Comparison of autogenous cancellous bone grafts obtained from the sternum and proximal portion of the tibia of llamas.

OBJECTIVE: To describe the anatomy of the sternum in llamas, define the surgical approach to the sternum for collection of cancellous bone graft tissue, and compare the histologic appearance of graft tissue obtained from the sternum with that obtained from the proximal portion of the tibia. DESIGN: Prospective study. ANIMALS: 12 llamas, 3 to 19 years old, that had been submitted for necropsy. PROCEDURE: Radiographs were taken of the sternum and left tibia of the llamas. Measurements of the sternum were determined from the radiographs and adjusted for magnification. Sternebrae volumes were estimated from these measurements. Anatomic dissections to the center of the fourth sternebra and the proximal portion of the tibia were made, and a surgical approach to the sternum was developed. Cancellous graft tissue was obtained from each site and submitted for histologic evaluation. RESULTS: Sternebrae 3, 4, and 5 were significantly larger in volume than the other sternebrae. The ventral aspect of the fourth sternebra was readily accessed for removal of graft tissue by making a 6-cm-long ventral midline incision centered 17 cm craniad to the xipnoid. Mean soft tissue thickness overlying the ventral aspect of the fourth sternebra was 3.1 cm. More tissue was obtained from the sternal (mean, 9.11 g) than from the tibial (mean, 5.16 g) sites. Sternal graft tissue consisted of trabecular bone spicules with predominantly hematopoietic marrow, whereas tibial tissue consisted of trabecular bone spicules with only fatty marrow. CONCLUSIONS AND CLINICAL RELEVANCE: The fourth sternebra in llamas is readily accessible for obtaining autogenous cancellous bone graft tissue that consists of predominantly hematopoietic marrow.     

Sternal bone biopsy in standing horses

OBJECTIVE: To develop a technique for sternal bone biopsy in standing horses. STUDY DESIGN: Experimental study. ANIMALS: Five adult horses. METHODS: Horses were restrained in a standing stocks and sedated. The sternal biopsy site, identified by ultrasonographic examination, was clipped and prepared for aseptic surgery and infiltrated with local anesthetic. An electric bone biopsy drill (Osteocore; Institut Straumann AG, CH-4437, Waldenburg, Switzerland)), which yielded 4-mm-diameter bone specimens, was used to obtain sternal biopsies through a small skin incision. Sections (7 microm) of the bone specimens were stained with toluidine blue and Goldner's green trichrome and observed microscopically to determine suitability for histological and histomorphometric evaluation. RESULTS: The most suitable sternal biopsy site was at the 4th or 5th sternebra. The surgical procedure was easy to perform and well tolerated by the horses, and adequate samples were obtained on the first attempt. The only complications were incisional edema in all horses and wound drainage in 1 horse. CONCLUSIONS: Sternal bone biopsy may be successfully performed in standing horses, and the technique described in this report yields architecturally intact bone specimens. CLINICAL RELEVANCE: The sternum is an accessible site for cancellous bone biopsy specimens in standing horses.     

Use of cancellous bone graft in treatment of navicular bone osteomyelitis in a foal

A 3-month-old Quarter Horse filly stepped on a fence staple and developed navicular bone osteomyelitis of the right hindfoot. A 1.5-cm spherical portion of medullary cavity containing purulent material was debrided and flushed with 0.9% NaCl solution. Cancellous bone was collected from a caudal sternebra and placed into the defect. The solar defect had filled with granulation tissue and was epithelialized 6 weeks after surgery. At 6-month follow-up evaluation, the navicular bone defect had healed and the foal was sound on the limb. Cancellous bone grafting may have merit for the treatment of navicular bone osteomyelitis in the horse.     

