Exercise-induced metacarpophalangeal joint adaptation in the Thoroughbred racehorse.

Repetitive bone injury and development of stress fracture is a common problem in humans and animals. The Thoroughbred racehorse is a model in which adaptive failure and associated development of stress fracture is common. We performed a histologic study of the distal end of the third metacarpal bone in two groups of horses: young Thoroughbreds that were actively racing (n = 10) and a group of non-athletic horses (n = 8). The purpose of this study was to determine whether development of articular microcracks was associated with specific alterations to subchondral plate osteocytes. Morphometric measurements were made in five regions of the joint surface: lateral condyle, lateral condylar groove, sagittal ridge, medial condylar groove, and medial condyle. The following variables were quantified: hyaline cartilage width; calcified cartilage width; the number of tidemarks; microcrack density at the articular surface; blood vessel density entering articular cartilage; the presence of atypical bone matrix in the subchondral plate; bone volume fraction; and osteocyte density. Adaptation of articular cartilage was similar in both groups of horses. Vascularization of articular cartilage was increased in the group of non-athletic horses. Microcracks, which typically had an oblique orientation to the joint surface, were co-localized with blood vessels, and resorption spaces. Microcracking was increased in the condylar grooves of athletic horses compared with the other joint regions and was also increased compared with the condylar groove regions of non-athletic horses. Coalescence of microcracks also led to development of an intracortical articular condylar stress fracture in some joints and targeted remodeling of affected subchondral plate. The subchondral plate of the condyles in athletic horses was sclerotic, and contained atypically stained bone matrix with increased numbers of osteocytes with atypical morphology. However, osteocyte numbers were not significantly different between groups. We conclude that differences in site-specific microdamage accumulation and associated targeted remodeling between athletic and non-athletic horses are much greater than differences in subchondral osteocyte morphology. However, the presence of atypical subchondral bone matrix in athletic horses was associated with extensive osteocyte loss. Although osteocyte mechanotransduction is considered important for functional adaptation, in this model, adaptation is likely regulated by multiple mechanotransduction pathways.

Comparison of radiofrequency treatment and mechanical debridement of fibrillated cartilage in an equine model.

OBJECTIVE: To compare a radiofrequency energy (RFE) prototype probe to mechanical debridement (MD) and a commercially available RFE system used for chondroplasty in the treatment of an experimentally created partial thickness cartilage lesion in horses. The study design was experimental, randomized complete block, n=8, using fifteen mature ponies. METHODS: Grade 2 to 3 cartilage lesions were prepared in both patellae. After 10 months duration, the injuries were used to study the effects of MD, a commercially available bipolar RFE device (CoVac 50; ArthroCare Corporation) and a prototype monopolar RFE device (Smith & Nephew Endoscopy). Six months after treatment the patellae were examined for chondrocyte viability and cartilage structure. RESULTS: Mean depth of cell death was significantly different among groups (controls, MD

The effects of radiofrequency energy probe speed and application force on chondrocyte viability.

OBJECTIVE: To determine the thermal effects of monopolar radiofrequency energy (mRFE) on bovine articular cartilage when it was moved at different speeds and using varying application forces. METHODS: Thirty-six fresh osteochondral sections divided into two groups (18 sections/group) were used in this study. The first group was tested at three speed rates of mRFE probe (1 mm/sec, 5 mm/sec and 10 mm/sec) at a constant force (50 g) applied to the probe tip. In the second group, three application forces of the probe tip were tested (25 g, 50 g and 75 g) at a constant speed (5 mm/sec) (n = 6/test). All tests were performed using a custom-built jig to control the mRFE (Vulcan EAS) probe during a 20-mm pass on each section. After treatment, viability of osteochondral sections was determined by confocal laser microscopy (CLM) combined with vital cell staining. RESULTS: There were not any significant differences in cartilage thickness of tested osteochondral sections among the different speeds or forces. During the mRFE probe treatments at different speeds, CLM demonstrated that probe application at the speed of 1 mm/sec caused significantly greater chondrocyte death than at the speeds of 5 and 10 mm/sec, whereas there were no significant differences in chondrocyte death among the variable application forces (p > 0.05). DISCUSSION: This in vitro study demonstrated that RFE thermal penetration correlated most closely with probe application speed than application force for this mRFE probe. CLINICAL RELEVANCE: Improper use of mRFE may cause thermal injury on articular cartilage.

