Search

This study examined the effects of 1400W, an inhibitor of inducible nitric oxide (iNOS), on contractile function and iNOS expression in reperfused skeletal muscle. The right extensor digitorum longus (EDL) muscle of 104 rats underwent a sham operation or 3-h ischemia followed by 3-h or 24-h reperfusion (I/R). Rats received 3 mg/kg 1400W, 10 mg/kg 1400W, or water subcutaneously. Results showed that EDL contractile function in both 1400W-treated groups significantly outperformed the controls at 24-h but not at 3-h reperfusion. Although iNOS expression increased in all three I/R groups during reperfusion, a significantly smaller increase was found in 1400W-treated muscles after 3-h reperfusion, and more dramatically so after 24-h reperfusion. Our results indicate that inhibition of iNOS preserved the contractile function in reperfused skeletal muscle, perhaps via downregulating iNOS expression. Protection by 1400W at 24-h reperfusion suggests that the role of iNOS in exaggerating reperfusion injury is more prominent in the later stages of injury.

This study investigated the dosage effects of nitric oxide synthase (NOS) inhibitor N G-monomethyl-l-arginine(l-NMMA) on intermittent pneumatic compression (IPC)-induced vasodilation in uncompressed upstream muscle and the effects of IPC on endothelial NOS (eNOS) expression in upstream muscle. Afterl-NMMA infusion, mean arterial pressure increased by 5% from baseline (99.5 ± 18.7 mmHg; P < 0.05). Heart rate and respiratory rate were not significantly affected. One-hour IPC application on legs induced a 10% dilation from baseline in 10- to 20-μm arterioles and a 10–20% dilation in 21- to 40-μm arterioles and 41- to 70-μm arteries in uncompressed cremaster muscle. IPC-induced vasodilation was dose dependently reduced, abolished, or even reversed by concurrently infusedl-NMMA. Moreover, expression of eNOS mRNA in uncompressed cremaster muscle was upregulated to 2 and 2.5 times normal at the end of 1- and 5-h IPC on legs, respectively, and the expression of eNOS protein was upregulated to 1.8 times normal. These increases returned to baseline level after cessation of IPC. The results suggest that eNOS plays an important role in regulating the microcirculation in upstream muscle during IPC.

The authors have shown that exogenous nitric oxide (NO) protects innervated skeletal muscle against reperfusion injury. This study further evaluated the effects of exogenous NO donor on denervated skeletal muscle. Forty-eight denervated rat cremaster muscles underwent 3 hr of ischemia, followed by 90 min of reperfusion, and received systemic infusion of 100 nmol/min s-nitroso-n-acetylcysteine (SNAC) or an equal amount of phosphate-buffered saline (PBS). Results showed that the average diameter in 10 to 20 microm arterioles was between 107 percent and 123 percent of baseline in the SNAC group, and between 55 percent and 84 percent in the PBS group during 90 min of reperfusion. These values in 21 to 40 microm and 41 to 70 microm arteries were between 100 percent and 110 percent in the SNAC group, and between 70 percent and 90 percent in the PBS group from 20 to 90 min of reperfusion. Compared to the PBS group, the SNAC group had a statistically significantly greater vessel diameter in both 10 to 20 microm (p

This study examined mRNA and protein expressions of neuronal (nNOS), inducible (iNOS), and endothelial nitric oxide synthases (eNOS) in peripheral nerve after ischemia-reperfusion (I/R). Sixty-six rats were divided into the ischemia only and I/R groups. One sciatic nerve of each animal was used as the experimental side and the opposite untreated nerve as the control. mRNA levels in the nerve were quantitatively measured by competitive PCR, and protein was determined by Western blotting and immunohistochemical staining. The results showed that, after ischemia (2 h), both nNOS and eNOS protein expressions decreased. After I/R (2 h of ischemia followed by 3 h of reperfusion), expression of both nNOS and eNOS mRNA and protein decreased further. In contrast, iNOS mRNA significantly increased after ischemia and was further upregulated (14-fold) after I/R, while iNOS protein was not detected. The results reveal the dynamic expression of individual NOS isoforms during the course of I/R injury. An understanding of this modulation on a cellular and molecular level may lead to understanding the mechanisms of I/R injury and to methods of ameliorating peripheral nerve injury.

We evaluated the efficacy of the continuous suture technique (CST) in arteries and veins with varying external diameters (ED). In study 1 a direct end-to-end anastomosis was performed in 5 groups of animals (n = 15 in each group): group 1, rabbit carotid artery (ED, 1.8-2.0 mm); group 2, rabbit femoral artery (ED, 1.4-1.6 mm); group 3, rat femoral artery (ED, 0.7-0.9 mm); group 4, rabbit femoral vein (ED, 2.0-2.2 mm); and group 5, rat femoral vein (ED, 1.0-1.2 mm). In study 2 a graft from the femoral vein was interposed into the carotid artery, with a ratio of the diameter of graft to artery of 1.3:1 in the rats (group 6, n = 12) and 1:1 in the rabbits (group 7, n = 12). In each animal the vessel on one side was repaired using CST and the opposite vessel using the interrupted suture technique. Vessel samples were harvested 1, 2, and 4 weeks after anastomosis. The CST significantly reduced anastomosis time by up to 47% in arteries and 41% in veins. Bleeding time and blood loss were also significantly reduced with CST. Similar results were found in study 2. The total thrombosis rate was 8%, but no significant patency difference was noted between the CST and the interrupted suture technique in any vessel category. We conclude that the CST is a reliable and time-saving procedure in microvascular anastomosis of arteries with diameters greater than 0.7 mm and of veins with diameters greater than 1.0 mm.

Ischemia/reperfusion (I/R) injury appears to be a significant neutrophil-dependent component and may be ameliorated by blocking leukocyte-endothelial adhesion Using a rat extensor digitorum longus (EDL) muscle model, the present study tested the hypothesis that in vivo administration of the function-blocking monoclonal antibody (mAb) LAM1-116 which recognizes L-selectin, a cell-surface adhesion receptor, could decrease I/R injury. In 46 rats, one EDL served as a normal control and the opposite EDL underwent 3 hr of ischemia followed by 3 hr of reperfusion after pretreatment with LAM1-116 mAb, control IgG, or saline. Myeloperoxidase (MPO) activity showed only a two-fold increase from normal in LAM1-116-treated I/R EDL while a 27-fold increase occurred in the IgG 2a and saline groups, with a statistically significant (p < 0.001) difference. A significantly (p < 0.05) lower wet weight ratio, improved fatigue contractile force, and less neutrophil infiltration were found in LAM1-116-treated EDL, when compared to those in control IgG- or saline-treated EDL. The results indicate that blockade of L-selectin by LAM1-116 mAb can effectively reduce neutrophil infiltration in reperfused skeletal muscle, thereby decreasing tis sue edema and improving muscle fatigue contractile force. These findings may be important in understanding I/R injury.

