RCBF-quantification with 99mTc-HMPAO-SPECT: theory and first results

The activity concentrations of 99mTc-HMPAO in brain after intravenous injection were evaluated in 25 patients using SPECT. With additional first pass studies of heart and brain with the short lived isotope 195mAu, the cardiac output and the mean cerebral transit times of the patients were measured a short time before the HMPAO injection. The time dependence of 99mTc-HMPAO activity in the brain was registered during the first 5 min after injection over both hemispheres. Using a simplified three compartment model it was possible to calculate the mean retention fraction of HMPAO in brain from the time activity curves. It could be shown that the regional cerebral blood flow in ml/min per 100 g can be calculated from the activity concentration of HMPAO in the brain, the cardiac output and the retention fraction. In 15 of the 25 cases the rCBF was measured using the planar 133Xe inhalation method as a reference. We found a significant correlation between the mean hemisphere CBF from our method and the mean hemisphere grey matter CBF from the Xenon method with a correlation coefficient of 0.73.     

The behavior of99mTc-hexamethylpropyleneamineoxime (99mTc-HMPAO) in blood and brain

^99mTc-hexamethylpropyleneamineoxime (^99mTc-HMPAO) is a reagent for scanning cerebral blood flow. We investigated how^99mTc-HMPAO changed in the blood and brain. The^99mTc-HMPAO, which was prepared by adding of^99mTcO _- ⁴ to HMPAO and Sn(II), consisted of primary and secondary complexes, reduced hydrolyzed^99mTc, and^99mTc0pertechnetate. The percentage of the primary complex in^99mTc-HMPAO decreased with time after preparation. The primary complex converted to the secondary one very rapidly in the presence of plasma. When^99mTc-HMPAO was injected into patients,^99mTc activity was immediately partitioned in the plasma fraction, with approximately 60% in whole blood. In plasma,^99mTc was found to be associated with proteins such as albumin and globulin.^99mTc trapped in red cells was not washed out with either plasma or saline. Biodistribution studies showed that the less lipophilic compounds of^99mTc-HMPAO could not pass through the blood brain barrier (BBB), and therefore did not accumulate in the brain. The results of gel chromatography and equilibrium dialysis indicated that no specific^99mTc binding protein was present in the brain. Considering the instability of^99mTc-HMPAO in vivo, we proposed that the speed at which the primary complex converted to the less lipophilic compounds was important in allowing^99mTc-HMPAO to pass through the BBB and to be fixed in the brain.     

Regional differences between <Superscript>99m</Superscript>Tc-ECD and <Superscript>99m</Superscript>Tc-HMPAO SPET in perfusion changes with age and gender in healthy adults

A number of studies using single-photon emission tomography (SPET) have shown perfusion changes with age in several cortical and subcortical areas, which might distort the results of perfusion imaging studies of neuropsychiatric disorders. Technetium-99m labelled ethyl cysteinate dimer (ECD) and hexamethylpropylene amine oxime (HMPAO) are both used as markers of cerebral perfusion, but have different pharmacokinetics and retention patterns. The aim of this study was to determine whether age and gender effects on perfusion SPET differ depending on whether ^99mTc-HMPAO or ^99mTc-ECD is used. Forty-five subjects (20 male and 25 female, mean age 52.8±6.6 years) were assigned to ^99mTc-HMPAO SPET (HMPAO group), and 39 subjects (24 male and 15 female, mean age 52.6±6.7 years) to ^99mTc-ECD SPET (ECD group). SPET images were obtained about 10 min after intravenous injection of approximately 800 MBq ^99mTc-HMPAO or ^99mTc-ECD using the same SPET scanner. Three-dimensional volumetric magnetic resonance imaging was performed to as7sess morphological changes in the grey matter. All image processing and statistical analyses were performed using SPM99 software. An area in the right anterior frontal lobe showed an increase in perfusion with age only in the HMPAO group, whereas areas in the bilateral retrosplenial cortex showed decreases in perfusion with age only in the ECD group; neither group showed corresponding changes in the grey matter. The present study shows that different effects of age on perfusion are observed depending on whether ^99mTc-HMPAO and ^99mTc-ECD is used. This suggests that the results of perfusion SPET are differently confounded depending on the tracer used, and that perfusion SPET with these tracers has limitations when used in research on subtle perfusion changes.     