Role of endochondral ossification of articular cartilage and functional adaptation of the subchondral plate in the development of fatigue microcracking of joints.

The mechanisms that regulate functional adaptation of the articular ends of long bones are poorly understood. However, endochondral ossification of articular cartilage and modeling/remodeling of the subchondral plate and epiphyseal trabeculae are important components of the adaptive response. We performed a histologic study of the distal end of the third metacarpal/metatarsal bone of Thoroughbreds after bones were bulk-stained in basic fuchsin and calcified sections were prepared. The Thoroughbred racehorse is a model of an extreme athlete which experiences particularly high cyclic strains in distal limb bones. The following variables were quantified: microcrack boundary density in calcified cartilage (N.Cr/B.Bd); blood vessel boundary density in calcified cartilage (N.Ve/B.Bd); calcified cartilage width (Cl.Cg.Wi); duplication of the tidemark; and bone volume fraction of the subchondral plate (B.Ar/T.Ar). Measurements were made in five joint regions (lateral condyle and condylar groove; sagittal ridge; medial condylar and condylar groove). N.Cr/B.Bd was site-specific and was increased in the condylar groove region; this is the joint region from which parasagittal articular fatigue (condylar) fractures are typically propagated. Formation of resorption spaces in the subchondral plate was co-localized with microcracking. N.Ve/B.Bd was also site-specific. In the sagittal ridge region, N.Ve/B.Bd was increased, Cl.Cg.Wi was decreased, and B.Ar/T.Ar was decreased, when compared with the other joint regions. Multiple tidemarks were seen in all joint regions. Cumulative athletic activity was associated with a significant decrease in B.Ar/T.Ar in the condylar groove regions. N.Cr/B.Bd was positively correlated with B.Ar/T.Ar (P < 0.05, r(s) = 0.29) and N.Ve/B.Bd was negatively correlated with B.Ar/T.Ar (P < 0.005, r2 = 0.14) and Cl.Cg.Wi (P < 0.05, r2 = 0.07). We conclude that endochondral ossification of articular cartilage and modeling/remodeling of the subchondral plate promote initiation and propagation of site-specific fatigue microcracking of the joint surface, respectively, in this model. Microcracking of articular calcified cartilage likely represents mechanical failure of the joint surface. Propagation of microcracks into the subchondral plate is a critical factor in the pathogenesis of articular condylar fatigue (stress) fracture. Functional adaptation of the joint likely protects hyaline cartilage from injury in the short-term but may promote joint degeneration and osteoarthritis with ongoing athleticism.

Up-regulation of site-specific remodeling without accumulation of microcracking and loss of osteocytes.

Functional adaptation of bone normally protects the skeleton from fracture during daily activity. Accumulation of microcracking and loss of osteocytes have been implicated in the regulation and initiation of targeted (reparative) remodeling of bone and, in certain situations, the development of fatigue or stress fracture. We performed a histologic study of the dorsal cortex of the mid-diaphysis of the third metacarpal (Mc-III) bone of Thoroughbred racehorses after bones were bulk-stained in basic fuchsin and transverse calcified sections were prepared. The Thoroughbred racehorse is an extreme athlete whose Mc-III bone experiences particularly high cyclic strains during training and racing. A group of non-athletic horses was also included in the experiment. The following variables were quantified: activation frequency (Ac.f); bone formation rate (BFR); resorption space density (Rs.N/T.Ar); microcrack density (Cr.Dn); microcrack mean length (Cr.Le); microcrack surface density (Cr.S.Dn); osteocyte density (Ot.N/T.Ar; Ot.N/B.Ar); and bone volume fraction (B.Ar/T.Ar). Ac.f and BFR were estimated using a mathematical algorithm. Using confocal microscopy, bones were examined for fine microcracks, diffuse matrix injury, and disruption of the osteocyte syncytium. Low values for Cr.Dn (#/mm2) were found in both groups (0.022+/-0.008 and 0.013+/-0.006 for racing Thoroughbreds and non-athletic horses, respectively). There was no significant relationship between Cr.Dn and Ot.N/T.Ar; Ot.N/B.Ar, B.Ar/T.Ar, and Ot.N/T.Ar; Ot.N/B.Ar, and remodeling (Ac.f, Rs.N/T.Ar) and Ot.N/T.Ar; Ot.N/B.Ar. Intense remodeling of the Mc-III dorsal cortex was found in the racing Thoroughbreds (Ac.f 12.8+/-7.4 #/mm2/year; BFR 31.5+/-15.6%; Rs.N/T.Ar 0.19+/-0.09 #/mm2) and was significantly increased compared with non-athletic horses. Overall, remodeling was weakly correlated with Cr.Dn (r2=0.15, P<0.05). Subtle matrix injury, not detectable by bright-field microscopy, was particularly evident adjacent to resorption spaces in Thoroughbred bone. In non-athletic horses, disruption of the dendritic cell processes of osteocytes associated with cement lines and interstitial fragments was more evident. Taken together, these findings suggest that site-specific (targeted) induction of remodeling during functional adaptation of bone in a high-strain skeletal site is not dependent on accumulation of microcracking or loss of osteocytes. We hypothesize that athleticism can directly influence bone turnover in this extreme athlete through pathways that do not involve classical linear microcracks.

