A simple but reliable method for measuring 3D Achilles tendon moment arm geometry from a single,static magnetic resonance scan

Current methods for measuring in vivo 3D muscle-tendon moment arms generally rely on the acquisition of magnetic resonance imaging (MRI) scans at multiple joint angles. However, for patients with musculoskeletal pathologies such as fixed contractures, moving a joint through its full range of motion is not always feasible. The purpose of this research was to develop a simple, but reliable in vivo 3D Achilles tendon moment arm (ATMA) technique from a single static MRI scan. To accomplish this, for nine healthy adults (5 males, 4 females), the geometry of a cylinder was fit to the 3D form of the talus dome, which was used to estimate the talocrural flexion/extension axis, and a fifth-order polynomial fit to the line of action of the Achilles tendon. The single static scan in vivo 3D ATMA estimates were compared to estimates obtained from the same subjects at the same ankle joint angles using a previously validated 3D dynamic MRI based in vivo ATMA measurement technique. The ATMA estimates from the single scan in vivo 3D method (52.5 mm ± 5.6) were in excellent agreement (ICC = 0.912) to the validated in vivo 3D method (51.5 mm ± 5.1). These data show reliable in vivo 3D ATMA can be obtained from a single MRI scan for healthy adult populations. The single scan, in vivo 3D ATMA technique provides researchers with a simple, but reliable method for obtaining subject-specific ATMAs for musculoskeletal modelling purposes.     

Plasticity of human Achilles tendon mechanical and morphological properties in response to cyclic strain

The purpose of the current study in combination with our previous published data (Arampatzis et al., 2007) was to examine the effects of a controlled modulation of strain magnitude and strain frequency applied to the Achilles tendon on the plasticity of tendon mechanical and morphological properties. Eleven male adults (23.9±2.2 yr) participated in the study. The participants exercised one leg at low magnitude tendon strain (2.97±0.47%), and the other leg at high tendon strain magnitude (4.72±1.08%) of similar frequency (0.5 Hz, 1 s loading, 1 s relaxation) and exercise volume (integral of the plantar flexion moment over time) for 14 weeks, 4 days per week, 5 sets per session. The exercise volume was similar to the intervention of our earlier study (0.17 Hz frequency; 3 s loading, 3 s relaxation) allowing a direct comparison of the results. Before and after the intervention ankle joint moment has been measured by a dynamometer, tendon–aponeurosis elongation by ultrasound and cross-sectional area of the Achilles tendon by magnet resonance images (MRI). We found a decrease in strain at a given tendon force, an increase in tendon–aponeurosis stiffness and tendon elastic modulus of the Achilles tendon only in the leg exercised at high strain magnitude. The cross-sectional area (CSA) of the Achilles tendon did not show any statistically significant (P>0.05) differences to the pre-exercise values in both legs. The results indicate a superior improvement in tendon properties (stiffness, elastic modulus and CSA) at the low frequency (0.17 Hz) compared to the high strain frequency (0.5 Hz) protocol. These findings provide evidence that the strain magnitude applied to the Achilles tendon should exceed the value, which occurs during habitual activities to trigger adaptational effects and that higher tendon strain duration per contraction leads to superior tendon adaptational responses.     

Structural Achilles tendon properties in athletes subjected to different exercise modes and in Achilles tendon rupture patients.

The prevalence of Achilles tendon (AT) injury is high in various sports, and AT rupture patients have been reported to have a 200-fold risk of sustaining a contralateral rupture. Tendon adaptation to different exercise modes is not fully understood. The present study investigated the structural properties of the AT in male elite athletes that subject their AT to different exercise modes as well as in Achilles rupture patients. Magnetic resonance imaging of the foot and leg, anthropometric measurements, and maximal isometric plantar flexion force were obtained in 6 male AT rupture patients and 25 male elite athletes (kayak/control group n = 9, volleyball n = 8 and endurance running n = 8). AT cross-sectional area (CSA) was normalized to body mass. Runners had a larger normalized AT CSA along the entire length of the tendon compared with the control group (P < 0.05). The volleyball subjects had a larger normalized CSA compared with the control group (P < 0.05) in the area of thinnest tendon CSA. No structural differences of the AT were found in the rupture subjects compared with the control group. Rupture subjects did not subject their AT to greater force or stress during a maximal voluntary isometric plantar flexion than the other groups. The CSA of the triceps surae musculature was the strongest predictor of AT CSA (r(s) = 0.569, P < 0.001). This study is the first to show larger CSA in tendons that are subjected to intermittent high loads. AT rupture patients did not display differences in structural or loading properties of the tendons.     