In vivo metabolism and kinetics of99mTc-HMPAO

The cerebral distribution of^99mTc-labeled d, l, hexamethyl-propylene-amine-oxime (^99mTc-HMPAO) as a function of rCBF and time was examined in rats and in man. The results of this study confirm that^99mTc-HMPAO is distributed in brain in proportion to rCBF. However, the rapid systemic breakdown of the tracer in blood results in considerable difficulties in the assessment of the arterial concentration of the parent compound; incomplete extraction of^99mTc-HMPAO from blood to brain and significant efflux from brain represent further limitations in the use of this tracer for quantification of rCFB. Despite these limitations^99mTc-HMPAO is of potential interest for a qualitative assessment of rCBF in specific clinical conditions.     

“Luxury perfusion” with99mTc-HMPAO and123I-IMP SPECT imaging during the subacute phase of stroke

To compare the merits of¹²³I-isopropyl-iodoam-phetamine (¹²³I-IMP) and^99mTc-HMPAO in showing abnormal brain uptake distribution during cerebral ischemia, we studied ten patients during the subacute phase of their stroke, a period where metabolism and blood flow are frequently uncoupled. SPECT imaging was performed using both radiopharmaceuticals in the 10 patients from 48 h to 4 weeks after onset of symptoms. Two patients out of the 10 had similar defects with¹²³I-IMP and^99mTc-HMPAO SPECT, the location of the defects corresponding to the area of infarction observed on CT. Six patients had normal^99mTc-HMPAO SPECT and abnormal¹²³I-IMP SPECT with defects in the area of infarction shown by CT. The remaining 2 patients had hyperactive abnormalities on^99mTc-HMPAO in areas corresponding to defects on the¹²³I-IMP images. Two of the patients with SPECT mismatches were studied again more than 1 month after onset. On reexamination,^99mTc-HMPAO SPECT which was previously normal or hyperactive became hypoactive with a focal area of decreased activity corresponding to the defect on¹²³I-IMP. Crossed cerebellar diaschisis was found in 7 patients with^99mTc-HMPAO and was absent for both¹²³I-IMP and^99mTc-HMPAO in 3. We suggest that SPECT with⁹⁹mTc-HMPAO could show transient hyperemia not demonstrated by¹²³I-IMP whereas in some cases cerebral infarction would be more difficult to demonstrate with^99mTc-HMPAO than with¹²³I-IMP. SPECT with both tracers is recommended to follow the evolution of strokes in terms of regional cerebral blood flow and tissue metabolism.     

A study of acetazolamide-induced changes in cerebral blood flow using99mTc HMPAO SPECT in patients with cerebrovascular disease

For semiquantification of SPECT studies we tried to calculate cerebral^99mTc-HMPAO uptake related to injected dose and estimated brain volume. The method was applied to SPECT investigations of 27 patients who had a least one ischaemic attack and a confirmed 80–100% stenosis of the corresponding internal carotid artery (ICA). Vascular reactivity was tested by parenteral administration of acetazolamide (AZ). Increase in HMPAO uptake after AZ was evident in both hemispheres, although the increase (AZ effect) was significantly lower in the affected hemisphere (+24% versus +28%). No interhemispheric uptake differences were seen in patients with largely normal SPECT studies, although local asymmetries in HMPAO deposition were visible. Patients with low density lesions on CT and with a well-demarcated lesion in the same location on SPECT revealed interhemispheric uptake differences, with lower uptake on the affected side. This was not due solely to alterations in the lesion, but also to reduced HMPAO uptake and AZ effect in the surrounding area. The AZ effect showed no correlation with angiographic findings, indicating no major haemodynamic influence of the ICA stenosis on cerebral hemisphere perfusion. Calculated cerebral HMPAO uptake changes after AZ administration were in good accordance with absolute cerebral blood flow measurements, and made interindividual comparisons possible. However, as changes in the area around an infarct or local reduction in vascular reserve may not be reproduced adequately by uptake calculations, visual inspection is still necessary.     

