Biocompatibility and osteoconduction of macroporous silk fibroin implants in cortical defects in sheep.

The goal of the presented study was to compare the biocompatibility and cellular responses to porous silk fibroin (SF) scaffolds produced in a water-based (UPW) or a solvent based process (HFIP) using two different SF sources. For that reason, four different SF scaffolds were implanted (n=6) into drill hole defects in the cancellous bone of the sheep tibia and humerus. The scaffolds were evaluated histologically for biocompatibility, cell-material interaction, and cellular ingrowth. New bone formation was observed macroscopically and histologically at 8 weeks after implantation. For semiquantitative evaluation, the investigated parameters were scored and statistically analyzed (factorial ANOVA). All implants showed good biocompatibility as evident by low infiltration of inflammatory cells and the absent encapsulation of the scaffolds in connective tissue. Multinuclear foreign body giant cells (MFGCs) and macrophages were present in all parts of the scaffold at the material surface and actively degrading the SF material. Cell ingrowth and vascularization were uniform across the scaffold. However, in HFIP scaffolds, local regions of void pores were present throughout the scaffold, probably due to the low pore interconnectivity in this scaffold type in contrast to UPW scaffolds. The amount of newly formed bone was very low in both scaffold types but was more abundant in the periphery than in the center of the scaffolds and for HFIP scaffolds mainly restricted to single pores.

An Ultrasound Assisted Anchoring Technique (BoneWelding Technology) for Fixation of Implants to Bone - A Histological Pilot Study in Sheep.

The BoneWelding((R)) Technology offers new opportunities to anchor implants within bone. The technology melted the surface of biodegradable polymer pins by means of ultrasound energy to mould material into the structures of the predrilled bone. Temperature changes were measured at the sites of implantation in an in vitro experiment. In the in vivo part of the study two types of implants were implanted in the limb of sheep to investigate the biocompatibility of the method. One implant type was made of PL-DL-lactide (PLA), the second one was a titanium core partially covered with PLA. Healing period was 2 and 6 months, with 3 sheep per group. Bone samples were evaluated radiologically, histologically and histomorphometrically for bone remodeling and inflammatory reactions. Results demonstrated mild and short temperature increase during insertion. New bone formed at the implant without evidence of inflammatory reaction. The amount of adjacent bone was increased compared to normal cancellous bone. It was concluded that the BoneWelding((R)) Technology proved to be a biocompatible technology to anchor biodegradable as well as titanium-PLA implants in bone.

Comparison of computer assisted surgery with conventional technique for treatment of abaxial distal phalanx fractures in horses: an in vitro study.

OBJECTIVE: To (1) evaluate and compare computer-assisted surgery (CAS) with conventional screw insertion (conventional osteosynthesis [COS]) for treatment of equine abaxial distal phalanx fractures; (2) compare planned screw position with actual postoperative position; and (3) determine preferred screw insertion direction. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Cadaveric equine limbs (n=32). METHODS: In 8 specimens each, a 4.5 mm cortex bone screw was inserted in lag fashion in dorsopalmar (plantar) direction using CAS or COS. In 2 other groups of 8, the screws were inserted in opposite direction. Precision of CAS was determined by comparison of planned and actual screw position. Preferred screw direction was also assessed for CAS and COS. RESULTS: In 4 of 6 direct comparisons, screw positioning was significantly better with CAS. Results of precision analysis for screw position were similar to studies published in human medicine. None of evaluated criteria identified a preferred direction for screw insertion. CONCLUSION: For abaxial fractures of the distal phalanx, superior precision in screw position is achieved with CAS technique compared with COS technique. CLINICAL RELEVANCE: Abaxial fractures of the distal phalanx lend themselves to computer-assisted implantation of 1 screw in a dorsopalmar (plantar) direction. Because of the complex anatomic relationships, and our results, we discourage use of COS technique for repair of this fracture type.

Computer-assisted surgery for screw insertion into the distal sesamoid bone in horses: an in vitro study.

