Laser Doppler imaging (LDI) has become a standard method for optical measurement of tissue perfus... more Laser Doppler imaging (LDI) has become a standard method for optical measurement of tissue perfusion, but is limited by low resolution and long measurement times. We have developed an analysis technique based on a laser speckle imaging method that generates rapid, high-resolution perfusion images. We have called it laser speckle perfusion imaging (LSPI). This paper investigates LSPI output and compares it to LDI using blood flow models designed to simulate human skin at various levels of pigmentation. Results show that LSPI parameters can be chosen such that the instrumentation exhibits a similar response to changes in red blood cell concentration (0.1%-5%, 200 L/min) and velocity (0-800 L/min, 1% concentration) and, given its higher resolution and quicker response time, could provide a significant advantage over LDI for some applications. Differences were observed in the LDI and LSPI response to tissue optical properties. LDI perfusion values increased with increasing tissue absorption, while LSPI perfusion values showed a slight decrease. This dependence is predictable, owing to the perfusion algorithms specific to each instrument, and, if properly compensated for, should not influence each instrument's ability to measure relative changes in tissue perfusion.
This work represents the first clinical data acquired with the endoscopic laser speckle imaging (... more This work represents the first clinical data acquired with the endoscopic laser speckle imaging (eLSPI) system, a new diagnostic tool developed for real-time imaging of tissue blood flow during endoscopic surgical procedures. eLSPI was used to image tissue perfusion in the medial compartment of the knee of five patients requiring arthroscopic knee surgery. The effectiveness of eLSPI as a diagnostic tool was tested by measuring changes in tissue perfusion resultant from tourniquet application, and intra-articular epinephrine. eLSPI produced real-time perfusion video images of tissue blood flow in the knee joint. Tourniquet applications produced consistent decreases in mean perfusion index measurements (29.3% AE 5.1% in meniscus; 39.5% AE 8.2% in synovium with an intra patient variability of 6%-9%). A dose-dependent vasoconstrictive response to the administration of intra-articular epinephrine was measured, with maximum dose producing a mean decrease in perfusion of 31.0%-9.3% in meniscus and 41.2%-10.9% in synovium. eLSPI consistently detects decreases in articular tissue blood flow resultant from tourniquet inflation or from the administration of increasing concentrations of epinephrine. These are the first in vivo results indicating physiologic changes in articular tissue as a function of two commonly applied practices in endoscopic joint surgery.
Normal and healing adult rabbit medial collateral ligaments (MCL) have been assessed for microvas... more Normal and healing adult rabbit medial collateral ligaments (MCL) have been assessed for microvascular anatomy using a quantitative image analysis methodology. MCL preparation by ink-gelatin perfusion enabled acceptable visualisation of microvascular channels within the tissue. Fifteen adult rabbits were studied; 3 normal rabbits formed an external time-zero control group; 12 animals received a standardised gap injury to the right MCL. The ligament injury was permitted to heal for 3, 6, 17 or 40 wk (3 animals in each healing group). Results confirmed that the normal MCL is hypovascular (about 1.46% vascularity by area) and that microvascular channels are highly organised and oriented longitudinally deep within the tissue. Healing MCL scar becomes twice as vascular as normal ligament early on, but returns to near normal values by 40 wk. Microvascular channels appear less organised in scar than in contralateral controls, but remodel with time. The directional scatter and spatial extent of ligament microvascular channels is quantifiable in normal and healing tissues.
In spite of growing evidence that knee problems are common conditions in young and active patient... more In spite of growing evidence that knee problems are common conditions in young and active patients, little information about the frequency of these problems currently exists. In this descriptive epidemiologic study, the frequency of knee problems, as well as the typical patient ...
This study was carried out to test the hypothesis that improvements in ligament scar mechanical b... more This study was carried out to test the hypothesis that improvements in ligament scar mechanical behavior during healing may be related, in part, to increases in collagen fibril diameters. Forty-eight adult female New Zealand White rabbits had standardized midsubstance gap injuries created in their right medial collateral ligaments (MCLs) and were allowed normal cage activity until sacrifice in groups of 12 at 3, 6, 14 or 40 weeks post-injury. Eight animals in each group had both MCLs tested biomechanically while 4 animals had transmission EM investigation of midsubstance collagen fibril diameters by a standardized protocol. Results of mechanical tests showed a three- to fourfold increase in scar strength and stiffness over the intervals of healing studied while there was no change in collagen mean fibril minimum diameters. These results demonstrate no correlation between material or structural properties of scar and collagen fibril diameters in this model of healing and suggest that other mechanisms for scar mechanical improvement under these conditions must be investigated.
Journal of orthopaedic research : official publication of the Orthopaedic Research Society, 2014
To determine if decreased vascular responsiveness in the medial collateral ligament (MCL) of ante... more To determine if decreased vascular responsiveness in the medial collateral ligament (MCL) of anterior cruciate ligament transected (ACL-t) rabbit knees is due to pericyte deficiency associated with angiogenesis. Vascular responses to potassium chloride (KCl), phenylephrine, acetylcholine, and sodium nitroprusside (SNP) were evaluated in ACL-t rabbit knees (n = 6) and control knees (n = 5) using laser speckle perfusion imaging. Ligament degeneration was determined by ultrasound imaging. Vascular and pericyte volume were measured using quantitative immunohistochemical volumetric analysis using CD31 and α-smooth muscle actin antibodies with co-localization analysis. Perfusion was increased in the ACL-t rabbits 2.5-fold. Responsiveness to phenylephrine, SNP, and acetylcholine was significantly decreased in the ACL knee while no change in KCl responses was seen. MCL ultrasound imaging revealed decreased collagen organization, increased ligament thickness, and increased water content in t...
Medical & Biological Engineering & Computing, 1997
At present, there are only two laser Doppler perfusion imaging systems (LDIs) manufactured for me... more At present, there are only two laser Doppler perfusion imaging systems (LDIs) manufactured for medical applications: a 'stepwise' and a 'continuous' scanning LDI. The stepwise scanning LDI has previously been investigated and compared with coloured microsphere determined standardised flow. The continuous scanning LDI is investigated and compared with the stepwise scanning LDI for its ability to measure in vivo, hypoaemic, ligament tissue blood flow changes. The continuous scanning system was supplied with two lasers, red and near infrared (NIR), allowing for additional assessment of the effect of wavelength on imaging ligament perfusJon. Perfusion images were obtained from surgically exposed rabbit medial collateral ligaments (MCL). Continuous and stepwise LDI scans were compared using correlation and linear regression analysis of image averages and standard deviations. Using the same method of analysis, LDI measurements using red and NIR lasers indicated a high degree of correlation, at least over the ranges of perfusion assessed, indicating that red and NIR lasers measure similar regions of flow in the rabbit MCL. These experiments confirm that both LDI techniques provide a valid in v/vo measure of dynamic changes in connective tissue pedusion and could have significant impact on the understanding and treatment of joint injury and arthritis.
