Denervation impairs healing of the rabbit medial collateral ligament.

Little is known about the contribution of innervation to ligament healing after traumatic disruption, although there is good evidence of an important role for the peripheral nervous system in the healing of fractures and skin injuries. Tissues such as ligament, with a low resting blood supply, are dependent on substantial increases in blood flow and vascular volume during the initial stages of repair. We hypothesized that this initial healing response would be strongly promoted by neurogenic inflammation. Since the saphenous nerve (a major sensory branch of the femoral nerve) supplies the medial half of the knee joint, we elected to use femoral nerve transection as a model to determine the role of sensory and autonomic innervation in the initial outcome of repair of the injured medial collateral ligament. Twelve adult, female NZW rabbits underwent right medial collateral ligament transection. Of these, six rabbits underwent right femoral nerve transection to disrupt the somatic sensory and autonomic nerve supply to the knee joint and six were kept neurologically intact (controls). At six weeks post-injury, the animals were assessed by laser Doppler perfusion imaging (LDI) to determine the local blood flow, at both the injury site and at the uninjured contralateral ligament. The animals were then killed, the knee joints were removed and the biomechanical characteristics of the healing bone-median collateral ligament (MCL)-bone complexes assessed. In a separate cohort of 16 rabbits, vascular volumes of the injured ligaments were measured by infusion of a carmine red/gelatin solution. At six weeks post-injury, in vivo measurement of perfusion with LDI revealed that normally innervated ligaments had an almost three-fold higher average blood flow. Carmine red/gelatin infusion revealed a 50% higher density of blood vessels as compared to denervated ligaments. The force required for ultimate failure was found to be 50% higher in normally innnervated MCL's as compared to denervated MCL's: 153.14 +/- 20.71 N versus 101.29 +/- 17.88 N (p < 0.05). Static creep was increased by 66% in denervated MCL's: 2.83 +/- 0.45% versus 1.70 +/- 0.12% (p < 0.05). Total creep was increased by 45% in denervated MCL's: 5.29 +/- 0.62% compared to 3.64 +/- 0.31% in innervated MCL's (p < 0.05). We conclude that intact innervation makes a critical contribution to the early healing responses of the MCL of adult rabbits.     

Natural history of healing in the repaired medial collateral ligament

The purpose of this study was to assess morphologically the healing of repaired medial collateral ligaments (MCLs) in a rabbit model. Healing ligaments were examined grossly and histologically at various intervals, from 3 days to 2 ½ years after injury, and compared with the appearances of normal age-, sex-, and activity-matched controls. Results show that all ligaments healed by bridging scar formation rather than true ligament regeneration. Increases in cellularity and temporary matrix disorganization along the entire length of the ligaments during healing suggest a combination of diffuse mechanical damage from their failure in tension and regional inflammatory injury (in excess of surgical exposure alone) from the processes of degradation and replacement. Substance that was not injured physically in this model demonstrated complete recovery, while that replaced by scar did not. Healing processes were similar to those of other highly specialized soft tissues (e.g., tendons), with short phases of hemorrhage and inflammation, an intermediate phase of proliferation, and a prolonged phase of remodeling. Failure of repairs to maintain anatomical apposition of torn ends may have contributed to the delay of these healing processes by increasing scar mass. Incomplete scar remodeling at 2 ½ years, however, suggests much slower MCL healing than previously reported and probably, therefore, a longer period for potential treatment influence.     

Strength of medial structures of the knee joint are decreased by isolated injury to the medial collateral ligament and subsequent joint immobilization.

