Culture of equine fibroblast-like synoviocytes on synthetic tissue scaffolds towards meniscal tissue engineering: a preliminary cell-seeding study.

Introduction. Tissue engineering is a new methodology for addressing meniscal injury or loss. Synovium may be an ideal source of cells for in vitro meniscal fibrocartilage formation, however, favorable in vitro culture conditions for synovium must be established in order to achieve this goal. The objective of this study was to determine cellularity, cell distribution, and extracellular matrix (ECM) formation of equine fibroblast-like synoviocytes (FLS) cultured on synthetic scaffolds, for potential application in synovium-based meniscal tissue engineering. Scaffolds included open-cell poly-L-lactic acid (OPLA) sponges and polyglycolic acid (PGA) scaffolds cultured in static and dynamic culture conditions, and PGA scaffolds coated in poly-L-lactic (PLLA) in dynamic culture conditions. Materials and Methods. Equine FLS were seeded on OPLA and PGA scaffolds, and cultured in a static environment or in a rotating bioreactor for 12 days. Equine FLS were also seeded on PGA scaffolds coated in 2% or 4% PLLA and cultured in a rotating bioreactor for 14 and 21 days. Three scaffolds from each group were fixed, sectioned and stained with Masson's Trichrome, Safranin-O, and Hematoxylin and Eosin, and cell numbers and distribution were analyzed using computer image analysis. Three PGA and OPLA scaffolds from each culture condition were also analyzed for extracellular matrix (ECM) production via dimethylmethylene blue (sulfated glycosaminoglycan) assay and hydroxyproline (collagen) assay. PLLA coated PGA scaffolds were analyzed using double stranded DNA quantification as areflection of cellularity and confocal laser microscopy in a fluorescent cell viability assay. Results. The highest cellularity occurred in PGA constructs cultured in a rotating bioreactor, which also had a mean sulfated glycosaminoglycan content of 22.3 µg per scaffold. PGA constructs cultured in static conditions had the lowest cellularity. Cells had difficulty adhering to OPLA and the PLLA coating of PGA scaffolds; cellularity was inversely proportional to the concentration of PLLA used. PLLA coating did not prevent dissolution of the PGA scaffolds. All cell scaffold types and culture conditions produced non-uniform cellular distribution. Discussion/Conclusion. FLS-seeding of PGA scaffolds cultured in a rotating bioreactor resulted in the most optimal cell and matrix characteristics seen in this study. Cells grew only in the pores of the OPLA sponge, and could not adhere to the PLLA coating of PGA scaffold, due to the hydrophobic property of PLA. While PGA culture in a bioreactor produced measureable GAG, no culture technique produced visible collagen. For this reason, and due to the dissolution of PGA scaffolds, the culture conditions and scaffolds described here are not recommended for inducing fibrochondrogenesis in equine FLS for meniscal tissue engineering.

Risk factors associated with cast complications in horses: 398 cases (1997-2006).

OBJECTIVE: To determine the frequency of and risk factors for complications associated with casts in horses. DESIGN: Multicenter retrospective case series. ANIMALS: 398 horses with a half-limb or full-limb cast treated at 1 of 4 hospitals. PROCEDURES: Data collected from medical records included age, breed, sex, injury, limb affected, time from injury to hospital admission, surgical procedure performed, type of cast (bandage cast [BC; fiberglass tape applied over a bandage] or traditional cast [TC; fiberglass tape applied over polyurethane resin-impregnated foam]), limb position in cast (flexed, neutral, or extended), and complications. Risk factors for cast complications were identified via multiple logistic regression. RESULTS: Cast complications were detected in 197 of 398 (49%) horses (18/53 [34%] horses with a BC and 179/345 [52%] horses with a TC). Of the 197 horses with complications, 152 (77%) had clinical signs of complications prior to cast removal; the most common clinical signs were increased lameness severity and visibly detectable soft tissue damage Cast sores were the most common complication (179/398 [45%] horses). Casts broke for 20 (5%) horses. Three (0.8%) horses developed a bone fracture attributable to casting Median time to detection of complications was 12 days and 8 days for horses with TCs and BCs, respectively. Complications developed in 71%, 48%, and 47% of horses with the casted limb in a flexed, neutral, and extended position, respectively. For horses with TCs, hospital, limb position in the cast, and sex were significant risk factors for development of cast complications. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that 49% of horses with a cast developed cast complications.

Multiple osteochondral autografts for treatment of a medial trochlear ridge subchondral cystic lesion in the equine tarsus.