Evaluation of a technique for collection of cancellous bone graft from the proximal humerus in horses

OBJECTIVES: To describe a technique for collecting cancellous bone graft from the proximal humerus in horses. STUDY DESIGN: Prospective evaluation of an experimental bone graft collection technique. ANIMAL POPULATION: Eight horses, 3-15 years, weighing 495-605 kg. METHODS: Horses were anesthetized and positioned in lateral recumbency. The lateral aspect of the proximal humerus was exposed by a 7-10-cm incision extending distally from the greater humeral tubercle, followed by sharp dissection through the omotransversarius muscle and between the infraspinatus and deltoideus muscles. A 12-mm cortical defect was incrementally created in the lateral proximal humerus. Human bone graft harvesting equipment (Acumed, Beaverton, OR) was drilled through this defect to collect a core of cancellous bone. In five horses additional cancellous bone was then collected with conventional instruments. Bone samples were weighed and histologically examined. Horses were monitored and graded for quality of anesthetic recovery, incisional complications, and postoperative lameness. RESULTS: Total mean (+/-SD) surgical time for harvesting bone with the Acumed system and traditional techniques (n=5) was 38+/-6 minutes (range, 32-47 minutes). Mean cancellous bone weight collected with the Acumed system was 3.6+/-0.8 g (range, 2.0-4.6 g), and cancellous bone collected conventionally was 25.6+/-7.5 g (range, 16.8-34.2 g). Minimal incisional complications or postoperative lameness were observed. Mortality was 12.5%; one horse fractured the operated humerus during anesthetic recovery. CONCLUSION: The Acumed system provided limited cancellous bone when used with the technique described. However, the quantity of cancellous bone collected with traditional harvesting instruments was comparable to other sites used in horses. The procedure was associated with minimal postoperative incisional complications or lameness, but because one horse suffered a catastrophic humeral fracture further research is required to assess the effects of this procedure on humeral breaking strength. CLINICAL RELEVANCE: Based on the risk of catastrophic fracture, this technique cannot be recommended for use in clinical cases, especially if an unassisted recovery from general anesthesia is planned.     

Evaluation of the Tibia as a Source of Autogenous Cancellous Bone in the Horse

A 1 cm defect was created in the proximal medial cortex of 12 tibiae, cancellous bone was removed from the site, and the bones were fractured by loading in torsion. The fractures did not occur through the donor sites and the fracture patterns and loads to fracture were similar for the treated tibiae and their untreated mates. Cancellous bone was harvested from the proximal medial aspect of both tibiae in nine adult horses. The soft tissue wounds were monitored for more than 10 days in seven horses and healing of the osseous defects was evaluated radiographically at regular intervals to month 6 in two horses. The site provided convenient access with the horses in lateral or dorsal recumbency, the anatomic landmarks were reliable, and the surgical procedure was simple and short. Complications occurred in only three incisions.     

Optimisation of bone marrow aspiration from the equine sternum for the safe recovery of mesenchymal stem cells

Reasons for performing study: Mesenchymal stem cell (MSC) therapy for orthopaedic disease is being used with increasing frequency; there is a need to define a safe, reliable and effective technique for the recovery of MSCs from the sternum of the horse. Objectives: To describe an optimised safe technique for obtaining bone marrow‐derived MSCs from the sternum of the Thoroughbred horse. Methods: The anatomical relationship of the sternum with the heart and internal anatomy was demonstrated in cadavers. Sternal anatomy was evaluated ultrasonographically and after midline sectioning. Sternebrae were examined histologically after aspiration to determine the effect of needle insertion. The quality of the aspirate was evaluated as the number of colony‐forming units from sequential and separately aspirated 5 ml aliquots and assessed for their multipotency using trilineage differentiation. Results: The optimal safe location for the needle was the 5th sternebra because it had a safe dorsoventral thickness and was cranial to the apex of the heart. This sternebra could be reliably identified ultrasonographically. Aspirates could also be obtained from the 4th and 6th sternebrae, although the former is between the front limbs and the latter closer to the heart. Minimal disruption of the internal bony architecture was seen after needle insertion through the thin outer cortex and the first 5 ml aliquot contained the greatest number of colony‐forming units of mesenchymal stem cells with trilineage capabilities. Conclusions: Accurate placement of a Jamshidi needle into the medullary cavity of the 4th–6th individual sternebrae is facilitated by the use of ultrasonography and enables aspiration of bone marrow reliably with minimal damage to the sternum and risk to the horse. Potential clinical relevance: Sternal marrow aspiration as described is a safe and reliable technique to obtain MSCs for orthopaedic cell‐based therapies.     