Previous study has demonstrated that application of intermittent pneumatic compression on legs can cause vasodilation in distant skeletal muscle at the microcirculation level. This study evaluated the influence of inflation rate and peak-pressure duration on the vasodilatory effects of intermittent pneumatic compression. The cremaster muscles of 50 male rats were exposed and divided into five groups of 10 each. A specially designed intermittent pneumatic-compression device was applied in a medial-lateral fashion to both legs of all rats for 60 minutes, with an inflation rate and peak-pressure duration of 0.5 and 5 seconds, respectively, in group A, 5 and 0 seconds in group B, 5 and 5 seconds in group C, 10 and 0 seconds in group D, and 10 and 5 seconds in group E. Diameters of arterial segments were measured in vessels of three size categories (10-20, 21-40, and 41-70 microm) for 120 minutes. The results showed that the greatest increase in diameter was produced by intermittent pneumatic compression with the shortest inflation rate (0.5 seconds). A moderate increase resulted from compression with an inflation rate of 5 seconds, and no effective vasodilation occurred during compression with the longest inflation rate (10 seconds). When the groups with different inflation rates but the same peak-pressure duration were compared, there was a significant difference between any two groups among groups A, C, and E and between groups B and D. When the groups with different peak-pressure durations but the same inflation rate were compared, compression with a peak-pressure duration of 5 seconds caused a generally similar degree of diameter change as did compression without inflation at peak pressure. The findings suggest that inflation rate plays an important role in the modulation of distant microcirculation induced by intermittent pneumatic compression whereas peak-pressure duration does not significantly influence the vasodilatory effects of the compression. This may be due to the fact that rapid inflation produces a significant increase in shear stress on the vascular wall, which stimulates vascular endothelium to release nitric oxide, causing systemic vasodilation.

Local delivery of antibiotics via a degradable carrier has the potential for high local antibiotic levels and avoids systemic toxicity. Intravenous access, renal function monitoring, and later surgical removal may not be required when degradable local delivery modalities are used. This study examined the in vivo elution of gentamicin from processed bovine collagen (Type I). Gentamicin impregnated collagen (3 mg/kg) was implanted into the femoral medullary canal of 45 adult white rabbits. The gentamicin was released into the bone and averaged greater than 600 micrograms/ml during the initial 48 hours. Local bone levels fell to 144.40 +/- 229.84 micrograms/ml at 5 days and were subsequently greater than or equal to 10.30 +/- 5.02 micrograms/ml through Day 28. Serum levels reached an average peak of 1.25 +/- 0.29 micrograms/ml 5 hours after implantation and fell below 1.0 microgram/ml at 12 hours after implantation. Serum levels subsequently averaged less than or equal to 0.63 +/- 0.09 microgram/ml through Day 28. Collagen impregnated with gentamicin proved to be an effective degradable carrier of gentamicin in the healthy rabbit; it provided local bone concentrations above the minimum inhibitory concentration of gentamicin and serum concentrations below levels associated with systemic toxicity as long as 28 days after implantation.

To determine the changes in capillary perfusion, which occur with elevated tissue pressure, and to highlight the relationship between systemic blood pressure and compartment pressure, we designed an experiment that allowed direct observation of the microcirculation of skeletal muscle under normal and increased compartment pressures. In each of 10 anesthetized rats, the cremaster muscle was exposed and suspended in a transparent pressure chamber. In vivo videomicroscopy was then performed and blood pressure was monitored via left carotid artery cannulation. Two sets of data for each animal were obtained: deltaP (mean arterial pressure compartment pressure) at which the muscle capillary blood flow was completely arrested, and the number of capillaries per 10,000 square micrometers of skeletal muscle with blood flowing at compartment pressures of 0, 15, 30, 45, and 60 mm Hg. Capillary blood flow stopped at a deltaP of 25.5 mm Hg +/- 14.3 SD. We found that capillary blood flow, as measured by the number of capillaries with blood flow per 10,000 square micrometers, decreased significantly (P < 0.05) as compartment pressure reached 15, 30, 45, and 60 mm Hg, when compared to 0 mm Hg; there was no vessel collapse at these pressures. These data show that increasing compartment pressure reduces the number of perfused capillaries per unit area, and that there is complete cessation of muscle capillary blood flow when the compartment pressure is within about 25 mm Hg of the mean arterial pressure.

The effects of a nitric oxide (NO) donor on microcirculation and contractile function of reperfused skeletal muscle were studied. Rat cremaster muscles underwent 5 hours of ischemia and 90 minutes of reperfusion and were divided into two groups systemically infused with S-nitroso-N-acetylcysteine (SNAC, 100 nmol/min) and phosphate-buffered saline (PBS), respectively. The results showed that the vessels in the SNAC group had more rapid and complete recovery than that in controls. A significant difference was found from 10 to 40 minutes and at 90 minutes in 10-20-microm arterioles, from 10 to 90 minutes in 20-40-microm arterioles, and at 10 and 90 minutes in 40-70-microm arteries. When compared to controls, SNAC-treated muscles showed larger fluorescein filling areas at 15, 30, 60, and 90 minutes and greater isometric tetanic contractile forces in response to stimulation frequencies of 40, 70, 100, and 120 Hz. The data indicate that supplementation of exogenous NO could effectively improve microcirculation and contractile function of skeletal muscle during early reperfusion.

The present study was conducted to elucidate the effects of tirilazad mesylate (U-74006F), a potent inhibitor of lipid peroxidation, on vessel diameter, capillary perfusion, and contractile function of rat cremaster muscle during a 90-minute reperfusion period that followed 4 hours of warm ischemia. Two groups of 32 animals were treated with either 3 mg/kg U-74006F or the vehicle (citrate buffer) alone 30 minutes before. ischemia, 90 minutes after ischemia, and immediately before reperfusion. With use of intravital videomicros-copy, the internal luminal diameters of preselected vessels were measured prior to ischemia and during reperfusion The area that filled with fluorescein was determined at 15-minute intervals for as long as 90 minutes of reperfusion, and contractile function was examined in vitro in an organ bath at that point. In the U-74006F group, after 90 minutes of reperfusion the vessel diameters returned completely to baseline and the diameters of all three categories of vessels at every time point from 10 to 90 minutes of reperfusion had significantly more rapid recovery than the controls. Although some evidence of more rapid fluorescence was noted in the U-74006F group, the two groups did not differ significantly at any time period of reperfusion. In response to tetanic stimulation, the muscles treated with U-74006F had a significantly greater contractile force at all stimulation frequencies than the control musclss. Our findings indicate that pretreatment with U-74006F can effectively decrease the rise of vascular resistance and preserve the contractile function of skeletal muscle during early reperfusion, thereby attenuating ischemia-reperfusion injury.