Degradation of bone structural properties by accumulation and coalescence of microcracks.

Failure of bone adaptation to protect the skeleton from fatigue fracture is common, and site-specific accumulation and coalescence of microcracking in regions of high strain during cyclic loading is considered a key factor that decreases the resistance of whole bones to fracture. We investigated the effect of cyclic fatigue loading on the monotonic structural properties of the rat ulna during accumulation and coalescence of microcracks. Cyclic end-loading of the ulna was performed at 4 Hz ex vivo at an initial peak strain of -6000 muepsilon to 20% loss of stiffness (n = 7) or 40% loss of stiffness (n = 7) bilaterally. A 0% loss of stiffness monotonically loaded control group (n = 7) was also included. Volumetric bone mineral density (vBMD), ultimate strength (F(u)), stiffness (S), and energy-to-failure (U) were determined in one ulna and in the contralateral ulna vBMD, cortical bone area (B.Ar), maximum and minimum second moments of inertia (I(MAX) and I(MIN)), microcrack density (Cr.Dn), microcrack mean length (Cr.Le), and microcrack surface density (Cr.S.Dn) were determined. In two additional groups of rats, cyclic end-loading of the ulna was also performed ex vivo unilaterally to 20% loss of stiffness (n = 10) and 40% loss of stiffness (n = 10) and then vBMD, F(u), S, U, B.Ar, I(MAX), and I(MIN) were determined bilaterally. Fatigue loading had incremental degradative effects on ulna structural properties. This decreased resistance to fracture was associated with accumulation and coalescence of branching arrays of microcracks within the cortex of the ulna. Microcracking was most prominent in the middiaphysis and corresponded to the region of the bone that fractured during monotonic structural testing. Fatigue loading influenced the relationship between bone cross-sectional geometry and vBMD and ulna structural properties. At 40% loss of stiffness, F(u), S, and U were all significantly correlated with cross-sectional bone geometry and vBMD, whereas this was not the case at 20% loss of stiffness and with the 0% loss of stiffness monotonic control ulnae. We also found a biologically significant individual animal effect. Larger ulnae required a higher number of load cycles for fatigue to develop, retained higher strength, and accumulated a greater amount of microcracking at the end of the cyclic fatigue testing. Small increases in bone size and density can substantially improve the resistance of whole bones to fracture as microcracking accumulates and coalesces during cyclic fatigue loading.

Response of the osteocyte syncytium adjacent to and distant from linear microcracks during adaptation to cyclic fatigue loading.