OBJECTIVE: To adapt the multiple osteochondral autograft technique for treatment of a subchondral cystic lesion (SCL) of the proximal medial trochlear ridge (MTR) of the equine talus and assess long term outcome. STUDY DESIGN: Case report. ANIMALS: Quarter horse stallion with SCL of the proximal MTR of the talus. METHODS: Osteochondral autograft techniques used in dogs and humans were adapted and optimized in equine cadavers. A horse with a SCL of the MTR of the talus was treated by osteochondral autografting. Three osteochondral autografts were harvested from the distal lateral trochlear ridge of the talus in the affected tarsus. No curettage or debridement of the osteochondral lesion was performed. Three recipient beds were predrilled and osteochondral autografts were press-fit into the proximal MTR of the talus. Outcome was assessed by radiography, telephone interview of the owner, and direct observation during training. RESULTS; Postoperative radiographs indicated excellent filling of the osteochondral defect and graft-articular surface congruency. No complications were encountered after surgery. At 10 months after surgery, the horse successfully re-entered reining training, and continued to be sound and performing athletically 2 years after surgery. CONCLUSION: Osteochondral autografting deserves consideration and further evaluation as a primary treatment option for SCL of the proximal MTR of the talus in horses. CLINICAL RELEVANCE: Based on the outcome of this case, osteochondral autografting appears to be a safe, valuable and realistic option for treating SCL of the proximal MTR of the talus in equine athletes.

An in vitro biomechanical comparison of dynamic condylar screw plate combined with a dorsal plate and double plate fixation of distal diaphyseal radial osteotomies in adult horses.

OBJECTIVE: To compare stiffness and strength of a dynamic condylar screw plate combined with dorsal broad dynamic compression plate (DCS-bDCP) fixation with double broad dynamic compression plate (dbDCP) fixation used to repair oblique distal fractures of adult equine radii. STUDY DESIGN: Experimental. SAMPLE POPULATION: Adult equine radii (n=10 pair). METHODS: An unconstrained three-dimensional loading-measurement system was used to determine stiffness of a 50 mm long intact, and then DCS-bDCP or dbDCP-plated osteotomized/ostectomized segment of radii when subjected to a nondestructive sequence of compression, torsion, and lateral-to-medial (LM), medial-to-lateral (ML), cranial-to-caudal (CrCa), and caudal-to-cranial (CaCr) bending. Uniform load over the entire length of construct identified its weakest characteristics during torsion and LM and CrCa bending to failure. RESULTS: No difference was observed between osteotomized/ostectomized DCS-bDCP and dbDCP construct stiffness for all 6 loading modes, and strength for all 3 failure loads. Ostectomized DCS-bDCP and dbDCP construct stiffness was significantly lower than osteotomized radii, the latter approaching intact for axial, LM, and CrCa bending. Most frequent failure was bone fracture through exit site of a screw located adjacent to osteotomy/ostectomy. CONCLUSIONS: DCS-DCP and dbDCP constructs had comparable strength and stiffness when repairing osteotomies/ostectomies in equine adult radius bone. Fracture reduction increased stiffness that approached intact bone for loads that placed the unplated side in compression. CLINICAL RELEVANCE: DCS-bDCP and dbDCP constructs are comparable in stiffness and strength when applied to oblique distal diaphyseal osteotomies/ostectomies in equine radius bone. However, the DCS's localized effect on distal epiphyseal structure because of additional bone removal remains to be investigated under in vivo articular loading conditions.

Development of an in vitro three dimensional loading-measurement system for long bone fixation under multiple loading conditions: a technical description.

The purpose of this investigation was to design and verify the capabilities of an in vitro loading-measurement system that mimics in vivo unconstrained three dimensional (3D) relative motion between long bone ends, applies uniform load components over the entire length of a test specimen, and measures 3D relative motion between test segment ends to directly determine test segment construct stiffness free of errors due to potting-fixture-test machine finite stiffness.Intact equine cadaveric radius bones, which were subsequently osteotomized/ostectomized and instrumented with bone plates were subjected to non-destructive axial, torsion, and 4-point bending loads through fixtures designed to allow unconstrained components of non-load associated 3D relative motion between radius ends. 3D relative motion between ends of a 50 mm long test segment was measured by an infrared optical tracking system to directly determine its stiffness. Each specimen was then loaded to ultimate failure in either torsion or bending. Cortical bone cross-section diameters and published bone biomechanical properties were substituted into classical mechanics equations to predict the intact test segment theoretical stiffness for comparison and thus loading-measurement system verification.Intact measured stiffness values were the same order of magnitude as theoretically predicted. The primary component of relative motion between ends of the test segment corresponded to that of the applied load with the other 3D components being evident and consistent in relative magnitude and direction for unconstrained loading of an unsymmetrical double plate oblique fracture configuration. Bone failure configurations were reproducible and consistent with theoretically predicted.The 3D loading-measurement system designed: a) mimics unconstrained relative 3D motion between radius ends that occurs in clinical situations, b) applies uniform compression, torsion, and 4-point bending loads over the entire length of the test specimen, c) measures interfragmentary 3D relative motion between test segment ends to directly determine stiffness thus being void of potting-fixture-test machine stiffness error, and d) has the resolution to detect differences in the 3D motion and stiffness of intact as well osteotomized-instrumented and ostectomized-instrumented equine radii.