Past studies of the healing of the medial collateral ligament (MCL) in animal models have been conducted over a variety of healing intervals, some as early as 1 week. One concern with testing at early healing intervals is the difficulty in identifying and isolating the tissues that carry load. The purpose of this study was to determine if isolation of the MCL and healing time are critical factors in the assessment of structural strength in this model. Furthermore, the effect of immobilization on these critical factors was investigated. Our approach was to calculate the load-sharing ratio between the MCL and the MCL plus capsule. A 4 mm gap was created in the midsubstance of both hindlimb MCLs of 52 female New Zealand White rabbits (n=104). Of these, 29 rabbits had their right hindlimb pin immobilized (immobilized group), leaving the left hindlimb non-immobilized. Testing was performed at 3 (n=12), 6 (n=22), and 14 (n=24) weeks. The remaining 23 rabbits, which had both limbs non-immobilized (non-immobilized group), were tested at 3 (n=10), 6 (n=12), 14 (n=12), and 40 (n=12) weeks. For both groups, half of the specimens at each healing interval were used to test the MCL alone and half to test the MCL plus capsule, except for 3 week immobilized joints where only the MCL plus capsule was tested. Additionally, MCL (n=12), MCL plus capsule (n=6), and capsule alone (n=5) were tested from normal animals. The load-sharing ratio at MCL failure for the normal joint was 89%, suggesting an MCL-dominated response. For the non-immobilized group, the load-sharing ratio was 24% at 3 weeks of healing, suggesting a capsule-dominated response. At and after 6 weeks of healing, an MCL-dominated response was observed, with the ratio being 68% or greater. Thus, at less than 6 weeks of healing, the structural strength capabilities of the joint may be better represented by the medial structures rather than the isolated MCL. Immobilization delayed the transition from a capsule-dominated response to an MCL-dominated response in this model.     

A quantitative analysis of matrix alignment in ligament scars: a comparison of movement versus immobilization in an immature rabbit model

This investigation quantified the alignment of fibrillar matrix in normal rabbit medial collateral ligaments (MCLs) and in healing MCLs from animals treated with or without knee immobilization. Twenty-four immature female rabbits were given complete midsubstance injuries to their right MCLs. Fifteen of them had that knee pin immobilized in flexion, while the remaining nine were allowed unrestricted cage activity. Animals were sacrificed in groups of three at intervals of 3, 6, or 14 weeks after injury, and both healing MCLs and unoperated contralateral controls were fixed in situ for subsequent removal, freeze-fracture, and preparation for scanning electron microscopy (SEM). A random sampling of SEM photographs followed by automated, statistically validated image processing was used to quantify alignment of matrix in all samples. Results showed that nonimmobilized MCL scars in this model do remodel over 14 weeks of healing, returning to normal alignment values in that time. Surprisingly, MCL scars in immobilized knees were even better, with mean matrix alignments falling statistically within normal MCL limits at all healing intervals studied. If not due to an unknown sampling or fixation artifact, these results suggest that gross knee flexion and extension is not a prerequisite for scar matrix alignment in this immature model of ligament healing.     

Denervation alters mRNA levels of repair-associated genes in a rabbit medial collateral ligament injury model.

Previous experiments revealed that denervation impairs healing of the MCL. This suggested the hypothesis that denervation would decrease repair-associated mRNA levels in the injured MCL when compared with normally innervated injured MCL. Adult, skeletally mature female rabbits were assigned to one of four groups: unoperated control, femoral nerve transection alone (denervated controls), MCL partial tear or denervated MCL partial tear. At three days, two weeks, six weeks or sixteen weeks post-surgery, cohorts of 6 rabbits from each experimental group were killed. Ligaments were harvested, RNA extracted and RT-PCR was performed using rabbitspecific primers. In the denervated injury group, mRNA levels for the angiogenesis-associated gene MMP-13, matrix components Collagen I and III, growth factor TGF-beta and angiogenesis inhibitors TIMP-3, and TSP-1 had all increased by two-weeks post-injury, in comparison to the non-denervated injury group (p < or = 0.01). An increased level of TSP-1 mRNA was also detected in the denervated injured group at sixteen weeks post injury (p < or = 0.01). Contrary to the initial hypothesis, denervation led to increased mRNA levels for many relevant molecules during the early stages of MCL healing. Thus, inappropriate timing of over-expression of some molecules may potentially contribute to the decreased quality of the scar tissue, particularly molecules such as TSP-1. Neuronal derived factors strongly influence the in vivo metabolic activity of ligament and scar fibroblasts in the initial phases of healing.     

Viscoelastic properties of the human medial collateral ligament under longitudinal, transverse and shear loading.