Cyclic loading induces fatigue in bone and initiates a complex, functionally adaptive response. We investigated the effect of a single period of fatigue on the histologic structure and biomechanical properties of bone. The ulnae of 40 rats were subjected to cyclic fatigue (-6000 microepsilon) unilaterally until 40% loss of stiffness developed, followed by 14 days of adaptation. The contralateral ulna served as a treatment control (n = 20 rats), and a baseline loaded/non-loaded group (n = 20 rats/group) was included. Bones from 10 rats/group were examined histologically and the remaining bones (10 rats/group) were tested mechanically. The following measurements were collected: volumetric bone mineral density (vBMD); ultimate force (Fu); stiffness (S); energy-to-failure (U); cortical area (Ct.Ar); microcrack density (Cr.Dn); microcrack mean length (Cr.Le); microcrack surface density (Cr.S.Dn); osteocyte density (Ot.N/T.Ar and Ot.N/TV); bone volume fraction (B.Ar/T.Ar); resorption space density (Rs.N/Ct.Ar); and maximum and minimum area moments of inertia (IMAX and IMIN). Using confocal microscopy, the bones were examined for diffuse matrix injury, canalicular disruption, and osteocyte disruption. The adapted bones had increased B.Ar, IMAX, and IMIN in the mid-diaphysis. Fatigue loading decreased structural properties and induced linear microcracking. At 14 days, adaptation restored structural properties and microcracking was partially repaired. There was a significant nonlinear relationship between Ot.N/T.Ar and B.Ar/T.Ar during adaptation. Disruption of osteocytes was observed adjacent to microcracks immediately after fatigue loading, and this did not change after the period of adaptation. In fatigue-loaded bone distant from microcracks, diffuse matrix injury and canalicular disruption were often co-localized and were increased in the lateral (tension) cortex. These changes were partially reversed after adaptation. Loss of canalicular staining and the presence of blind-ends in regions with matrix injury were suggestive of rupture of dendritic cell processes. Taken together, these data support the general hypothesis that the osteocyte syncytium can respond to cyclic loading and influence targeted remodeling during functional adaptation. Changes in the appearance of the osteocyte syncytium were found in fatigue-loaded bone with and without linear microcracks. We hypothesize that the number of dendritic cell processes that experience load-related disruption may determine osteocyte metabolic responses to loading and influence targeted remodeling.

Bromodeoxyuridine immunohistochemistry of epon-embedded undecalcified bone in a canine fracture healing model.

We evaluated bromodeoxyuridine (BrdU) immunohistochemistry of paraffin-, methyl methacrylate (MMA)-, and epon-araldite (epon)-embedded canine bone specimens to establish an optimal technique for studying cell kinetics of fracture healing in a canine tibial gap model. Dogs were sacrificed 4 months after tibial ostectomy and 1 hr after i.v. injection of BrdU (100 mg/kg). BrdU immunohistochemical staining with a peroxidase-labeled streptavidin-biotin (LAB-SA) method was performed on thin sections of tibia fixed in 70% ethanol and embedded in paraffin, MMA, or epon. Thin section of small intestines fixed in 70% ethanol and 10% neutral buffered formalin and embedded in paraffin, MMA, or epon were BrdU-stained and served as a model for proliferating tissue. Good and consistent BrdU immunostaining without detachment of bone sections was obtained for epon-embedded undecalcified bone sections. BrdU-positive cells were easily identifiable, in contrast to negligible background staining. BrdU-labeled osteoprogenitor cells and osteoblasts were observed around and on the surface of woven bone in external and internal callus of the ostectomy gap. Nuclei of osteocytes were not labeled. In contrast to the epon-embedded specimens, BrdU immunostaining of paraffin-embedded decalcified and MMA-embedded undecalcified bone specimens was unsatisfactory. The results of this study suggest that BrdU immunohistochemistry of ethanol-fixed, epon-embedded, undecalcified canine bone sections is a technique suitable for study of fracture healing with the described methodology.

A study of fracture callus material properties: relationship to the torsional strength of bone.

This study was designed to quantitate the local material properties of fracture callus during gap healing and to relate these local properties to the torsional strength of bone in a canine model under external fixation. Bilateral tibial transverse osteotomies were performed in 32 dogs and stabilized using unilateral external skeletal fixators with a 2-mm gap. Dogs were divided into four equal groups and euthanized at either 2, 4, 8, or 12 weeks. The torsional properties of one bone of each pair were determined. In both bones of each pair, the indentation stiffness, calcium content, and histomorphometric properties of six sites of periosteal callus, six sites of endosteal callus, four sites of cortical bone, and two sites of gap tissue were determined. Each of the four types of tissue had a specific structural or material property change during the study period. The indentation stiffness of periosteal callus increased up to 8 weeks and then plateaued. Endosteal callus stiffness peaked at 8 weeks and then decreased by 12 weeks. Gap tissue stiffness increased linearly over time. Cortical bone stiffness decreased over time. Indentation stiffness was significantly associated with the calcium content of periosteal callus (R2 = 0.50, p less than 0.0001) and gap tissue (R2 = 0.66, p less than 0.0001). The local stiffnesses of gap tissue and periosteal callus were significantly associated with the maximum torque (gap, R2 = 0.50, p less than 0.0001; periosteal, R2 = 0.34, p less than 0.05) and the torsional stiffness (gap, R2 = 0.44, p less than 0.0001; periosteal, R2 = 0.65, p less than 0.0001) of the bone.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrophic nonunion can be predicted with dual energy x-ray absorptiometry in a canine ostectomy model.

