and Papio) and goat joint. The findings indicate that the joint of the goat seems to be superior to the joint of Macaca and Papio as a research model.

Re-expansion of balloon-expandable stents after growth.

OBJECTIVES: The purpose of this study was to evaluate the feasibility of re-expansion of balloon expandable intravascular stents and to examine the gross and histologic effects of re-expansion on vascular integrity. BACKGROUND: Intravascular stents have been used successfully as an adjunct to balloon dilation of congenital pulmonary artery branch stenosis and postoperative stenosis of the pulmonary arteries in children. However, use of rigid stents in children could result in development of relative stenosis at the site of stent implantation with subsequent growth of the child. METHODS: Stainless steel "iliac" stents were placed in the thoracic aorta of 10 normal juvenile swine by a transcatheter technique. Angiography and re-expansion were performed at a mean of 11 weeks (n = 9) and again at 18 weeks (n = 5). After euthanasia, the aortic specimens were removed for gross and histologic examination. RESULTS: Stents were successfully implanted in 10 swine. Re-expansion was successfully performed in each animal at 11 weeks and at 18 weeks. Aortic growth produced a relative constriction of the aorta of 20% +/- 10% (mean +/- SD) at the site of stent implantation at both 11 and 18 weeks. Re-expansion produced a significant increase in mean stent diameter from 10.1 +/- 1 mm to 12.3 +/- 1.2 mm at 11 weeks and from 11.2 +/- 0.7 to 13.5 +/- 1.1 mm at 18 weeks after implantation (p < 0.001). Balloon dilation produced a relative increase in stent diameter of 21% +/- 7% at 11 weeks and 18% +/- 4% at 18 weeks. Stent re-expansion was accompanied by plastic deformation of the neointima without neointimal dissection. Where neointima was thick, there was no evidence of neointimal abrasion, but where neointima was thin, areas of localized neointimal abrasion were observed with focal fibrin and platelet adherence to the stent struts. There was no evidence of medial or adventitial hemorrhage or dissection produced by re-expansion. CONCLUSIONS: Re-expansion of intravascular stents is feasible after growth in juvenile swine without significant injury to neointima, media or adventitia. The results of this study support careful and selective use of intravascular stents as an adjunct to balloon dilation of congenital stenoses in children.

Intermittent gastric dilatation after gastropexy in a dog.

Gastroperitoneal adhesions, which developed after tube gastrostomy in a 3-year-old dog, caused an inverted L configuration of the pyloric antrum and duodenum, resulting in periodic episodes of gastric dilatation. The dog had undergone tube gastrostomy for treatment of gastric dilatation/volvulus, but gastropexy adhesions broke down 27 months later, necessitating a second pexy procedure. Adhesions then developed, constricting gastric outflow and trapping gas in the stomach and proximal duodenum. When the ventral row of adhesions was surgically dissected, the angle between the pyloric antrum and the duodenum was straightened, facilitating flow of digesta. Gastropexy rarely causes the degree of adhesion formation and the complications reported in this dog.

Cardiopulmonary reference standards in the pregnant sheep (Ovis aries): a comparative study of ovine and human physiology in obstetrics.

1. Complete blood counts, respiratory rate, tidal volume, arterial and mixed venous blood gases; heart rate, cardiac output, and arterial, pulmonary artery, central venous and pulmonary wedge pressures were determined in 10 pregnant adult sheep of 42.7-65.7 kg body weight. 2. Arithmetic means, standard deviations, and coefficients of variation were calculated to develop reference values; in addition, the 95% confidence limits for ranges were established. 3. Comparison of derived data with that from non-pregnant sheep shows changes similar to those seen when examining pregnant and non-pregnant humans. 4. These results indicate the pregnant sheep is an acceptable model for human obstetrical research.

Relationship of oxygen dose to angiogenesis induction in irradiated tissue.

This study was accomplished in an irradiated rabbit model to assess the angiogenic properties of normobaric oxygen and hyperbaric oxygen as compared with air-breathing controls. Results indicated that normobaric oxygen had no angiogenic properties above normal revascularization of irradiated tissue than did air-breathing controls (p = 0.89). Hyperbaric oxygen demonstrated an eight- to ninefold increased vascular density over both normobaric oxygen and air-breathing controls (p = 0.001). Irradiated tissue develops a hypovascular-hypocellular-hypoxic tissue that does not revascularize spontaneously. Results failed to demonstrate an angiogenic effect of normobaric oxygen. It is suggested that oxygen in this sense is a drug requiring hyperbaric pressures to generate therapeutic effects on chronically hypovascular irradiated tissue.