Functional outcomes of anterior cruciate ligament (ACL) injury are generally poorer than those of... more Functional outcomes of anterior cruciate ligament (ACL) injury are generally poorer than those of medial collateral ligament (MCL) tears. Following ligament damage, all phases of ligament healing require an adequate blood supply. We hypothesized that the differences in healing properties of the ACL and MCL would reflect their vascular responses to joint injury. This paper examines the long-term changes in blood flow and vascular volume of rabbit knee ligaments after direct injury, and under conditions of chronic joint instability induced by section of the posterior cruciate ligament (PCL). Standardized injuries were surgically induced in adult rabbit knee ligaments: partial MCL transection, partial ACL transection, or complete PCL transection (joint instability). Sixteen weeks later the blood flow and vascular volume of the ACL and MCL were measured and compared to control and sham-operated animals. Direct ligament injury induced significant increases in standardized blood flow and vascular volume of both ACL and MCL after 16 weeks; however, the vascular volume of the ACL was not higher than the control levels in the MCL. We conclude that direct injury to both the anterior cruciate and MCLs induces long-term physiological responses. Joint laxity is a common sequel to PCL injury. Chronic joint laxity failed to induce adaptive vascular responses in the ACL, while the MCL shows significant amplification of blood supply. Although both MCL and ACL showed increased weight after PCL transection, the lack of a long-term vascular response in the ACL may be a major factor in its the diminished healing potential.
Anterior cruciate ligament (ACL) injury induces maladaptive vascular responses that degrade media... more Anterior cruciate ligament (ACL) injury induces maladaptive vascular responses that degrade medial collateral ligament (MCL) function. The purpose of this study was to determine if early or delayed ACL reconstruction can prevent or reverse the abnormal changes in vascular function that occur in the uninjured MCL after ACL injury. Twenty-four rabbits were divided into four groups (n = 6); control, ACL-deficient (ACL-X), immediate ACL reconstructed (ACL-IR) and delayed ACL reconstructed (ACL-DR). After 8 weeks, MCLs were assessed for blood flow, responses to acetylcholine (ACh) and phenylephrine (Phe) and autoregulatory responses, using laser speckle perfusion imaging. In ACL-X knees, blood flow in the MCL increased by 2.5-fold compared to control. MCL hyperemia was diminished in ACL-DR knees and was unaltered in ACL-IR knees. MCL vasculature was unresponsive to ACh and Phe in ACL-X. These responses were partially restored by ACL reconstruction. Autoregulatory responses were not significantly different between groups. ACL-DR decreased hyperemia in the MCL and partially attenuated abnormal MCL vascular responses. ACL-IR was more effective at preventing MCL hyperemia and preserving vascular responsiveness to ACh and Phe. This suggests that the vascular alterations in the uninjured rabbit MCL are largely caused by abnormal mechanical loading resulting from ACL deficiency and can be prevented through early reconstruction. Early ACL reconstruction could limit the progression of microvascular dysfunction of the MCL, and preserve physiological joint homeostasis. This might prevent joint degeneration and delay the progression of osteoarthritis.
Loss of the posterior cruciate ligament (PCL) of the knee has a significant impact on joint stabi... more Loss of the posterior cruciate ligament (PCL) of the knee has a significant impact on joint stability and biomechanical function. Changes in joint biomechanics may result in mal-adaptive tissue degeneration and functional alteration of supporting ligaments. This study examines the effects of joint laxity on the vascular physiology of the intact anterior cruciate (ACL) and medial collateral (MCL) ligaments after PCL transection in rabbits. One-year-old female New Zealand white rabbits were assigned to control (n = 12). sham-operated (n = 12) or PCL transected (2. 6 or 16 weeks, n = 12 per time point) groups. Half of the animals (n = 6 per group) were used for ACL and MCL blood flow determination using coloured microsphere infusion (ml/min/100 g), and half were used for vascular volume determination (given as vascular index, pllg). In the MCL, PCL transection induced large, significant (4-S-fold) increases in blood flow (peak at 2 weeks) and vascular index (peak at 6 weeks) compared to sham-operated animals that returned towards control values by 16 weeks. In contrast, the ACL showed no increase in blood flow in lax joints, and a relatively small (2-fold) increase in vascular index at 6 weeks only. The wet weight and water content of both the MCL and ACL were significantly increased in PCL-deficient joints. We conclude that joint laxity (instability) subsequent to loss of the PCL in rabbits impacts the vascular physiology of intact supporting ligaments, inducing both vasomotor and angiogenic responses in the MCL. Changes in wet weight and water content of both the MCL and ACL demonstrate prolonged physiological adaptation of intact structures in lax joints.
Little is known about the contribution of innervation to ligament healing after traumatic disrupt... more Little is known about the contribution of innervation to ligament healing after traumatic disruption, although there is good evidence of an important role for the peripheral nervous system in the healing of fractures and skin injuries. Tissues such as ligament, with a low resting blood supply, are dependent on substantial increases in blood flow and vascular volume during the initial stages of repair. We hypothesized that this initial healing response would be strongly promoted by neurogenic inflammation. Since the saphenous nerve (a major sensory branch of the femoral nerve) supplies the medial half of the knee joint, we elected to use femoral nerve transection as a model to determine the role of sensory and autonomic innervation in the initial outcome of repair of the injured medial collateral ligament. Twelve adult, female NZW rabbits underwent right medial collateral ligament transection. Of these, six rabbits underwent right femoral nerve transection to disrupt the somatic sensory and autonomic nerve supply to the knee joint and six were kept neurologically intact (controls). At six weeks post-injury, the animals were assessed by laser Doppler perfusion imaging (LDI) to determine the local blood flow, at both the injury site and at the uninjured contralateral ligament. The animals were then killed, the knee joints were removed and the biomechanical characteristics of the healing bone-median collateral ligament (MCL)-bone complexes assessed. In a separate cohort of 16 rabbits, vascular volumes of the injured ligaments were measured by infusion of a carmine red/gelatin solution. At six weeks post-injury, in vivo measurement of perfusion with LDI revealed that normally innervated ligaments had an almost threefold higher average blood flow. Carmine redlgelatin infusion revealed a 50% higher density of blood vessels as compared to denervated ligaments. The force required for ultimate failure was found to be 50'% higher in normally innnervated MCL's as compared to denervated MCL's: 153.14 3~ 20.71 N versus 101.29 f 17.88 N (p < 0.05). Static creep was increased by 66% in denervated MCL's: 2.83 & 0.45% versus 1.70 & 0.12'%1 (p < 0.05). Total creep was increased by 45% in denervated MCL's: 5.29 & 0.6% compared to 3.64 * 0.31% in innervated MCL's (p < 0.05). We conclude that intact innervation makes a critical contribution to the early healing responses of the MCL of adult rabbits.
