Change in salt intake affects blood pressure of chimpanzees: implications for human populations.

BACKGROUND: Addition of up to 15.0 g/d salt to the diet of chimpanzees caused large rises in blood pressure, which reversed when the added salt was removed. Effects of more modest alterations to sodium intakes in chimpanzees, akin to current efforts to lower sodium intakes in the human population, are unknown. METHODS AND RESULTS: Sodium intakes were altered among 17 chimpanzees in Franceville, Gabon, and 110 chimpanzees in Bastrop, Tex. In Gabon, chimpanzees had a biscuit diet of constant nutrient composition except that the sodium content was changed episodically over 3 years from 75 to 35 to 120 mmol/d. In Bastrop, animals were divided into 2 groups; 1 group continued on the standard diet of 250 mmol/d sodium for 2 years, and sodium intake was halved for the other group. Lower sodium intake was associated with lower systolic, diastolic, and mean arterial blood pressures in Gabon (2-tailed P<0.001, unadjusted and adjusted for age, sex, and baseline weight) and Bastrop (P<0.01, unadjusted; P=0.08 to 0.10, adjusted), with no threshold down to 35 mmol/d sodium. For systolic pressure, estimates were -12.7 mm Hg (95% confidence interval, -16.9 to -8.5, adjusted) per 100 mmol/d lower sodium in Gabon and -10.9 mm Hg (95% confidence interval, -18.9 to -2.9, unadjusted) and -5.7 mm Hg (95% confidence interval, -12.2 to 0.7, adjusted) for sodium intake lower by 122 mmol/d in Bastrop. Baseline systolic pressures higher by 10 mm Hg were associated with larger falls in systolic pressure by 4.3/2.9 mm Hg in Gabon/Bastrop per 100 mmol/d lower sodium. CONCLUSIONS: These findings from an essentially single-variable experiment in the species closest to Homo sapiens with high intakes of calcium and potassium support intensified public health efforts to lower sodium intake in the human population.

Effects of mechanical stimulation on the biomechanics and histology of stem cell-collagen sponge constructs for rabbit patellar tendon repair.

The objective of this study was to determine how mechanical stimulation affects the biomechanics and histology of stem cell-collagen sponge constructs used to repair central rabbit patellar tendon defects. Autogenous tissue-engineered constructs were created for both in vitro and in vivo analyses by seeding mesenchymal stem cells from 10 adult rabbits at 0.14x10(6) cells/construct in type I collagen sponges. Half of these constructs were mechanically stimulated once every 5 min for 8 h/day to a peak strain of 4% for 2 weeks. The other half remained in an incubator without mechanical stimulation for 2 weeks. Samples allocated for in vitro testing revealed that mechanically stimulated constructs had 2.5 times the linear stiffness of nonstimulated constructs. The remaining paired constructs for in vivo studies were implanted in bilateral full-thickness, full-length defects in the central third of rabbit patellar tendons. Twelve weeks after surgery, repair tissues were assigned for biomechanical (7 pairs) and histologic (3 pairs) analyses. Maximum force, linear stiffness, maximum stress, and linear modulus for the stimulated (vs. nonstimulated) repairs averaged 70% (vs. 55%), 85% (vs. 55%), 70% (vs. 50%), and 50% (vs. 40%) of corresponding values for the normal central third of the patellar tendons. The average force-elongation curve for the mechanically stimulated repairs also matched the corresponding curve for the normal patellar tendons, up to 150% of the peak in vivo force values recorded in a previous study. Construct and repair linear stiffness and linear modulus were also positively correlated (r = 0.6 and 0.7, respectively). Histologically both repairs showed excellent cellular alignment and mild staining for decorin and collagen type V, and moderate staining for fibronectin and collagen type III. This study shows that mechanical stimulation of stem cell-collagen sponge constructs can significantly improve tendon repair biomechanics up to and well beyond the functional limits of in vivo loading.

Effects of cell-to-collagen ratio in stem cell-seeded constructs for Achilles tendon repair.

