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An in vivo animal model was used to evaluate overuse and overuse plus intrinsic tendon injury or extrinsic tendon compression in the development of rotator cuff injury. Forty-four male Sprague-Dawley rats were divided into groups of 22. Each left shoulder received an intrinsic or extrinsic injury plus overuse (treadmill running), and each right shoulder received only overuse. Eleven rats from each group were sacrificed at 4 and 8 weeks. Supraspinatus tendons were evaluated histologically or geometrically and biomechanically. Ten rats constituted a cage-activity control group. Both supraspinatus tendons of the experimental groups had increases in cellularity and collagen disorganization and changes in cell shape compared with control tendons. Tendons with injury plus overuse exhibited a worse histologic grade than those with overuse alone. The cross-sectional area of both supraspinatus tendons of the experimental rats was significantly more than in control tendons. The area of the injury plus overuse tendons was increased on average compared with overuse-alone tendons. Biomechanically, the tissue moduli of overuse/intrinsic injury tendons at 4 weeks and those of the overuse/extrinsic injury tendons at 8 weeks were significantly lower than in control tendons. Tissue moduli of the overuse/injury tendons were significantly lower than in the overuse-alone tendons at 8 weeks. This study demonstrated that damage to the supraspinatus tendon can be caused by overuse and intrinsic injury, overuse and extrinsic compression, and overuse alone.
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PMID:The effects of overuse combined with intrinsic or extrinsic alterations in an animal model of rotator cuff tendinosis. 985 Jul 82

This study was designed to determine the effects of a single injection of a species-specific preparation of cytokines into rabbit patellar tendons and to compare the results with a known model of tendinitis, the collagenase-injection model. New Zealand White rabbits were divided into two groups and two time periods (4 and 16 weeks) and injected in the midsubstance of the right patellar tendon with either cytokines or collagenase under ultrasound guidance to confirm intratendinous needle placement. The left patellar tendon was injected with 0.025 ml of saline solution and served as a control. The rabbits were returned to cage activity after injection. At death, two rabbits in each group underwent histological analysis; the remaining eight animals in each time frame were evaluated biomechanically and then biochemically with use of the patella/whole patellar tendon/tibia complex. Histologic results at 4 weeks in the tendons injected with cytokines demonstrated increased cellularity, which was resolving by 16 weeks. The matrix appeared unchanged. The tendons injected with collagenase demonstrated increased angiogenesis of the matrix, hypercellularity, and fibrosis around the tendon at 4 weeks. At 16 weeks, myxoid changes, focal fibrosis, and collagen-bundle disarray with persistent increase in cellularity were noted. Biomechanically, a significant decrease in ultimate load at 16 weeks was seen in the tendons injected with cytokines but no change was seen in cross-sectional area. The tendons injected with collagenase demonstrated a significant increase in cross-sectional area at 4 and 16 weeks compared with those injected with cytokines. Biochemically, there was no significant difference in collagen content between the two groups at 4 or 16 weeks but the tendons injected with collagenase demonstrated a significant increase in crosslinking at 16 weeks. Our conclusion is that the tendons injected with the cytokine preparation represent a model of mild, seemingly reversible tendon injury. The cytokine preparation produces no matrix damage or evidence of collagen degradation and is species specific.
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PMID:Cytokine-induced tendinitis: a preliminary study in rabbits. 1022 32