Previous work has shown that innervation participates in normal ligament healing. The present stu... more Previous work has shown that innervation participates in normal ligament healing. The present study was performed to determine if exogenous nerve growth factor (NGF) would improve the healing of injured ligament by promoting reinnervation, blood flow, and angiogenesis. Two groups of 30 Sprague-Dawley rats underwent unilateral medial collateral ligament transection (MCL). One group was given 10 microg NGF and the other was given PBS via osmotic pump over 7 days after injury. After 7, 14, and 42 days, in vivo blood flow was measured using laser speckle perfusion imaging (LSPI). Morphologic assessments of nerve density, vascularity, and angiogenesis inhibitor production were done in three animals at each time point by immunohistochemical staining for the pan-neuronal marker PGP9.5, the endothelial marker vWF, and the angiogenesis inhibitor thrombospondin-2 (TSP-2). Ligament scar material and structural mechanical properties were assessed in seven rats at each time point. Increased nerve density was promoted by NGF at both 14 and 42 days. Exposure to NGF also led to increased ligament vascularity, as measured by histologic assessment of vWF immunohistochemistry, although LSPI-measured blood flow was not significantly different from controls. NGF treatment also led to decreased expression of TSP-2 at 14 days. Mechanical testing revealed that exposure to NGF increased failure load by 40%, ultimate tensile strength by 55%, and stiffness by 30% at 42 days. There were no detectable differences between groups in creep properties. The results suggest that local application of NGF can improve ligament healing by promoting both reinnervation and angiogenesis, and results in scars with enhanced mechanical properties.
In this study, the short-term effects of immobilization on joint damage and medial collateral lig... more In this study, the short-term effects of immobilization on joint damage and medial collateral ligament (MCL) healing were investigated in unstable, anterior cruciate ligament (ACL)-deficient knees in rabbits. Forty-six 12-month-old female New Zealand white rabbits were separated into three groups. Animals from each group had surgery on their right knees: group I, sham controls (n = 9); group II, complete transection of the ACL and removal of a 4 mm segment (gap injury) of MCL midsubstance with no immobilization of the limb (n = 19); and group III, same injuries to the ACL and MCL (as group II) but with immobilization of the limb (n = 18). No surgical repair of disrupted ligaments was performed. Left knees served as unoperated contralateral controls. All animals were allowed unrestricted cage activity until sacrifice in subgroups at 3, 6, and 14 weeks of healing when biomechanical properties of all MCLs were measured. All knee joints were systematically examined for gross evidence of damage to articular cartilage, menisci, and periarticular soft tissues. To monitor relative in vivo loads on injured limbs during healing, hindlimb weight bearing was assessed at biweekly intervals. Results indicated that animals in both groups II and III bore relatively lower loads (compared to preinjury values) on their injured hindlimbs. Mechanical testing of MCLs showed only minor changes in sham controls, while group II and III healing MCLs demonstrated significantly lower force and stress at MCL complex failure compared to contralateral controls. In specific comparisons of group III to group II animals, we noted that immobilization prevented joint damage over the early intervals studied. In addition, immobilization resulted in MCL laxity similar to contralateral control values but inhibited development of structural strength and stiffness in healing MCLs. These results suggest that in the rabbit, short-term immobilization of an ACL-deficient knee offers some advantages to the joint and to certain low load behaviors of the healing MCL, but it also results in a smaller quantity of scar tissue that is less able to resist higher loads. Longer-term studies involving remobilization are necessary before the effects of brief immobilization on joint damage and MCL healing in this ACL-deficient model can be fully defined.
Journal of Manipulative and Physiological Therapeutics, 1999
Background: Evidence exists that conservative rehabilitation protocols fail to achieve full recov... more Background: Evidence exists that conservative rehabilitation protocols fail to achieve full recovery of muscle strength and function after joint injuries. The lack of success has been attributed to the high amount of muscle inhibition found in patients with pathologic conditions of the knee joint. Clinical evaluation shows that anterior knee pain is typically associated with sacroiliac joint dysfunction, which may contribute to the muscle inhibition observed in this patient group. Objective: To assess whether sacroiliac joint manipulation alters muscle inhibition and strength of the knee extensor muscles in patients with anterior knee pain. Design and Setting: The effects of sacroiliac joint manipulation were evaluated in patients with anterior knee pain. The manipulation consisted of a high-velocity low-amplitude thrust in the side-lying position aimed at correcting sacroiliac joint dysfunction. Before and after the manipulation, torque, muscle inhibition, and muscle activation for the knee extensor muscles were measured during isometric contractions using a Cybex dynamometer, muscle stimulation, and electromyography, respectively. Participants: Eighteen patients (mean age, 30.5 ± 13.0 years) with either unilateral (n = 14) or bilateral (n = 4) anterior knee pain. Results: Patients showed substantial muscle inhibition in the involved and the contralateral legs as estimated by the interpolated twitch technique. After the manipulation, a decrease in muscle inhibition and increases in knee extensor torques and muscle activation were observed, particularly in the involved leg. In patients with bilateral anterior knee pain, muscle inhibition was decreased in both legs after sacroiliac joint adjustment. Conclusions: Spinal manipulation might offer an interesting alternative treatment for patients with anterior knee pain and muscle inhibition. Because this clinical outcome study was of descriptive nature rather than a controlled design, biases might have occurred. Thus the results have to be verified in a randomized, controlled, double-blinded trial before firm conclusions can be drawn or recommendations can be made.
Testing environment is an important factor in the outcome of mechanical tests on connective tissu... more Testing environment is an important factor in the outcome of mechanical tests on connective tissue. The purpose of this investigation was to determine the effect of ligament water content on ligament mechanical behaviour by altering the test environment. Water content of medial collateral ligament (MCLs) from 19 three-month-old New Zealand White rabbits was varied in subsets of ligaments pairs by means of immersion in 2,10 or 25% sucrose or 0.9% phosphate-buffered saline (PBS) solutions for 1 h. One knee joint was cycled 50 times in the designated solution (experimental), while the contralateral knee (uncycled control) was simultaneously soaked in the same tank. Following cycling, the water contents of both test and control ligaments were determined. Water contents of 22 normal MCLs were determined immediately post-sacrifice and served as 'normal water content' controls. Normalized peak cyclic load changes were used as a measure of the viscoelastic behaviour of each MCL. Results demonstrated that only ligaments soaked (but not cycled) in a 10% sucrose solution had water contents @S&2.5%) which were statistically similar to the 22 fresh normal MCLs (63.9 f 6.0%). Ligaments soaked in PBS (74.0& 1.3%) or 2% sucrose (69.2 f 2.3%) had significantly higher water contents compared to fresh normal MCLs. Ligaments with higher water contents (e.g. soaked in PBS or 2% sucrose) demonstrated greater cyclic load relaxation compared to ligaments with lower contents (e.g. soaked in 25 or 10% sucrose). Different fluid test environments can significantly alter ligament water content and, in turn, significantly affect ligament viscoelastic behaviour.