OBJECTIVE: To compare the precision of computer-assisted surgery with a conventional technique (CV) using a special guiding device for screw insertion into the distal sesamoid bone in horses. STUDY DESIGN: In vitro experimental study. SAMPLE POPULATION: Cadaveric forelimb specimens. METHODS: Insertion of a 3.5 mm cortex screw in lag fashion along the longitudinal axis of intact (non-fractured) distal sesamoid bones was evaluated in 2 groups (8 limbs each): CV and computer-assisted surgery (CAS). For CV, the screw was inserted using a special guiding device and fluoroscopy, whereas for CAS, the screw was inserted using computer-assisted navigation. The accuracy of screw placement was verified by radiography, computed tomography, and specimen dissection. RESULTS: Surgical precision was better in CAS compared with CV. CONCLUSION: CAS improves the accuracy of lateromedial screw insertion, in lag fashion, into the distal sesamoid bone. CLINICAL RELEVANCE: The CAS technique should be considered for improved accuracy of screw insertion in fractures of the distal sesamoid bone.

An animal model in sheep for biocompatibility testing of biomaterials in cancellous bones.

BACKGROUND: The past years have seen the development of many synthetic bone replacements. To test their biocompatibility and ability for osseointegration, osseoinduction and -conduction requires their placement within bone preferably in an animal experiment of a higher species. METHODS: A suitable experimental animal model in sheep with drill holes of 8 mm diameter and 13 mm depth within the proximal and distal humerus and femur for testing biocompatibility issues is introduced. RESULTS: This present sheep model allows the placing of up to 8 different test materials within one animal and because of the standardization of the bone defect, routine evaluation by means of histomorphometry is easily conducted. This method was used successfully in 66 White Alpine Sheep. When the drill holes were correctly placed no complications such as spontaneous fractures were encountered. CONCLUSION: This experimental animal model serves an excellent basis for testing the biocompatibility of novel biomaterials to be used as bone replacement or new bone formation enhancing materials.

Clinical evaluation of the CRIF 4.5/5.5 system for long-bone fracture repair in cattle.

OBJECTIVE: To report clinical evaluation of the clamp rod internal fixator 4.5/5.5 (CRIF 4.5/5.5) in bovine long-bone fracture repair. STUDY DESIGN: Retrospective study. ANIMALS: Cattle (n=22) with long-bone fractures. METHODS: Records for cattle with long-bone fractures repaired between 1999 and 2004 with CRIF 4.5/5.5 were reviewed. Quality of fracture repair, fracture healing, and clinical outcome were investigated by means of clinical examination, medical records, radiographs, and telephone questionnaire. RESULTS: Successful long-term outcome was achieved in 18 cattle (82%); 4 were euthanatized 2-14 days postoperatively because of fracture breakdowns. Two cattle had movement of clamps on the rod. Moderate to severe callus formation was evident in 11 cattle 6 months postoperatively. CONCLUSIONS: Movement of clamps on the rod was recognized as implant failure unique to the CRIF. This occurred in cattle with poor fracture stability because of an extensive cortical defect. The CRIF system may not be ideal to treat metacarpal/metatarsal fractures because its voluminous size makes skin closure difficult, thereby increasing the risk of postoperative infections. CLINICAL RELEVANCE: CRIF cannot be recommended for repair of complicated long-bone fractures in cattle.

Assessment of stiffness and strength of 4 different implants available for equine fracture treatment: a study on a 20 degrees oblique long-bone fracture model using a bone substitute.

OBJECTIVE: To compare the mechanical properties of 4 stabilization methods for equine long-bone fractures: dynamic compression plate (DCP), limited contact-DCPlate (LC-DCP), locking compression plate (LCP), and the clamp-rod internal fixator (CRIF--formerly VetFix). STUDY DESIGN: In vitro mechanical study. SAMPLE POPULATION: Bone substitute material (24 tubes) was cut at 20 degrees to the long axis of the tube to simulate an oblique mid-shaft fracture. METHODS: Tubes were divided into 4 groups (n=6) and double plated in an orthogonal configuration, with 1 screw of 1 implant being inserted in lag fashion through the "fracture". Thus, the groups were: (1) 2 DCP implants (4.5, broad, 10 holes); (2) 2 LC-DCP implants (5.5, broad, 10 holes); (3) 2 LCP implants (4.5/5.0, broad, 10 holes) and 4 head locking screws/plate; and (4) 2 CRIF (4.5/5.0) and 10 clamps in alternating position left and right of the rod. All constructs were tested in 4-point bending with a quasi-static load until failure. The implant with the interfragmentary screw was always positioned on the tension side of the construct. Force, displacement, and angular displacement at the "fracture" line were determined. Construct stiffness under low and high loads, yield strength, ultimate strength, and maximum angular displacement were determined. RESULTS: None of the implants failed; the strength of the bone substitute was the limiting factor. At low loads, no differences in stiffness were found among groups, but LCP constructs were stiffer than other constructs under high loads (P=.004). Ultimate strength was lowest in the LCP group (P=.01), whereas yield strength was highest for LCP constructs (409 N m, P=.004). CRIF had the lowest yield strength (117 N m, P=.004); no differences in yield strength (250 N m) were found between DCP and LC-DCP constructs. Differences were found for maximum angular displacement at the "fracture" line, between groups: LPC

Comparison of computer-assisted surgery with conventional technique for the treatment of axial distal phalanx fractures in horses: an in vitro study.