Elastic extracellular matrix protein tenascin-C (TN) has very restricted expression in normal tissues, but is expressed in large quantities during embryogenesis and hyperplastic processes. To examine the importance of mechanical stress on the regulation of TN expression in vivo, the effects of various mechanical loading states (immobilization and three forms of subsequent remobilization) on the expression of TN were studied immunohistochemically at the bone-tendon attachment of the rat quadriceps muscle. This osteotendinous junction (OTJ) was selected as study site, since it receives its mechanical stimuli only from muscle contracting activity, which is easy to block by cast immobilization. TN was expressed abundantly in the normal OTJ. Following the removal of the mechanical stress from the junction by cast-immobilization of three weeks, the immunoreactivity of TN was almost completely absent. Normal mechanical stress in the form of free remobilization of eight weeks (free cage activity) resulted in a slight increase in TN expression, but could not restore the expression of TN to the level of the healthy contralateral leg. After the application of the increased mechanical stress (intensified remobilization of the eight weeks by low- or high-intensity treadmill running), the distribution and immunoreactivity of TN reached the level of the healthy contralateral limb in the low-intensity running group or even exceeded that in the high-intensity running group. High TN expression was seen around the chondrocytes and fibroblasts of the OTJ as well as around the collagen fibers of the tendon belly. We conclusively show that mechanical strain regulates the expression of TN in vivo, and propose that mechanical stress is a major regulator of TN expression in fibroblasts and chondrocytes. This may be an important aspect of the regulation of TN expression during embryogenesis, tendon degeneration, wound healing, bone formation, and in the other normal or regenerative morphogenetic processes TN is postulated to take part in.
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PMID:Mechanical loading regulates tenascin-C expression in the osteotendinous junction. 1046 31

The ability of Staphylococcus aureus to recognize several extracellular matrix or plasma proteins (e.g., fibrinogen, fibronectin, and collagen) promotes bacterial attachment to artificial surfaces. Whereas most S. aureus clinical isolates elaborate a wide repertoire of bacterial surface receptors' called adhesins, exhibiting specific binding of individual host proteins, S. epidermidis is lacking most of such protein adhesins. To document the interactions between S. epidermidis and various surface-adsorbed proteins, we first compared promotion of bacterial attachment by seven purified human proteins immobilized onto poly(methyl methacrylate) (PMMA) coverslips. Only two of them, namely fibronectin and fibrinogen, exhibited adhesion-promoting activities. In the presence of native heparin or two functionalized dextrans (CMDBS for Carboxy Methyl, Benzylamide sulfonate/sulfate), a dose-dependent inhibition of S. epidermidis adhesion to fibronectin-coated, but not to fibrinogen-coated surfaces was observed. The inhibitory effects of each CMDBS were much stronger than that of native heparin. In contrast, a control highly negatively charged, dextran exclusively substituted with carboxy methyl groups exerted no inhibition on S. epidermidis adhesion. To evaluate how CMDBS could interfere with S. epidermidis attachment to coverslips coated in vivo with extracellular matrix components, we also tested PMMA surfaces retrieved from tissue cages subcutaneously implanted in guinea pigs. Each CMDBS, but not heparin, strongly inhibited S. epidermidis adhesion to explanted coverslips, even in the presence of tissue cage fluid. In conclusion, fibronectin plays an important role in promoting S. epidermidis attachment to implanted biomaterials. Furthermore, S. epidermidis adhesion to fibronectin-coated or implanted biomaterials can be efficiently blocked in vitro by CMDBS.
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PMID:Inhibition by heparin and derivatized dextrans of Staphylococcus epidermidis adhesion to in vitro fibronectin-coated or explanted polymer surfaces. 1067 17

The inframammary fold is a defining element in the shape and structure of the female breast. It should be preserved whenever possible in ablative procedures and recreated accurately when the breast is reconstructed after mastectomy. To date, no accurate anatomic description of this essential structure exists. Previous studies have suggested that the fold is produced by a supporting ligament running from the dermis in the fold region to a variety of locations on the rib cage. This clinic's experience with mastectomy, augmentation mammaplasty, and breast reconstruction does not support the existence of a ligamentous structure. To define the structure of the inframammary fold, 10 female and 2 male cadavers were studied. The anterior chest wall was removed en bloc and frozen in orthostatic position. Parasagittal sections were made of the inframammary fold with the chest wall intact. After decalcification of the ribs and routine histologic preparation, thin sections were stained with Gomori's trichrome. On light microscopic examination, no demonstrable ligamentous structure of dense regular connective tissue could be identified in the fold region in any of the 12 specimens. Superficial and deep fascial layers were uniformly observed anterior to the pectoralis major and serratus anterior muscles. The superficial fascia was connected to the dermis in the fold region in a variety of configurations. In some cases, the deep fascia fused with the superficial fascia and dermis at the fold level. In other cases, bundles of collagen fibers arising from the superficial fascial layer were found to insert into the dermis at the inframammary fold, slightly inferior to it, or both. These bundles were observed consistently in sections from the sternum to the middle axillary line. They were distinct from Cooper's suspensory ligaments, which are seen more superiorly in the glandular tissue.
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PMID:Inframammary fold: a histologic reappraisal. 1069 60