This study evaluated the influence of the dose and administration methods of enoxaparin, a low-molecular-weight heparin, on the patency rate of crushed rat femoral arteries following anastomosis. An impact crush with a 25-kg magnitude was applied to a 2-mm segment of 100 rat femoral arteries, followed by anastomosis. The arteries were divided into five groups: group 1 received systemic enoxaparin alone with a relatively high dose (45 IU) twice a day for 3 days; groups 2 and 3 received topical irrigation with a lower (15 IU/mL) concentration and a higher (45 IU/mL) concentration, respectively; group 4 received systemic and topical application at a lower (15 IU) dose and concentration (15 IU/mL); and group 5 received systemic and topical application at a higher (45 IU) dose and concentration (45 IU/mL). The results of this study demonstrate the following: (1) topical irrigation with enoxaparin at a concentration of 45 IU/mL-three times higher than that recommended for clinical use adjusted by body weight (15 IU/mL)-is effective for antithrombotic action; (2) a combination of systemic and local application does not offer additional benefit in the patency rate when compared to local irrigation alone; (3) systemic administration alone does not prevent thrombus formation; and (4) enoxaparin is potentially useful to enhance the patency rate in compromised microvessels.

Calcitonin gene-related peptide is a potent intrinsic vasodilator, can induce prostacyclin release, and may inhibit membrane lipid peroxidation. This study examines the effect of calcitonin gene-related peptide on vessel diameters, capillary perfusion, and contractile function of skeletal muscle after 4 or 5 hours of ischemia and during immediate reperfusion using the rat cremaster muscle model. Forty-two male rats were used; half of these received 0.2 ml of 10(-7) M calcitonin gene-related peptide after 0, 15, and 30 minutes of reperfusion, while the other half received normal saline as a control. By means of intravital videomicroscopy, the diameters of 10 vessels per muscle were measured prior to ischemia and during reperfusion. The fluorescein filling area was determined at 15, 30, and 60 minutes of reperfusion. After 1 hour of reperfusion, muscle function was examined in vitro by quantifying the contractile response to electric field stimulation of the muscles in an organ bath system. There was a significant increase in the diameter of the arterioles, but not the small arteries, at every time point from 10 to 60 minutes of reperfusion. The fluorescein filling area was increased in treated muscles at every time point. Contractile function was not significantly preserved. In light of the ability of calcitonin gene-related peptide to relieve vasospasm and improve capillary perfusion, it may be useful in reducing reperfusion injury in the future.

An absorbable staple for meniscal fixation was developed and evaluated in an in vivo canine study. The staple consisted of two rigid barbed legs, made of a copolymer of polyglycolic acid and polylactic acid, connected by a flexible suture made of the same absorbable material. The staple was developed for arthroscopic use, eliminating the risk of nerve or vascular injuries associated with suture fixation. Eighty-two mongrel dogs underwent meniscal repair in this study, which was performed in two sequential parts. An incision was made in the medial meniscus at the peripheral third and was fixed with either the staple or with a single 3-0 PDS suture. The animals from the first part of the study, in which the biocompatibility of a prototype staple design, was evaluated were sacrificed at 3 days, 2 weeks, 6 weeks, 3 months, and 9 months. Those from the second part of the study, in which an improved staple design of the same material, was evaluated were sacrificed at 6 weeks, 4 months, and 1 year. The operated menisci were either examined histologically or were mechanically tested for tensile strength. Absorption of the staple began by 3 months and was almost complete by 1 year. Mechanically, the staple provided greater tensile strength augmentation of the meniscus than suture fixation for up to 4 months. In the long term, there was no difference between the staple and suture in enhancement of healing.

Local delivery of antibiotics by a degradable carrier has the potential for high local antibiotic levels and avoids systemic toxicity. Intravenous access, renal function monitoring, and subsequent surgical removal may not be required when degradable local delivery modalities are used. This study examined the in vivo elution of gentamicin from processed bovine collagen (type I) in 66 adult White rabbits. Collagen impregnated with gentamicin (3 mg/kg) was implanted into the vastus lateralis, and data were collected from 15 minutes to 28 days after implantation. Local tissue biopsies were taken a minimum of 2 mm from the implantation site. The gentamicin was released into the local tissue and averaged more than 3,800 micrograms/ml during the initial 4 hours after implantation. Local levels fell to 6.90 +/- 5.22 micrograms/ml at 24 hours and subsequently were 2.70 +/- 1.75 micrograms/ml or more through day 28. Serum levels reached an average peak of 4.04 +/- 1.75 micrograms/ml at 5 hours after implantation, decreased after the initial 24 hours, and subsequently were less than 0.41 +/- 0.20 microgram/ml through day 28. Collagen impregnated with gentamicin proved to be an effective degradable carrier of gentamicin in the healthy rabbit; it provided local tissue concentrations above the minimum inhibitory concentration and serum concentrations below levels associated with systemic toxicity for 28 days after implantation.

The effect of lymphotoxin (LT) on the functional recovery of crushed peripheral nerves was studied. Using a specially designed compression device, a 5 mm segment of the right sciatic nerve of rats was subjected to a 100 g crush load with a 2 hr duration. The rats in the experimental and control groups received two doses of LT (20 micrograms/kg each) or the same volume of saline, respectively, administered intraperitoneously 24 hr and 1 hr before the procedure. Walking track tests and histologic examinations were performed at intervals up to 56 days after the crush. Motor functional recovery in the LT pretreated group started at day 7 while the crushed limb in the control group remained totally dysfunctional. The sciatic functional index improved faster in the LT group than in the control group during the second week after the crush and reached a significant difference (P < 0.05) at day 18. Subsequently, both groups had a similar evolution. Histologic results paralleled the functional findings. In conclusion, LT can promote motor functional recovery of crushed rat peripheral nerve in the early stage of regeneration.

This study introduces a new rat thrombosis model in which a 2-mm segment of femoral artery is crushed by an impact load. Ninety-five femoral arteries were divided into five groups. The vessels in Groups I and 2 underwent only crush injuries with a 15-kg and 25-kg load, respectively. Group 3 vessels were not crushed, but did undergo vessel anastomosis by a standard microsurgical technique. The vessels in Groups 4 and 5 were crushed by a 25-kg load, and then divided and anastomosed. During the procedure, the vessel lumina were topically irrigated with saline (Group 4) or heparin solution (Group 5). Thrombosis and thrombolysis were evaluated at set time points up to 56 days after operation. While all vessels in Groups 1 and 3 remained patent, the rate of occlusive thrombus formation in Group 2 significantly (p < 0.001) dropped from 85 percent at day 1 to 11 percent at day 7. The intima and media in Groups 4 and 5 were severely disrupted and often occluded the lumen, Group 5 had a significantly (p < 0.01 to 0.001) lower rate of occlusive thrombus formation (40 to 45 percent) at days 1 and 7 than Group 4 (90 percent). Histology in Groups 2, 4, and 5 at day 1 showed no intimnal and almost no medial tissue left in the crushed area. The adventitia and remaining external elastic lamina were adherent to thrombus in the occluded vessels or covered by fibrin and platelet mesh in the patent vessels. The results documented that spontaneous thrombolysis occurs in thrombosed arteries following crush injury alone, but not in thrombosed arteries after crush injury followed by suture anastomoses. The degree of disruption of the internal elastic lamina and the presence of sutures appear to contribute to occlusive thrombus formation following crush injury and anastomoses. Topical irrigation with heparin solution, at the concentration routinely used clinically, significantly reduces the thrombosis rate at the anastomosis site in crushed vessels, but does not promote thrombolysis.