Ligament viscoelasticity controls viscous dissipation of energy and thus the potential for injury or catastrophic failure. Viscoelasticity under different loading conditions is likely related to the organization and anisotropy of the tissue. The objective of this study was to quantify the strain- and frequency-dependent viscoelastic behavior of the human medial collateral ligament (MCL) in tension along its longitudinal and transverse directions, and under shear along the fiber direction. The overall hypothesis was that human MCL would exhibit direction-dependent viscoelastic behavior, reflecting the composite structural organization of the tissue. Incremental stress relaxation testing was performed, followed by the application of small sinusoidal strain oscillations at three different equilibrium strain levels. The peak and equilibrium stress-strain curves for the longitudinal, transverse and shear tests demonstrate that the instantaneous and long-time stress-strain response of the tissue differs significantly between loading conditions of along-fiber stretch, cross-fiber stretch and along-fiber shear. The reduced relaxation curves demonstrated at least two relaxation times for all three test modes. Relaxation resulted in stresses that were 60-80% of the initial stress after 1000 s. Incremental stress relaxation proceeded faster at the lowest strain level for all three test configurations. Dynamic stiffness varied greatly with test mode and equilibrium strain level, and showed a modest but significant increase with frequency of applied strain oscillations for longitudinal and shear tests. Phase angle was unaffected by strain level (with exception of lowest strain level for longitudinal samples) but showed a significant increase with increasing strain oscillation frequency. There was no effect of test type on the phase angle. The increase in phase and thus energy dissipation at higher frequencies may protect the tissue from injury at faster loading rates. Results suggest that the long-time relaxation behavior and the short-time dynamic energy dissipation of ligament may be governed by different viscoelastic mechanisms, yet these mechanisms may affect tissue viscoelasticity similarly under different loading configurations.     

Subject-specific finite element analysis of the human medial collateral ligament during valgus knee loading.

The objectives of this study were (1) to develop subject-specific experimental and finite element (FE) techniques to study the three-dimensional stress-strain behavior of ligaments, with application to the human medial collateral ligament (MCL), and (2) to determine the importance of subject-specific material properties and initial (in situ) strain distribution for prediction of the strain distribution in the MCL under valgus loading. Eight male knees were subjected to varus-valgus loading at flexion angles of 0 degrees, 30 degrees, and 60 degrees. Three-dimensional joint kinematics and MCL strains were recorded during kinematic testing. Following testing, the MCL of each knee was removed to allow measurement of the in situ strain distribution and to perform material testing. A FE model of the femur-MCL-tibia complex was constructed for each knee to simulate valgus loading at each flexion angle, using subject-specific bone and ligament geometry, material properties, and joint kinematics. A transversely isotropic hyperelastic material model was used to represent the MCL. The MCL in situ strain distribution at full extension was used to apply in situ strain to each MCL FE model. FE predicted MCL strains during valgus loading were compared to experimental measurements using regression analysis. The subject-specific FE predictions of strain correlated reasonably well with experimentally measured MCL strains (R(2)=0.83, 0.72, and 0.66 at 0 degrees, 30 degrees, and 60 degrees, respectively). Despite large inter-subject variation in MCL material properties, MCL strain distributions predicted by individual FE models that used average MCL material properties were strongly correlated with subject-specific FE strain predictions (R(2)=0.99 at all flexion angles). However, predictions by FE models that used average in situ strain distributions yielded relatively poor correlations with subject-specific FE predictions (R(2)=0.44, 0.35, and 0.33 at flexion angles of 0 degrees, 30 degrees, and 60 degrees, respectively). The strain distribution within the MCL was nonuniform and changed with flexion angle. The highest MCL strains occurred at full extension in the posterior region of the MCL proximal to the joint line during valgus loading, suggesting this region may be most vulnerable to injury under these loading conditions. This work demonstrates that subject-specific FE models can predict the complex, nonuniform strain fields that occur in ligaments due to external loading of the joint.     

Bovine xenograft collateral ligament replacement in the dog

A Study of the use of glutaraldehyde-stabilized bovine xenograft material as a collateral ligament replacement in 16 dogs has been done. Six xenograft implant complexes harvested 4 months postoperatively failed in tension at 772.2 ± 463.5 versus 799.7 ± 162.7 N ( ± 1 SD) for controls (p > 0.05, paired t test). Histologic evaluation in 10 dogs after implants of up to 1 year duration demonstrated a progressive invasion of the xenograft by host tissues. Xenograft remnants were easily identifiable at 1 year. The host tissues invaded in parallel to the passive collagen scaffolding of the xenograft and consisted of vessels and fibroblastic elements that produced collagen of host origin.     