Visualization of pressure-dependent luxury perfusion in a patient with subacute cerebral infarction

Luxury perfusion characterized by depressed metabolism compared with CBF might be changed by decreasing cerebral perfusion pressure during the sitting position. A 77-yr-old man with subacute cerebral infarction was studied with brain X-ray computed tomography (CT), raise-up test with^99mTc-d,1-hexamethylpropyleneamine oxime (HMPAO) brain single photon emission tomography (SPECT) and positron emission tomography (PET). Brain X-ray CT revealed a low-density area in the left middle cerebral artery (MCA) anterior area. Raise-up^99mTc-HMPAO brain SPECT revealed decreased uptake in the left MCA anterior area in the sitting position and subsequent supine^99mTc-HMPAO brain SPECT revealed hot accumulation there. PET study in the supine position demonstrated some differences between CBF and the cerebral metabolic rate for oxygen in the left MCA anterior area, indicating luxury perfusion. CBF in the area of luxury perfusion might be decreased during the sitting or standing position and increased during the supine position by dysautoregulation of the cerebral vessels in the luxury perfusion during the subacute infarct.     

An evaluation of99mTc-HMPAO uptake in cerebral gliomas —a comparison with X-ray CT

Nineteen patients with biopsy-proven cerebral gliomas were studied with^99mTc-HMPAO single photon emission tomography (SPELT) imaging and X-ray computed tomography (CT). The uptake of^99mTc-HMPAO was correlated with tumour size and morphology as shown by X-ray CT, and overall patient survival. It appears that uptake of^99mTc-HMPAO is associated with larger, ill-defined tumours and was an adverse factor in patient survival. In those tumours with normal or increased uptake,^99mTc-HMPAO imaging is useful in distinguishing the tumour margin from surrounding oedema.     

Can technetium 99m bisdiethylphosphinoethanebis-t butylisocyanide (99mTc-DEPIC) be used for routine radionuclide ventriculography?

Radionuclide ventriculography is a useful investigation in the evaluation of cardiac function. Generally, in vivo technetium 99m-labelled red blood cells (RBC) yield good quality images in ventriculography. However, it is widely believed that some drugs have an adverse effect on RBC labelling. Zanelli et al. (1987) developed a radiopharmaceutical (technetium 99m bisdiethylphosphinoethanebis-t-butylisocyanide,^99mTc-DEPIC) to obtain better results in patients using such drugs. We untertook a prospective study of 6 patients with cardiovascular and/or pulmonary disease using several kinds of drugs to evaluate imaging of the cardiac blood pool with^99mTc-DEPIC and in vivo labelled^99mTc-RBC. After injection, blood samples were taken, and gated equilibrium blood pool studies were performed. The radiochemical purity of the injected^99mTc-DEPIC varied from 76.4 to 93.6% (mean 86.4%, SD 5.7%). The protein (pre-albumin) binding was 100%. Biological half-life in blood varied from 3.3 to 4.7 h (mean 4.1 h, SD 0.5 h). For^99mTc-RBC no significant blood disappearance was seen for 8 h. The percentage of RBC-bound^99mTc varied from 96.9% to 98.3% (mean 97.0%, SD 0.5%) and was stable for at least 8 h. The heart-to-lung, heart-to-spleen, and heart-to-liver ratios were higher for^99mTc-RBC than for^99mTc-DEPIC. Furthermore,^99mTc-DEPIC showed a significant decline of the ejection fraction with time. Visually, the images with^99mTc-RBC were superior to those with^99mTc-DEPIC, especially a few hours after injection. According to our findings, in vivo labelling of^99mTc-RBC is still the method of choice for routine radionuclide ventriculography. The decline of the ejection fraction, the short blood half-life, and the intense liver uptake make^99mTc-DEPIC less suitable for this purpose.     

Evaluation of a new leukocyte labeling procedure with 99mTc-HMPAO

A technique for labeling leukocytes with ^99mTc using hexamethylpropylene-amineoxime (HMPAO) was evaluated in vitro. The labeling procedure resulted in a cell bound fraction of radioactivity of 56% after ^99mTc incubation and 96% after 1 washing of cells. In the final cell suspension 84% of the radioactivity was attached to the polymorphonuclear (PMN) leukocytes constituting 94% of white cells. Only 5% was bound to residual red blood cells. The stability evaluated in autologous plasma showed a decline of cell bound activity from 96% to 84% over 3 h. The chemotactic function of PMN leukocytes was unaffected by the labeling procedure. These findings demonstrate that HMPAO, albeit cell unspecific, is efficient for labeling PMN leukocytes with ^99mTc. The stability of the labeling procedure is high and the technique does not affect cell function. No other current ^99mTc leukocyte labeling technique possesses all these qualities.     

