Comparison of Chondrogenic Potential in Equine Mesenchymal Stromal Cells Derived from Adipose Tissue and Bone Marrow

Objective— To compare the chondrogenic potential of adult equine mesenchymal stem cells derived from bone marrow (MSCs) or adipose tissue (ASCs). Study Design— In vitro experimental study. Animals— Adult Thoroughbred horses (n=11). Methods— BM (5 horses; mean [±SD] age, 4±1.4 years) or adipose tissue (6 horses; mean age, 3.5±1.1 years) samples were obtained. Cryopreserved MSCs and ASCs were used for pellet cultures in stromal medium (C) or induced into chondrogenesis±transforming growth factor‐3 (TGFβ₃) and bone morphogenic factor‐6 (BMP‐6). Pellets harvested after 3, 7, 14, and 21 days were examined for cross‐sectional size and tissue composition (hematoxylin and eosin), glycosaminoglycan (GAG) staining (Alcian blue), collagen type II immunohistochemistry, and by transmission electron microscopy. Pellet GAG and total DNA content were measured using dimethylmethylene blue and Hoechst DNA assays. Results— Collagen type II synthesis was predominantly observed in MSC pellets from Day 7 onward. Unlike ASC cultures, MSC pellets had hyaline‐like matrix by Day 14. GAG deposition occurred earlier in MSC cultures compared with ASC cultures and growth factors enhanced both MSC GAG concentrations (P<.0001) and MSC pellet size (P<.004) after 2 weeks in culture. Conclusion— Equine MSCs have superior chondrogenic potential compared with ASCs and the equine ASC growth factor response suggests possible differences compared with other species. Clinical Relevance— Elucidation of equine ASC and MSC receptor profiles will enhance the use of these cells in regenerative cartilage repair.     

Assessment of Humeral Length in Dogs After Repair of Salter–Harris Type IV Fracture of the Lateral Part of the Humeral Condyle

Objective— To evaluate the effect of fracture and subsequent repair on future bone growth of the humerus after Salter–Harris type IV fracture of the lateral part of the humeral condyle (LPHC).
Study Design— Prospective study.
Animals— Dogs (n=11).
Methods— Dogs that had LPHC fracture and an open distal humeral physis repaired (1992–2006) were re-examined and radiographed at ≥12 months of age and humeral length was measured.
Result— Measurements from 11 dogs showed a significant ( P =.02) increase in length of the humeral diaphysis of the affected leg compared with that of the intact limb (median, 1.2%; range, 1.3–3.4%). Condylar deformity secondary to growth disturbance was not observed.
Conclusion— Shortening or growth deformity was not observed after fracture and repair even if a transcondylar screw was placed through the distal humeral growth plate. A mild overgrowth of the humeral diaphysis was observed, although likely considered clinically unimportant.
Clinical Relevance— Fracture of the LPHC and subsequent repair in dogs >3 months of age do not impair growth of the humeral diaphysis. A transcondylar humeral screw placed through the humeral physis will not result in shortening of the humeral diaphysis. Implant removal to allow for further growth is therefore not indicated.     

Clinical Use of Recombinant Human Bone Morphogenic Protein-2 in a Whooping Crane (Grus americana)

Objective— To report use of recombinant human bone morphogenetic protein-2 (rhBMP-2) as adjunctive therapy for treatment of a comminuted, open, proximal humeral fracture in an avian species.
Study Design— Clinical report.
Animals— A 3.5-month-old male whooping crane ( Grus americana ).
Methods— An open, severely comminuted humeral facture was stabilized with an intramuscular (IM) pin/type IA external skeletal fixator with tie-in configuration. rhBMP-2 was applied in a calcium phosphate matrix (CPM) paste directly to the fracture site as a bone graft substitute. Radiographic evidence of bone healing was monitored for 14 weeks.
Results— Substantial bony callus was evident at 4 weeks and at 8 weeks there was bridging callus with obvious bony remodeling. The fixation was destabilized at 9 weeks by IM pin removal, bone healing progressed and the fixator was removed at 11 weeks. By 14 weeks both cortices had been re-established with continued callus remodeling evident.
Conclusion— rhBMP-2, applied in a CPM paste, was used as a bone graft substitute in the treatment of a comminuted, open humeral fracture in a whooping crane.
Clinical Relevance— Use of rhBMP-2/CPM should be considered in treatment of avian fractures.     