The objective of the present study was to test the hypotheses that implantation of cell-seeded constructs in a rabbit Achilles tendon defect model would 1) improve repair biomechanics and matrix organization and 2) result in higher failure forces than measured in vivo forces in normal rabbit Achilles tendon (AT) during an inclined hopping activity. Autogenous tissue-engineered constructs were fabricated in culture between posts in the wells of silicone dishes at four cell-to-collagen ratios by seeding mesenchymal stem cells (MSC) from 18 adult rabbits at each of two seeding densities (0.1 x 10(6) and 1 x 10(6) cell/mL) in each of two collagen concentrations (1.3 and 2.6 mg/mL). After 5 days of contraction, constructs having the two highest ratios (0.4 and 0.8 M/mg) were damaged by excessive cell traction forces and could not be used in subsequent in vivo studies. Constructs at the lower ratios (0.04 and 0.08 M/mg) were implanted in bilateral, 2 cm long gap defects in the rabbit's lateral Achilles tendon. At 12 weeks after surgery, both repair tissues were isolated and either failed in tension (n = 13) to determine their biomechanical properties or submitted for histological analysis (n = 5). No significant differences were observed in any structural or mechanical properties or in histological appearance between the two repair conditions. However, the average maximum force and maximum stress of these repairs achieved 50 and 85% of corresponding values for the normal AT and exceeded the largest peak in vivo forces (19% of failure) previously recorded in the rabbit AT. Average stiffness and modulus were 60 and 85% of normal values, respectively. New constructs with lower cell densities and higher scaffold stiffness that do not excessively contract and tear in culture and that further improve the repair stiffness needed to withstand various levels of expected in vivo loading are currently being investigated.

The effect of autologous mesenchymal stem cells on the biomechanics and histology of gel-collagen sponge constructs used for rabbit patellar tendon repair.

The objective of this study was to introduce mesenchymal stem cells (MSCs) into a gel-sponge composite and examine the effect the cells have on repair biomechanics and histology 12 weeks postsurgery. We tested two related hypotheses-adding MSCs would significantly improve repair biomechanics and cellular organization, and would result in higher failure forces than peak in vivo patellar tendon (PT) forces recorded for an inclined hopping activity. Autogenous tissue-engineered constructs were created by seeding MSCs from 15 adult rabbits at 0.1 x 10(6) cells/mL in 2.6 mg/mL of collagen gel in collagen sponges. Acellular constructs were created using the same concentration of collagen gel in matching collagen sponges. These cellular and acellular constructs were implanted in bilateral full-thickness, full-length defects in the central third of patellar tendons. At 12 weeks after surgery, repair tissues were assigned for biomechanical (n = 12 pairs) and histological (n = 3 pairs) analyses. Maximum force and maximum stress for the cellular repairs were about 60 and 50% of corresponding values for the normal central third of the PT, respectively. Likewise, linear stiffness and linear modulus for these cellular repairs averaged 75 and 30% of normal PT values, respectively. By contrast, the acellular repairs exhibited lower percentages of normal PT values for maximum force (40%), maximum stress (25%), linear stiffness (30%), and linear modulus (20%). Histologically, both repairs showed strong staining for collagen types III and V, fibronectin, and decorin. The cellular repairs also showed cellular alignment comparable to that of normal tendon. This study shows that introducing autogenous mesenchymal stem cells into a gel-collagen sponge composite significantly improves tendon repair compared to the use of a gel-sponge composite alone in the range of in vivo loading.

Effects of cell-to-collagen ratio in mesenchymal stem cell-seeded implants on tendon repair biomechanics and histology.

Autogenous tissue-engineered constructs were fabricated at four cell-to-collagen ratios (0.08, 0.04, 0.8, and 0.4 M/mg) by seeding mesenchymal stem cells (MSCs) from 16 adult rabbits at one of two seeding densities (0.1 x 10(6) and 1 x 10(6) cells/mL) in one of two collagen concentrations (1.3 and 2.6 mg/mL). The highest two ratios (0.4 and 0.8 M/mg) were damaged by excessive cell contraction and could not be used in subsequent in vivo studies. The remaining two sets of constructs were implanted into bilateral full-thickness, full-length defects created in the central third of the patellar tendon (PT). At 12 weeks after surgery, repair tissues were assigned for biomechanical (n = 13) and histological (n = 3) analyses. A second group of rabbits (n = 6) received bilateral acellular implants with the same two collagen concentrations. At 12 weeks, repair tissues were also assigned for biomechanical (n = 4) and histological (n = 2) analyses. No significant differences were observed in any structural or material properties or in histological appearance among the two cell-seeded and two acellular repair groups. Average maximum force and maximum stress of the repairs were approximately 30% of corresponding values for the central one-third of normal PT and higher than peak in vivo forces measured in rabbit PT from one of our previous publications. However, average repair stiffness and modulus were only 30 and 20% of normal PT values, respectively. Current repairs achieved higher maximum forces than in previous studies and without ectopic bone, but will need to achieve sufficient stiffness as well to be effective in the in vivo range of loading.