OBJECTIVES: To compare the precision obtained with computer-assisted screw insertion for treatment of mid-sagittal articular fractures of the distal phalanx (P3) with results achieved with a conventional technique. STUDY DESIGN: In vitro experimental study. SAMPLE POPULATION: Thirty-two cadaveric equine limbs. METHODS: Four groups of 8 limbs were studied. Either 1 or 2 screws were inserted perpendicular to an imaginary axial fracture of P3 using computer-assisted surgery (CAS) or conventional technique. Screw insertion time, predetermined screw length, inserted screw length, fit of the screw, and errors in placement were recorded. RESULTS: CAS technique took 15-20 minutes longer but resulted in greater precision of screw length and placement compared with the conventional technique. CONCLUSION: Improved precision in screw insertion with CAS makes insertion of 2 screws possible for repair of mid-sagittal P3 fractures. CLINICAL RELEVANCE: CAS although expensive improves precision in screw insertion into P3 and consequently should yield improved clinical outcome.

Biocompatibility and resorption of a brushite calcium phosphate cement.

A hydraulic calcium phosphate cement with beta-tricalcium phosphate (TCP) granules embedded in a matrix of dicalcium phosphate dihydrate (DCPD) was implanted in experimentally created defects in sheep. One type of defect consisted of a drill hole in the medial femoral condyle. The other, partial metaphyseal defect was located in the proximal aspect of the tibia plateau and was stabilized using a 3.5 mm T-plate. The bone samples of 2 animals each per group were harvested after 2, 4, 6 and 8 weeks. Samples were evaluated for cement resorption and signs of immediate reaction, such as inflammation, caused by the cement setting in situ. Differences regarding these aspects were assessed for both types of defects using macroscopical, radiological, histological and histomorphometrical evaluations. In both defects the brushite matrix was resorbed faster than the beta-TCP granules. The resorption front was followed directly by a front of new bone formation, in which residual beta-TCP granules were embedded. Cement resorption occurred through (i) extracellular liquid dissolution with cement disintegration and particle formation, and (ii) phagocytosis of the cement particles through macrophages. Signs of inflammation or immunologic response leading to delayed new bone formation were not noticed at any time. Cement degradation and new bone formation occurred slightly faster in the femur defects.

An animal model for interface tissue formation in cemented hip replacements.

OBJECTIVE: To create a model in sheep for investigation of early changes related to the formation of an interface membrane in hip prosthesis. STUDY DESIGN: Experimental study. ANIMALS: Twenty-four female adult Swiss Alpine sheep. METHODS: Sheep were divided into 2 groups of 12 for unilateral cemented total hip arthroplasty. In Group I, the prosthesis was fixed with retrograde cement gun injection to achieve a complete cement mantle, whereas in Group II a primary cement mantle defect was produced. Groups I and II were further divided into 2 sub-groups with study end points of 2 and 8.5 months after surgery. Radiographs were evaluated postoperatively and at euthanasia for migration of the femoral component and bone resorption. Histologic sections were evaluated semiquantitatively for changes in cell types and numbers, and bone reactions; and quantitatively for size of interface membrane and new bone formation. RESULTS: Radiographically, there tended to be an increase in bone resorption and periosteal bone formation throughout the femoral shaft in Group II compared with Group I, but this was only statistically significant at the region of the femoral neck (R5) at both time periods (P<.05). Semiquantitative histologic evaluation revealed significant increases (P<.05) in cellularity, numbers of fibroblasts, giant cells, macrophages, and mononuclear cells, in Group II primarily at 2 months after surgery. This was also true for interface membrane formation and bone remodeling. Quantitative data showed an increased in the size of the interface membrane and area of bone formation at 8.5 months in Group II. CONCLUSIONS: The cement defect model offered controlled and repeatable production of an interface membrane. The results suggest that a primary cement mantle defect could be a possible trigger for implant instability, eliciting a cascade of biomechanical and molecular events in bone tissue leading to aseptic loosening. CLINICAL RELEVANCE: The results show the effect of defects in the cement mantle in promoting interface membrane formation. Long-term and biochemical studies are required to evaluate the relevance of this interface membrane formation.