Uphill running improves rat Achilles tendon tissue mechanical properties and alters gene expression without inducing pathological changes

Overuse Achilles tendinopathy is a common and challenging problem in sports medicine. Little is known about the etiology of this disorder, and the development of a good animal model for overuse tendinopathy is essential for advancing insight into the disease mechanisms. Our aim was to test a previously proposed rat model for Achilles tendon overuse. Ten adult male Sprague-Dawley rats ran on a treadmill with 10° incline, 1 h/day, 5 days/wk (17-20 m/min) for 12 wk and were compared with 12 control rats. Histological, mechanical, and gene-expression changes were measured on the Achilles tendons after the intervention, and local tendon glucose-uptake was measured before and after the intervention with positron emission tomography. No differences were detected between runners and controls in tissue histology or in glucose uptake, indicating that tendon pathology was not induced. Greater tendon tissue modulus (P < 0.005) and failure stress/body weight (P < 0.02) in runners compared with controls further supported that tendons successfully adapted to uphill running. Several genes of interest were regulated after 12 wk of running. Expression of collagen III and insulin-like growth factor I was increased, while collagen I was unchanged, and decreases were seen in noncollagen matrix components (fibromodulin and biglycan), matrix degrading enzymes, transforming growth factor-β1, and connective tissue growth factor. In conclusion, the tested model could not be validated as a model for Achilles tendinopathy, as the rats were able to adapt to 12 wk of uphill running without any signs of tendinopathy. Improved mechanical properties were observed, as well as changes in gene-expression that were distinctly different from what is seen in tendinopathy and in response to short-term tendon loading.     

Quadriceps tendon allografts as an alternative to Achilles tendon allografts: a biomechanical comparison

Quadriceps tendon with a patellar bone block may be a viable alternative to Achilles tendon for anterior cruciate ligament reconstruction (ACL-R) if it is, at a minimum, a biomechanically equivalent graft. The objective of this study was to directly compare the biomechanical properties of quadriceps tendon and Achilles tendon allografts. Quadriceps and Achilles tendon pairs from nine research-consented donors were tested. All specimens were processed to reduce bioburden and terminally sterilized by gamma irradiation. Specimens were subjected to a three phase uniaxial tension test performed in a custom environmental chamber to maintain the specimens at a physiologic temperature (37 ± 2 °C) and misted with a 0.9 % NaCl solution. There were no statistical differences in seven of eight structural and mechanical between the two tendon types. Quadriceps tendons exhibited a significantly higher displacement at maximum load and significantly lower stiffness than Achilles tendons. The results of this study indicated a biomechanical equivalence of aseptically processed, terminally sterilized quadriceps tendon grafts with bone block to Achilles tendon grafts with bone block. The significantly higher displacement at maximum load, and lower stiffness observed for quadriceps tendons may be related to the failure mode. Achilles tendons had a higher bone avulsion rate than quadriceps tendons (86 % compared to 12 %, respectively). This was likely due to observed differences in bone block density between the two tendon types. This research supports the use of quadriceps tendon allografts in lieu of Achilles tendon allografts for ACL-R.     

An improved radiological method for the evaluation of Achilles tendon xanthomatosis

Xanthomatous infiltration of tendons is a clinical feature common to many cases of hyperlipidemia. The xanthomas can be detected and only grossly assessed by palpation. This report describes a radiological technique used to assess these lesions at the Achilles tendon level. The “mammography technique” applied to the study of Achilles tendons was used in 32 normolipemic subjects and 32 hyperlipidemic patients. Both tendons could be observed in their entire length and their thickness, greatly increased in xanthomatosis, could be accurately measured. The results of this survey suggest that the radiological approach may provide a useful tool for the routine evaluation and follow-up of tendon xanthomas.     

Achilles tendon rupture: avoiding tendon lengthening during surgical repair and rehabilitation

Achilles tendon rupture is a serious injury for which the best treatment is still controversial. Its primary goal should be to restore normal length and tension, thus obtaining an optimal function. Tendon elongation correlates significantly with clinical outcome; lengthening is an important cause of morbidity and may produce permanent functional impairment. In this article, we review all factors that may influence the repair, including the type of surgical technique, suture material, and rehabilitation program, among many others.     