Septic Arthritis of the Distal Interphalangeal Joint in 12 Horses

The medical records of 12 horses with septic arthritis of a distal interphalangeal joint were reviewed to determine clinical features and response to treatment. Sepsis was caused by trauma or an injection that resulted in an open or contaminated distal interphalangeal joint. All horses were severely lame. Treatment included broad-spectrum parenterally administered antimicrobial drugs (ten horses), percutaneous through-and-through joint lavage (eight horses), indwelling drains (three horses), immobilization of the limb in a cast (three horses), intraarticular injection of sodium hyaluronate (one horse), intraarticular injection of antimicrobial drugs (five horses), curettage of the distal phalanx (one horse), and cancellous bone grafting to promote fusion (one horse). Five horses were euthanatized. Ankylosis of the affected joint developed in five horses, four of which are pasture sound. Two horses treated medically are sound although one underwent subsequent palmar digital neurectomy for treatment of navicular syndrome.     

Histologic features of the healing of bone graft donor sites in dogs

Healing of cancellous bone graft donor sites in the proximal tibial metaphysis of 12 healthy adult dogs was studied histologically. Cancellous bone was curetted from the metaphysis of the proximal end of the tibia, via a 1-cm diameter circular opening in the medial cortex. A hematoma and fibrovascular tissue filled the bone defect at 2 weeks. At 4 and 8 weeks, endosteal callus, composed initially of cartilage and woven bone and later of lamellar bone, filled the marrow cavity. At 12 weeks, the normal structural arrangement of lamellar bone and hematopoietic marrow was reestablished in the marrow cavity. The medial cortex defect was filled only with lamellar trabecular bone. It was concluded that, in adult dogs, a second cancellous bone graft could be collected from the proximal portion of the tibial metaphysis 12 weeks or more after an initial collection.     

Evaluation of the proximal portion of the femur as an autogenous cancellous bone donor site in dogs.

The proximal portion of the femur was evaluated as a source of autogenous cancellous bone in dogs. Bilateral oval cortical defects were created in the lateral subtrochanteric area of the femur in 16 dogs. Cancellous bone was removed and the weight recorded. Cancellous bone was similarly harvested from the proximal portion of the humerus in 7 of these dogs. Subtrochanteric femoral defects in 11 dogs were randomly assigned to receive cancellous bone graft obtained from the femur (n = 4) or the humerus (n = 7). Subtrochanteric defects in 5 dogs were not grafted. Radiographic assessment of subtrochanteric defects was performed at 4-week intervals, and histologic assessment at 4, 8, 16, and 24 weeks after surgery. Nongrafted donor sites healed by ingrowth of trabecular bone during the first 12 weeks after surgery. By week 24, the lateral cortical wall had reformed, but remodeling was incomplete. Donor sites grafted with cancellous bone healed similarly, but with more rapid healing and more complete remodeling evident by week 24. Although the mean weight of cancellous bone harvested from the proximal portion of the femur (0.82 +/- 0.22 g) was significantly (P less than 0.05) less than that harvested from the proximal portion of the humerus (1.38 +/- 0.29 g), there was no qualitative histologic or radiographic difference in bony healing of grafted defects. We determined that the proximal portion of the femur can be safely used to provide moderate amounts of cancellous bone, and that a second bone graft can be collected from the same subtrochanteric donor site after 12 weeks.     

Arthrodesis of the Distal Interphalangeal Joint in Two Horses Using Three Parallel 5.5-mm Cortical Screws

The distal interphalangeal joint was successfully arthrodesed in two horses using three parallel 5.5-mm cortical screws and an autogenous cancellous bone graft. The screws were directed from the palmar proximal border of the second phalanx dorso-distally across the joint space and into the third phalanx. The technique was first developed on a normal horse. The second horse, a clinical case, ruptured its deep digital flexor tendon with complete luxation of the distal interphalangeal joint. Bony fusion of the distal interphalangeal joint occurred in both horses, but both also had residual lameness at a walk. Twenty-one months after the arthrodesis procedure, the clinical patient died from complications related to a subsolar abscess in the operated limb.     