Clinical Applications of Demineralized Bone Matrix: A Retrospective and Case‐Matched Study of Seventy‐Five Dogs

Objectives— To evaluate the outcome in dogs treated with demineralized bone matrix (DBM) as an adjunct to orthopedic procedures. Study Design— Retrospective and case‐match study. Animals— Dogs (n=75). Methods— Medical records (1999–2006) and radiographs of dogs that had orthopedic procedures (comminuted fractures, tibial plateau leveling osteotomy [TPLO] where correction for tibial rotation created an osteotomy gap, arthrodeses, open corrective osteotomies) where DBM was used were reviewed for signalment, quantity of DBM implanted, duration of exercise restriction, radiographic healing, and complications. Dogs that had TPLO and correction of tibial torsion (n=15), or arthrodesis (n=16) were compared with case‐matched controls. Data were analyzed using Kruskal–Wallis test, ANOVA, Tukey's HSD test, and logistic regression analysis. Results— Mean (±SD) healing time for orthopedic surgeries with DBM augmentation were 15±6.97 (weeks) and complication rate was 19% (14 dogs). Dogs with a TPLO gap filled with DBM were allowed to return to normal exercise 2 weeks earlier than dogs with a well‐apposed TPLO site. Radiographic healing, duration of exercise restriction, and timing of destabilization were similar in dogs undergoing carpal and tarsal arthrodesis whether they received DBM, autogenous graft, or both. Conclusions— DBM can be used to treat uncomplicated bone defects associated with comminuted fracture repairs, open osteotomies, and arthrodeses in dogs. Under these circumstances, clinicians might expect similar clinical outcomes without the possibility of side effects associated with the harvest of autogenous cancellous bone. Clinical Relevance— DBM is safe for use in dogs.     

Acceleration of Second and Fourth Metatarsal Fracture Healing with Recombinant Human Bone Morphogenetic Protein‐2/Calcium Phosphate Cement in Horses

Objective— To compare the efficacy of recombinant human bone morphogenetic protein‐2 (rhBMP‐2)/calcium phosphate (CP) to autogenous cancellous bone graft (CBG) and to no treatment on bone healing, in surgically induced osteotomies and ostectomies of the accessory metatarsal bones in an equine model. Study Design— Experimental. Animals— Adult horses (n=9). Methods— Segmental ostectomies of the second metatarsal bone (MT2) and osteotomies of the fourth metatarsal bone (MT4) were performed bilaterally in 9 horses. There were a total of 35 defects (1 MT4 was previously fractured) created and supplemented randomly either with no treatment (untreated control), rhBMP‐2/CP cement, or matrix (CPC or CPM), or CBG. Radiography was performed every 2 weeks until study endpoint at 12 weeks. After euthanasia, bone healing was evaluated using radiography, mechanical testing, and histology. Data were analyzed with ANOVA followed by the Duncan's Multiple Range Test or nonparametric analyses. Results— At 12 weeks, radiographic scores for union were significantly greater for the rhBMP‐2 (P<.0001) and CBG (P=.004) groups compared with the untreated control group, for both MT2 ostectomies and MT4 osteotomies. The rhBMP‐2 treated MT2 had greater maximum torque to failure in torsion than CBG and control limbs at 12 weeks (P=.011). Histologic analysis demonstrated increased bone formation and more mature bone at the ostectomy site for MT2 in the rhBMP‐2 and CBG groups compared with the untreated control group. Conclusion— Injection of rhBMP‐2/CP into surgically induced ostectomies and osteotomies of the accessory metatarsal bones might accelerate early bone healing in the horse. Clinical Relevance— RhBMP‐2/CP may be as effective if not superior to CBG as an adjuvant treatment to accelerate healing of bone defects.