Cerebral blood flow measurement in patients with impaired consciousness: usefulness of 99mTc-HMPAO single-photon emission tomography in clinical practice

The relationship between impairment of consciousness and quantitative cerebral blood flow (CBF) was investigated. The mean CBF of the whole brain was measured by the Patlak-plot method using technetium-99m hexamethylpropylene amine oxime single-photon emission tomography (^99mTc-HMPAO SPET) in patients with the following diseases: cerebral infarction, intraparenchymal haemorrhage, subarachnoid haemorrhage, brain tumour and cerebral contusion. The clinical symptoms were evaluated according to the severity of impaired consciousness, aphasia and dementia. Four hundred and eighty-five CBF measurements were performed. Patients with alert consciousness showed an age-related decline in mean CBF. Patients with aphasia showed a significant reduction in mean CBF compared with those without aphasia. Impaired consciousness was proportional to reduction in mean CBF regardless of types of pathology, and the size of lesion did not influence the mean CBF. Patients with dementia showed a significant reduction in mean CBF proportional to the severity of dementia. The quantitative measurement of CBF using ^99mTc-HMPAO SPET is reliable in clinical evaluations. Received 9 March and in revised form 23 May 1998     

Targeting murine heart and brain: visualisation conditions for multi-pinhole SPECT with <Superscript>99m</Superscript>Tc- and <Superscript>123</Superscript>I-labelled probes

Purpose  The study serves to optimise conditions for multi-pinhole SPECT small animal imaging of ¹²³I- and ^99mTc-labelled radiopharmaceuticals with different distributions in murine heart and brain and to investigate detection and dose range thresholds for verification of differences in tracer uptake. Methods  A Triad 88/Trionix system with three 6-pinhole collimators was used for investigation of dose requirements for imaging of the dopamine D₂ receptor ligand [¹²³I]IBZM and the cerebral perfusion tracer [^99mTc]HMPAO (1.2–0.4 MBq/g body weight) in healthy mice. The fatty acid [¹²³I]IPPA (0.94 ± 0.05 MBq/g body weight) and the perfusion tracer [^99mTc]sestamibi (3.8 ± 0.45 MBq/g body weight) were applied to cardiomyopathic mice overexpressing the prostaglandin EP₃ receptor. Results  In vivo imaging and in vitro data revealed 45 kBq total cerebral uptake and 201 kBq cardiac uptake as thresholds for visualisation of striatal [¹²³I]IBZM and of cardiac [^99mTc]sestamibi using 100 and 150 s acquisition time, respectively. Alterations of maximal cerebral uptake of [¹²³I]IBZM by >20% (116 kBq) were verified with the prerequisite of 50% striatal of total uptake. The labelling with [^99mTc]sestamibi revealed a 30% lower uptake in cardiomyopathic hearts compared to wild types. [¹²³I]IPPA uptake could be visualised at activity doses of 0.8 MBq/g body weight. Conclusion  Multi-pinhole SPECT enables detection of alterations of the cerebral uptake of ¹²³I- and ^99mTc-labelled tracers in an appropriate dose range in murine models targeting physiological processes in brain and heart. The thresholds of detection for differences in the tracer uptake determined under the conditions of our experiments well reflect distinctions in molar activity and uptake characteristics of the tracers. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

99mTc-MDP retention in osteoporosis: Relationship to other indices of bone cell activity and response to calcium and vitamin D therapy

Serum calcium, albumin, phosphorus, and alkaline phosphatase, urinary creatinine and retention of ^99mTc-methylene bisphosphonate (^99mTc-MDP) were measured in 61 subjects with osteoporosis and the values compared with those obtained in normal subjects. ^99mTc-MDP retention was inversely related with urinary creatinine output in normal subjects. In osteoporotic subjects urinary creatinine output was lower and ^99mTc-MDP retention higher even when urinary creatinine output was taken into account. Other measurements were similar. In 21 subjects these measurements together with urinary hydroxyproline were performed before and after treatment with calcium and vitamin D. ^99mTc-MDP and alkaline phosphatase fell; urinary hydroxyproline was unchanged. A single 24 h urine measurement after ^99mTc-MDP injection is a valuable method of predicting whether calcium and vitamin D therapy will be useful in a particular case of osteoporosis.     

