Anterior Cruciate Ligament Reconstruction In Rabbits Research Articles

Hyperbaric oxygen therapy enhances graft healing and mechanical properties after anterior cruciate ligament reconstruction: An experimental study in rabbits.

Hyperbaric oxygen therapy (HBOT) has proven successful in wound healing. However, its potential effects on anterior cruciate ligament (ACL) injuries remain uncertain. This study aimed to investigate the impact of HBOT on graft healing following ACL reconstruction in rabbits. Male New Zealand rabbits underwent ACL reconstruction and were randomly divided into two groups: the HBOT group and the ambient air group. The HBOT group received 100% oxygen at 2.5 atmospheres absolute for 2 h daily for 5 consecutive days, starting from the first day after surgery. The ambient air group was maintained in normal room air throughout the entire period. After 12 weeks following the surgery, animals were euthanized, and their knees were harvested for analysis. The HBOT group demonstrated superior graft maturation and integration in comparison to the ambient air group, as evidenced by lower graft signal intensity on magnetic resonance imaging, decreased femoral and tibial tunnel size, and higher bone mineral density values on high-resolution peripheral quantitative computed tomography scans. Additionally, biomechanical testing indicated that the HBOT group had greater load to failure and stiffness values than the ambient air group. In conclusion, the adjuvant use of HBOT improved ACL graft maturation and integration, reduced tunnel widening, and enhanced the biomechanical properties of the graft. These results may provide important insights into the potential clinical application of HBOT as a therapeutic intervention to enhance graft healing after ACL reconstruction, paving the way for further research in this area.

Establishment of near and non isometric anterior cruciate ligament reconstruction with artificial ligament in a rabbit model.

BackgroundTunnel position deicide the isometry of graft attachment in synthetic anterior cruciate ligament (ACL) reconstruction. Near-isometric tunnel position may have advantage in graft integration and knee function in ACL reconstruction (ACLR) with polyethylene terephthalate (PET) ligament. Few studies focused on tunnel position isometry when conduct ACLR with an animal model. This study aimed to establish a preclinical rabbit model of near and non isometric ACLR with PET ligament and investigate the advantage of near-isometric ACLR compared to non-isometric ACLR.MethodsNine hind limbs of rabbit were used in tunnel position study. Two femoral(anatomic, nonanatomic) tunnels and three tibial(anterior, middle, posterior) tunnels were used to measure tunnel position isometry during knee full range of motion. The tunnel position combination with minimal isometry was considered as near-isometric tunnel position. Then, 48 rabbits divided into two groups were conducted near or non isometric ACLR with PET ligament with graft fixation angle of 30° and constant tension of 5N. PET ligament isometry, range of motion(ROM) restriction, knee laxity were recorded after operation and followed up with macroscopic observation, microcomputed tomography (micro-CT) analysis, histology assessment and biomechanical test at 4 and 8 weeks postoperatively.ResultsThe tunnel combination with minimal isometry was femoral anatomic position and tibial posterior position(5.19 ​± ​1.78%) and considered as near-isometric tunnel position. ROM restriction were observed in non-isometric group (22.50 ​± ​14.14°) while none in near-isometric group. However, no ROM restriction observed at 8 weeks in both group. Knee laxity compared to contralateral knee were better in near-isometric group than non-isometric group (stable/slack/total 10/2/12 VS 3/9/12, p ​= ​0.012) at 8 weeks postoperatively. Supeiror PET ligament integration were also observed in near-isometric group through macroscopic observation, micro-CT analysis, histology assessment at both 4 and 8 weeks. The failure load in the Near-Isometric group at 8 weeks were higher than timezero reconstruction with statistical difference (156.8N ​± ​25.98N vs.102.6 ​± ​22.96N, p ​= ​0.02).ConclusionA rabbit model of ACLR based on tunnel position isometry was successfully established in this study. The near-isometric tunnel position in rabbit model was femoral anatomic position and tibial posterior position. A near-isometric ACLR with PET ligament did not cause ROM restriction and had a better graft integration and follow-up stability than non-isometric ACLR with ROM restriction.The Translational Potential of this ArticleThe study demonstrate the establishmentof near-isometric tunnel position and non-isometric tunnel position with significant difference of ROM restriction and graft-bone integration. The described tunnel positions with differential isometry in a rabbit ACLR provides a reproducible and translational small animal model and enables preclinical research between tunnel position isometry and its affection on variable grafts, graft integration and knee function.