This study demonstrated that dual energy x-ray absorptiometry can be used to distinguish between normal union and atrophic nonunion, with high sensitivity and high negative predictive value, by 8 weeks after surgery in a canine model. Eighteen adult mixed-breed dogs were divided into two equal groups: normal union and atrophic nonunion. In the normal union group, a 5 mm mid-diaphyseal transverse ostectomy was performed in the right tibia, and the bone was stabilized with a unilateral external fixator. In the atrophic nonunion group, a 5 mm mid-diaphyseal ostectomy was performed; the distal 1.5 cm of the bone ends, including the periosteum, were frozen twice to -20 degrees C using liquid nitrogen and thawed slowly twice; and the bone was stabilized with a unilateral external fixator. The members of the research team were blinded to the group assignments until after all dogs were killed and all data were acquired. Radiography and dual energy x-ray absorptiometry of both tibiae were performed at weeks 1, 2, 3, 4, 6, 8, 10, 12, 14, and 16 after surgery. All dogs were killed at 16 weeks, and the torsional stiffness and maximum torque of both tibiae of five dogs in each group were determined. Mechanically, the tibiae in the normal union group had significantly higher maximum torque (43-fold higher) and torsional stiffness (86-fold higher) than the tibiae in the atrophic nonunion group. The sensitivity and negative predictive value of dual energy x-ray absorptiometry for predicting nonunion were 100% by 8 weeks after surgery. The specificity and positive predictive value reached 78 and 82%, respectively, by 16 weeks. Radiographic scores were significantly higher for the union group than for the nonunion group beginning at 2 weeks after surgery. The earliest time after surgery that radiography classified an ostectomy as a nonunion was significantly later (4.6 +/- 1.2 weeks) than for dual energy x-ray absorptiometry (2.6 +/- 1.4 weeks). Before these results can be extrapolated to human applications, further animal studies must be performed to evaluate clinically or experimentally induced fractures, or both, rather than the well defined ostectomies that were performed in this study.

Mechanical characteristics of proximal femoral reconstruction after 50% resection.

Six techniques of proximal femoral replacement were compared in vitro with the use of compression, bending, and torsional testing in a canine model. One femur of each pair was osteotomized in the midshaft region, and the proximal portion was replaced with one of six techniques. These techniques included (a) a segmental proximal femoral endoprosthesis cemented into the distal femur with no allograft (technique ES); (b) a long-stem endoprosthesis press-fit into an allograft and cemented into the distal femur with a transverse osteotomy (technique AT); (c) the same construct as technique AT, but with a step-cut at the osteotomy (technique AS); (d) a long-stem endoprosthesis interlocked into an allograft and cemented into the distal femur with a transverse osteotomy (technique AI); (e) a short-stem endoprosthesis cemented into an allograft combined with one plate laterally stabilizing the allograft to the distal femur with a transverse osteotomy (technique AP1); and (f) the same construct as technique AP1, but with an additional plate cranially (technique AP2). A long-stem endoprosthesis cemented into the contralateral intact femur served as the control. Techniques that involved a long-stem endoprosthesis and cementing distally (AT, AS, and AI) were more resistant in torsion than the plated replacement techniques (AP1 and AP2). The segmental replacement construct (ES) was equal to or stronger than all other techniques under each testing condition. In torsion, the addition of a step-cut (AS) significantly lowered angular displacement of the reconstruction when compared with the reconstruction with a transverse osteotomy (AT) (p < 0.05). Bones with one-plate fixation (AP1) were significantly weaker in torsional stiffness and maximum torque and in mediolateral bending (p < 0.05) than all other techniques. The addition of a second plate (AP2) increased the mechanical properties of the construct so that it was greater than the one-plate method and was equal to (bending and compression) or still weaker (torsion) than the other techniques. The results indicate that segmental replacement methods and allograft/endoprosthetic composites that involve long-stem endoprostheses fixed with cement are mechanically superior to methods that involve short-stem endoprostheses with single or double plating at the osteotomy sites.

Dual energy x-ray absorptiometry of implanted femora after cemented and press-fit total hip arthroplasty in a canine model.