Sympathetic-derived neuropeptide Y (NPY) helps regulate inflammatory responses in injury and dise... more Sympathetic-derived neuropeptide Y (NPY) helps regulate inflammatory responses in injury and disease, is a vasoconstrictor, and stimulates angiogenesis. Rupture of the anterior cruciate ligament (ACL) is a common clinical presentation that results in tissue inflammation, hyperemia, and angiogenesis in the intact medial collateral ligament (MCL). This study is the first to examine the vasoregulatory role of NPY in ACL-deficient knee joints by using the newly developed technique of laser speckle perfusion imaging (LSPI). MCL blood flow was measured in two groups of adult rabbits: unoperated control ( n = 6), and 6-wk ACL transected ( n = 5). Under anesthesia, the MCL was surgically exposed and tissue blood flow was imaged at high resolution using LSPI. NPY was applied to the MCL vasculature in topical boluses of 100 μl (dose range 10−14 to 10−9 mol), and the α-adrenoceptor agonist phenylephrine was applied in doses of 10−14, 10−10, and 10−7 mol. In control rabbits, topical administrat...
An image analysis technique has been developed to quantitatively describe the fine vascular patte... more An image analysis technique has been developed to quantitatively describe the fine vascular patterns observed in ligament tissue. The longitudinal orientational distribution and total vessel volume of India-ink-perfused blood vessel segments in normal and healing ligaments were determined. The methods involved special vascular preparation of adult rabbit knee medial collateral ligaments (MCL) by India-ink perfusion. Black and white microscope images of ink-perfused tissue sections were subjected to a thresholding procedure to binarize digitized ligament images, which were then skeletonized and analyzed for directional distribution based on the least-squares technique. Analysis of medial collateral ligaments in New Zealand White rabbits using this method has shown that scarred tissue is more vascular and has a more chaotic angular distribution of blood-vessel segments than normal ligament tissue.
Objectives. This study was aimed at investigating muscle strength and quadriceps inhibition in pa... more Objectives. This study was aimed at investigating muscle strength and quadriceps inhibition in patients with unilateral anterior knee pain syndrome. Design. Functional assessments were made before an arthroscopic knee surgery and 6 weeks and 6 months following the surgical intervention. Background. Traumatic knee injuries have been associated with severe muscle inhibition of the knee extensor muscles. Muscle inhibition is a serious hindrance in the rehabilitation process and prevents full functional recovery of the affected joint. Methods. 30 patients participated in the study. Isometric quadriceps strength was measured using a KinCom dynamometer. Muscle inhibition was assessed using the interpolated twitch technique which requires applying a single electrical twitch to the femoral nerve during a maximal isometric knee extensor contraction. Pain was assessed with a 100 mm visual analogue pain scale. Results. Pre-surgery, substantial muscle inhibition and pain was observed in the affected leg compared with the contralateral leg. Over the 6 month period there was a decrease in pain and muscle inhibition, although the decrease in muscle inhibition failed to reach statistical significance. Muscle strength showed a decrease 6 weeks post-surgery followed by an increase 6 months following surgery compared with pre-surgical values. Conclusions. The arthroscopic intervention was successful in reducing pain in patients with unilateral anterior knee pain syndrome. However, muscle inhibition was still substantial 6 months following surgery and was significantly higher in the affected and contralateral limb than in normal subjects. Relevance Clinical evidence indicates that conservative rehabilitation programs are not entirely successful in improving muscle strength and function in patients with severe joint injuries. The lack of success has typically been attributed to the high amount of muscle inhibition found in these patients. The present study confirmed a persisting muscle inhibition 6 months following knee surgery not only in the affected but also in the contralateral leg. Future studies should be aimed at designing exercise protocols that successfully eliminate muscle inhibition and, thus, potentially allow for full restoration of muscle function.
Previous work has shown a progressive, age-related loss of knee joint innervation in the C57BL6Ni... more Previous work has shown a progressive, age-related loss of knee joint innervation in the C57BL6Nia mouse. We did three experiments to describe further the loss and determine whether it might contribute to the development of knee osteoarthrosis in this model. Immunocytochemistry showed that the percentage of neurons expressing substance P and calcitonin gene-related peptide increased with age, indicating a relatively selective loss of mechanoreceptors. Histological examination of knee joints of mice at various ages showed that loss of joint innervation always preceded histological changes of cartilage degeneration. The mice usually developed a mild form of osteoarthrosis, but surgical ablation of joint innervation caused the development of severe patellofemoral osteoarthrosis. The ndings are consistent with the hypothesis that an age-related loss of joint innervation may contribute to the development of osteoarthrosis.
Background and Objective: Laser speckle perfusion imaging (LSPI) is a minimally invasive optical ... more Background and Objective: Laser speckle perfusion imaging (LSPI) is a minimally invasive optical measure of relative changes in blood flow, providing real-time, high resolution, two-dimensional maps of vascular structure. Standard LSI imaging uses a light-reflective geometry that limits the measurement to a thin surface layer of 0.2-1 mm. The objective of this study was to test a new LSI instrument geometry with the laser source opposed to the image capture plane (light transmissive). Captured light then travels the entire tissue thickness (10-15 mm), sampling much deeper regions of interest than conventional optical imaging techniques. Study Design: Reflective-light (conventional) and transmissive-light LSI modes were used to measure finger joint blood flow during a timed tourniquet occlusion of the brachial artery in volunteer participants. Results: There was greatly increased visibility of vessels underlying the skin in the light-transmissive mode LSI mode. Established LSI algorithms were shown to still work in the light-transmissive mode, despite decorrelation due to finite laser coherence length and the light passing through a tissue thickness of 10-15 mm. Conclusion: Transmissive LSI can be used to measure blood flow deep (10-15 mm) into tissues. This could be useful for non-invasive measurements of finger joint synovial blood flow in diagnosing and treating peripheral vascular disorders, such as rheumatoid arthritis.