Ultrastructural comparison of medial collateral ligament repair after single or multiple applications of GaAlAs laser in rats

BACKGROUND AND OBJECTIVES: To examine single versus multiple applications of a gallium aluminum arsenide (GaAlAs) laser on the ultrastructural morphology of surgically injured medial collateral ligaments (MCLs) in rats. STUDY DESIGN/MATERIALS AND METHODS: Sixteen rats were studied with 12 receiving right MCL transection and 4 receiving sham injury. Group 1 (n = 4) received one session of laser (31.6 J/cm(2)) immediately after injury. Group 2 (n = 4) received 9 doses of transcutaneous laser (3.5 J/cm(2)). The controls (Group 3, n = 4) received one session of placebo laser, while the sham Group 4 (n = 4) received no treatment. Ultrastructural analyses were done with electron microscopy at 3 weeks. RESULTS: The mass-averaged diameters of collagen fibril in the core and periphery of MCLs treated with multiple laser were larger than the control and those with single laser treatment (P < 0.05). However, the sham injured group had larger fibrils than all other groups (P < 0.05). CONCLUSIONS: The repairing MCLs had smaller collagen fibrils than the sham injured ligaments. Multiple laser treatments enhanced the collagen growth in the repairing MCLs at 3 weeks after injury, which are superior to a single treatment with similar dosage.     

Cytochemical evidence for a proteoglycan-associated filamentous network in ligament extracellular matrix.

The purpose of this investigation was to examine the extracellular matrix of rabbit ligament before and after digestion with glycosaminoglycan degrading enzymes. In order to preserve and enhance the visibility of negatively charged tissue components, particularly the glycosaminoglycan-containing proteoglycans, the cationic stains ruthenium red (RR) and ruthenium hexamine trichloride (RHT) were used. Cross-sections of the midsubstance of 10-month-old (mature) rabbit medial collateral ligaments fixed using conventional procedures revealed a sparse population of stellate-shaped cells that did not appear to be interconnected. Similar tissue fixed in either RR or RHT showed an extensive network of thin, electron-dense "seams" that interconnected cells and appeared to irregularly subdivide the extracellular matrix (ECM). These seams mainly consisted of a meshwork of microfilaments throughout which small granules were dispersed. Numerous 14-nm microfibrils, as well as mature elastic fibers were also present within the seams. The size and shape of the microfilaments, together with their threadlike, beaded appearance suggested that they could be Type VI collagen. The seam granules were easily removed with chondroitinase ABC, chondroitinase AC II, and mild (0.18 M) salt treatment. Only chondroitinase ABC succeeded in removing additional granules, tentatively identified as proteodermatan sulphate molecules, that were periodically located at d band sites along the Type I collagen fibrils. These results suggest that the seam granules are not dermatan sulphate containing proteoglycans, and further, that these proteoglycans may be sequestered into specific zones within the ECM through loose association with the seam microfilaments. While the functional significance of the seams remains unknown and their specific composition clearly requires further study, it is likely that they represent important functional (e.g., viscoelastic) or biological (e.g., nutritional) subdivisions of ligament substance.     

The early effects of joint immobilization on medial collateral ligament healing in an ACL-deficient knee: a gross anatomic and biomechanical investigation in the adult rabbit model.