The effect of running, strength, and vibration strength training on the mechanical, morphological, and biochemical properties of the Achilles tendon in rats.

Compared with muscle or bone, there is a lack of information about the relationship between tendon adaptation and the applied loading characteristic. The purpose of the present study was to analyze the effect of different exercise modes characterized by very distinct loading patterns on the mechanical, morphological, and biochemical properties of the Achilles tendon. Sixty-four female Sprague-Dawley rats were divided into five groups: nonactive age-matched control (AMC; n = 20), voluntary wheel running (RT; n = 20), vibration strength-trained (LVST; n = 12), high-vibration strength-trained (HVST; n = 6), and high strength-trained (HST; n = 6) group. After a 12-wk-long experimental period, the Achilles tendon was tested mechanically and the cross-sectional area, the soleus and gastrocnemius muscle mass, and mRNA concentration of collagen I, collagen III, tissue inhibitor of metalloproteinase-1 (TIMP-1), transforming growth factor-beta, connective tissue growth factor, and matrix metalloproteinase-2 was determined. Neither in the LVST nor in the HVST group could any adaptation of the Achilles tendon be detected, although the training had an effect on the gastrocnemius muscle mass in the LVST group (P < 0.05). In the HST group, the highest creep was found, but the effect was more pronounced compared with the LVST group (P < 0.05) than with the AMC group. That indicates that this was rather induced by the low muscle mass rather than by training. However, the RT group had a higher TIMP-1 mRNA concentration in the Achilles tendon in contrast to AMC group (P < 0.05), which suggests that this exercise mode may have an influence on tendon adaptation.     

Elastic properties of human Achilles tendon are correlated to muscle strength.

The purpose of this study was to investigate whether the mechanical properties of the Achilles tendon were correlated to muscle strength in the triceps surae in humans. Twenty-four men and twelve women exerted maximal voluntary isometric plantar flexion (MVIP) torque. The elongation (DeltaX) and strain of the Achilles tendon (epsilon), the proximal part of which is the composite of the gastrocnemius tendon and the soleus aponeurosis, at MVIP were determined from the displacement of the distal myotendinous junction of the medial gastrocnemius using ultrasonography. The Achilles tendon force at MVIP (F) was calculated from the MVIP torque and the Achilles tendon moment arm. There were no significant differences in either the F-DeltaX or F-epsilon relationships between men and women. DeltaX and epsilon were 9.8 +/- 2.6 mm and 5.3 +/- 1.6%, respectively, and were positively correlated to F (r = 0.39, P < 0.05; r = 0.39, P < 0.05), which meant that subjects with greater muscle strength could store more elastic energy in the tendon. The regression y-intercepts for the F-DeltaX (P < 0.01) and F-epsilon (P < 0.05) relationship were significantly positive. These results might indicate that the Achilles tendon was stiffer in subjects with greater muscle strength, which may play a role in reducing the probability of tendon strain injuries. It was suggested that the Achilles tendon of subjects with greater muscle strength did not impair the potential for storing elastic energy in tendons and may be able to deliver the greater force supplied from a stronger muscle more efficiently. Furthermore, the difference in the Achilles tendon mechanical properties between men and women seemed to be correlated to the difference in muscle strength rather than gender.     

Achilles tendon loading during walking: application of a novel optic fiber technique

An optic fiber (? 0.5 mm) was utilized for the study of Achilles tendon forces (ATF) in eight volunteers who walked over a 10 m force platform at three speeds (1.1 ± 0.1 m × s⁻¹, 1.5 ± 0.1 m × s⁻¹ and 1.8 ± 0.2 m × s⁻¹). The presented ATF-time curves showed great intersubject variation in magnitudes of the sudden release of force after initial contact and in the peak ATF's (1430 ± 500 N). This intersubject variation in the peak force decreased only by 4% when cross-sectional area of the tendon was considered. Measured ground reaction forces and plantar pressures confirmed that the subjects walked quite normally during recordings. The peak ATF was found to be rather insensitive to speed in contrast to the rate of ATF development which increased 32% ( p < 0.5) from slow to fast walking speed. It is concluded that the optic fiber technique can be applied to study loading of the musculo-tendinous complex during normal locomotion such as walking. Accepted: 13 October 1997     

Region-specific mechanical properties of the human patella tendon.