Laser Doppler imaging (LDI) has become a standard method for optical measurement of tissue perfus... more Laser Doppler imaging (LDI) has become a standard method for optical measurement of tissue perfusion, but is limited by low resolution and long measurement times. We have developed an analysis technique based on a laser speckle imaging method that generates rapid, high-resolution perfusion images. We have called it laser speckle perfusion imaging (LSPI). This paper investigates LSPI output and compares it to LDI using blood flow models designed to simulate human skin at various levels of pigmentation. Results show that LSPI parameters can be chosen such that the instrumentation exhibits a similar response to changes in red blood cell concentration (0.1%-5%, 200 L/min) and velocity (0-800 L/min, 1% concentration) and, given its higher resolution and quicker response time, could provide a significant advantage over LDI for some applications. Differences were observed in the LDI and LSPI response to tissue optical properties. LDI perfusion values increased with increasing tissue absorption, while LSPI perfusion values showed a slight decrease. This dependence is predictable, owing to the perfusion algorithms specific to each instrument, and, if properly compensated for, should not influence each instrument's ability to measure relative changes in tissue perfusion.
This work represents the first clinical data acquired with the endoscopic laser speckle imaging (... more This work represents the first clinical data acquired with the endoscopic laser speckle imaging (eLSPI) system, a new diagnostic tool developed for real-time imaging of tissue blood flow during endoscopic surgical procedures. eLSPI was used to image tissue perfusion in the medial compartment of the knee of five patients requiring arthroscopic knee surgery. The effectiveness of eLSPI as a diagnostic tool was tested by measuring changes in tissue perfusion resultant from tourniquet application, and intra-articular epinephrine. eLSPI produced real-time perfusion video images of tissue blood flow in the knee joint. Tourniquet applications produced consistent decreases in mean perfusion index measurements (29.3% AE 5.1% in meniscus; 39.5% AE 8.2% in synovium with an intra patient variability of 6%-9%). A dose-dependent vasoconstrictive response to the administration of intra-articular epinephrine was measured, with maximum dose producing a mean decrease in perfusion of 31.0%-9.3% in meniscus and 41.2%-10.9% in synovium. eLSPI consistently detects decreases in articular tissue blood flow resultant from tourniquet inflation or from the administration of increasing concentrations of epinephrine. These are the first in vivo results indicating physiologic changes in articular tissue as a function of two commonly applied practices in endoscopic joint surgery.
Normal and healing adult rabbit medial collateral ligaments (MCL) have been assessed for microvas... more Normal and healing adult rabbit medial collateral ligaments (MCL) have been assessed for microvascular anatomy using a quantitative image analysis methodology. MCL preparation by ink-gelatin perfusion enabled acceptable visualisation of microvascular channels within the tissue. Fifteen adult rabbits were studied; 3 normal rabbits formed an external time-zero control group; 12 animals received a standardised gap injury to the right MCL. The ligament injury was permitted to heal for 3, 6, 17 or 40 wk (3 animals in each healing group). Results confirmed that the normal MCL is hypovascular (about 1.46% vascularity by area) and that microvascular channels are highly organised and oriented longitudinally deep within the tissue. Healing MCL scar becomes twice as vascular as normal ligament early on, but returns to near normal values by 40 wk. Microvascular channels appear less organised in scar than in contralateral controls, but remodel with time. The directional scatter and spatial extent of ligament microvascular channels is quantifiable in normal and healing tissues.
In spite of growing evidence that knee problems are common conditions in young and active patient... more In spite of growing evidence that knee problems are common conditions in young and active patients, little information about the frequency of these problems currently exists. In this descriptive epidemiologic study, the frequency of knee problems, as well as the typical patient ...
This study was carried out to test the hypothesis that improvements in ligament scar mechanical b... more This study was carried out to test the hypothesis that improvements in ligament scar mechanical behavior during healing may be related, in part, to increases in collagen fibril diameters. Forty-eight adult female New Zealand White rabbits had standardized midsubstance gap injuries created in their right medial collateral ligaments (MCLs) and were allowed normal cage activity until sacrifice in groups of 12 at 3, 6, 14 or 40 weeks post-injury. Eight animals in each group had both MCLs tested biomechanically while 4 animals had transmission EM investigation of midsubstance collagen fibril diameters by a standardized protocol. Results of mechanical tests showed a three- to fourfold increase in scar strength and stiffness over the intervals of healing studied while there was no change in collagen mean fibril minimum diameters. These results demonstrate no correlation between material or structural properties of scar and collagen fibril diameters in this model of healing and suggest that other mechanisms for scar mechanical improvement under these conditions must be investigated.
Journal of orthopaedic research : official publication of the Orthopaedic Research Society, 2014
To determine if decreased vascular responsiveness in the medial collateral ligament (MCL) of ante... more To determine if decreased vascular responsiveness in the medial collateral ligament (MCL) of anterior cruciate ligament transected (ACL-t) rabbit knees is due to pericyte deficiency associated with angiogenesis. Vascular responses to potassium chloride (KCl), phenylephrine, acetylcholine, and sodium nitroprusside (SNP) were evaluated in ACL-t rabbit knees (n = 6) and control knees (n = 5) using laser speckle perfusion imaging. Ligament degeneration was determined by ultrasound imaging. Vascular and pericyte volume were measured using quantitative immunohistochemical volumetric analysis using CD31 and α-smooth muscle actin antibodies with co-localization analysis. Perfusion was increased in the ACL-t rabbits 2.5-fold. Responsiveness to phenylephrine, SNP, and acetylcholine was significantly decreased in the ACL knee while no change in KCl responses was seen. MCL ultrasound imaging revealed decreased collagen organization, increased ligament thickness, and increased water content in t...
Medical & Biological Engineering & Computing, 1997
At present, there are only two laser Doppler perfusion imaging systems (LDIs) manufactured for me... more At present, there are only two laser Doppler perfusion imaging systems (LDIs) manufactured for medical applications: a 'stepwise' and a 'continuous' scanning LDI. The stepwise scanning LDI has previously been investigated and compared with coloured microsphere determined standardised flow. The continuous scanning LDI is investigated and compared with the stepwise scanning LDI for its ability to measure in vivo, hypoaemic, ligament tissue blood flow changes. The continuous scanning system was supplied with two lasers, red and near infrared (NIR), allowing for additional assessment of the effect of wavelength on imaging ligament perfusJon. Perfusion images were obtained from surgically exposed rabbit medial collateral ligaments (MCL). Continuous and stepwise LDI scans were compared using correlation and linear regression analysis of image averages and standard deviations. Using the same method of analysis, LDI measurements using red and NIR lasers indicated a high degree of correlation, at least over the ranges of perfusion assessed, indicating that red and NIR lasers measure similar regions of flow in the rabbit MCL. These experiments confirm that both LDI techniques provide a valid in v/vo measure of dynamic changes in connective tissue pedusion and could have significant impact on the understanding and treatment of joint injury and arthritis.