In this study, the short-term effects of immobilization on joint damage and medial collateral ligament (MCL) healing were investigated in unstable, anterior cruciate ligament (ACL)-deficient knees in rabbits. Forty-six 12-month-old female New Zealand white rabbits were separated into three groups. Animals from each group had surgery on their right knees: group I, sham controls (n = 9); group II, complete transection of the ACL and removal of a 4 mm segment (gap injury) of MCL midsubstance with no immobilization of the limb (n = 19); and group III, same injuries to the ACL and MCL (as group II) but with immobilization of the limb (n = 18). No surgical repair of disrupted ligaments was performed. Left knees served as unoperated contralateral controls. All animals were allowed unrestricted cage activity until sacrifice in subgroups at 3, 6, and 14 weeks of healing when biomechanical properties of all MCLs were measured. All knee joints were systematically examined for gross evidence of damage to articular cartilage, menisci, and periarticular soft tissues. To monitor relative in vivo loads on injured limbs during healing, hindlimb weight bearing was assessed at biweekly intervals. Results indicated that animals in both groups II and III bore relatively lower loads (compared to preinjury values) on their injured hindlimbs. Mechanical testing of MCLs showed only minor changes in sham controls, while group II and III healing MCLs demonstrated significantly lower force and stress at MCL complex failure compared to contralateral controls. In specific comparisons of group III to group II animals, we noted that immobilization prevented joint damage over the early intervals studied. In addition, immobilization resulted in MCL laxity similar to contralateral control values but inhibited development of structural strength and stiffness in healing MCLs. These results suggest that in the rabbit, short-term immobilization of an ACL-deficient knee offers some advantages to the joint and to certain low load behaviors of the healing MCL, but it also results in a smaller quantity of scar tissue that is less able to resist higher loads. Longer-term studies involving remobilization are necessary before the effects of brief immobilization on joint damage and MCL healing in this ACL-deficient model can be fully defined.     

Comparison of single and multiple applications of GaAlAs laser on rat medial collateral ligament repair

BACKGROUND AND OBJECTIVES: To examine single versus multiple applications of the gallium aluminum arsenide (GaAlAs) laser on the healing of surgically injured medial collateral ligaments (MCLs) in rats. STUDY DESIGN/MATERIALS AND METHODS: Sixteen rats were studied, with 12 receiving surgical transection to their right MCL and 4 receiving a sham injury. Group 1 (n = 4) received a single dose of GaAlAs laser therapy (wavelength 660 nm, average power 8.8 mW, pulse 10 kHz, dosage 31.6 J/cm(2)) directly to their MCL during surgery. Group 2 (n = 4) received 9 doses of GaAlAs laser therapy applied transcutaneously on alternate days (wavelength 660 nm, average power 8.8 mW, pulse 10 kHz, dosage 3.5 J/cm(2)). The controls (Group 3, n = 4) received one session of placebo laser at the time of surgery, with the laser equipment shut down, while the sham injured Group 4 (n = 4) received no treatment. Biomechanical tests for structural stiffness, ultimate tensile strength (UTS), and load-relaxation were done at 3 weeks after injury. The stiffness and UTS data were normalized by expressing as a percentage of the left side of each animal before statistical analysis. RESULTS: The load-relaxation data did not show any differences between the groups (P = 0.18). The normalized stiffness levels of Groups 2 (81.08+/-11.28%) and 4 (92.66+/-13.19%) were significantly higher (P = 0.025) than that of the control Group 3 (58.99+/-15.91%). The normalized UTS of Groups 2 (81.38+/-5.68%) and 4 (90.18+/-8.82%) were also significantly higher (P = 0.012) than that of the control (64.49+/-9.26%). Although, Group 1 had higher mean stiffness and UTS values than the control, no statistically significant difference was found between these two groups. CONCLUSIONS: Multiple laser therapy improves the normalized strength and stiffness of repairing rat MCLs at 3 weeks after injury. The multiple treatments seem to be superior to a single treatment when the cumulative dosages are comparable between the two modes of application.     

Effects of a therapeutic laser on the ultrastructural morphology of repairing medial collateral ligament in a rat model

BACKGROUND AND OBJECTIVES: Low energy laser therapy has been shown to enhance mechanical strength of healing medial collateral ligament (MCL) in rats. The present study investigated its effects on the ultrastructural morphology and collagen fibril profile of healing MCL in rats. STUDY DESIGN/MATERIALS AND METHODS: Thirty-two mature male Sprague-Dawley (SD) rats were used. Twenty-four underwent surgical transection to their right MCLs and eight received only skin wound. Immediately after surgery, eight of the MCL transected rats were treated with a single dose of laser therapy at 63.2 J cm(-2), eight were treated with a single dose of laser therapy at 31.6 J cm(-2), the rest had no treatment and served as control. At 3 and 6 weeks after surgery, the MCLs were harvested and examined with electron microscopy for collagen fibril size, distribution, and alignment. RESULTS: Significant differences (P < 0.001) were found in fibril diameters from the same anatomical site and time period among different groups. The mass-averaged diameters of the laser-treated (64.99-186.29 nm) and sham (64.74-204.34 nm) groups were larger than the control group (58.66-85.89 nm). The collagen fibrils occupied 42.55-59.78, 42.63-53.94, and 36.92-71.64% of the total cross-sectional areas in the laser-treated, control and sham groups, respectively. Mode obliquity was 0.53-0.84 among the three groups. CONCLUSIONS: Single application of low energy laser therapy increases the collagen fibril size of healing MCLs in rats.     