An experimental animal model of aseptic loosening of hip prostheses in sheep to study early biochemical changes at the interface membrane.

BACKGROUND: Aseptic loosening of hip prosthesis as it occurs in clinical cases in human patients was attributed to wear particles of the implants, the response of the tissue dominated by macrophages and the production of inflammatory mediators and matrix degrading enzymes; however, the cascade of events initiating the process and their interaction regarding the time course is still open and discussed controversially. Therefore, the goal of this study was to establish an experimental animal model in sheep allowing to follow the cascade of early mechanical and biochemical events within the interface membrane and study the sequence of how they contribute to the pathological bone resorption necessary for aseptic loosening of the implant. METHODS: A cemented modular system (Biomedtrix) was used as a hip replacement in 24 adult Swiss Alpine sheep, with one group receiving a complete cement mantle as controls (n = 12), and the other group a cement mantle with a standardized, lateral, primary defect in the cement mantle (n = 12). Animals were followed over time for 2 and 8.5 months (n = 6 each). After sacrifice, samples from the interface membranes were harvested from five different regions of the femur and joint capsule. Explant cell cultures were performed and supernatant of cultures were tested and assayed for nitric oxide, prostaglandin E2, caseinolytic and collagenolytic activity. RNA extraction and quantification were performed for inducible nitric oxide synthase, cyclooxygenase-2, interleukin 1, and interleukin 6. Overall differences between groups and time periods and interactions thereof were calculated using a factorial analysis of variance (ANOVA). RESULTS: The development of an interface membrane was noticed in both groups at both time points. However, in the controls the interface membrane regressed in thickness and biological activity, while both variables increased in the experimental group with the primary cement mantle defect over time. Nitric oxide (NO) and PGE2 concentrations were higher in the 8.5 months group (P < 0.0001) compared to the 2 months group with a tendency for the unstable group to have higher concentrations. The same was true for collagenolytic activity (P = 0.05), but not for caseinolytic activity that decreased over time (P < 0.0001). CONCLUSION: In this study, a primary cement mantle defect of the femoral shaft elicited biomechanical instability and biochemical changes over time in an experimental animal study in sheep, that resembled the changes described at the bone cement-interface in aseptic loosening of total hip prosthesis in humans. The early biochemical changes may well explain the pathologic bone resorption and formation of an interface membrane as is observed in clinical cases. This animal model may aid in future studies aiming at prevention of aseptic loosening of hip prosthesis and reflect some aspects of the pathogenesis involved.

Mechanical comparison of 3 different clamp and 2 different rod types of a new veterinary internal fixation system, 4.5/5.5-mm VetFix.

OBJECTIVE: To compare the gripping force of a standard clamp with click-on and T clamps of the 4.5/5.5-mm VetFix (AO Research Institute, Davos, Switzerland), mounted on smooth and roughened rods. STUDY DESIGN: In vitro mechanical study. METHODS: Rods were mounted on a clamp-holding device. Each clamp type was tested on a materials-testing machine in groups of 6 specimens for torsional and axial load stability on smooth and roughened rods (total, 72 specimens). The variable evaluated was the discontinuity point (DP) as a measure for gliding resistance of the clamp on the rod. Means (+/-SD) were calculated for all groups separately. For statistical comparison, a multiple regression model was fitted with level of significance set at P =.05. RESULTS: The direction of torsion had a significant effect on DP for both asymmetric clamps (standard, click-on). In torsion and axial load, the roughened rod had significantly higher DP levels than the smooth rod for the standard clamp. Rods differed only in axial load for the T clamp, again favoring the roughened rod. The click-on clamp did not reach 10% of the values achieved by the other clamps, independent from the rods and loading conditions. Compared with the T clamp, standard clamp DP values were significantly higher in torsion on the roughened rod as well as in axial load on both rod types. CONCLUSIONS: The roughened rod with a standard clamp was mechanically superior. For the T clamp, the small loss of gripping force may be compensated for by the possibility of inserting 2 more screws. CLINICAL RELEVANCE: The roughened rod may enhance the bending, compressive, and torsional properties of the 4.5/5.5-mm VetFix system by reducing the risk of clamp gliding on the rod. The standard clamp is the clamp of choice, except for the metaphyseal or epiphyseal region, at which use of a T clamp may be indicated to increase the number of screws inserted at the end of the rod.