Functional outcomes of anterior cruciate ligament (ACL) injury are generally poorer than those of... more Functional outcomes of anterior cruciate ligament (ACL) injury are generally poorer than those of medial collateral ligament (MCL) tears. Following ligament damage, all phases of ligament healing require an adequate blood supply. We hypothesized that the differences in healing properties of the ACL and MCL would reflect their vascular responses to joint injury. This paper examines the long-term changes in blood flow and vascular volume of rabbit knee ligaments after direct injury, and under conditions of chronic joint instability induced by section of the posterior cruciate ligament (PCL). Standardized injuries were surgically induced in adult rabbit knee ligaments: partial MCL transection, partial ACL transection, or complete PCL transection (joint instability). Sixteen weeks later the blood flow and vascular volume of the ACL and MCL were measured and compared to control and sham-operated animals. Direct ligament injury induced significant increases in standardized blood flow and vascular volume of both ACL and MCL after 16 weeks; however, the vascular volume of the ACL was not higher than the control levels in the MCL. We conclude that direct injury to both the anterior cruciate and MCLs induces long-term physiological responses. Joint laxity is a common sequel to PCL injury. Chronic joint laxity failed to induce adaptive vascular responses in the ACL, while the MCL shows significant amplification of blood supply. Although both MCL and ACL showed increased weight after PCL transection, the lack of a long-term vascular response in the ACL may be a major factor in its the diminished healing potential.
Anterior cruciate ligament (ACL) injury induces maladaptive vascular responses that degrade media... more Anterior cruciate ligament (ACL) injury induces maladaptive vascular responses that degrade medial collateral ligament (MCL) function. The purpose of this study was to determine if early or delayed ACL reconstruction can prevent or reverse the abnormal changes in vascular function that occur in the uninjured MCL after ACL injury. Twenty-four rabbits were divided into four groups (n = 6); control, ACL-deficient (ACL-X), immediate ACL reconstructed (ACL-IR) and delayed ACL reconstructed (ACL-DR). After 8 weeks, MCLs were assessed for blood flow, responses to acetylcholine (ACh) and phenylephrine (Phe) and autoregulatory responses, using laser speckle perfusion imaging. In ACL-X knees, blood flow in the MCL increased by 2.5-fold compared to control. MCL hyperemia was diminished in ACL-DR knees and was unaltered in ACL-IR knees. MCL vasculature was unresponsive to ACh and Phe in ACL-X. These responses were partially restored by ACL reconstruction. Autoregulatory responses were not significantly different between groups. ACL-DR decreased hyperemia in the MCL and partially attenuated abnormal MCL vascular responses. ACL-IR was more effective at preventing MCL hyperemia and preserving vascular responsiveness to ACh and Phe. This suggests that the vascular alterations in the uninjured rabbit MCL are largely caused by abnormal mechanical loading resulting from ACL deficiency and can be prevented through early reconstruction. Early ACL reconstruction could limit the progression of microvascular dysfunction of the MCL, and preserve physiological joint homeostasis. This might prevent joint degeneration and delay the progression of osteoarthritis.
Loss of the posterior cruciate ligament (PCL) of the knee has a significant impact on joint stabi... more Loss of the posterior cruciate ligament (PCL) of the knee has a significant impact on joint stability and biomechanical function. Changes in joint biomechanics may result in mal-adaptive tissue degeneration and functional alteration of supporting ligaments. This study examines the effects of joint laxity on the vascular physiology of the intact anterior cruciate (ACL) and medial collateral (MCL) ligaments after PCL transection in rabbits. One-year-old female New Zealand white rabbits were assigned to control (n = 12). sham-operated (n = 12) or PCL transected (2. 6 or 16 weeks, n = 12 per time point) groups. Half of the animals (n = 6 per group) were used for ACL and MCL blood flow determination using coloured microsphere infusion (ml/min/100 g), and half were used for vascular volume determination (given as vascular index, pllg). In the MCL, PCL transection induced large, significant (4-S-fold) increases in blood flow (peak at 2 weeks) and vascular index (peak at 6 weeks) compared to sham-operated animals that returned towards control values by 16 weeks. In contrast, the ACL showed no increase in blood flow in lax joints, and a relatively small (2-fold) increase in vascular index at 6 weeks only. The wet weight and water content of both the MCL and ACL were significantly increased in PCL-deficient joints. We conclude that joint laxity (instability) subsequent to loss of the PCL in rabbits impacts the vascular physiology of intact supporting ligaments, inducing both vasomotor and angiogenic responses in the MCL. Changes in wet weight and water content of both the MCL and ACL demonstrate prolonged physiological adaptation of intact structures in lax joints.
Little is known about the contribution of innervation to ligament healing after traumatic disrupt... more Little is known about the contribution of innervation to ligament healing after traumatic disruption, although there is good evidence of an important role for the peripheral nervous system in the healing of fractures and skin injuries. Tissues such as ligament, with a low resting blood supply, are dependent on substantial increases in blood flow and vascular volume during the initial stages of repair. We hypothesized that this initial healing response would be strongly promoted by neurogenic inflammation. Since the saphenous nerve (a major sensory branch of the femoral nerve) supplies the medial half of the knee joint, we elected to use femoral nerve transection as a model to determine the role of sensory and autonomic innervation in the initial outcome of repair of the injured medial collateral ligament. Twelve adult, female NZW rabbits underwent right medial collateral ligament transection. Of these, six rabbits underwent right femoral nerve transection to disrupt the somatic sensory and autonomic nerve supply to the knee joint and six were kept neurologically intact (controls). At six weeks post-injury, the animals were assessed by laser Doppler perfusion imaging (LDI) to determine the local blood flow, at both the injury site and at the uninjured contralateral ligament. The animals were then killed, the knee joints were removed and the biomechanical characteristics of the healing bone-median collateral ligament (MCL)-bone complexes assessed. In a separate cohort of 16 rabbits, vascular volumes of the injured ligaments were measured by infusion of a carmine red/gelatin solution. At six weeks post-injury, in vivo measurement of perfusion with LDI revealed that normally innervated ligaments had an almost threefold higher average blood flow. Carmine redlgelatin infusion revealed a 50% higher density of blood vessels as compared to denervated ligaments. The force required for ultimate failure was found to be 50'% higher in normally innnervated MCL's as compared to denervated MCL's: 153.14 3~ 20.71 N versus 101.29 f 17.88 N (p < 0.05). Static creep was increased by 66% in denervated MCL's: 2.83 & 0.45% versus 1.70 & 0.12'%1 (p < 0.05). Total creep was increased by 45% in denervated MCL's: 5.29 & 0.6% compared to 3.64 * 0.31% in innervated MCL's (p < 0.05). We conclude that intact innervation makes a critical contribution to the early healing responses of the MCL of adult rabbits.