In vitro comparison of equine cancellous bone graft donor sites and tibial periosteum as sources of viable osteoprogenitors

OBJECTIVE: To compare the osteogenic potential of cancellous bone of conventional graft sites with that of one nonconventional site (fourth coccygeal vertebra) and to investigate the tibial periosteum as a donor site with respect to osteogenic potential. STUDY DESIGN: In vitro osteogenic cell culture system. SAMPLE POPULATION: Eight adult horses. METHODS: Cancellous bone or tibial periosteum was aseptically collected and cut into bone chips or periosteal strips of 1 to 2 mm(3) for primary explant cultures. After 2 weeks, primary tissue cultures that yielded a population of osteogenic cells were counted and subcultured at 1 x 10(5) cells/35-mm dish in osteogenic media. After 7 to 10 days, subcultures were stained with Von Kossa (VK) to assess mineralized bone nodule formation. VK-positive bone nodules were counted as osteoprogenitors and compared among 3 donor sites, which provided consistent primary osteogenic cells (tuber coxae, fourth coccygeal vertebra, periosteum) using ANOVA (P <.05). RESULTS: Sternal and tibial bone yielded viable osteogenic cells from 25% and 50% of horses, respectively, whereas yields from tuber coxae, coccygeal vertebra, and periosteum were 75%, 100%, and 100%, respectively. Tuber coxae and periosteum had significantly greater numbers of osteoprogenitors compared with fourth coccygeal vertebra. CONCLUSIONS: Among the conventional donor sites, tuber coxae most consistently yielded viable osteogenic cells with an acceptable percentage of osteoprogenitors. Sternal and tibial sites were unreliable in providing osteogenic cells. Two new donor sites, the fourth coccygeal vertebra and tibial periosteum, were tissues with good osteogenic potential. CLINICAL RELEVANCE: When a source of transplantable viable osteoprogenitor cells is desired, use of the tuber coxae as a conventional donor site is warranted. Use of tibial periosteum or fourth coccygeal vertebra as reliable sources of transplantable osteoprogenitors should be considered.     

The effect of intra-articular methylprednisolone acetate and exercise on equine carpal subchondral and cancellous bone microhardness

Dorsal carpal osteochondral injury is a major cause of lameness in horses undergoing high intensity training. Intra-articular corticosteroid treatment is used commonly to manage exercise-associated articular pain, but its use remains highly controversial in the equine athlete. This project, therefore, aimed to compare the mechanical properties of intra-articular MPA and diluent-treated middle carpal subchondral and cancellous bone in horses undergoing a short-term treadmill exercise programme. It was hypothesised that subchondral and cancellous bone mechanical properties are influenced by intra-articular administration of methylprednisolone acetate (MPA). Eight 2-year-old female horses had MPA or diluent administered into contralateral middle carpal joints at 14 day intervals, for a total of 4 treatments per horse. Horses underwent a standard treadmill exercise protocol until euthanasia (Day 70). Standard sites were located on the dorsal aspect of third, radial and intermediate carpal bones. Osteochondral samples from each test site were divided into subchondral bone and cancellous bone portions. These were dried, resin-embedded and gold-coated. Microhardness measurements were obtained at each test site. No significant effect of intra-articular treatment was detected. At each site, cancellous bone trabecular struts had an 18-19% higher microhardness value than the overlying subchondral bone. These findings indicate that intra-articular administration of MPA at this dose has no effect on subchondral or cancellous bone adaptation to short-term exercise and, therefore, on the propensity of carpal bones to injury. Further investigation into the calcified cartilage layer, effect of different corticosteroid preparations and diffusion of medication are required.     