The effect of a press-fit and cemented titanium alloy endoprosthesis on the measurement of bone mineral density (BMD) of the proximal femur in a canine model with dual energy x-ray absorptiometry (DXA) was determined. Seven regions of interest, corresponding to zones 1-7 according to Gruen et al., were measured. Eight unpaired femora were scanned with DXA before implantation (five separate scans per femur), after press-fitting with a titanium alloy femoral component (five separate scans per femur); and after cementing with the same component (five separate scans per femur). When the titanium alloy endoprosthesis was press-fit, the BMD of five of seven regions of the proximal femur increased significantly (range, 2.7-23.1%; mean, 11.2%), although the mean precision error of this measurement was not altered (before implantation, 1.1%; after press-fit 1.8%). Cementing of the implant caused a variable effect on BMD, resulting in a decrease in distal regions where cortical bone was relatively thin compared with the cement mantle (regions 3, 4, and 5) and in an increase in the other regions. Cementing of the implant significantly increased the mean precision error of measurement to 7.2% (range, 1.1-12.6%).

Mechanical evaluation of six types of reconstruction following 25, 50, and 75% resection of the proximal femur.

The structural stiffness and the stiffness of the osteotomy site after six types of reconstruction of the proximal femur were compared by testing in axial compression, mediolateral bending, and axial torsion in a canine model. An osteotomy was carried out for 25, 50, or 75% of the length of each femur, and the proximal portion was replaced by one of five allograft/endoprosthetic composites or a segmental replacement. The reconstructions included (a) a composite press-fit proximally and cemented distally, (b) a composite cemented proximally and distally, (c) a composite cemented proximally and fixed with two plates at the allograft-host bone interface, (d) a composite cemented proximally and secured distally with bicortical screws, (e) a composite secured proximally and distally with bicortical screws, and (f) a segmental prosthesis cemented into the distal femur. The results showed that the segmental reconstruction and the reconstruction with double-plate fixation and a cemented endoprosthesis were structurally stiffer and had greater stiffness of the osteotomy site than the other reconstructions. In comparison, reconstructions that involved cement alone or cement and press-fit techniques generally were more compliant than the others, both structurally and at the osteotomy site.

Thermal response and torque resistance of five cortical half-pins under simulated insertion technique.

A model was developed that can quantitate heat generation during placement of half-pins in cortical bone. Five half-pins were tested to assess differences in insertion torque, heat generation, and micro-damage at the pin-bone interface. Thin thermocouple probes were placed 0.5 mm from the track of the pin and within the pin to measure its temperature during insertion. Scanning electron microscopy was used to view the pin-bone interface to assess the microdamage during placement. The design of the tip of the pin influenced insertion torque and heat generation. Higher heat generation was measured when a thermocouple was placed within the pin itself and less was measured when thermocouple probes were placed within bone samples 0.5 mm from the impending pin track. Furthermore, insertion torque and thermal responses were related, but there were no significant differences in microdamage to bone when different pins and drilling/tapping techniques were used. Due to the significant heat generation at the pin-bone interface, proper cooling with saline irrigation should be applied during pin insertion regardless of the design of the pin. The microdamage observed at the surface of the pin track may have significant implications with regard to loosening of pins, but such effects must be studied with in vivo models.

In vitro and in vivo study on the effect of autogenous cancellous bone and intramedullary polymethylmethacrylate on allograft construct strength.

An in vitro study was performed to compare the effects of augmenting interlocking nails of one of two diameters (5 or 6 mm) with intramedullary polymethylmethacrylate. Subsequently, an in vivo study was performed to compare the effects of augmenting the interlocking nail with five combinations of intramedullary polymethylmethacrylate and autogenous cancellous bone applied to the periosteal surface or within the medullary canal. Dogs were killed 6 months after the procedure for biomechanical evaluation of the femora in axial compression, mediolateral and craniocaudal bending, and torsion. Results from the in vitro study at the proximal osteotomy indicated the 6-mm interlocking nail with intramedullary polymethylmethacrylate had greater stiffness than the 5-mm interlocking nail without it (p < 0.05). At the distal osteotomy, regardless of the diameter of the interlocking nail, the addition of intramedullary polymethylmethacrylate increased stiffness (p < 0.05). Results from the in vivo study indicated greater global construct stiffness with an interlocking nail alone, an interlocking nail augmented with intramedullary polymethylmethacrylate and cancellous bone at the periosteal surface, and an interlocking nail augmented with cancellous bone within the medullary canal and at the periosteal surface (p < 0.05). At the osteotomy level, the interlocking nail augmented with intramedullary polymethylmethacrylate and cancellous bone at the periosteal surface had greater stiffness than did an interlocking nail alone or an interlocking nail augmented with either intramedullary polymethylmethacrylate, cancellous bone within the medullary canal, or cancellous bone at the periosteal surface (p < 0.05) but produced the same results as an interlocking nail augmented with cancellous bone within the medullary canal and at the periosteal surface. The results suggest that augmenting interlocking nail fixation with intramedullary polymethylmethacrylate by itself offers no advantage but that a combination of intramedullary polymethylmethacrylate and cancellous bone at the periosteal surface improves structural properties at 6 months.