Ischemia-reperfusion injury represents a complex series of vascular and cellular events that resembles an acute inflammatory reaction within the reperfused tissue. This article provides an overview of glucocorticosteroid effects on cells and tissues involved in inflammatory reaction following ischemia-reperfusion of muscle and cutaneous tissue. Glucocorticosteroids exert a variety of effects that influence the microcirculation. These effects include leukocyte recruitment, reduction of vascular permeability, inhibition of formation of cytokines or other mediators, and modulation of enzyme systems involved in inflammation. The current view is that glucocorticosteroids act through cytoplasmic receptors by controlling the transcription of certain genes encoding regulatory proteins, but the exact mechanisms of glucocorticoid action on ischemia-reperfusion are not completely understood. Potential mechanisms may involve modulation of neutrophil and endothelial function, inhibition of formation of arachidonic acid products, and attenuation of lipid peroxidation of biological membranes through membrane stabilization and scavenging of toxic free radicals.

This study evaluated the effects of dexamethasone (DXM) on contractile function of reperfused extensor digitalis longus (EDL) muscles following 3-hour ischemia and 24-hour reperfusion. The rats were divided into four groups: normal muscle, ischemia with saline treatment, ischemia/reperfusion with saline treatment, and ischemia/reperfusion with DXM treatment groups. DXM (0.6 mg kg[-1]) or saline (3.0 ml kg[-1]) was administered at 3 hours prior to ischemia. Results showed that although contractile force in all three treated groups was significantly lower than that of normal EDL, the average isometric tetanic contractile force in the DXM-treated group was significantly greater than that in the saline-treated ischemia and ischemia/reperfusion groups. A significant difference was also seen at the peak force and at 5 seconds of the fatigue trains, and with a longer fatigue half-time (FT1/2) in the DXM-treated group than in the other two groups. Histologically, edema, inflammation and necrosis of muscle fiber were less severe in the DXM-treated group than in the saline-treated group. The results indicate that pretreatment with DXM appears to attenuate, but does not completely reverse, the contractile function deficit of ischemic skeletal muscle during the first 24 hours of reperfusion.

This study was designed to observe the effect of the nonsteroidal antiinflammatory piroxicam on a controlled muscle strain injury in the rabbit model. The tibialis an terior tendons of 90 New Zealand White rabbits were detached at their distal insertions, and the right tendon was stretched to the yield point of the deformation curve. One group of 50 rabbits received piroxicam treat ment and the others received no treatment. At 1, 2, 4, and 7 days the parameters of muscle function, tensile strength, and histology were examined. Muscle con tractile force was significantly greater in the piroxicam- treated group at Day 1, but no difference was noted at any other time period. Tensile strength was not signifi cantly different at any time period in either group. His tology revealed delayed degradation of damaged tissue and slowed regeneration of muscle tissue at the injury site in the piroxicam-treated group.Piroxicam and other antiinflammatories are fre quently given to athletes being treated for muscle strain injuries to control pain through their effect on the in flammatory process. This study demonstrates that pi roxicam does not adversely influence the recovery of contractile and tensile strength in a followup period of 1 week. Therefore, antiinflammatory treatment may be beneficial early in the course of muscle injury.

The pathogenesis and treatment of rupture of the Achilles tendon remain a source of controversy. This study presents the results of a biomechanical, functional, and morphological evaluation of a group of rats that had division and repair of the Achilles tendon. A total of 46 rats were used: 18 for biomechanical testing, 18 for functional evaluation, and 10 for histology. Morphological examination revealed an early inflammatory response with loose connective tissue formation that was replaced gradually by fibroblasts and a collagenous matrix. The functional evaluation (Achilles functional index [AFI]) was made from measurements of the hind pawprints of walking rats. Division and repair of the Achilles tendon produced a significant functional impairment (mean [+/- SEM] AFI = -87 +/- 8; p < 0.001), which gradually improved with healing time. The load to failure for the repaired tendons consistently improved with healing time, in a manner similar to the functional recovery. The average deformation (repair/control) varied considerably and was not related to healing time. The stiffness of the repaired tendons increased with healing time and was 60% of the corresponding control side by day 15. The major finding of this study was a strong correlation between the AFI and the failure load of the healing tendon-bone constructs (250-300 g group, r = 0.97, p < 0.001; 325-375 g group, r = 0.96, p < 0.001).

Controversy exists regarding the treatment of Achilles tendon ruptures. The aim of this study was to determine whether surgical repair of the rat Achilles tendon offered any biomechanical, functional, or morphological advantage over no repair. Thirty-two male Sprague-Dawley rats were randomly allocated into four groups: (1) sham operated (skin incision only), (2) no repair (complete division of the Achilles tendon and plantaris tendon without repair), (3) internal splint (plantaris left intact), and (4) Achilles repair (with a modified Kessler-type suture). Functional performance was determined from the measurements of hindpaw prints utilizing the Achilles Functional Index. On day 15, the animals were killed, and biomechanical and histological evaluations were performed on both the injured and uninjured Achilles tendon constructs.All groups subjected to Achilles tendon division had a significant initial functional impairment that gradually improved so that by day 15 there were no functional or failure load impairments in any group. The injured tendons in all three groups subjected to Achilles tendon division had a 13-fold increase in the cross-sectional area and were less stiff and more deformable than uninjured and sham-operated tendons on day 15 ( P < .001). The magnitude of the biomechanical and morphological changes at postoperative day 15 and the initial impairment and rate of functional recovery were similar for no repair, internal splint, and Achilles repair groups. In summary, this study demonstrates that surgical repair of the Achilles tendon in the rat does not offer any advantage over nonoperative management.

The effect of temperature on the mechanical failure properties of rabbit skeletal muscle (tibialis anterior and extensor digitorum longus) was examined. For all tests, one leg was maintained at 25°C and the contralateral leg at 40°C. Muscles were pulled to failure according to assignment into one of three groups: 1) passive failure at 10 cm/sec, 2) passive failure at 1 cm/sec, or 3) active (muscle is stimulated to contract as it is pulled) failure at 10 cm/sec. Load to failure was higher in the cold muscle for all groups tested. Total deformation was the same except in Group 1, when the warm muscle had a greater deformation. Energy absorbed before failure was greater in the cold muscle in Groups 2 and 3. Stiffness was higher in cold muscles for all muscles except the extensor digitorum longus in Group 1. In this study, temperature had a significant effect on the tensile properties; these thermal effects were de pendent on both loading rate and contractile state. Comparing loading rates, warm muscle tested at 10 cm/sec had higher failure loads than that tested at 1 cm/sec. Comparing stimulated versus unstimulated muscle (Group 1 versus Group 3), the stimulated tibialis anterior muscle absorbed more energy than unstimu lated ones. Stimulated extensor digitorum longus mus cles had higher failure loads, absorbed more energy, and were stiffer than nonstimulated muscles. This study offers experimental data to support the theory that warming muscles can aid in injury prevention and im provement in athletic performance.