The present study investigated the mechanical properties of tendon fascicles from the anterior and posterior human patellar tendon. Collagen fascicles from the anterior and posterior human patellar tendon in healthy young men (mean +/- SD, 29.0 +/- 4.6 yr, n = 6) were tested in a mechanical rig. A stereoscopic microscope equipped with a digital camera recorded elongation. The fascicles were preconditioned five cycles before the failure test based on pilot data on rat tendon fascicle. Human fascicle length increased with repeated cycles (P < 0.05); cycle 5 differed from cycle 1 (P < 0.05), but not cycles 2-4. Peak stress and yield stress were greater for anterior (76.0 +/- 9.5 and 56.6 +/- 10.4 MPa, respectively) than posterior fascicles (38.5 +/- 3.9 and 31.6 +/- 2.9 MPa, respectively), P < 0.05, while yield strain was similar (anterior 6.8 +/- 1.0%, posterior 8.7 +/- 1.4%). Tangent modulus was greater for the anterior (1,231 +/- 188 MPa) than the posterior (583 +/- 122 MPa) fascicles, P < 0.05. In conclusion, tendon fascicles from the anterior portion of the human patellar tendon in young men displayed considerably greater peak and yield stress and tangent modulus compared with the posterior portion of the tendon, indicating region-specific material properties.     

Collagen self-assembly and the development of tendon mechanical properties

The development of the musculoskeleton and the ability to locomote requires controlled cell division as well as spatial control over deposition of extracellular matrix. Self-assembly of procollagen and its final processing into collagen fibrils occurs extracellularly. The formation of crosslinked collagen fibers results in the conversion of weak liquid-like embryonic tissues to tough elastic solids that can store energy and do work. Collagen fibers in the form of fascicles are the major structural units found in tendon. The purpose of this paper is to review the literature on collagen self-assembly and tendon development and to relate this information to the development of elastic energy storage in non-mineralizing and mineralizing tendons. Of particular interest is the mechanism by which energy is stored in tendons during locomotion. In vivo, collagen self-assembly occurs by the deposition of thin fibrils in recesses within the cell membrane. These thin fibrils later grow in length and width by lateral fusion of intermediates. In vitro, collagen self-assembly occurs by both linear and lateral growth steps with parallel events seen in vivo; however, in the absence of cellular control and enzymatic cleavage of the propeptides, the growth mechanism is altered, and the fibrils are irregular in cross section. Results of mechanical studies suggest that prior to locomotion the mechanical response of tendon to loading is dominated by the viscous sliding of collagen fibrils. In contrast, after birth when locomotion begins, the mechanical response is dominated by elastic stretching of crosslinked collagen molecules.     

Digital photography provides a fast,reliable, and noninvasive method to estimate anthocyanin pigment concentration in reproductive and vegetative plant tissues

Anthocyanin pigments have become a model trait for evolutionary ecology as they often provide adaptive benefits for plants. Anthocyanins have been traditionally quantified biochemically or more recently using spectral reflectance. However, both methods require destructive sampling and can be labor intensive and challenging with small samples. Recent advances in digital photography and image processing make it the method of choice for measuring color in the wild. Here, we use digital images as a quick, noninvasive method to estimate relative anthocyanin concentrations in species exhibiting color variation. Using a consumer‐level digital camera and a free image processing toolbox, we extracted RGB values from digital images to generate color indices. We tested petals, stems, pedicels, and calyces of six species, which contain different types of anthocyanin pigments and exhibit different pigmentation patterns. Color indices were assessed by their correlation to biochemically determined anthocyanin concentrations. For comparison, we also calculated color indices from spectral reflectance and tested the correlation with anthocyanin concentration. Indices perform differently depending on the nature of the color variation. For both digital images and spectral reflectance, the most accurate estimates of anthocyanin concentration emerge from anthocyanin content‐chroma ratio, anthocyanin content‐chroma basic, and strength of green indices. Color indices derived from both digital images and spectral reflectance strongly correlate with biochemically determined anthocyanin concentration; however, the estimates from digital images performed better than spectral reflectance in terms of r² and normalized root‐mean‐square error. This was particularly noticeable in a species with striped petals, but in the case of striped calyces, both methods showed a comparable relationship with anthocyanin concentration. Using digital images brings new opportunities to accurately quantify the anthocyanin concentrations in both floral and vegetative tissues. This method is efficient, completely noninvasive, applicable to both uniform and patterned color, and works with samples of any size.     