Direct angioplasty of the basilar artery in baboons.

The lower portion of the basilar artery is a site of significant disease, both aneurysmal and stenotic, but has been considered inaccessible via traditional surgical approaches. A baboon model was used to evaluate the feasibility of a direct surgical reconstruction via a transoral-transclival approach, utilizing temporary occlusion of the basilar artery. An arteriotomy was performed and either repaired primarily or a venous patch graft was sutured in place. All animals survived the surgery; one suffered fatal postoperative upper airway obstruction, but the other four were neurologically intact at the time of death. Patency at the site of repair was confirmed both angiographically and histologically. We conclude that temporary clamping in conjunction with direct reconstruction of the basilar artery is feasible in this primate model. Future studies will focus on further applications of this technique.

Cryogenically preserved adult and juvenile goat mandibular condylar transplantation.

Loss of a functional temporomandibular joint (TMJ) has long been a clinical challenge in both children and adults. Although reconstruction to date has been performed with various prosthetic devices or autogenous costochondral grafts, these procedures have a potential for complications and morbidity. Our studies were performed to determine the feasibility of healing, growth, and long-term function of TMJ reconstruction techniques with cryogenically preserved mandibular allografts in the goat model. This species was chosen because the surgical anatomy and biomechanics of the goat TMJ are very similar to those of the human TMJ. The positive results of the studies and their relevance will be described.

Evaluation of Gore-Tex graft material in the repair of right ventricular outflow tract defect.

Following right ventriculotomy, Gore-Tex PTFE vascular grafts were placed in eight neutered male, 6- to 8-week-old, 8- to 12-kg pigs. Ten to 14 months after surgery each pig was evaluated by right heart catheterization. The swine were sacrificed and the hearts were evaluated grossly and microscopically. Comparison of the mean derived cardiovascular hemodynamic parameters in this group with published data on swine and humans indicated normal cardiovascular physiology. Since there was no gradient across the patched areas, it appears that the patches had no adverse effects on the cardiovascular system of growing pigs over an approximately 1-year time period. In addition, the Gore-Tex appeared to be satisfactory for the repair of right ventricular outflow enlargement. Its relative ease of handling, configuration, and lack of aneurysm formation were advantages over other available materials. However, focal calcification and chronic inflammatory reaction did indicate the possibility of long-term prosthetic failure.

Simulated nasoalveolar palatal defects: a canine model to study bone grafts.

A simple, surgical technique to create a simulated nasoalveolar palatal defect in the canine is described. The procedure has been performed in 60 canines, and we have not experienced closure of the created clefts. The surgically created clefts had to fulfill five criteria: (1) bilateral maxillary alveolar clefts had to exist in each research animal; (2) each cleft had to have a 1-cm bony width; (3) a demonstrable oronasal communication had to be present; (4) each cleft had to be lined by healthy epithelialized mucosa; (5) there had to be functional teeth adjacent to each side of every cleft.

Glutaraldehyde-fixed bovine pericardium for right ventricular outflow enlargement.

Right ventricular outflow tract enlargement using glutaraldehyde-fixed bovine pericardium was evaluated experimentally using the pig model. Twelve piglets (8-16 kg) underwent closure of surgically created, longitudinal right ventriculotomies using premeasured, round, bovine pericardial patches. Half of the animals had pulmonary artery banding to increase right sided pressures. The animals were studied an average of 5.1 months following surgery. Cardiac catheterization was performed on eight animals. Pressure measurements revealed no gradient across any outflow patch, and angiography showed no stenosis or dilatation. There was marked right ventricular hypertension in the banded animals. The hearts from all twelve animals were examined grossly and microscopically. There was endothelialization of all patches with no aneurysm formation. Measurements revealed that most patches had become oval shaped. There was no difference in patch size changes between banded and non-banded animals. The patches showed no significant size change in the longitudinal plane (average from 20.8 mm to 20.3 mm, 6.3% change); however, there was a significant decrease in the transverse size (20.8 mm to 17.3 mm, 16.0%) (p less than 0.001). The change in patch size was probably due to reshaping to conform to the longitudinal ventriculotomy and caused no narrowing. In this study, glutaraldehyde-fixed bovine pericardium appeared to be satisfactory for right ventricular outflow enlargement. Favorable qualities include ease of handling, hemostasis, and no aneurysm formation.