Role of angiogenesis after muscle derived stem cell transplantation in injured medial collateral ligament

We performed this study to investigate the therapeutic role of vascular endothelial growth factor (VEGF) in medial collateral ligament (MCL) healing. Murine muscle derived stem cells (MDSCs) obtained via the preplate technique were retrovirally transduced to express: (1) VEGF and nLacZ (MDSC‐VEGF), (2) soluble fms‐like tyrosine kinase‐1 (sFLT1, a VEGF‐specific antagonist) and nLacZ (MDSC‐sFLT1), and (3) nLacZ (MDSC‐nLacZ). After transecting the MCL of immunodeficient rats, 5 × 10⁵ cells of each of the transduction groups list above were transplanted into the MCL injury site. A control group was injected with phosphate‐buffered saline (PBS) only. Immunohistochemical staining demonstrated that there were more Isolectin B4 and β‐galactosidase double positive cells in the rats transplanted with MDSC‐VEGF transduced cells than the other groups at week 1. Capillary density was significantly higher in the MDSC‐VEGF group than the other groups at week 2; however, there were no significant differences in the biomechanical assessment between the MDSC‐VEGF and MDSC‐nLacZ groups. On the other hand, the MDSC‐sFLT1 group revealed a lower capillary density than the other two groups and the functional ligament healing of the MDSC‐sFLT1 group was significantly decreased compared to the other groups when assessed biomechanically. The findings of the present study suggest that angiogenesis plays a critical role in the healing process of injured MCL. © 2011 Orthopaedic Research Society. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 30:627–633, 2012     

Ulnar collateral ligament strain of the thumb metacarpophalangeal joint: biomechanical comparison of two postoperative immobilization techniques

Background

The aim of this study was to compare postoperative immobilization techniques of the thumb metacarpophalangeal (MP) ulnar collateral ligament (UCL) in a cadaver model of a noncompliant patient.

Methods

A cadaveric model with fresh-frozen forearms was used to simulate pinch under two immobilization conditions: (1) forearm-based thumb spica splint alone and (2) forearm-based thumb spica splint with supplemental transarticular MP Kirschner wire fixation. Pinch was simulated by thumb valgus loading and flexor pollicis longus (FPL) loading. Ulnar collateral ligament displacements were measured and strain values calculated. Statistical analysis was performed using a repeated measures analysis of variance model.

Results

With valgus thumb loading, we noted a significantly lower UCL strain in the splint and pin group compared to splint immobilization alone. Increased load was associated with a statistically significant increase in UCL strain within each immobilization condition. FPL loading resulted in negative displacement, or paradoxical shortening, of the UCL in both immobilization groups.

Conclusions

While immobilized, valgus thumb force, as opposed to MP flexion, is a likely contributor to UCL strain during simulated pinch representing noncompliance during the postoperative period. Supplemental thumb MP pin fixation more effectively protects the UCL from valgus strain. UCL shortening with FPL loading likely represents paradoxical MP extension due to flexion of the distal phalanx against the distal splint, suggesting attempted thumb flexion with splint immobilization alone does not jeopardize UCL repair.

Clinical Relevance

This study provides a foundation to aid clinical decision-making after UCL repair. It reinforces the practice of surgeons who routinely pin their MP joints, but also brings to attention that the use of temporary MP pin fixation may be considered in difficult cases, such as those with potential noncompliance or tenuous repair.     