A mechanical comparison of gluteus medius attachment methods in a canine model.

Seven techniques of gluteus medius attachment were compared in vitro in a canine model. Four methods were used for tendon to tendon attachment: (a) two modified Bunnell-Mayer sutures, (b) two Kessler locking loop sutures, (c) two horizontal mattress sutures, and (d) two sutures with a looped suture technique. The two methods of tendon to bone attachment were repair with a 1 cm spiked staple and repair with a spiked washer and screw, and the method of bone to bone attachment involved two figure-eight cerclage wires. The contralateral limb served as a control. Compared with the bone to bone attachment, the four tendon to tendon attachments and the two tendon to bone attachments had significantly less strength (31 and 30% of the ultimate load for bone to bone repair, respectively) and tensile stiffness (24 and 39% of the tensile stiffness for bone to bone attachment, respectively) (p < 0.0001). The control specimens were significantly stronger and stiffer than all specimens (p < 0.05) except those that had bone to bone fixation. There were no significant differences among the four tendon to tendon suture repairs with regard to either strength or stiffness; the values ranged from 28-45% of those of the controls. Among the tendon to bone repairs, fixation with a spiked washer and screw was significantly stronger than that with a spiked staple (p = 0.032), but there was no difference between these two techniques with regard to stiffness.

The effect of recombinant human bone morphogenetic protein-2 on femoral reconstruction with an intercalary allograft in a dog model.

This study compared the effect of augmentation of allograft host bone junctions with recombinant human bone morphogenetic protein-2 (rhBMP-2) on an absorbable collagen sponge (ACS), autogenous cancellous bone graft (CBG), and a collagen sponge alone in a canine intercalary femoral defect model repaired with a frozen allograft. Outcome assessment included serial radiographs, dual energy X-ray absorptiometry scans, and gait analyses, and mechanical testing and histology of post-mortem specimens. The distal junction healed more quickly and completely with rhBMP-2 than ACS alone based on qualitative radiography and histologic evaluations. The primary tissue in the unhealed gaps in the ACS group was fibrous connective tissue. The proximal allograft host bone junction had complete bone union in the three treatment groups. There was significantly greater new bone callus formation at both junctions with rhBMP-2 than with CBG or ACS alone that resulted in increased bone density around the allograft host bone junctions. All dogs shifted their weight from the treated leg to the contralateral pelvic limb immediately after surgery. Weight bearing forces were redistributed equally between the pelvic limbs at 12 weeks after surgery with rhBMP-2, at 16 weeks after surgery with CBG, and at 24 weeks after surgery with ACS alone. Bending and compressive stiffnesses of the whole treated femora were equal to the contralateral control femora in all treatment groups, whereas torsional rigidities of the whole treated femora for the CBG and ACS groups were significantly less than the control. Both the proximal and distal junctions the treated with rhBMP-2 had torsional stiffnesses and strengths equal to intact control bones. Ultimate failure torques of the proximal junctions of the CBG group and of both junctions of the ACS group were significantly less than the BMP-treated bones. Augmentation of the allograft host bone junctions with rhBMP-2 on an ACS gave results for all parameters measured that equaled or exceeded autogenous graft in this canine intercalary femoral defect model.

Comparison of three methods of gluteal muscle attachment to an allograft/endoprosthetic composite in a canine model.