Biomineralizaion of hydroxyapatite on polyethylene terephthalate artificial ligaments promotes graft-bone healing after anterior cruciate ligament reconstruction: An in vitro and in vivo study.

The purpose of the present study is to modify the polyethylene terephthalate ligament with hydroxyapatite via biomineralization and to investigate its effect on graft-bone healing. After biomineralization of hydroxyapatite, the surface characterization of polyethylene terephthalate ligament was examined by scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, and water contact angle measurements. The compatibility and osteoinduction, along with the underlying signaling pathway involved of hydroxyapatite-polyethylene terephthalate ligament, were evaluated in vitro. Moreover, a rabbit anterior cruciate ligament reconstruction model was established, and the polyethylene terephthalate or hydroxyapatite-polyethylene terephthalate artificial ligament was implanted into the knee. The micro-computed tomography analysis, histological, and immunohistochemical examination as well as biomechanical test were performed to investigate the effect of hydroxyapatite coating in vivo. The results of scanning electron microscopy, Fourier transform infrared spectroscopy, and X-ray diffraction showed that the hydroxyapatite was successfully deposited on the polyethylene terephthalate ligament. Water contact angle of the hydroxyapatite-polyethylene terephthalate group was significantly smaller than that of the polyethylene terephthalate group. The in vitro study showed that hydroxyapatite coating significantly improved adhesion and proliferation of MC3T3-E1 cells. The osteogenic differentiation of cells was also enhanced through the activation of ERK1/2 pathway. The micro-computed tomography, histological, and immunohistochemical results showed that biomineralization of hydroxyapatite significantly promoted new bone and fibrocartilage tissue formation at 12 weeks postoperatively. Moreover, the failure load and stiffness in the hydroxyapatite-polyethylene terephthalate group were higher than that in the polyethylene terephthalate group. Therefore, biomineralizaion of hydroxyapatite enhances the biocompatibility and osseointegration of the polyethylene terephthalate artificial ligament, thus promoting graft-bone healing for anterior cruciate ligament reconstruction through the activation of ERK1/2 pathway.

Evaluation of a bioengineered ACL matrix's osteointegration with BMP-2 supplementation.

A poly (l-lactic) acid bioengineered anterior cruciate ligament (ACL) matrix has previously demonstrated the ability to support tissue regeneration in a rabbit ACL reconstruction model. The matrix was designed for optimal bone and ligament regeneration by developing a matrix with differential pore sizes in its bone and ligament compartments. Building upon past success, we designed a new bioengineered ACL matrix that is easier to install and can be used with endobutton fixation during ACL reconstruction. To achieve this, a new braiding procedure was developed to allow the matrix to be folded in half, making two-limbs, while maintaining its bone and ligament compartments. The osteointegration of the matrix with and without bone morphogenetic protein 2 (BMP-2) supplementation was evaluated in a rabbit ACL reconstruction model. Two doses of BMP-2 were evaluated, 1 and 10 μg, and delivered by saline injection into the bone tunnel at the end of surgery. A fibrous matrix-to-bone interface with occasional Sharpey’s fibers was the primary mode of osteointegration observed. The matrix was also found to support a fibrocartilage matrix-to-bone interface. In some cases, the presence of chondrocyte-like cells was observed at the aperture of the bone tunnel and the center of the matrix within the bone tunnel. Treatment with BMP-2 was associated with a trend towards smaller bone tunnel cross-sectional areas, and 1 μg of BMP-2 was found to significantly enhance osteoid seam width in comparison with no BMP-2 or 10 μg of BMP-2 treatment. Regenerated tissue was well organized within the bioengineered ACL matrix and aligned with the poly (l-lactic) acid fibers. Disorganized tissue was found between the two-limbs of the bioengineered ACL matrix and hypothesized to be due to a lack of structural scaffolding. This study suggests that the bioengineered ACL matrix can undergo similar modes of osteointegration as current autografts and allografts, and that BMP-2 treatment may enhance osteoblastic activity within the bone tunnels.

A LbL-Assembled Bioactive Coating Modified Nanofibrous Membrane for Rapid Tendon-Bone Healing in ACL Reconstruction.