Osteonecrosis of the femoral head was induced in dogs by a process of deep freezing, accompanied by stripping of the soft-tissue attachments from the femoral neck and intertrochanteric area, in an effort to develop an experimental model to study treatment modalities for avascular necrosis of the hip. Immediate uniform necrosis was created in a defined area. Thereafter, spontaneous healing originated mainly from the adjacent viable bone by migration of undifferentiated mesenchymal tissue into the necrotic bone, genesis of fibrosis, and, finally, formation of new bone. Osteogenesis occurred primarily through intramembranous ossification without a preexisting template. Quantitative measurements showed a difference in the rates of ingrowth and revascularization of the necrotic area during the fibrotic and osteogenic phases of the healing. The de novo osteogenesis was slower than the migration of fibrosis. These findings indicate that, without osteotomy, we can reproducibly create necrosis of bone in a defined area of the proximal femur and induce a reparative process that incompletely heals the defect. Although it does not fully simulate the human disorder of osteonecrosis, our experimental surgical model provides a basis for further laboratory investigation into the management of avascular necrosis of the femoral head.

Diametric growth at the anastomotic site, following microvascular anastomoses in pediatric patients, remains a problem, when nonabsorbable suture material is used. This study investigated the long-term effects of femoral artery anastomoses with nonabsorbable sutures in 20 growing rats with a mean weight of 101 g. The right femoral artery was anastomosed with 10-0 polypropylene monofilament suture and the left with identical nylon suture, using 5 to 6 interrupted sutures for each end-to-end anastomosis. At a 6-month harvest, the mean body weight had increased by 729 percent and the diameter of the femoral artery by 240 percent. The patency rate at the anastomotic sites was 97.5 percent and the complication rate was 10 percent, including one occlusion and three aneurysm formations at the sites. Angiography, diameter measurement, and histology demonstrated no stenosis or thrombus formation in all animals. Histology also showed normal vessel lumen size and well-organized re-endothelialization, without intimal hyperplasia at the anastomosis sites. With an interrupted suture technique, nonabsorbable sutures do not compromise growth at the anastomosis site or long-term patency of the anastomosed vessels. This study indicates that interrupted nonabsorbable suture materials are able to create excellent long-term results in microvascular anastomoses in rapidly growing hosts.

The selection of suture materials is an important factor in further improving the results of microsurgical operations. In this study, two kinds of nonabsorbable suture materials, 10-0 polypropylene and 10-0 nylon monofilament, were compared in end-to-end anastomosis of 66 femoral arteries of adult rats. Both suture types were of identical size (70 μm needle/28 μm suture) and each artery received eight sutures. The vessels were harvested at various intervals from 2 hr to 180 days postoperatively and were evaluated by pathology, radiology, and tensile strength test. The results show that both sutures are capable of achieving excellent long-term patency (100%) of anastomosed sites. Polypropylene suture was equivalent to nylon in mechanical integrity of the anastomosis sites but was superior in handling, knotting, and biocompatibility. These physical and biological properties of polypropylene sutures may offer the benefits of diminished early and late complications at anastomosis sites and reduced operation time. Enhancement of contrast against surrounding tissue may make polypropylene a superior alternative to nylon sutures for microsurgical use. © 1993 Wiley-Liss Inc.

Ultrasonic driven tools have been developed to facilitate the removal of bone cement during revision arthroplasty. The effect on whole bone strength of cement removal by ultrasonic tools was examined in a canine femur model. Paired, fresh-frozen canine femora were divided into two groups. In group A, one femur from each pair was subjected to cement extraction with ultrasonic tools. In group B, one femur from each pair was subjected to manual cement extraction. Contralateral femora from each pair served as controls to determine the strength of intact femora. Torsional fractures were produced using a servocontrolled hydraulic testing machine (Minneapolis Testing System, Minneapolis, MN). Maximum torque, maximum angle, and energy capacity to failure were determined. Results were recorded as a reduction in percent value of the tested specimen versus the contralateral control. When comparing femora with cement removal by ultrasonic tools to the contralateral control femur, there were no statistical differences in ultimate torque (P = .83), maximum angle (P = .89), and energy capacity (P = .74) by analysis of variance. In addition, there were no significant differences between the group with ultrasonic tool cement removal and the group with manual tool removal. The authors conclude that in this canine model, removal of cement with ultrasonically driven tools has no adverse effects on whole bone strength.

To evaluate the effect of nerve-graft polarity on function, a 1-cm segment of sciatic nerve was excised and reoriented in three groups of 20 adult Sprague-Dawley rats. In one group, the nerve was cut and anastomosed in the original orientation to act as a control. In the second group, the nerve-graft polarity was reversed 180 degrees. The final group underwent reversal of polarity 180 degrees and was rotated 180 degrees, (i.e., the posterior tibial nerve fascicles proximally were opposite to the peroneal nerve fascicles distally). Functional recovery was evaluated using Bain's modification of de Medinaceli's sciatic functional index (SFI). Rat-track analysis was performed over a 120-day period. Histologic correlation was also performed at the time of sacrifice. From our study, we concluded that reversing nerve-graft polarity, with or without rotation, does not influence subsequent function in this model.

The effect of extracorporeal shock wave lithotripsy on interfacial strength between prosthesis and bone in cementless arthroplasty was examined using a rabbit model. Paired femora, each implanted with fiber mesh porous coated titanium implants, were harvested from rabbits 15 weeks after implantation. In group I, one femur from each pair was exposed to lithotripsy treatment consisting of 2,000 shocks at 20 kV. In group II, one femur from each pair was exposed to 2,000 shocks at 26 kV. Contralateral femora from each pair served as controls in both groups. Mechanical pushout tests were conducted on the implants using a 1321 Instron testing machine at a constant rate of 1 mm/minute. Shock waves generated at 20 kV were found to have no significant decrease on either the prosthesis/bone interfacial strength or energy to failure of cementless implants. Shock waves generated at 26 kV produced a mean 17.45% decrease in the prosthesis/bone interfacial strength, which approached statistical significance (P = .062), and a 7.84% mean decrease in the energy to failure (P = .268). However, in four of the seven group II specimens, cortical fractures occurred. These findings suggest that lithotripsy will not aid in the removal of uncemented porous coated devices and lithotripsy inadvertently focused at an uncemented device will not disrupt significantly the prosthesis-bone interface.