Previous work has shown that innervation participates in normal ligament healing. The present stu... more Previous work has shown that innervation participates in normal ligament healing. The present study was performed to determine if exogenous nerve growth factor (NGF) would improve the healing of injured ligament by promoting reinnervation, blood flow, and angiogenesis. Two groups of 30 Sprague-Dawley rats underwent unilateral medial collateral ligament transection (MCL). One group was given 10 microg NGF and the other was given PBS via osmotic pump over 7 days after injury. After 7, 14, and 42 days, in vivo blood flow was measured using laser speckle perfusion imaging (LSPI). Morphologic assessments of nerve density, vascularity, and angiogenesis inhibitor production were done in three animals at each time point by immunohistochemical staining for the pan-neuronal marker PGP9.5, the endothelial marker vWF, and the angiogenesis inhibitor thrombospondin-2 (TSP-2). Ligament scar material and structural mechanical properties were assessed in seven rats at each time point. Increased nerve density was promoted by NGF at both 14 and 42 days. Exposure to NGF also led to increased ligament vascularity, as measured by histologic assessment of vWF immunohistochemistry, although LSPI-measured blood flow was not significantly different from controls. NGF treatment also led to decreased expression of TSP-2 at 14 days. Mechanical testing revealed that exposure to NGF increased failure load by 40%, ultimate tensile strength by 55%, and stiffness by 30% at 42 days. There were no detectable differences between groups in creep properties. The results suggest that local application of NGF can improve ligament healing by promoting both reinnervation and angiogenesis, and results in scars with enhanced mechanical properties.
In this study, the short-term effects of immobilization on joint damage and medial collateral lig... more In this study, the short-term effects of immobilization on joint damage and medial collateral ligament (MCL) healing were investigated in unstable, anterior cruciate ligament (ACL)-deficient knees in rabbits. Forty-six 12-month-old female New Zealand white rabbits were separated into three groups. Animals from each group had surgery on their right knees: group I, sham controls (n = 9); group II, complete transection of the ACL and removal of a 4 mm segment (gap injury) of MCL midsubstance with no immobilization of the limb (n = 19); and group III, same injuries to the ACL and MCL (as group II) but with immobilization of the limb (n = 18). No surgical repair of disrupted ligaments was performed. Left knees served as unoperated contralateral controls. All animals were allowed unrestricted cage activity until sacrifice in subgroups at 3, 6, and 14 weeks of healing when biomechanical properties of all MCLs were measured. All knee joints were systematically examined for gross evidence of damage to articular cartilage, menisci, and periarticular soft tissues. To monitor relative in vivo loads on injured limbs during healing, hindlimb weight bearing was assessed at biweekly intervals. Results indicated that animals in both groups II and III bore relatively lower loads (compared to preinjury values) on their injured hindlimbs. Mechanical testing of MCLs showed only minor changes in sham controls, while group II and III healing MCLs demonstrated significantly lower force and stress at MCL complex failure compared to contralateral controls. In specific comparisons of group III to group II animals, we noted that immobilization prevented joint damage over the early intervals studied. In addition, immobilization resulted in MCL laxity similar to contralateral control values but inhibited development of structural strength and stiffness in healing MCLs. These results suggest that in the rabbit, short-term immobilization of an ACL-deficient knee offers some advantages to the joint and to certain low load behaviors of the healing MCL, but it also results in a smaller quantity of scar tissue that is less able to resist higher loads. Longer-term studies involving remobilization are necessary before the effects of brief immobilization on joint damage and MCL healing in this ACL-deficient model can be fully defined.
Journal of Manipulative and Physiological Therapeutics, 1999
Background: Evidence exists that conservative rehabilitation protocols fail to achieve full recov... more Background: Evidence exists that conservative rehabilitation protocols fail to achieve full recovery of muscle strength and function after joint injuries. The lack of success has been attributed to the high amount of muscle inhibition found in patients with pathologic conditions of the knee joint. Clinical evaluation shows that anterior knee pain is typically associated with sacroiliac joint dysfunction, which may contribute to the muscle inhibition observed in this patient group. Objective: To assess whether sacroiliac joint manipulation alters muscle inhibition and strength of the knee extensor muscles in patients with anterior knee pain. Design and Setting: The effects of sacroiliac joint manipulation were evaluated in patients with anterior knee pain. The manipulation consisted of a high-velocity low-amplitude thrust in the side-lying position aimed at correcting sacroiliac joint dysfunction. Before and after the manipulation, torque, muscle inhibition, and muscle activation for the knee extensor muscles were measured during isometric contractions using a Cybex dynamometer, muscle stimulation, and electromyography, respectively. Participants: Eighteen patients (mean age, 30.5 ± 13.0 years) with either unilateral (n = 14) or bilateral (n = 4) anterior knee pain. Results: Patients showed substantial muscle inhibition in the involved and the contralateral legs as estimated by the interpolated twitch technique. After the manipulation, a decrease in muscle inhibition and increases in knee extensor torques and muscle activation were observed, particularly in the involved leg. In patients with bilateral anterior knee pain, muscle inhibition was decreased in both legs after sacroiliac joint adjustment. Conclusions: Spinal manipulation might offer an interesting alternative treatment for patients with anterior knee pain and muscle inhibition. Because this clinical outcome study was of descriptive nature rather than a controlled design, biases might have occurred. Thus the results have to be verified in a randomized, controlled, double-blinded trial before firm conclusions can be drawn or recommendations can be made.
Testing environment is an important factor in the outcome of mechanical tests on connective tissu... more Testing environment is an important factor in the outcome of mechanical tests on connective tissue. The purpose of this investigation was to determine the effect of ligament water content on ligament mechanical behaviour by altering the test environment. Water content of medial collateral ligament (MCLs) from 19 three-month-old New Zealand White rabbits was varied in subsets of ligaments pairs by means of immersion in 2,10 or 25% sucrose or 0.9% phosphate-buffered saline (PBS) solutions for 1 h. One knee joint was cycled 50 times in the designated solution (experimental), while the contralateral knee (uncycled control) was simultaneously soaked in the same tank. Following cycling, the water contents of both test and control ligaments were determined. Water contents of 22 normal MCLs were determined immediately post-sacrifice and served as 'normal water content' controls. Normalized peak cyclic load changes were used as a measure of the viscoelastic behaviour of each MCL. Results demonstrated that only ligaments soaked (but not cycled) in a 10% sucrose solution had water contents @S&2.5%) which were statistically similar to the 22 fresh normal MCLs (63.9 f 6.0%). Ligaments soaked in PBS (74.0& 1.3%) or 2% sucrose (69.2 f 2.3%) had significantly higher water contents compared to fresh normal MCLs. Ligaments with higher water contents (e.g. soaked in PBS or 2% sucrose) demonstrated greater cyclic load relaxation compared to ligaments with lower contents (e.g. soaked in 25 or 10% sucrose). Different fluid test environments can significantly alter ligament water content and, in turn, significantly affect ligament viscoelastic behaviour.