Racing prognosis of horses following surgically repaired olecranon fractures.

Olecranon fracture is a common orthopedic problem in juvenile horses. Prognosis for complete fracture healing when various methods of internal fixation are used is good; however, the impact of olecranon fracture stabilization on the likelihood that foals will start on a racecourse is unknown. Medical records of foals undergoing internal fixation for an olecranon fracture were reviewed. The dam's foaling records were obtained and lifetime racing records were then retrieved for both the affected foals and 1 of their siblings. Twenty-two of 24 repaired fractures healed completely, subsequently, 16 of the foals started in at least 1 race. Statistical calculations suggest that when compared with their siblings, the occurrence of olecranon factures requiring internal fixation in juvenile racehorses will not significantly reduce the likelihood that they will race; however, the siblings had significantly more lifetime race starts and higher career earnings.

Intracranial abscess caused by Rhodococcus equi infection in a foal.

CASE DESCRIPTION: A 4-month-old Missouri Fox Trotter colt was examined for a 5-week history of head tilt after treatment for suspected pulmonary Rhodococcus equi infection. CLINICAL FINDINGS: Computed tomography revealed osteolysis of the occipital, temporal, and caudal portion of the parietal bones of the left side of the cranium. A soft tissue mass compressing the occipital region of the cerebral cortex and cerebellum was associated with the osteolytic bone. TREATMENT AND OUTCOME: A rostrotentorial-suboccipital craniectomy approach was performed to remove fragmented occipital bone, debulk the intracranial mass, and obtain tissue samples for histologic examination and bacterial culture. All neurologic deficits improved substantially within 3 days after surgery. Bacterial culture of the resected soft tissue and bone fragments yielded R equi. CLINICAL RELEVANCE: Intracranial surgery in veterinary medicine has been limited to dogs and cats; however, in select cases, extrapolation of surgical techniques used in humans and small animals can assist with intracranial procedures in horses.

Heel bulb lacerations in horses: 101 cases (1988-1994).

OBJECTIVE: To determine clinical history, structures involved, treatment, and outcome of lacerations of the heel bulb and proximal phalangeal region (pastern) in horses. DESIGN: Retrospective study. ANIMALS: 101 horses. PROCEDURES: Medical records of horses with lacerations of the heel bulb and pastern were reviewed, and follow-up information was obtained. RESULTS: 75 horses were Quarter Horses. Most horses were not treated with antimicrobial drugs prior to referral. Mean +/- SD time from injury to referral was 24 +/- 45 hours (range, 1 to 168 hours). Lacerations were most frequently caused by contact with wire or metal objects. In 17 horses, lacerations involved synovial structures; the distal interphalangeal joint was most commonly affected. One horse was euthanatized after initial examination. Wound treatment consisted of cleansing, lavage, debridement, lavage of affected synovial structures, suturing of fresh wounds, and application of a foot bandage or cast. Fifty-six horses were treated with systemically administered antimicrobial drugs. Follow-up information was collected for 61 horses. Fifty-one horses returned to their intended use and had no further complications; 10 horses had complications associated with the wound, and of those horses, 5 were euthanatized and 1 horse died from an unrelated cause. Horses with lacerations that involved synovial structures had worse outcomes than horses with lacerations that did not involve synovial structures. CONCLUSIONS AND CLINICAL RELEVANCE: Horses that sustain heel bulb lacerations can successfully return to their intended use. Involvement of the distal interphalangeal joint is associated with poor prognosis.

Use of a hand-assisted laparoscopic technique for removal of a uterine leiomyoma in a standing mare.

A 20-year-old Thoroughbred mare was evaluated because of a 2-year history of infertility. The mare had normal estrous cycles and had been bred 7 times by different stallions. Ultrasonographic examination revealed a homogeneous hyperechoic intramural mass in the tip of the right uterine horn; the mass was also detected via hysteroscopy Unilateral ovariectomy and partial hysterectomy were performed by use of a hand-assisted laparoscopic technique. Leiomyoma was diagnosed via histologic examination of the mass. Unilateral ovariectomy and partial hysterectomy are recommended in mares with leiomyoma in a uterine horn, especially if the tumor is associated with infertility. The hand-assisted laparoscopic technique allows direct visualization of abdominal structures and accurate placement of ligatures without applying tension on the broad ligament, and eliminates the risks and costs of general anesthesia.