Pentoxifylline improves microvascular blood flow in conditions of vascular insufficiency. The clinical benefits of pentoxifylline have been attributed to its effects on the cellular elements of whole blood, although a few studies suggest it may also be a vasodilator. The purpose of this study was to determine whether pentoxifylline has a vasodilator effect on the luminal diameter of small arteries preconstricted with norepinephrine and on resting small arteries in the rat cre master muscle.Intravital videomicroscopy was used in order to observe directly the vasodi lator capacity of topically applied pentoxifylline. The results reveal that pentox ifylline (100 μM and more) can significantly dilate small arteries preconstricted with norepinephrine. Pentoxifylline had no effect on the diameter of resting small arteries.These results suggest that vasodilation may play a role in the ability of pen toxifylline to improve arterial blood flow.

An end-to-side anastomosis is often a necessity in replantation surgery. This study examines the effects of elliptical versus slit arteriotomy and the angle of anastomosis on vessel patency and aneurysm formation. Forty male rats were divided into four groups of ten rats each. The right and left common iliac arteries were selected as a model. The results showed no significant statis tical difference in patency rates among our four experimental groups. The end-to-side anastomotic technique of choice uses the elliptical arteriotomy placed at a 90° angle. It provides good visualization and easy suture placement. © Wiley-Liss, Inc.

Sixteen embalmed hemipelves were used to determine the optimal acetabular screw placement to provide maximal screw pull-out strength in unicortical and bicortical screw fixation. The anterior column, superior ilium, posterior column, ischium, and pubis regions of the pelvis were tested using 6.5-mm titanium alloy screws and a hydraulic servo-controlled 1321 Instron testing machine. Force vs displacement data were acquired. Bicortical fixation was stronger than unicortical fixation in the four zones compared. This difference was significant in the superior ilium, posterior column, and ischium. The anterior column could not accept unicortical screws due to inadequate bone depth, which ranged between only 6 mm and 10 mm. Bicortical fixation was significantly greater in the superior ilium, posterior column, and ischium than in the anterior column or pubis. Unicortical fixation was greatest in the superior ilium. This information may aid decisions concerning the positioning of screws to augment acetabular component fixation.

Although tissue is denervated during replantation of a severed part, tissue transfer, or muscle transplantation, there are few studies concerning the effects of acute denervation on muscle microcirculation. We have described a surgical procedure that totally denervates the rat cremaster muscle. Histological examination of the denervated tissue has given convincing evidence of nerve degeneration and skeletal muscle atrophy, accompanied by electrophysiological evidence of total denervation. The diameters of each component of the microcirculation were measured before and after denervation. Arterioles and arteries ranging in size from 10 to 70 microns in diameter were found to increase significantly in size immediately after acute denervation. Larger arteries and veins did not undergo significant diametrical increases. These findings suggest that total acute denervation significantly increases the diameter of small arteries and arterioles, thereby decreasing the resistance in the arterial bed and increasing blood flow. Since this phenomenon is of limited duration (20 min), it would appear to be ineffective in enhancing reperfusion and oxygenation at the time of reattachment of amputated parts or during vascularized tissue transfers, until methods of prolonging it for several hours or more are found.

This experimental study correlated varying methods of nerve repair with the degree of functional return using the rat sciatic nerve as a model. Six groups of adult male Sprague-Dawley rats (300 to 350 g) had their left sciatic nerve crushed (Group 1), transected and not repaired (Group 6), or transected and repaired using four methods that varied the degree of axonal alignment (Groups 2, 3, 4, and 5). Functional return was evaluated weekly using the sciatic functional index (SFI) and histology. Group 1 (crush) had a significantly better functional return than the other groups (p less than .001). Group 6 (no repair) had a significantly worse functional return than the other groups (p less than .001). Groups 2, 3, 4, and 5, regardless of the precision of axonal alignment, experienced the same degree of functional return (p less than .05). In all groups, with the exception of Group 1 (crush), the degree of functional return was poor. As multiple modifications in conventional microsurgical neurorrhaphy have not resulted in a significant functional improvement, factors not addressed by current techniques apparently compromise clinical results. Further improvement in the results of neurorrhaphy will require modifications in our techniques beyond presently conventional microsurgical methods.

This study investigated the effects of inhibition of NF-kappaB activation on microcirculation and inducible NOS expression in reperfused rat cremaster muscle. The muscle from 16 rats underwent 5-h ischemia and 90-min reperfusion. Each rat received NF-kappaB inhibitor pyrrolidine dithiocarbamate (PDTC, 150 mg/kg) or phosphate-buffered saline 15 min before reperfusion. Results showed that PDTC treatment had a significant overall increase in muscle blood flow during reperfusion. Blood flow more rapidly recovered to and over baseline in the PDTC-treated group than in controls, with a significant difference at 10-30 min and 70-90 min. Expression of iNOS mRNA had a 167-fold increase from normal in controls, but was significantly (P < 0.05) reduced to a 63-fold increase in PDTC-treated muscles. In addition, PDTC treatment significantly (P < 0.05) decreased a reperfusion-induced increase in activated NF-kappaB p65 and nuclear p65 protein. Our results suggest that NF-kappaB is involved in I/R injury and that inhibition of NF-kappaB p65 activation affords protection against I/R injury, perhaps via downregulating expression of iNOS transcription.

Evaluation of graft-host bone interactions after failed vascularized fibular grafting of femoral head necrosis may elucidate the reasons for failure of the procedure. According to the authors' study, the vascularized fibula implanted into the femoral head before collapse has the potential for restructuring the major segment of the affected head and delaying joint degeneration for many years if circumferential graft-host union is established. Asymmetric bone healing and non-union between the graft and the necrotic subchondral bone in the weight-bearing area lead to failure, progression of symptoms, and subsequent early hip replacement.

Inducible nitric oxide synthase (iNOS) participates in many pathological events, and selective inhibition of iNOS has been shown to reduce ischemia-reperfusion (I/R) injury in different tissues. To further confirm its role in this injury process, I/R injury was observed in denervated cremaster muscles of iNOS-deficient (iNOS−/−) and wild-type mice. After 3-h ischemia and 90-min reperfusion, blood flow in reperfused muscle was 80 ± 8.5% (mean ± SE) of baseline at 10-min reperfusion and completely returned to the preischemia baseline after 20 min in iNOS−/− mice. In contrast, blood flow was 32 ± 7.4% at 10 min and increased to 60 ± 20% of the baseline level at 90 min in wild-type mice ( P < 0.001 vs. iNOS−/− mice at all time points). The increased muscle blood flow in iNOS−/− mice was associated with significantly less vasospasm in all three sizes of arterial vessel size categories. The weight ratio to the contralateral muscle not subjected to I/R was greater in wild-type mice (173 ± 11%) than in iNOS−/− mice (117 ± 3%; P < 0.01). Inflammation and neutrophil extravasation were also more severe in wild-type mice. Western blot analysis demonstrated an absence of iNOS protein band in iNOS−/− mice and upregulation of iNOS protein expression in wild-type mice. Our results confirm the importance of iNOS in I/R injury. Upregulated iNOS exacerbates I/R injury and appears to be a therapeutic target in protection of tissues against this type of injury.