Overuse activity has been implicated as an etiologic factor in injury to the rotator cuff and to the supraspinatus tendon in particular. Due in part to the lack of an appropriate animal model, expex85ental studies have not addressed this issue. With the use of a rat model, we measured the effects of an overuse running regimen on 36 Sprague-Dawley rats after 4 (n = 12), 8 (n = 12), or 16 (n = 12) weeks of exercise and compared them with a control group of rats (n = 10) who were allowed normal cage activity. The histologic characteristics, the gross morphologic characteristics, and the mechanical properties of the tendon tissue were evaluated. The supraspinatus tendons in the exercised animals demonstrated significant changes as a result of overuse at all time points compared with the normal group. There was an increase in cellularity and a loss of the normal collagen fiber organization consistent with what has been seen in human tendinopathy. The tendons from the exercise groups were larger than normal in cross-sectional analysis at 4 weeks (129% of control, P < .01) and continued to increase in size with time to 16 weeks (164% of control, P = .01). The mechanical properties of the tendons deteriorated in response to overuse exercise with a decreased modulus of elasticity ranging from 52% to 61% of control (P = .07 at 4 weeks, P < .05 at 8 and 16 weeks) and a decreased maximum stress of failure ranging from 51% to 63% of control (P < .007). These findings support overuse activity as an etiologic factor in the development of supraspinatus tendinopathy and begin to describe the changes in the tendons as a result of such activity. This model can now be used to study the effect of various treatment modalities on these injuries.
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PMID:Neer Award 1999. Overuse activity injures the supraspinatus tendon in an animal model: a histologic and biomechanical study. 1081 Jun 84

Adolescent meat-type poultry and cage layers exhibit a high incidence of bone problems that include bone weakness, deformity, breakage, and infection and osteoporosis-related mortalities. These problems include economic and welfare issues. To improve bone quality in poultry, it is essential to understand the physiological basis of bone maturity and strength in poultry. A complex array of factors that include structural, architectural, compositional, physiological, and nutritional factors interactively determine bone quality and strength. Bone is approximately 70% mineral, 20% organic, and 10% water. Collagen is the major organic matrix that confers tensile strength to the bone, whereas hydroxyapatite provides compressional strength. In recent years, the roles of different collagen crosslinks have been shown to be important in the increase of bone mechanical strength. Similarly, age-related glyco-oxidative modifications of collagen have been shown to increase the stiffness of collagen. These posttranslational modifications of matrix can affect bone quality as it would be affected by the changes in the mineralization process. Our studies show that the growth in the tibia continued until 25 wk of age, which correlated with the increase in the content of hydroxylysylpridinoline (HP) and lysylpyridinoline (LP), the collagen crosslinks. The tibia from 5-wk-old chicks were strong but brittle because of low collagen crosslinks and high mineral content. Bone maturity may relate to its crosslink content. Compared to crosslink content, bone density and ash content showed moderate increases during growth. The bones from younger turkeys were more susceptible to corticosteroid-induced stunting of growth, which also resulted in decreased bone strength. This review discusses how different factors can compromise bone strength by reducing growth, altering shape, affecting mineralization, and affecting collagen crosslinking.
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PMID:Factors regulating bone maturity and strength in poultry. 1090 Dec 6