Age-related changes in brain perfusion of normal subjects detected by 99mTc-HMPAO SPECT

Previous functional imaging data generally show impairment in global cerebral blood flow (CBF) with age. Conflicting data, however, concerning age-related changes in regional CBF (rCBF) have been reported. We examined the relative rCBF in a sample of healthy subjects of various ages, to define and localize any age-related CBF reduction. Twenty-seven healthy subjects (17 male, 10 female; mean age 49 ± 15, range 26–71, median 47 years) were studied by ^99mTc-HMPAO brain SPECT. The younger age group consisted of subjects below, the older group above 47 years of age, respectively. Analysis was performed by applying three preformed templates, each containing delineated regions of interest (ROIs), to three transaxial brain slices at approximately 4, 6, and 7 cm above the orbitomeatal line (OML). The average number of counts for each ROI was normalized to mean uptake of the cerebellum and of the whole brain slice. Globally, ^99mTc-HMPAO uptake ratio normalized to cerebellum was significantly decreased in older subjects, affecting both hemispheres. A slight left-to-right asymmetry was observed in HMPAO uptake of the whole study group. It did not, however, change with age. Regionally, both cortical and subcortical structures of older subjects were involved: uptake ratio to cerebellum was significantly lower (after correction for multiple testing) in the left basal ganglia and in the left superior temporal, right frontal and bilateral occipital cortices at 4 cm above the OML. At 6 cm above the OML, reduced uptake ratios were identified in the left frontal and bilateral parietal areas. At 7 cm, reduced uptake was detected in the right frontal and left occipital cortices. Most of these differences were reduced when uptake was normalized to whole slice, whereas an increase in uptake ratios was observed in the cingulate cortex of the elderly. An inverse correlation between age and HMPAO uptake ratios normalized to cerebellum was observed in a number of brain regions. These findings suggest that advancing age has a differential effect on cerebral perfusion reflected in brain ^99mTc-HMPAO uptake. Received: 8 November 1996 Accepted: 26 August 1997     

A quantitative approach to technetium-99m ethyl cysteinate dimer: a comparison with technetium-99m hexamethylpropylene amine oxime

To develop non-invasive regional cerebral blood flow (rCBF) measurements using technetium-99m ethyl cysteinate dimer (^99mTc-ECD) and single-photon emission tomography (SPET), the same graphical analysis as was described in our previous reports using technetium-99m hexamethylpropylene amine oxime (^99mTc-HMPAO) was applied to time-activity data for the aortic arch and brain hemispheres after intravenous injection of^99mTc-ECD. Hemispherical brain perfusion indices (BPI) for^99mTc-ECD showed a highly significant correlation (n = 22,r = 0.935,P = 0.0001) with those for^99mTc-HMPAO in 11 patients who underwent both tracer studies. Using both linear regression line equations between^99mTc-ECD BPI and^99mTc-HMPAO BPI and between^99mTc-HMPAO BPI and mean cerebral blood flow (CBF) values obtained from a xenon-133 inhalation SPET method in a previous study,^99mTc-ECD BPI was converted to¹³³Xe CBF values (y = 2.60x + 19.8). Then raw SPET images of^99mTc-ECD were converted to rCBF maps using Lassen's correction algorithm. In this algorithm, the correction factor a was fixed to 1.5, 2.6 and infinite. In the comparison of rCBF values for^99mTc-ECD SPET with those for^99mTc-HMPAO SPET in 396 regions of interest in the aforementioned 11 patients, the fixed correction factor of 2.6 gave nearly the same rCBF values for^99mTc-ECD (50.1 ± 16.9 ml/100 g/min, mean ± SD) as for^99mTc-HMPAO (49.9 ± 17.3 ml/100 g/min). In conclusion, the same non-invasive method as has been used in^99mTc-HMPAO studies is applicable to a^99mTc-ECD study for the measurement of rCBF without any blood sampling.     