N(omega)-Propyl-L-arginine (NPA) is reported to be a highly selective inhibitor of neuronal nitric oxide synthase (nNOS). This in vivo study observed its role in ischemia/reperfusion (I/R) injury in rat skeletal muscle. Our results showed that NPA infusion significantly increased vessel diameters and blood flow in reperfused cremaster muscle, and slightly increased contractile function in reperfused extensor digitorum longus (EDL) muscle. In addition, NPA treatment slightly increased I/R-mediated downregulation of nNOS and eNOS mRNA and protein levels. Although NPA showed a beneficial role in I/R injury, our in vivo data do not support NPA as a selective nNOS inhibitor. Also, our data do not provide any insight into the mechanism of NPA. Thus, the in vivo mechanism of action of NPA needs to be further identified, and the role of nNOS in skeletal muscle I/R still remains to be determined.

Purpose To investigate whether inhibition of inducible nitric oxide synthase (iNOS) improves microcirculation in denervated and reperfused skeletal muscle. Methods The cremaster muscles of 52 rats received iNOS inhibitor 1400W (3 mg/kg) or phosphate buffered saline (PBS) and underwent either 3 hours of ischemia and 1.5 hours of reperfusion or a sham operation. During reperfusion the vessel diameters were measured by using intravital videomicroscopy and overall muscle blood flow was measured with laser Doppler flowmetry. The expression of NOS messenger RNA (mRNA) and protein was determined by using real-time reverse-transcription polymerase chain reaction and Western blot, respectively. Results 1400W treatment significantly increased the mean blood flow of the reperfused muscle compared with controls, and this was associated with significantly less vasospasm in 10 to 20 μm, 21 to 40 μm, and 41 to 70 μm arterioles. The expression of iNOS mRNA and protein in controls increased 23-fold and 6-fold from normal, respectively, but was reduced to only a 2-fold increase in the 1400W-treated muscles. The ischemia/reperfusion (I/R)-induced decrease of endothelial NOS (eNOS) and neuronal NOS (nNOS) expression in controls was not significantly changed after 1400W treatment. Conclusions Our data support a nitric oxide-mediated mechanism in reperfusion injury and show the importance of inhibition of iNOS in reducing reperfusion injury in denervated skeletal muscle. Our results suggest potential benefits via inhibition of iNOS to improve clinical outcomes not only for hand surgeons who work in the microsurgery field, but also for other physicians whose work involves ischemia/reperfusion injury.

To determine the role of inhibition of complement activation in the contractile function of skeletal muscle ischemia-reperfusion (I/R) injury, the rat extensor digitorum longus (EDL) muscles underwent 3 h ischemia and received human C1-esterase inhibitor (C1-INH, 100 IU/kg), a synthetic C1q A chain peptide with a similar inhibitory effect on activated C1 (peptide, 5 mg/kg), or human serum albumin control. Results showed a significant overall increase in tetanic contractile forces of the reperfused EDL in both C1-INH and peptide groups compared to controls. Maximum improvement occurred with peptide treatment at 120-Hz stimulation, with an increase in force from 38 +/- 4% of normal in controls to 52 +/- 4% in peptide-treated rats. There were no significant differences between C1-INH and peptide groups. Plasma C3 and C4 activities were significantly increased in both treated groups, suggesting inhibition of complement activation. Our results suggest that complement activation is involved in I/R injury, and inhibition of complement activation may therefore represent a potential therapeutic approach to reducing or preventing I/R injury.

This study evaluated the effects of the selective inducible nitric oxide synthase (iNOS) inhibitor N-[3-(aminomethyl)benzyl]acetamidine (1400W) on the microcirculation in reperfused skeletal muscle. The cremaster muscles from 32 rats underwent 5 h of ischemia followed by 90 min of reperfusion. Rats received either 3 mg/kg 1400W or PBS subcutaneously before reperfusion. We found that blood flow in reperfused muscles was

The aim of the current study was to test the hypothesis that the induction of an underlying immunologic condition in rabbits may enhance the development of steroid-induced osteonecrosis. Thirty-five adult rabbits were divided into four groups. Group I: 10 rabbits were immunized at 15-day intervals for 2 months by murine antibodies to deoxyribonucleic acid autoantibodies. Four weeks after the end of the immunization, the animals received injections of methylprednisolone for 7 days and then prednisolone per os for 8 months. Group II: 10 animals only received immunizations according to the protocol used in Group I. Group III: 10 animals only were treated with corticosteroids according to the protocol used in Group I. Group IV: five animals were used as controls. Various changes were observed in the proximal metaphysis and diaphysis of the femur in eight of 10 animals in Group I (80%) and in five of 10 animals in Group II (50%) when compared with the animals in Group III and Group IV. The most common feature was evidence of new and old hemorrhage through the sinusoids, exudative reaction and thrombus formation in veins and small arteries. Focal necrotic areas of bone marrow showed an accumulation of cell debris, residue of hemorrhage, and disappearance of marrow elements. These findings suggest that (1) corticosteroids may potentiate the effects of a preexisting condition to increase the risk of osteonecrosis; (2) immunologic reaction may be an important factor in the pathogenesis of necrotic lesions; and (3) repeated intramedullary hemorrhage and thrombus formation may represent early major pathologic findings in bone necrosis.

This study was conducted to elucidate the role of the cytokine interleukin-1 beta on peripheral nerve recovery following crush injuries of two different magnitudes. Eighty-eight female rats were divided into four groups. A 5-mm segment of the right sciatic nerve was subjected to a 100-g crush load for 2 hours in the rats in Groups A1 and B1 or to a 15,000-g crush load for 10 minutes in the rats in Groups A2 and B2. The rats in Groups A1 and A2 received 10 microg/100 g body weight human recombinant interleukin-1 beta intraperitoneally 48, 24, and 1 hours before the nerve injury. The rats in Groups B1 and B2 were treated with an equal volume of normal saline solution with identical schedule guidelines. Walking-track tests (sciatic functional index) performed at intervals until 56 days after the crush and measurements of the contractile force of the extensor digitorum longus muscle made until 28 days were used to evaluate functional recovery of the nerve. During the second week after injury, the rats treated with interleukin-1 beta (A1) had an earlier recovery on the walking track than did those treated with saline solution (B1); this difference reached significance (p < 0.05) at day 11. Although Group A2 demonstrated a trend toward earlier recovery compared with Group B2, there was no significant difference between the two groups. After low or high-load crush injury, tetanic contractile forces were greater in the rats treated with human recombinant interleukin-1 beta than in those treated with saline solution. The results suggest that treatment with human recombinant interleukin-1 beta before crush injury can promote function in the peripheral nerve after the injury. However, the mechanisms that underlie the observed beneficial effects are not completely understood and only speculations can be made.