Treatment of radial fractures in adult horses: an analysis of 15 clinical cases

Fifteen cases of radial fractures in adult horses weighing more than 300 kg are discussed. Four of the horses were destroyed on humane grounds immediately because of a poor prognosis and expense of internal fixation; and two horses at five days and five weeks, respectively after treatment by cast application was initiated. Internal fixation was used in nine horses but of these only two horses recovered completely and resumed their former activities. In eight cases, two plates were applied, one lateral or medial and the other cranial. The internal fixation techniques of all nine horses were scrutinised and suggestions made for the future treatment of radial fractures. These suggestions are (1) the use of ASIF 5.5 mm cortical bone screws using the total width or thickness of the bone in each case. (2) Application of the dynamic condylar screw with its plate in distal or proximal fractures to allow more support. (3) Incorporation of a cancellous bone graft to the fracture. (4) Possible table recovery to prevent breakdown of the fixation during recovery. It is important that bone plates are applied over the total length of the bone. The fact that only two out of 15 horses survived and recovered completely underlines the problems associated with treatment of radial fractures in the adult horse.     

Use of rotational periosteal flaps in primary repair of sinofacial fractures in seven horses

ABSTRACT

Case history: Medical records were reviewed of horses (n?=?7) undergoing surgery for fracture of one or more facial bones extending into the paranasal sinuses that was repaired primarily within 24 hours of the time of injury using a rotational periosteal flap, between April 2009 and May 2017. A kick from another horse was the cause of the injury of three horses, and one horse was injured when it collided with a tree. The cause of the injury of three horses was unknown.

Clinical findings and treatment: Fractures were of the right maxillary bone in two horses, the left maxillary bone in two horses, the left frontal and left nasal bones in two horses, and the right frontal bones in one horse. The fracture of all but one horse was accompanied by an open wound. The fracture of all seven horses was reduced, stabilised, and covered with a rotational, periosteal flap. Surgery was carried out while standing in six horses, and while anesthetised in one horse. All horses had a deficit in the fractured facial bones after the fracture was reduced. Four horses had complications following surgery, but all horses were reported to have excellent cosmetic outcomes and had retuned to their previous level of activity, as reported by their owners.

Clinical relevance: Covering a primarily repaired sinofacial fracture of a horse with a rotational periosteal flap resulted in good cosmetic outcomes, and may be especially beneficial if the fracture is accompanied by loss of bone.     

A Modified Cloward''s Technique for Arthrodesis of the Normal Metacarpophalangeal Joint in the Horse

A modified Cloward's technique was performed for arthrodesis of one metacarpophalangeal (MCP) joint in eight horses. Dorsal arthrotomies were performed medial and lateral to the common digital extensor tendon and two 16 mm holes were drilled through the joint. A perforated cylindrical stainless steel basket filled with cancellous bone was impacted into each hole. The limbs were supported in casts for 8 weeks. The joints were examined and radiographed at 4 weeks, 8 weeks, 6 months, and 10 months. One horse was euthanatized at week 14 to assess the progress of the arthrodesis. In the other seven horses, there was clinical fusion at month 6. Dynamographic evaluations were performed 11 months after surgery at the walk and trot. The maximum vertical forces exerted during weight bearing by treated and control limbs were compared. No difference was detected at the walk; however, a significant difference was present at the trot (p less than 0.05). It was calculated that at the trot the horses placed 90% as much force on the treated limb as on the control limb. Eleven months after surgery, the baskets contained compact and cancellous bone. Ingrowth of bone occurred through all openings, completely filling the baskets and fusing the joints.     

Use of an external skeletal fixation device and bone graft for arthrodesis of the metacarpophalangeal joint in horses

Arthrodesis of the metacarpophalangeal joint was attempted in 9 adult horses, using a new external skeletal fixation device and autogenous cancellous bone graft. Four horses had excellent results, 2 horses had fair results, arthrodesis failed in 2 horses, and 1 horse fractured the third metacarpal bone. The advantages of the technique included minimal surgical trauma to the area of injury, immediate rigid fixation of the metacarpophalangeal joint, and access for treatment of soft-tissue lesions at the site of injury. Although pin loosening and pin-track infection were complications when the device was in place, all pin tracks healed without complications after device removal.     