Sympathetic-derived neuropeptide Y (NPY) helps regulate inflammatory responses in injury and dise... more Sympathetic-derived neuropeptide Y (NPY) helps regulate inflammatory responses in injury and disease, is a vasoconstrictor, and stimulates angiogenesis. Rupture of the anterior cruciate ligament (ACL) is a common clinical presentation that results in tissue inflammation, hyperemia, and angiogenesis in the intact medial collateral ligament (MCL). This study is the first to examine the vasoregulatory role of NPY in ACL-deficient knee joints by using the newly developed technique of laser speckle perfusion imaging (LSPI). MCL blood flow was measured in two groups of adult rabbits: unoperated control ( n = 6), and 6-wk ACL transected ( n = 5). Under anesthesia, the MCL was surgically exposed and tissue blood flow was imaged at high resolution using LSPI. NPY was applied to the MCL vasculature in topical boluses of 100 μl (dose range 10−14 to 10−9 mol), and the α-adrenoceptor agonist phenylephrine was applied in doses of 10−14, 10−10, and 10−7 mol. In control rabbits, topical administrat...
An image analysis technique has been developed to quantitatively describe the fine vascular patte... more An image analysis technique has been developed to quantitatively describe the fine vascular patterns observed in ligament tissue. The longitudinal orientational distribution and total vessel volume of India-ink-perfused blood vessel segments in normal and healing ligaments were determined. The methods involved special vascular preparation of adult rabbit knee medial collateral ligaments (MCL) by India-ink perfusion. Black and white microscope images of ink-perfused tissue sections were subjected to a thresholding procedure to binarize digitized ligament images, which were then skeletonized and analyzed for directional distribution based on the least-squares technique. Analysis of medial collateral ligaments in New Zealand White rabbits using this method has shown that scarred tissue is more vascular and has a more chaotic angular distribution of blood-vessel segments than normal ligament tissue.
Objectives. This study was aimed at investigating muscle strength and quadriceps inhibition in pa... more Objectives. This study was aimed at investigating muscle strength and quadriceps inhibition in patients with unilateral anterior knee pain syndrome. Design. Functional assessments were made before an arthroscopic knee surgery and 6 weeks and 6 months following the surgical intervention. Background. Traumatic knee injuries have been associated with severe muscle inhibition of the knee extensor muscles. Muscle inhibition is a serious hindrance in the rehabilitation process and prevents full functional recovery of the affected joint. Methods. 30 patients participated in the study. Isometric quadriceps strength was measured using a KinCom dynamometer. Muscle inhibition was assessed using the interpolated twitch technique which requires applying a single electrical twitch to the femoral nerve during a maximal isometric knee extensor contraction. Pain was assessed with a 100 mm visual analogue pain scale. Results. Pre-surgery, substantial muscle inhibition and pain was observed in the affected leg compared with the contralateral leg. Over the 6 month period there was a decrease in pain and muscle inhibition, although the decrease in muscle inhibition failed to reach statistical significance. Muscle strength showed a decrease 6 weeks post-surgery followed by an increase 6 months following surgery compared with pre-surgical values. Conclusions. The arthroscopic intervention was successful in reducing pain in patients with unilateral anterior knee pain syndrome. However, muscle inhibition was still substantial 6 months following surgery and was significantly higher in the affected and contralateral limb than in normal subjects. Relevance Clinical evidence indicates that conservative rehabilitation programs are not entirely successful in improving muscle strength and function in patients with severe joint injuries. The lack of success has typically been attributed to the high amount of muscle inhibition found in these patients. The present study confirmed a persisting muscle inhibition 6 months following knee surgery not only in the affected but also in the contralateral leg. Future studies should be aimed at designing exercise protocols that successfully eliminate muscle inhibition and, thus, potentially allow for full restoration of muscle function.
Previous work has shown a progressive, age-related loss of knee joint innervation in the C57BL6Ni... more Previous work has shown a progressive, age-related loss of knee joint innervation in the C57BL6Nia mouse. We did three experiments to describe further the loss and determine whether it might contribute to the development of knee osteoarthrosis in this model. Immunocytochemistry showed that the percentage of neurons expressing substance P and calcitonin gene-related peptide increased with age, indicating a relatively selective loss of mechanoreceptors. Histological examination of knee joints of mice at various ages showed that loss of joint innervation always preceded histological changes of cartilage degeneration. The mice usually developed a mild form of osteoarthrosis, but surgical ablation of joint innervation caused the development of severe patellofemoral osteoarthrosis. The ndings are consistent with the hypothesis that an age-related loss of joint innervation may contribute to the development of osteoarthrosis.
Background and Objective: Laser speckle perfusion imaging (LSPI) is a minimally invasive optical ... more Background and Objective: Laser speckle perfusion imaging (LSPI) is a minimally invasive optical measure of relative changes in blood flow, providing real-time, high resolution, two-dimensional maps of vascular structure. Standard LSI imaging uses a light-reflective geometry that limits the measurement to a thin surface layer of 0.2-1 mm. The objective of this study was to test a new LSI instrument geometry with the laser source opposed to the image capture plane (light transmissive). Captured light then travels the entire tissue thickness (10-15 mm), sampling much deeper regions of interest than conventional optical imaging techniques. Study Design: Reflective-light (conventional) and transmissive-light LSI modes were used to measure finger joint blood flow during a timed tourniquet occlusion of the brachial artery in volunteer participants. Results: There was greatly increased visibility of vessels underlying the skin in the light-transmissive mode LSI mode. Established LSI algorithms were shown to still work in the light-transmissive mode, despite decorrelation due to finite laser coherence length and the light passing through a tissue thickness of 10-15 mm. Conclusion: Transmissive LSI can be used to measure blood flow deep (10-15 mm) into tissues. This could be useful for non-invasive measurements of finger joint synovial blood flow in diagnosing and treating peripheral vascular disorders, such as rheumatoid arthritis.
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Papers by Robert Bray