The effects of initial end contact on medial collateral ligament healing: a morphological and biomechanical study in a rabbit model

In this investigation, the effect of initial end contact on medial collateral ligament (MCL) healing was studied in the rabbit model. Sixty-eight 1-year-old New Zealand white rabbits were separated into two groups. In one group, a 4 mm saggital Z-plasty was performed in the right MCL midsubstance (contact group), and in the other group, an analogous 4 mm midsubstance segment was removed (gap group). Left knees were unoperated to serve as internal contralateral MCL controls. Animals had unrestricted cage activity until sacrifice in groups of eight at 3, 6, 14, and 40 weeks postoperatively. Early results demonstrated that contact and gap injuries healed with what appeared to be scar tissue both morphologically and biomechanically. In both groups, laxities recovered to their contralateral values within 6 weeks and biomechanical viscoelastic behaviors recovered to 68-92% of contralaterals by 14-40 weeks. Despite these similarities, contacts showed morphological and biomechanical evidence of improved healing over gaps. Contact scars remodeled more quickly, recovered laxity more quickly, and were generally closer to contralaterals than gaps in terms of their structural strength, stiffness, and material behaviors, after 40 weeks of healing. With the exception of appearances and failure stress, all measured properties of contact healing MCLs were statistically indistinguishable from contralateral MCLs at 40 weeks of healing. These advantages of contact healing in this model support speculations that there are differences in the early rate and possibly in the later quality of ligament healing when cut rabbit MCL ends are in proximity. Longer-term studies to define end points and mechanisms of healing are required.     

The organization of collagen in cryofractured rabbit articular cartilage: a scanning electron microscopic study

Adult rabbit articular cartilage was prepared for scanning electron microscopy using, in order, glutaraldehyde fixation, enzymatic removal of proteoglycan, dehydration in ethanol, cryofracture in liquid nitrogen, and critical-point drying. Enzymes were effective in fixed material. Fixation, cryofracture, alignment of fracture surfaces with "split lines," and retention of subchondral bone were found to be necessary steps for the preservation of collagen detail. The fibrous framework was found to be similar to that proposed by Benninghoff and favored by more recent phase-contrast microscopic studies. Vertical fibers extending from subchondral bone and a network of tangentially oriented superficial fibrils converge in the transitional zone. No random layer is seen. Pericellular capsules interdigitate with the vertical fibers. When cartilage is prepared in a manner that minimizes tissue damage, scanning electron microscopy provides useful, unique information.     

Therapeutic low energy laser improves the mechanical strength of repairing medial collateral ligament

BACKGROUND AND OBJECTIVES: Low energy laser therapy has been shown to enhance collagen production but its effect on tissue strength is not well reported. We tested the effects of therapeutic laser on the strength of healing medial collateral ligaments (MCLs) in rats. STUDY DESIGN/MATERIALS AND METHODS: Twenty-four rats received surgical transection to their right MCL and eight received sham operation. After surgery, 16 received a single dose of gallium aluminum arsenide laser to their transected MCL for 7.5 minutes (n = 8) or 15 minutes (n = 8) and eight served as control with placebo laser, while the sham group didn't receive any treatment. The MCLs were biomechanically tested at either 3 or 6 weeks post-operation. RESULTS: The normalized ultimate tensile strength (UTS) and stiffness of laser and sham groups were larger than control (P < 0.001). The UTS of laser and sham groups were comparable. Laser and sham groups had improved in stiffness from 3 to 6 weeks (P < 0.001). CONCLUSIONS: A single dose of low energy laser therapy improves the UTS and stiffness of repairing MCL at 3 and 6 weeks after injury.     

The biomechanical and morphological changes in the medial collateral ligament of the rabbit after immobilization and remobilization

The effects of immobilization and remobilization on the biomechanical and morphological properties of the femur-medial collateral ligament-tibia complex and each of its components were investigated in the rabbit. Specimens that had been obtained after periods of unilateral immobilization of the knee and remobilization were evaluated for structural properties. In addition, the mechanical properties of the substance of the medial collateral ligament and the histological characteristics of both the substance of the ligament and its sites of insertion were evaluated. After immobilization, there were significant reductions in the ultimate load and energy-absorbing capabilities of the bone-ligament complex, and an increased number of failures occurred by tibial avulsion. The tissue of the medial collateral ligament also became less stiff as a result of immobilization. Histologically, the femoral and tibial insertion sites showed increased osteoclastic activity, resorption of bone, and disruption of the normal attachment of the bone to the ligament. With remobilization, the ultimate load and energy-absorbing capabilities of the bone-ligament complex improved but did not return to normal. Failure by tibial avulsion became less frequent, and the stress-strain characteristics of the medial collateral ligament returned to normal. Histologically, the sites of insertion of the ligament also showed evidence of recovery.