Effect of beta-tricalcium phosphate in surgically created subchondral bone defects in male horses

beta-Tricalcium phosphate (TCP) was evaluated as a synthetic bone grafting material in horses. Sixteen horses were randomly assigned to 4 groups of 4 horses. In each horse, an 11-mm by 10-mm circular defect with a smooth margin was created surgically in the weight-bearing surface of both third metacarpal bones (McIII) and the left third metatarsal bone. The defect in the left McIII was filled with TCP, the defect in the right McIII was filled with a mixture of 50% autogenous cancellous bone (ACB) and 50% TCP, and the defect in the left third metatarsal bone was not filled to serve as a control. Nuclear imaging and radiographic evaluation of surgical sites were performed at postsurgical week (PSW) 4 and 13 in group-I horses and at PSW 26, 39, and 52 in groups-II, -III, and -IV horses, respectively. Horses in groups I, II, III, and IV were euthanatized at PSW 13, 26, 39, and 52, respectively, and specimens were harvested for histologic evaluation. Results were compared among treatment methods, as well as among time intervals. The greatest activity at defect sites was seen in group-I horses at PSW 4. Activity counts in limbs of group-I horses at PSW 13 and counts in limbs of group-II horses at PSW 26 were not significantly (P less than or equal to 0.05) different. Activity counts in groups-III and -IV horses at PSW 39 and 52, respectively, were less than those in groups I and II. However, there was no significant difference in activity counts between limbs implanted with TCP or ACB:TCP.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro heterogeneity of osteogenic cell populations at various equine skeletal sites

Bone cell cultures were evaluated to determine if osteogenic cell populations at different skeletal sites in the horse are heterogeneous. Osteogenic cells were isolated from cortical and cancellous bone in vitro by an explant culture method. Subcultured cells were induced to differentiate into bone-forming osteoblasts. The osteoblast phenotype was confirmed by immunohistochemical testing for osteocalcin and substantiated by positive staining of cells for alkaline phosphatase and the matrix materials collagen and glycosaminoglycans. Bone nodules were stained by the von Kossa method and counted. The numbers of nodules produced from osteogenic cells harvested from different skeletal sites were compared with the use of a mixed linear model. On average, cortical bone sites yielded significantly greater numbers of nodules than did cancellous bone sites. Between cortical bone sites, there was no significant difference in nodule numbers. Among cancellous sites, the radial cancellous bone yielded significantly more nodules than did the tibial cancellous bone. Among appendicular skeletal sites, tibial metaphyseal bone yielded significantly fewer nodules than did all other long bone sites. This study detected evidence of heterogeneity of equine osteogenic cell populations at various skeletal sites. Further characterization of the dissimilarities is warranted to determine the potential role heterogeneity plays in differential rates of fracture healing between skeletal sites.     

Paranasal sinus surgery through a frontonasal flap in sedated, standing horses

OBJECTIVE: To report experience with paranasal sinus surgery through a frontonasal flap in sedated, standing horses. STUDY DESIGN: Treatment of 10 horses with naturally occurring paranasal sinus disease through a frontonasal bone flap created with the horses standing. ANIMALS: Ten adult horses. METHODS: After restraint and sedation, local anesthetic was injected subcutaneously along the proposed incision line over the conchofrontal sinus and was instilled into the sinuses through a small hole created in the frontal bone. A 3-sided, rectangular, cutaneous incision that extended through the periosteum was created over the frontal and nasal bones. The incision was extended into the conchofrontal sinus using a bone saw, and the base of the flap, on the midline of the face, was fractured. The sinuses were explored, and the horse was treated for the disease encountered. The flap was repositioned; subcutaneous tissue and skin were sutured separately. RESULTS: The horses had few signs of discomfort during creation of the bone flap and during disease treatment. Diseases encountered included inspissated exudate in the ventral conchal sinus (five horses), feed and exudate throughout the sinuses (one horse), occlusion of the nasomaxillary aperature (one horse), polyp (one horse), osteoma (one horse), and progressive ethmoidal hematoma (one horse). CONCLUSION: In selected cases, surgery of the paranasal sinuses can be performed safely on sedated and standing horses through a frontonasal bone flap. CLINICAL RELEVANCE: Performing surgery through a frontonasal bone flap with the horse standing and sedated, rather than anesthetized, eliminates risks and expense of general anesthesia.