Discrepant99mTc-ECD images of CBF in patients with subacute cerebral infarction: A comparison of CBF,CMRO2 and99mTc-HMPAO imaging

Three patients with subacute ischemic cerebral infarction examined by SPECT with^99mTc-ECD and PET within the same day showed signs of luxury perfusion in the subacute phase, which is between 9 to 20 days after the onset. A^99mTc-HMPAO SPECT study was also performed within 2 days of the ECD-SPECT study. ECD-SPECT images of three patients displayed a focal decreased uptake in the infarcted lesions, while in infarcted foci, there was almost equivalent or increased CBF compared to normal and unaffected areas, decreased CMRO₂, and high HMPAO uptake. The ECD-SPECT results were similar to those of CMRO₂ rather than CBF, though the HMPAO-SPECT image was similar to that of CBF. In one patient, HMPAO images revealed hyperfixation of the tracer. In the chronic phase and in the acute phase before 5 days after the onset, there were no discrepancies among the ECD-SPECT, CBF, HMPAO-SPECT, and CMRO₂ images. These observations indicated that^99mTc-ECD is a good indicator of damaged brain tissues in subacute ischemic infarction. They also suggested that^99mTc-ECD is a potential agent with which to evaluate cerebral tissue viability in some pathological states of cerebrovascular disease. The characteristics may be suitable for confirming the effects of thrombolytic therapy in acute ischemia, because these conditions often show signs of luxury perfusion when the therapy is successful.     

The Clinical Usefulness of 99mTc HMPAO Leukocyte/99mTc Phytate Bone Marrow Scintigraphy for Diagnosis of Prosthetic Knee Infection: A Preliminary Study

Purpose

The preferred radionuclide imaging procedure for diagnosing prosthetic joint infection is combined radiolabeled leukocyte/^99mTc sulfur colloid bone marrow scintigraphy, which has an accuracy of over 90 %. Unfortunately, sulfur colloid is no longer available in South Korea. In this study, we evaluated the usefulness of ^99mTc phytate, a substitute for ^99mTc sulfur colloid, when combined with radiolabeled leukocyte scintigraphy in suspected prosthetic knee infections.

Methods

Eleven patients (nine women, two men; mean age 72 ± 6 years) with painful knee prostheses and a suspicion of infection underwent both ^99mTc HMPAO leukocyte scintigraphy (LS) and ^99mTc phytate bone marrow scintigraphy (BMS). The combined images were interpreted as positive for infection when radioactivity in the LS at the site of clinical interest clearly exceeded that of the BMS (discordant); they were interpreted as negative when the increased activity in the LS was consistent with an increased activity in the BMS (concordant). The final diagnosis was made with microbiological or intraoperative findings and a clinical follow-up of at least 12 months.

Results

Five of eleven patients were diagnosed as having an infected prosthesis. The overall sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy of the combined LS/BMS were 100 %, 83 %, 83 %, 100 % and 91 %, respectively.

Conclusion

We find that combined ^99mTc HMPAO LS/^99mTc phytate BMS shows comparable diagnostic performance to other studies utilizing sulfur colloid. Combined ^99mTc HMPAO LS/^99mTc phytate BMS is therefore expected to be an acceptable alternative to combined radiolabeled LS/^99mTc sulfur colloid BMS for diagnosing prosthetic knee infections.     

Diagnosis of brain death with technetium 99m hexamethylpropylene amine oxime

We report on the utility of technetium 99m hexamethylpropylene amine oxime (^99mTc-HMPAO) to diagnose brain death following cardiac surgery on a 49-year-old man with triple-vessel coronary artery disease. The imaging parameters and criteria to diagnose irreversible brain damage (brain death) with ^99m-TcHMPAO are outlined. Brain imaging with this tracer seems to be more reliable than classic radionuclide angiography and has a potential value to confirm the diagnosis of brain death at an early stage. In addition, it may be used to evaluate the perfusion pattern to other vital organs with potential for transplantation.Partially presented as a poster demonstration to the 19th Annual Meeting of the BNMS. Offprint requests to: D.C. Costa     