Effects of freeze-thaw on the biomechanical and structural properties of the rat Achilles tendon

Rodent models are commonly used to investigate tendon healing, with the biomechanical and structural properties of the healed tendons being important outcome measures. Tendon storage for later testing becomes necessary when performing large experiments with multiple time-points. However, it is unclear whether freezing rodent tendons affects their material properties. Thus the aim of this study was to determine whether freezing rat Achilles tendons affects their biomechanical or structural properties. Tendons were frozen at either −20 °C or −80 °C directly after harvesting, or tested when freshly harvested. Groups of tendons were subjected to several freeze-thaw cycles (1, 2, and 5) within 3 months, or frozen for 9 months, after which the tendons were subjected to biomechanical testing. Additionally, fresh and thawed tendons were compared morphologically, histologically and by transmission electron microscopy. No major differences in biomechanical properties were found between fresh tendons and those frozen once or twice at −20 °C or −80 °C. However, deterioration of tendon properties was found for 5-cycle groups and both long-term freezing groups; after 9 months of freezing at −80 °C the tear resistance of the tendon was reduced from 125.4 ± 16.4N to 74.3 ± 18.4N (p = 0.0132). Moreover, tendons stored under these conditions showed major disruption of collagen fibrils when examined by transmission electron microscopy. When examined histologically, fresh samples exhibited the best cellularity and proteoglycan content of the enthesis. These properties were preserved better after freezing at −80 °C than after freezing at −20 °C, which resulted in markedly smaller chondrocytes and less proteoglycan content. Overall, the best preservation of histological integrity was seen with tendons frozen once at −80 °C. In conclusion, rat Achilles tendons can be frozen once or twice for short periods of time (up to 3 months) at −20 °C or −80 °C for later testing. However, freezing for 9 months at either −20 °C or −80 °C leads to deterioration of certain parameters.     

Contribution of PTHrP to mechanical strain-induced fibrochondrogenic differentiation in entheses of Achilles tendon of miniature pigs

Background

Fibrochondrocytes are involved in entheses repair, but their response to mechanical strain (MS) is ill known.

Objective

To determine if parathyroid hormone-related protein (PTHrP) expression in fibrochondrocytes from fibrocartilaginous entheses is modulated by MS, and to further observe the regulatory effects of human (h) PTHrP on fibrochondrocyte differentiation.

Methods

Fibrochondrocytes from fibrocartilaginous entheses of Guizhou miniature pig?s Achilles tendon were submitted or not to MS (4%, 8% or 12% cyclic tensile strain; 1 Hz). Fibrochondrocytes were also exposed to: cyclopamine (Indian hedgehog (Ihh) inhibitor) (10 μM), hPTHrP (10 nM) or cyclopamine/hPTHrP (cyclopamine 10uM+hPTHrP 10 nM). Types I, II and X collagen and PTHrP expressions were measured by real-time RT-PCR and Western blot.

Result

Under 4% strain load for 12 h, types I and II collagen mRNA expressions were increased (+324% and +659%, P<0.001), while type X collagen was decreased (−89%, P<0.001). At 12%, types I and II collagen mRNA expressions were decreased (−62% and −62%, P<0.001), while type X collagen was increased (+375%, P<0.05). Under 4% strain load, PTHrP mRNA expression was increased in relation with strain duration (from 3 to 12 h: +168%, P<0.001), while at 12%, PTHrP expression decreased with time (from 3 to 12 h: −81%, P<0.001). Using cyclopamine for 24 h, PTHrP mRNA expression was significantly decreased (−88%, P<0.05), types I and II collagen were decreased (−90% and −82%, P<0.001), and type X collagen was increased (+261%, P<0.001).

Conclusions

Dynamic MS modulate PTHrP expressions. Thus, PTHrP might play an important role in fibrochondrocyte differentiation, indirectly revealing a role in entheses? formation and repair.     

Effect of habitual running on human Achilles tendon load-deformation properties and cross-sectional area.