IntroductionIn anterior cruciate ligament (ACL) reconstruction, hamstring tendon autograft is a well-accepted surgical choice as an alternative ACL graft. But the main disadvantage of autograft is its inefficient healing with host bone-tunnel which will leading to surgery failure.MethodsA biomimetic nanofibrous membrane for tendon-bone integration is fabricated in this work, which is composed of polycaprolactone (PCL) electrospinning membrane and chitosan/hyaluronic acid (CS/HA) multilayers film.ResultsBy using layer-by-layer (LbL) self-assembly this functional CS/HA multilayer films are deposited on the surface of PCL nanofiber to enable the local delivery of stromal cell-derived factor-1 α (SDF-1α) and bone morphogenetic protein-2 (BMP-2) in tendon-bone interface. This membrane can promote cell proliferation and recruitment, as well as inducing the osteogenic differentiation and recruitment of BMSCs.ConclusionFurther in vivo studies demonstrate that to wrap the tendon autograft using the membrane may afford superior tendon-bone integration and inhibit scar tissue formation in a rabbit ACL reconstruction model. More importantly, the biomechanical properties of the tendon-bone interface have been improved. This study shows that this biomimetic nanofibrous membrane is effective for improving tendon-bone healing after ACL reconstruction surgery.

Effect of Autologous Platelet-Rich Plasma and Gelatin Sponge for Tendon-to-Bone Healing After Rabbit Anterior Cruciate Ligament Reconstruction

To investigate platelet-rich plasma (PRP) combined with gelatin sponge (GS) to improve tendon-bone interface healing and structure formation. Characterization of the GS scaffold was performed with a scanning electron microscope, and the release curve after loading with PRP was evaluated. A real-time reverse transcription quantitative polymerase chain reaction assay was performed to test the levels of tendon-to-bone healing-related gene expression. Finally, 18 New Zealand white rabbits were randomly divided into 3 groups and underwent semitendinosus autograft anterior cruciate ligament reconstruction: autograft group without PRP, PRP group, and PRP-GS group. All rabbits were killed 8weeks after the operation. Magnetic resonance imaging scans, biomechanical testing, and histologic evaluation were performed. An enzyme-linked immunosorbent assay and cell counting kit-8 assay showed that the GS could control the release of PRP and prolong its bioactivity time, as well as promote bone marrow mesenchymal stem cell proliferation. In the PRP-GS group, the levels of related genes were upregulated compared with the PRP group (P < .05). Lower signal in the magnetic resonance images indicated fibrocartilage formation in the 2 groups with PRP. In addition, histologic staining showed that the tendon-bone connection had a greater fibrocartilaginous transition region in the PRP-GS group, and the histologic scores were higher (vs the PRP group, P= .039). The maximum failure load and stiffness were higher in the PRP-GS group than in the other 2 groups. GS loading with PRP could prolong the bioactivity time of PRP and promote bone marrow mesenchymal stem cell proliferation and osteogenic gene expression invitro. It also promoted the early healing process at the tendon-bone junction in a rabbit anterior cruciate ligament reconstruction model. GS is a natural material and offers satisfactory biocompatibility. Using GS as a scaffold to control the release of bioactive factors in bone tunnels may be useful, but additional studies in human subjects will be necessary to evaluate its clinical prospects.

Silk fibroin coating through EDC/NHS crosslink is an effective method to promote graft remodeling of a polyethylene terephthalate artificial ligament.

Anterior cruciate ligament reconstruction using polyethylene terephthalate artificial ligaments is one of the research hotspots in sports medicine but it is still challenging to achieve biological healing. The purpose of this study was to modify polyethylene terephthalate ligament with silk fibroin through ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysuccinimide (NHS) crosslink and to investigate the performance of graft remodeling in vitro and in vivo. After silk fibroin coating, changes in the surface properties of ligament were characterized by scanning electron microscopy, attenuated total reflectance-Fourier transform infrared spectroscopy and water contact angle measurements. The compatibility of polyethylene terephthalate ligament with silk fibroin coating was investigated in vitro. The results showed the silk fibroin coating significantly improved adhesion, proliferation and extracellular matrix secretion of fibroblast cells. Moreover, a rabbit anterior cruciate ligament reconstruction model was established to evaluate the effect of ligament with silk fibroin coating in vivo. The gross observation and histological results showed that the silk fibroin coating significantly inhibited inflammation response and promoted new tissue regeneration with fusiform cells infiltration in and around the graft. Furthermore, the expressions of collagen I protein and mRNA in the silk fibroin-coated polyethylene terephthalate group were much higher than those in the control group according to the immunohistochemical and real-time polymerase chain reaction results. Therefore, silk fibroin coating through EDC/NHS crosslink promotes the biocompatibility and remodeling process of polyethylene terephthalate artificial ligament in vitro and in vivo. It can be considered as a potential solution to the problem of poor remodeling of artificial ligaments after anterior cruciate ligament reconstruction in the clinical applications.