Finding an optimal method for imaging lymphatic vessels of the upper limb

Lymphoscintigraphy involves interstitial injection of radiolabelled particulate materials or radioproteins. Although several variations in the technique have been described, their place in clinical practice remains controversial. Traditional diagnostic criteria are based primarily on lymph node appearances but in situations such as breast cancer, where lymph nodes may have been excised, these criteria are of limited use. In these circumstances, lymphatic vessel morphology takes on greater importance as a clinical endpoint, so a method that gives good definition of lymphatic vessels would be useful. In patients with breast cancer, for example, such a method, used before and after lymph node resection, may assist in predicting the development of breast cancer-related lymphoedema. The aim of this study was to optimise a method for the visualisation of lymphatic vessels. Subcutaneous (sc) and intradermal (id) injection sites were compared, and technetium-99m nanocolloid, a particulate material, was compared with ^99mTc-human immunoglobulin (HIG), which is a soluble macromolecule. Twelve normal volunteers were each studied on two occasions. In three subjects, id ^99mTc-HIG was compared with sc ^99mTc-HIG, in three id ^99mTc-nanocolloid was compared with sc ^99mTc-nanocolloid, in three id ^99mTc-HIG was compared with id ^99mTc-nanocolloid and in three sc ^99mTc-HIG was compared with sc ^99mTc-nanocolloid. Endpoints were quality of lymphatic vessel definition, the time after injection at which vessels were most clearly visualised, the rate constant of depot disappearance (k) and the systemic blood accumulation rate as measured by gamma camera imaging over the liver or cardiac blood pool. Excellent definition of lymphatic vessels was obtained following id injection of either radiopharmaceutical, an injection route that was clearly superior to sc. Differences between radiopharmaceuticals were less clear, although after id injection, ^99mTc-HIG gave images that were marginally but significantly better than those given by ^99mTc-nanocolloid. Image quality correlated inversely with time after injection at which the best image was obtained, consistent with the notion that good vessel definition was dependent on a narrow bolus width. k was approximately three times higher after id injection than after sc injection but it was not significantly different between radiopharmaceuticals for either injection route. Intradermal ^99mTc-HIG gave a cardiac blood pool signal that, over the first 60 min, increased about five times faster than that with sc ^99mTc-HIG, but no clear difference was observed in the rate of increase in hepatic activity between id ^99mTc-nanocolloid and sc ^99mTc-nanocolloid. We conclude that id injection provides rapid access of radiotracers to lymphatic vessels, which is ideal for imaging lymphatic vessel morphology. ^99mTc-HIG is marginally superior to nanocolloid for this purpose and, in drainage basins from which lymph nodes have been excised, is not handicapped by a potentially inferior ability, compared with radiocolloid, to image lymph nodes.     

Early distribution of intravenously injected mesenchymal stem cells in rats with acute brain trauma evaluated by (99m)Tc-HMPAO labeling

Introduction

Stem cell tracking is essential for evaluation of its migration, transplantation and therapeutic response. The aim of this study was to evaluate early distribution of intravenously transplanted rat bone marrow mesenchymal stem cells (BMSCs) in rats with acute cerebral trauma by labeling with ^99mTc-hexamethylpropyleneamine oxime (^99mTc-HMPAO).

Methods

^99mTc-HMPAO-labeled BMSCs were injected intravenously to trauma rats (n=14) and sham-operated controls (n=13). Gamma camera images were acquired at 4 h after injection, and then organs were removed for gamma counting. Confocal microscope was used to confirm the migration of ^99mTc-BMSCs by co-labeling with PKH26. Cytometric analysis was performed to evaluate apoptotic or necrotic change until the seventh day after labeling.

Results

^99mTc-BMSCs were distributed mostly to lungs, liver and spleen at 4 h, and uptake of these organs was not significantly different between traumatic rats and controls. Meanwhile, the cerebral uptake of ^99mTc-BMSCs was significantly higher in the traumatic rats than in controls (0.40% vs. 0.20%; P=.0002). Additionally, ^99mTc-BMSCs' uptake of traumatic hemisphere was significantly higher than that of contralateral ones (0.27% vs. 0.13%; P=.0001) in traumatic rats. Regardless of radiolabeling, BMSCs migrated to traumatic regions, but not to nontraumatic hemispheres. However, gamma camera failed to demonstrate ^99mTc-BMSCs in traumatic hemispheres. No significant apoptotic or necrotic change was observed until 7 days after radiolabeling.

Conclusions

Early distribution of BMSCs in traumatic brain disease could be monitored by ^99mTc-labeling, which does not induce cellular death. However, our data showed that the amount of migrated ^99mTc-BMSCs was not enough to be demonstrated by clinical gamma camera.