Characterization of in vivo Achilles tendon forces in rabbits during treadmill locomotion at varying speeds and inclinations.

The objective of this study was to test the hypothesis that increasing the speed and inclination of the treadmill increases the peak Achilles tendon forces and their rates of rise and fall in force. Implantable force transducers (IFT) were inserted in the confluence of the medial and lateral heads of the left gastrocnemius tendon in 11 rabbits. IFT voltages were successfully recorded in 8 animals as the animals hopped on a treadmill at each of two speeds (0.1 and 0.3 mph) and inclinations (0 degrees and 12 degrees). Instrumented tendons were isolated shortly after sacrifice and calibrated. Contralateral unoperated tendons were failed in uniaxial tension to determine maximum or failure force, from which safety factor (ratio of maximum force to peak in vivo force) was calculated for each activity. Peak force and the rates of rise and fall in force significantly increased with increasing treadmill inclination (p<0.001). Safety factors averaged 30.8+/-7.5 for quiet standing, 7.0+/-2.9 for level hopping, and 5.2+/-0.7 for inclined hopping (mean+/-SEM). These in vivo force parameters will help tissue engineers better design functional tissue engineered constructs for rabbit Achilles tendon and other tendon repairs. Force patterns can also serve as input data for mechanical stimulation of tissue-engineered constructs in culture. Such approaches are expected to help accelerate tendon repair after injury.

Effects of initial graft tension on knee stability after anterior cruciate ligament reconstruction using hamstring tendons: a cadaver study.

PURPOSE: Tension degradation within hamstring grafts and anterior knee laxity were analyzed in a cadaveric anterior cruciate ligament (ACL) reconstruction model undergoing cyclic motion. It was hypothesized that suture fixation of a hamstring graft would lose tension during cycling initially and then stabilize, and that anterior knee laxity would increase as tension was lost. Hamstring grafts fixed under 3 different loads were evaluated to determine how initial graft tension affected knee laxity after cyclic motion. TYPE OF STUDY: Cadaveric biomechanical analysis. METHODS: Eighteen pairs of fresh-frozen hamstring tendons were tested on 2 cadaveric knees undergoing ACL reconstruction. The hamstring pairs were separated equally and randomly into one of 3 tension groups: 68 N (15 lb), 45 N (10 lb), and 23 N (5 lb). The loads were applied to the graft at 30 degrees of flexion, and the grafts were secured to the tibia with a suture and post technique. The knee was then cycled 1,000 times using an Instron machine (Instron, Canton, MA) through a range of motion between 0 degrees to 90 degrees. Constant monitoring and recording of graft tension was performed. A KT-1000 (Medmetrics, San Diego, CA) was performed (1) on the intact knee, (2) after ACL excision, (3) after ACL reconstruction and initial graft fixation, and (4) at the completion of the 1,000 cycles. An analysis of variance test was used to evaluate data. RESULTS: The tension within the grafts after 1,000 cycles decreased to 34.5 N (7.6 lb), 16.8 N (3.7 lb), and 15.4 N (3.4 lb) from the preloads of 68, 45, and 23 N, respectively (P <.05 in all cases). This represented an average decrease of 50.2% of the initial tension after 1,000 cycles. Manual-maximum KT testing of the intact knees was 5.8 +/- 0.3 mm, and after ACL excision was 13.2 +/- 0.9 mm. KT testing revealed 6.0 +/- 0.9 mm, 8.1 +/- 1.9 mm, and 8.9 +/- 1.1 mm of anterior translation after fixation in the tension groups of 68, 45, and 23 N, respectively. After 1,000 cycles, the translation increased to 7.8 +/- 1.0 mm, 10.5 +/- 1.9 mm, and 10.3 +/- 1.5 mm, respectively. CONCLUSIONS: This study showed that initial graft tension decreases with cyclic loading, resulting in increased knee laxity. To restore anterior translation to within 3 mm of the native ACL condition after cyclic loading, approximately 68 N of initial tension is required using this fixation technique.

The effect of suture anchor design and orientation on suture abrasion: An in vitro study.