This study used radiography, gait analysis, gluteal muscle mass, mechanical testing, and qualitative histology to compare three methods of gluteal muscle attachment to an allograft/endoprosthetic composite of the proximal 25% of the femur in an in vivo canine model. The three methods of gluteal muscle attachment were identical to those used clinically in human patients for hip revision and proximal femoral limb salvage: the host gluteal tendon sutured to the allograft tendon (tendon group), the host greater trochanter with intact gluteal tendons secured to the allograft with a cable-grip system (grip group), and periosteally vascularized proximal femoral bone onlay with intact tendons wrapped around the allograft (wrap group). On the basis of radiographs taken every 2 months, the tendon group had more graft fractures than did the grip or wrap group. Radiographic union of the graft-host bone junction occurred more rapidly and there was less graft resorption in the wrap group than in the other two groups. In all dogs, peak vertical ground-reaction forces in the treated limb decreased immediately after surgery and then slowly increased over the length of the study. The dogs in the wrap group regained normal weight-bearing on the treated limb more quickly than did those in the other groups. The constructs in the tendon group were weaker and less stiff immediately after surgery than were those in the other groups or in intact controls. Histologic analysis confirmed that the wrap technique resulted in complete union of the host bone-allograft junction more often than did the other techniques. The wrap method had the best functional outcome after 9 months when an allograft/endoprosthetic composite was used during total hip arthroplasty in this canine model.

Effect of intramedullary polymethylmethacrylate and autogenous cancellous bone on healing of frozen segmental allografts.

This study was designed to compare bone mineral density, periosteal callus production, new bone formation, bone porosity, histologic appearance, and union of mid-diaphyseal segmental allografts of the femur stabilized with an interlocking nail technique in a canine model 6 months after the procedure. An in vivo study was performed to compare the effects of augmenting interlocking nail fixation with an interlocking nail alone, intramedullary polymethylmethacrylate, intramedullary polymethylmethacrylate and autogenous cancellous bone applied to the periosteal surface of the host-allograft junction, autogenous cancellous bone applied to the endosteal surface of the allograft, autogenous cancellous bone applied to the periosteal surface of the host-allograft junction, and autogenous cancellous bone applied to the periosteal surface of the host-allograft junction and to the endosteal surface of the allograft. There were no differences among treatments for bone mineral density at any time interval. Callus area 4 weeks after the procedure was greater along the lateral and cranial surfaces for treatments with periosteal cancellous bone (p < 0.05). New bone within the allograft segment did not differ among treatments and was reduced compared with the host-allograft junctions (p < 0.05). The amount and quality of bone tissue at the host-allograft junctions were greatest with treatments of intramedullary polymethylmethacrylate and autogenous cancellous bone applied to the periosteal surface of the host-allograft junction and of autogenous cancellous bone applied to the periosteal surface of the host-allograft junction and to the endosteal surface of the allograft (p < 0.05). The rate of bone union was lower, and there was a greater gap (non-bone tissue) remaining between host and allograft bone with treatment involving just intramedullary polymethylmethacrylate than with other treatments (p < 0.05). The results suggest that augmenting interlocking nail fixation with intramedullary polymethylmethacrylate by itself offers no advantage but that a combination of intramedullary polymethylmethacrylate and cancellous bone at the periosteal surface or of cancellous bone within the medullary canal and at the periosteal surface improves the quality of healing at 6 months.

Comparison of allograft/endoprosthetic composites with a step-cut or transverse osteotomy configuration.

This study was designed to compare the biomechanical and functional characteristics of allograft/endoprosthetic composites of the proximal 25% of the femur repaired with either a transverse or a step-cut osteotomy, using a canine model (10 dogs, five with each type of osteotomy). Serial radiography and weight-bearing studies were performed monthly, and mechanical testing was done 6 months after surgery. The femora were tested in torsion and compared with the contralateral control (insertion of a femoral component but no osteotomy). At 6 months, the composites with a step-cut osteotomy had 36% greater structural stiffness than the composites with a transverse osteotomy (p < 0.005) and 121% greater maximum torque at failure than the controls (p < 0.005), without greater structural stiffness. Evaluation of peak vertical ground reaction forces revealed significantly greater weight-bearing on the experimental limb in dogs with a transverse osteotomy. The results of this relatively short-term study were mixed. Despite the increased structural stiffness of the allograft/endoprosthetic composite with a step-cut osteotomy, the dogs with this type of reconstruction had decreased weight-bearing throughout the course of the study. The step-cut osteotomy may augment the stability of the allograft/endoprosthetic composite, allowing faster healing (as demonstrated by the results of mechanical testing), but in some way, not understood, may cause pain in the reconstructed limb. Longer term studies are needed to answer these questions and to determine whether alteration of the traditional transverse osteotomy has any advantage.