Enhanced tendon\u2013bone healing with acidic fibroblast growth factor delivered in collagen in a rabbit anterior cruciate ligament reconstruction model

BackgroundThe objective of the present study was to investigate the effectiveness of acidic fibroblast growth factor delivered in collagen (aFGF/collagen) for promoting tendon–bone interface healing after anterior cruciate ligament (ACL) reconstruction in rabbits.MethodsACL reconstructions were performed in the right hind limbs of New Zealand rabbits. Each left long digital extensor tendon was harvested as an autograft, and collagen incorporating different concentrations of aFGF or same amount of collagen alone was applied at the tendon–bone interface after ACL reconstruction. The control group underwent ACL reconstruction only. There were high and low aFGF/collagen groups, collagen alone group, and control group (n = 21 rabbits per group). Histological and biomechanical analyses were performed at 4, 8, and 12 weeks postoperatively to evaluate the effect of aFGF/collagen on tendon–bone interface healing.ResultsResults of biomechanical tests showed that at both 8 and 12 weeks postoperatively, the elastic modulus and stiffness in both the high and low aFGF/collagen treatment groups were significantly higher than those in the control group and collagen alone group, with that in the high aFGF/collagen concentration group being the highest. Histological analysis showed that at 8 weeks, tightly organized Sharpey-like fibers were observed in both aFGF/collagen groups with new bone growth into the tendon in the high concentration group. At 12 weeks postoperatively, a fibrocartilage transition zone was observed in the bone tunnels in both aFGF/collagen groups, especially in the high aFGF/collagen group.ConclusionApplication of the aFGF/collagen composite could enhance early healing at the tendon–bone interface after ACL reconstruction, especially with the use of a high aFGF/collagen concentration.

Exogenous stromal derived factor-1 releasing silk scaffold combined with intra-articular injection of progenitor cells promotes bone-ligament-bone regeneration

Anterior cruciate ligament (ACL) is one of the most difficult tissues to heal once injured. Ligament regeneration and tendon-bone junction healing are two major goals of ACL reconstruction. This study aimed to investigate the synergistic therapeutic effects of Stromal cell-derived factor 1 (SDF-1)-releasing collagen-silk (CSF) scaffold combined with intra-articular injection of ligament-derived stem/progenitor cells (LSPCs) for ACL regeneration and the amelioration in the long-term complication of osteoarthritis (OA). The stem cell recruitment ability of CSF scaffold and the multipotency, particularly the tendon forming ability of LSPCs from rabbits were characterized in vitro, while the synergistic effect of the CSF scaffold and LSPCs for ACL regeneration and OA amelioration were investigated in vivo at 1, 3, and 6 months with a rabbit ACL reconstruction model. The CSF scaffold was used as a substitute for the ACL, and LSPCs were injected into the joint cavity after 7 days of the ACL reconstruction. CSF scaffold displayed a controlled release pattern for the encapsulated protein for up to 7 days with an increased stiffness in the mechanical property. LSPCs, which exhibited highly I Collagen and CXCR4 expression, were attracted by SDF-1 and successfully relocated into the CSF scaffold at 1 month in vivo. At 3 and 6 months post-treatment, the CSF scaffold combined with LSPCs (CSFL group) enhanced the regeneration of ACL tissue, and promoted bone tunnel healing. Furthermore, the OA progression was impeded efficiently. Our findings here provided a new strategy that using stem cell recruiting CSF scaffold with tissue-specific stem cells, could be a promising solution for ACL regeneration. Statement of SignificanceIn this study, we developed a silk scaffold with increased stiffness and SDF-1 controlled release capacity for ligament repair. This advanced scaffold transplantation combined with intra-articular injection of LSPCs (which was isolated from rabbit ligament for the first time in this study) promoted the regeneration of both the tendinous and bone tunnel portion of ACL. This therapeutic strategy also ameliorated cartilage degeneration and reduced the severity of arthrofibrosis. Hence, combining LSPCs injection with SDF-1-releasing silk scaffold is demonstrated as a therapeutic strategy for ACL regeneration and OA treatment in the clinic. [Display omitted]