PURPOSE: To evaluate the effects of suture anchor design and orientation on suture abrasion in a cyclic model. TYPE OF STUDY: In vitro. METHODS: Biomechanical studies have shown suture breakage to be a predominant mode of failure in a suture anchor repair construct. It is possible that suture abrasion during knot tying or in vivo cyclic loading may contribute to early failure. This study specifically investigates suture abrasion caused by 17 commonly used suture anchors and demonstrates the effects of suture anchor angulation and rotation on suture abrasion. To eliminate target tissue as a source of failure, all anchors were implanted into a solid block of sawbones material and tested with No. 2 Ethibond Excel sutures (Ethicon, Somerville, NJ). The testing model focused on 3 variables: suture anchor type, suture pull angle (SA) and angle of anchor rotation (RA). Abrasion testing was then performed on a servohydraulic materials testing system by continually cycling the suture back and forth through each anchor with an excursion of 4 cm at a rate of 0.5 Hz under a load of 10 N until suture failure occurred. RESULTS: Sutures performed significantly better when cycled in line with the anchor at 0 degrees SA with 0 degrees RA than they did at 45 degrees SA with 0 degrees RA or 45 degrees SA with 90 degrees RA. We found no significant difference between anchors tested at 45 degrees SA with 0 degrees RA and 45 degrees SA with 90 degrees RA. For tests performed using metallic suture anchors, all constructs failed by fraying of the suture. Constructs using biopolymer anchors and nonabsorbable polymeric anchors experienced a mixture of suture and anchor eyelet failures. CONCLUSIONS: In addition to the statistically significant detrimental effects of suture anchor angulation and rotation on suture abrasion, suture anchor eyelet design may also influence suture abrasion. Surgeons should be aware of the effects of anchor angulation, suture position in the eyelet, and design and composition of the eyelet to maximize the durability of the construct.

In vivo forces used to develop design parameters for tissue engineered implants for rabbit patellar tendon repair.

Previous studies in tissue engineering have shown that suspending undifferentiated mesenchymal stem cells in collagen gels and wrapping them about a suture causes alignment of cells and contraction of constructs in culture in a form that is suitable for implantation for tendon repair. Little is known about the patterns of these in vivo signals that might improve tendon repair biomechanics. Three hypotheses were tested in this study using the rabbit patellar tendon (PT) model: (1) peak in vivo forces and the rates of rise and fall in these forces will increase significantly with increasing levels of activity; (2) the PTs safety factor for all activities will be in the range of values found for tendons (2.5-3); (3) rabbits will not "favor" the operated limb at the time of evaluation but maintain similar vertical ground reaction forces in both limbs during quiet standing (QS). In vivo rabbit PT forces were measured during QS and while the animal hopped on a treadmill whose speed (0.04 and 0.13 m/s) and inclination (0 degrees and 12 degrees) were controlled. Implantable force transducers were surgically placed in one PT and data collected three days post surgery in each of eight New Zealand White rabbits. Peak tensile forces increased significantly with inclination of the treadmill and the rates of rise and fall in tendon force increased significantly with both speed and inclination (p<0.001). Such design criteria should be useful in mechanically stimulating cell-gel constructs for tendon repair.

A survey of the tension applied to a doubled hamstring tendon graft for reconstruction of the anterior cruciate ligament.

PURPOSE: Currently there is no consensus regarding the amount of tension to apply to a graft when reconstructing the anterior cruciate ligament (ACL). We undertook a study to determine whether sports trained orthopedic surgeons tension hamstring tendon grafts maximally during ACL reconstruction, and also whether surgeons tend to load their grafts within a narrow range of tensions. TYPE OF STUDY: Cross-sectional study. METHODS: One fresh-frozen cadaveric knee with appropriately placed femoral and tibial tunnels and five pairs of preconditioned semitendinosus and gracilis tendons were used. Custom-made computer software and a custom-made, load measurement device was employed. Thirteen orthopedic sports medicine physicians from our community took part in the study. Surgeons were asked to tension the graft as they would in surgery and were then asked to tension the graft maximally. RESULTS: The mean and standard deviation of the normal tension (14.8 +/- 7.2 lb) was significantly less (P =.005) than the mean maximal tension (22.3 +/- 6.9 lb). CONCLUSIONS: This study shows that most ACL surgeons do not tension their graft maximally. Moreover, graft tensioning is highly variable among sports medicine orthopedists. These findings revisit the question as to whether tension should be more accurately measured and controlled for intraoperatively.