Intermittent pneumatic compression has been established as a method of clinically preventing deep vein thrombosis, but the mechanism has not been documented. This study observed the effects of intermittent pneumatic compression of legs on the microcirculation of distant skeletal muscle. The cremaster muscles of 80 male rats were exposed, a specially designed intermittent pneumatic-compression device was applied to both legs for 60 minutes, and the microcirculation of the muscles was assessed by measurement of the vessel diameter in three categories (10-20, 21-40, and 41-70 microm) for 120 minutes. The results showed significant vasodilation in arterial and venous vessels during the application of intermittent pneumatic compression, which disappeared after termination of the compression. The vasodilation reached a maximum 30 minutes after initiation of the compression and could be completely blocked by an inhibitor of nitric oxide synthase, NG-monomethyl-L-arginine (10 micromol/min). A 120-minute infusion of NG-monomethyl-L-arginine, beginning coincident with 60 minutes of intermittent pneumatic compression, resulted in a significant decrease in arterial diameter that remained at almost the same level after termination of the compression. The magnitude of the decrease in diameter in the group treated with intermittent pneumatic compression and NG-monomethyl-L-arginine was comparable with that in the group treated with NG-monomethyl-L-arginine alone. The results imply that the production of nitric oxide is involved in the positive influence of intermittent pneumatic compression on circulation. It is postulated that the rapid increase in venous velocity induced by intermittent pneumatic compression produces strong shear stress on the vascular endothelium, which stimulates an increased release of nitric oxide and thereby causes systemic vasodilation.

The authors investigated the effect of NG-monomethyl-L-arginine acetate (L-NMMA), a nitric oxide synthase (NOS) inhibitor, on the contractile function of skeletal muscle following ischemia/reperfusion (I/R) injury. The extensor digitorum longus (EDL) muscles of 50 rats were divided into seven groups. Contractile function in non-ischemic EDL did not change statistically significantly with L-NMMA infusion. I/R (1.5 hr I and 3 hr R) significantly decreased EDL contractile function, with an average maximal twitch force of 56 percent of the contralateral normal muscle force and isometric tetanic contractile forces between 47 and 84 percent at four different stimulation frequencies. Following L-NMMA administration at three different dosages, contractile function of I/R muscle decreased in a dose-dependent manner. The highest dosage of L-NMMA (10 micromol/min) reduced the average maximal twitch force to 15 percent and the isometric tetanic contractile forces to between 10 to 23 percent. Histologic evaluation revealed increased edema, neutrophil infiltration, and muscle-fiber necrosis in L-NMMA-infused EDL, compared to the controls. 1) Skeletal muscle contractile function was dose-dependently decreased with the administration of L-NMMA during I/R. 2) The concentrations of L-NMMA used in this study did not influence the function of non-ischemic EDL. These findings suggest that reduction of NO production during I/R is damaging to skeletal muscle function and would impair successful functional outcomes in microsurgical replantation.

This in vivo double-blind study evaluated the effect of recombinant human glial growth factor 2 (rhGGF2), a Schwann cell mitogen, on the recovery of motor function of rat sciatic nerve following crush injury. Seventy three rats were divided into three groups. Group I ( n = 5), sham operated; Groups II ( n = 34) and III ( n = 34) received a 100 g crush load for 2 h over a 5 mm segment of the sciatic nerve. Group III was treated with 1 mg\kg rhGGF2, via subcutaneous injection one day before nerve crush and daily for the following four days. Group II received an equivalent volume of saline as a control. Motor functional recovery was assessed by calculating the sciatic functional index (SFI) and the recovery rate of tetanic contractile force of the extensor digitorum longus (EDL) muscle. Recovery of nerve function was evident at day 11 after crush in the rhGGF2-treated animals, whereas the nerves in controls were still paralyzed. The rhGGF2-treated animals showed a significant improvement of the SFI between days 11–21 postoperatively when compared to controls. The isometric tetanic contractile force was stronger in the rhGGF2-treated group than in controls, with a significant difference at 40 to 70 Hz stimulus frequencies on day 4. Correlation analysis showed that tetanic contractile force had a linear correlation with the SFI. Histologic assessment indicated that the rhGGF2-treated animals showed less severe degeneration and earlier robust remyelination of axons than controls. The results suggest that treatment with rhGGF2 is effective in promoting nerve regeneration as seen in measurements of functional recovery and qualitative assessment of nerve morphology. The mechanism of GGFs protective effect may be related to its direct action on Schwann cells, stimulating their mitosis as well as inducing neurotrophic factors essential to neuronal maintenance and repair.

The ultimate goal of replantation and microsurgical reconstructive operations is to regain or improve impaired function of the tissue. However, the data related to the influence of NO on tissue function are limited. This study evaluated the effects of the NO donor S-nitroso- N-acetylcysteine (SNAC) on contractile function of skeletal muscle during reperfusion. Forty-nine rats were divided into six groups. The extensor digitorum longus (EDL) muscles in groups I and II were not subjected to ischemia-reperfusion but were treated with a low (100 nmol/min) or high (1 μmol/min) dose of SNAC. In groups III- V, the EDL underwent 3 h of ischemia and 3 h of reperfusion and was also treated with low (100 nmol/min) or high doses (1 or 5 μmol/min) of SNAC. Group VI was a phosphate-buffered saline (PBS)-treated control group. Twenty additional animals were used to document systemic effects of SNAC and PBS only. SNAC or PBS was infused for 6.5 h, beginning 30 min before ischemia and continuing throughout the duration of reperfusion. Contractile testing compared the maximal twitch force, isometric tetanic contractile forces, fatigue, and fatigue half time of the experimental EDL and the contralateral nontreated EDL. The findings indicate that 1) SNAC does not influence contractile function of EDL muscle not subjected to ischemia-reperfusion, 2) SNAC significantly protects the contractile function of ischemic skeletal muscle against reperfusion injury in the early reperfusion period, and 3) the protective role of SNAC is critically dosage dependent; protection is lost at higher doses. The conclusion from this study is that supplementation with exogenous NO exerts a protective effect on the tissue against reperfusion injury.

This study compared the effects of bone preparation on tibial stem stability in total knee arthroplasty. Six pairs of fresh-frozen tibias underwent implantation of an 18 x 75-mm press-fit stem (minus the tibial tray). The standard press-fit technique was performed creating a cavity 17 mm in diameter. For the bone compaction technique, stainless-steel dilators were machined ranging in diameter from 6 to 16 mm in 2-mm increments. The technique consisted of using the dilators and a mallet to create a compacted tunnel in a sequential manner to 16 mm in diameter followed by implantation of the stem. The specimens were mounted on a materials testing machine and loads were applied in both planes: anteroposterior and mediolateral. A preconditioning load of 100 N was applied followed by a second 100-N load from which the stiffness (N/mm) of implantation was calculated. The stability of the tibial stems implanted by compaction averaged 84.7% greater when compared with the press-fit stems (P.017 by multivariate analysis).