Differences in placement of calcium phosphate-hybridized tendon grafts within the femoral bone tunnel during ACL reconstruction do not influence tendon-to-bone healing

BackgroundCalcium phosphate (CaP)-hybridization of tendon grafts has been shown to improve tendon-to-bone healing. The purpose of this study was to clarify the influence of different tendon graft placement methods on tendon-to-bone healing using CaP-hybridized tendon grafts in anterior cruciate ligament (ACL) reconstructions in rabbits.MethodsWe compared two methods of tendon graft placement within the femoral bone tunnel: suspension of the tendon graft within the bone tunnel (suspension group) and implantation of the tendon graft coherent with the bone socket (coherence group). CaP-hybridized tendon grafts were used in both groups. Fifty-six male Japanese white rabbits were used for this study. The results of biomechanical tests (n = 9) and histological analyses (n = 5) were evaluated at 2 and 4 weeks after surgery.ResultsThe ultimate failure load, stiffness, stress, soft tissue remaining in bone tunnel after biomechanical testing, and direct bonding area at tendon–bone interface did not differ significantly between the suspension and coherence groups at either 2 or 4 weeks after surgery (p > 0.05). In both groups, the ultimate failure load, stress, soft tissue remaining in the bone tunnel, and direct bonding area at interface at 4 weeks after surgery were significantly greater than those at 2 weeks after surgery (p < 0.05).ConclusionsTendon-to-bone healing in both groups progressed until the endpoint of 4 weeks. There was no influence of the CaP-hybridized tendon graft placement method on tendon-to-bone healing at 4 weeks after ACL reconstruction in rabbits. Thus, the CaP-hybridized tendon grafts were unaffected by differences in their placement within the bone tunnel and became equally anchored to the bone tunnel during the early postoperative period. The tendon graft placement method may not influence tendon-to-bone healing in ACL reconstruction when CaP-hybridized tendon grafts are used.

Three-Dimension-Printed Porous Poly(Propylene Fumarate) Scaffolds with Delayed rhBMP-2 Release for Anterior Cruciate Ligament Graft Fixation.

Anterior cruciate ligament (ACL) ruptures reconstructed with tendon grafts are commonly fixed with bioabsorbable implants, which are frequently complicated by incomplete bone filling upon degradation. Bone regeneration after ACL reconstruction could be enhanced by utilizing tissue engineering techniques and three-dimensional (3D) printing to create a porous bioabsorbable scaffold with delayed delivery of recombinant-human bone morphogenetic protein 2 (rhBMP-2). The first aim of this study was to design a 3D poly(propylene fumarate) (PPF) porous scaffold that maintained suitable pullout strength for future testing in a rabbit ACL reconstruction model. Our second aim was to determine the release kinetics of rhBMP-2 from PPF scaffolds that utilized both calcium-phosphate coatings and growth factor delivery on microspheres, both of which have been shown to decrease the initial burst release of rhBMP-2 and increase bone regeneration. To determine the degree of scaffold porosity that maintained suitable pullout strength, tapered scaffolds were fabricated with increasing porosity (0%, 20%, 35%, and 44%) and pullout testing was performed in a cadaveric rabbit ACL reconstruction model. Scaffolds were coated with carbonate hydroxyapatite (synthetic bone mineral [SBM]), and radiolabeled rhBMP-2 was delivered in four different experimental groups as follows: Poly(lactic-co-glycolic acid) microspheres only, microspheres and collagen (50:50), collagen only, and saline solution only. rhBMP-2 release was measured at day 1, 2, 4, 8, 16, and 32. The microsphere delivery groups had a smaller burst release and released a smaller percentage of rhBMP-2 over the 32 days than the collagen and saline only groups. In conclusion, a porous bioabsorbable scaffold with suitable strength for a rabbit ACL reconstruction was developed. Combining a synthetic bone mineral coating with microspheres had an additive effect, decreasing the initial burst release and cumulative release of rhBMP-2. Future studies need to evaluate this scaffold's fixation strength and bone filling capabilities in vivo compared to traditional bioabsorbable implants.