Whether the cross-sectional area (CSA) and mechanical properties of the human Achilles tendon change in response to habitual exercise remains largely unexplored. The present study evaluated the CSA and contraction-induced displacement of the aponeurosis-tendon complex of the triceps surae in 11 untrained subjects before (tests 1 and 2) and after (test 3) approximately 9 mo of regular running ( approximately 78 training sessions). Displacement of the tendon-aponeurosis complex obtained by ultrasonography; electromyography of the gastrocnemius, soleus, and dorsiflexor muscles; and joint angular rotation were recorded during graded isometric plantarflexion ramps. Tendon CSA and moment arm were measured by using MRI, and tendon force was calculated from joint moments and tendon moment arm. A treadmill test was used to determine submaximal oxygen consumption (Vo2) at a given speed and maximal Vo2. The total running duration was approximately 43 h, distributed over 34 wk. Maximal Vo2 increased 8.6% (P < 0.01), and submaximal Vo2 decreased 6.2% (P < 0.05). Tendon-aponeurosis displacement during maximal voluntary contraction was unchanged (tests 1-3, 5.2 +/- 0.6, 5.2 +/- 0.5, and 5.3 +/- 0.4 mm, respectively) and yielded a structural stiffness of 365 +/- 50, 358 +/- 40, and 384 +/- 52 N/mm for tests 1-3, respectively (P > 0.05). Tendon CSA also remained unchanged (tests 1-3, 34.2 +/- 2.2, 33.9 +/- 2.2, and 33.8 +/- 2.1 mm2, respectively). In conclusion, a total training stimulus of approximately 9 mo of running in previously untrained subjects was adequate to induce significant cardiovascular improvements, although it did not result in any changes in the mechanical properties of the triceps surea tendon-aponeurosis complex or in the dimensions of Achilles tendon.     

Tendon mineralization is progressive and associated with deterioration of tendon biomechanical properties,and requires BMP-Smad signaling in the mouse Achilles tendon injury model

Ectopic tendon mineralization can develop following tendon rupture or trauma surgery. The pathogenesis of ectopic tendon mineralization and its clinical impact have not been fully elucidated yet. In this study, we utilized a mouse Achilles tendon injury model to determine whether ectopic tendon mineralization alters the biomechanical properties of the tendon and whether BMP signaling is involved in this condition. A complete transverse incision was made at the midpoint of the right Achilles tendon in 8-week-old CD1 mice and the gap was left open. Ectopic cartilaginous mass formation was found in the injured tendon by 4 weeks post-surgery and ectopic mineralization was detected at 8 to 10 weeks post-surgery. Ectopic mineralization grew over time and volume of the mineralized materials of 25-weeks samples was about 2.5 fold bigger than that of 10-weeks samples, indicating that injury-induced ectopic tendon mineralization is progressive. In vitro mechanical testing showed that max force, max stress and mid-substance modulus in the 25-weeks samples were significantly lower than the 10-weeks samples. We observed substantial increases in expression of bone morphogenetic protein family genes in injured tendons 1 week post-surgery. Immunohistochemical analysis showed that phosphorylation of both Smad1 and Smad3 was highly increased in injured tendons as early as 1 week post-injury and remained high in ectopic chondrogenic lesions 4-weeks post-injury. Treatment with the BMP receptor kinase inhibitor (LDN193189) significantly inhibited injury-induced tendon mineralization. These findings indicate that injury-induced ectopic tendon mineralization is progressive, involves BMP signaling and associated with deterioration of tendon biomechanical properties.     

Unrestrained video-assisted plethysmography: a noninvasive method for assessment of lung mechanical function in small animals.

The assessment of lung mechanical function in small animals, particularly mice, is essential for investigations into the pathophysiology of pulmonary disease. The most accurate and specific methods for making this assessment are highly invasive and so provide data of questionable relevance to normality. By contrast, present noninvasive methods based on unrestrained plethysmography have no direct link to the mechanical properties of the lung. There is thus a need for a completely noninvasive method for determining lung mechanical function in small animals. In the present study, we demonstrate an extension of unrestrained plethysmography in which changes in lung volume are estimated via orthogonal video imaging of the thorax. These estimates are combined with the pressure swings recorded as mice breathe inside a heated and humidified chamber to yield an estimate of specific airway resistance (sRaw). We used this new technique, which we term "unrestrained video-assisted plethysmography" (UVAP), to measure sRaw in 11 BALB/c mice exposed to aerosols of saline, methacholine, and albuterol and obtained mean values of 0.71, 1.23 and 1.10 cmH(2)O x s, respectively. Mean breathing frequency was 4.3, 3.4, and 3.6 breaths/s, respectively, while the corresponding mean tidal volumes were 0.36, 0.44 and 0.37 ml, respectively. We conclude that UVAP, a noninvasive method, is able to provide usefully accurate estimates of sRaw and breathing pattern parameters in mice.