There are drawbacks to using threaded cylindrical cages (e.g., limited area for bone ingrowth and metal precluding radiographic visualization of bone healing). To somewhat offset these drawbacks, a barbell-shaped cage has been designed. The central core of the barbell can be wrapped with collagen sheets infiltrated with bone morphogenetic protein. The obvious theoretical advantages of a barbell cage have to be weighed against potential biomechanical disadvantages. Our purpose was to compare the biomechanical properties of an anterior lumbar interbody reconstruction using 18-mm-diameter threaded cylindrical cages, with a reconstruction using barbell cages (18-mm diameter and 6 mm wide at both cylindrical ends, with a round 4-mm-diameter bar joining the two ends). Twelve cadaveric lumbar motion segments were tested. Three L5-S1 segments received two threaded cylindrical cages, and three L5-S1 segments received two barbell cages. Three L3-L4 segments received one threaded cylindrical cage, and three L3-L4 segments received one barbell cage. A series of biomechanical loading sequences were carried out on each motion segment, and stiffness curves were obtained. After the biomechanical testing, an axial compressive load was applied to the motion segments until failure. They were then radiographed and bisected through the disc, and the subsidence (or penetration) of the cage(s) in the cancellous bone of the vertebral bodies was measured. There was no difference in terms of stiffness between the motion segments with the threaded cylindrical cage(s) inserted and those with the barbell cage(s) inserted (p > 0.15). The average values of subsidence was 0.96 mm for the threaded cylindrical cage group and 0.80 mm for the barbell cage group (difference not significant: p = 0.38). The results suggest that a reconstruction using barbell cages is a biomechanically acceptable alternative to one using threaded cylindrical cages.
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PMID:Anterior lumbar interbody fusion using a barbell-shaped cage: a biomechanical comparison. 1158 37

The homozygous ablation of the gene encoding the PTH/PTHrP receptor (PPR(-/-)) leads to early lethality and limited developmental defects, including an acceleration of chondrocyte differentiation. In contrast to the findings in homozygous PTHrP-ablated (PTHrP(-/-)) animals, these PPR(-/-) mice show an increase in cortical bone, a decrease in trabecular bone, and a defect in bone mineralization. Opposite observations are made in Jansen's metaphyseal chondrodysplasia, a disorder caused by constitutively active PPR mutants, and in transgenic animals expressing one of these receptor mutants (HKrk-H223R) under control of the type alpha1(I) collagen promoter. Expression of the Jansen transgene under the control of the type alpha1(II) collagen promoter was, furthermore, shown to delay chondrocyte differentiation and to prevent the dramatic acceleration of chondrocyte differentiation in PTHrP(-/-) mice, thus rescuing the early lethality of these animals. In the present study we demonstrated that the type alpha1(II) collagen promoter Jansen transgene restored most of the bone abnormalities in PPR(-/-) mice, but did not prevent their perinatal lethality. These findings suggested that factors other than impaired gas exchange due to an abnormal rib cage contribute to the early death of PPR(-/-) mice.
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PMID:Partial rescue of PTH/PTHrP receptor knockout mice by targeted expression of the Jansen transgene. 1171 97

This study used a rat model to investigate the microstructural organization of collagen through the transition from scar to intact residual segments of a healing medial collateral ligament (MCL). Twenty-two male retired breeder Sprague-Dawley rats were randomly separated into two groups. Eleven underwent surgical transections of both MCLs and were allowed unrestricted cage activity until euthanized two weeks post surgery. The remaining eleven rats were used as normal controls. All 44 MCLs were harvested including intact femoral and tibial insertions and prepared for scanning electron microscopy (SEM) imaging. At harvest the scar region in the healing ligaments was more translucent than the normal tissue. Ligaments were viewed from femoral to tibial insertions at magnifications of 100X through 20,000X. Tissue away from the scar region in the transected MCLs was indistinguishable from normal tissue in uninjured ligaments. Collagen fibers and fibrils in these tissues were more aligned along the longitudinal axis of the ligament than in the scar tissue. Continuity of collagen fibers and fibrils were consistently observed from the residual portions of the transected ligament through the scar region. Bifurcations/fusions, but no anastomoses, in fibers and fibrils were observed in both normal and scar tissues of ligaments. Qualitatively, bifurcations were encountered more frequently in scar tissue. In the transition region, larger diameter fibers from the residual tissue bifurcated into smaller diameter fibrils in the scar. This connection between larger diameter and smaller diameter fibers and fibrils indicates that bifurcations/fusions are likely to be the dominant way in which force is transmitted from a region with larger fibrils (residual ligament) into and through a region with smaller fibrils (scar).
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PMID:Microstructural morphology in the transition region between scar and intact residual segments of a healing rat medial collateral ligament. 1171 67


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