Mechanical Loading Improves Tendon-Bone Healing in a Rabbit Anterior Cruciate Ligament Reconstruction Model by Promoting Proliferation and Matrix Formation of Mesenchymal Stem Cells and Tendon Cells

Background/Aims: This study investigated the effect of mechanical stress on tendon-bone healing in a rabbit anterior cruciate ligament (ACL) reconstruction model as well as cell proliferation and matrix formation in co-culture of bone-marrow mesenchymal stem cells (BMSCs) and tendon cells (TCs). Methods: The effect of continuous passive motion (CPM) therapy on tendon-bone healing in a rabbit ACL reconstruction model was evaluated by histological analysis, biomechanical testing and gene expressions at the tendon-bone interface. Furthermore, the effect of mechanical stretch on cell proliferation and matrix synthesis in BMSC/TC co-culture was also examined. Results: Postoperative CPM therapy significantly enhanced tendon-bone healing, as evidenced by increased amount of fibrocartilage, elevated ultimate load to failure levels, and up-regulated gene expressions of Collagen I, alkaline phosphatase, osteopontin, Tenascin C and tenomodulin at the tendon-bone junction. In addition, BMSC/TC co-culture treated with mechanical stretch showed a higher rate of cell proliferation and enhanced expressions of Collagen I, Collagen III, alkaline phosphatase, osteopontin, Tenascin C and tenomodulin than that of controls. Conclusion: These results demonstrated that proliferation and differentiation of local precursor cells could be enhanced by mechanical stimulation, which results in enhanced regenerative potential of BMSCs and TCs in tendon-bone healing.

Adipose-Derived Regenerative Cells Promote Tendon-Bone Healing in a Rabbit Model

To evaluate the therapeutic effect of adipose-derived regenerative cell (ADRC) administration on tendon-bone healing in a rabbit anterior cruciate ligament (ACL) reconstruction model. ACL reconstruction with semitendinosus tendon autograft was performed in the right knees of adult white rabbits. Eighty rabbits were divided into 2 groups: the treatment group, in which the graft was coated with ADRCs mixed in a fibrin glue carrier during surgery, and the control group, in which the graft was coated with fibrin glue only. At 2, 4, 6, 8, and 12 weeks postoperatively, 8 rabbits were killed in each group. Three were used for histologic evaluation at the tendon-bone interface and 5 for biomechanical examination. On histologic analysis, chondroid cells appeared more orderly and more regular in size and shape and Sharpey-like fibers, which connected the tendon graft and bone tissue, appeared earlier in ADRC-treated tissues than in control tissues. On biomechanical analysis, the ultimate failure load in the ADRC-treated group was significantly greater than that in the control group at 2 weeks (29.5 ± 7.2 N v 20.9 ± 2.7 N, P= .016) and 4 weeks (32.3 ± 3.9 N v 22.8 ± 5.4 N, P= .016). Stiffness was significantly higher in the ADRC-treated group than in the control group at 6 weeks (21.7 ± 5.9 N/mm v 12.6 ± 4.9 N/mm, P= .037). Although the ultimate failure load and stiffness of the ADRC-treated limbs were higher than those of the limbs in the control group at 8 and 12 weeks, these differences were not significant. Local administration of ADRCs promoted the early healing process at the tendon-bone junction, both histologically and mechanically, in a rabbit ACL reconstruction model. ADRCs could be used to enhance graft healing in ACL reconstruction.

Experimental study of tendon graft fixation in anterior cruciate ligament reconstruction with cortical press-fit bolt in rabbits

To explore the histological outcomes of tendon-bone healing in anterior cruciate ligament (ACL) reconstruction with cortical press-fit bolt (CPB). Twenty-four healthy female or male New Zealand White rabbits (2-3 months old) underwent bilateral ACL reconstruction with extensor digitorum longus tendon. A random method was used to decide one knee would receive the routine ACL reconstruction (control group) and another cortical press-fit bolt fixation (experimental group). After general anesthesia, extensor digitorum longus tendon was harvested and ACL reconstruction performed. All animals were sacrificed at 4, 8 and 12 weeks postoperation. Radiological and histological examinations were made at each timepoint. The specimens were stained with different methods to observe the pathological changes of tendon graft, bone tunnel and cortical press-fit bolt. More revascularization and massive new bone were found in tendon-bone junction of experimental group at 4, 8 and 12 weeks postoperation. The circum-graft new vessel proportion of the experimental and control groups were 0.48 ± 0.12 and 0.26 ± 0.05 respectively (P < 0.05). In the experimental group, more cartilage cells were present in tendon-bone junction at 12 weeks and the circum-graft new bone areas in two groups were 0.41 ± 0.11 and 0.21 ± 0.10 mm(2) respectively (P < 0.05). Cortical press-fit blot may improve tendon-bone healing after ACL reconstruction in rabbits. The application prospects of this procedure are promising.

Augmentation of Tendon-to-Bone Healing with a Magnesium-Based Bone Adhesive

Background Healing of an anterior cruciate ligament graft in a bone tunnel occurs by formation of fibrous scar tissue, which is weaker than the normal fibrocartilaginous insertion. Hypothesis We hypothesized that a magnesium-based bone adhesive would improve tendon-to-bone healing in a rabbit anterior cruciate ligament reconstruction model. Study Design Controlled laboratory study. Methods Thirty-five New Zealand White rabbits underwent bilateral anterior cruciate ligament reconstructions with semitendinosus autografts. A total of 12.5 g of bone adhesive was placed in the intraosseous tunnel around the graft in one limb, while the tunnels in the contralateral limb received no implant. Sixteen animals each were sacrificed at 3 weeks and at 6 weeks (12 biomechanical testing/4 histology). Outcomes included semiquantitative histologic analyses for new cartilage formation and fibrous tissue formation in the tendon-bone interface, microcomputed tomography to quantify new bone formation along the bone tunnel, and biomechanical testing of load-to-failure and stiffness. Three animals were sacrificed at time 0 to confirm adequate tunnel fill with the bone adhesive on microcomputed tomography. Results All specimens had adequate tunnel fill with the bone adhesive at time 0. Application of the bone adhesive resulted in more cartilage formation and less fibrous tissue formation at the tendon-bone interface at 6 weeks compared with controls (P < .05). There was significantly more bone formation in the tibia of the treated limbs at 6 weeks (P = .01). The load-to-failure was significantly higher in the treated group at 6 weeks (71.8 ± 31.8 N vs 43.4 ± 14.8 N; P = .04). There were no differences in stiffness at either time point, and there were no differences at 3 weeks in any outcome variable. Conclusion The magnesium-based bone adhesive improves tendon-to-bone healing based on histologic and biomechanical testing at 6 weeks in a rabbit model of anterior cruciate ligament reconstruction. Clinical Relevance Further studies are needed to investigate the clinical potential of this bone adhesive to enhance healing and decrease recovery time in soft-tissue ligament reconstruction.

Augmentation of Tendon Healing in an Intraarticular Bone Tunnel with Use of a Bone Growth Factor

We hypothesized that an exogenous bone growth factor could augment healing of a tendon graft in a bone tunnel in a rabbit anterior cruciate ligament-reconstruction model. Seventy rabbits underwent bilateral anterior cruciate ligament reconstructions with a semitendinosus tendon graft. One limb received a collagen sponge carrier vehicle containing a mixture of bone-derived proteins while the contralateral limb was treated with either no sponge or a sponge without bone-derived proteins. The reconstruction was evaluated at 2, 4, or 8 weeks with histologic, biomechanical, and magnetic resonance imaging analysis. Histologic analysis demonstrated that specimens treated with bone-derived proteins had a more consistent, dense interface tissue and closer apposition of new bone to the graft, with occasional formation of a fibrocartilaginous interface, when compared with control specimens. The treated specimens had significantly higher load-to-failure rates than did control specimens. Treatment with bone-derived proteins resulted in an average increase in tensile strength of 65%. The treated specimens were stronger than control specimens at each time point, but the difference was greatest at 8 weeks. On the basis of signal characteristics and new bone formation, magnetic resonance imaging was useful for predicting which limb was treated, the site of failure, and the limbs with higher load-to-failure values. This study demonstrates the potential for augmenting tendon healing in an intraarticular bone tunnel using an osteoinductive growth factor.