Comparison of bone SUV obtained from different SPECT/CT systems.

OBJECTIVE: In bone scintigraphy, it is difficult to compare quantitative values, such as standardized uptake value (SUV), obtained from 2 different single-photon emission computed tomography combined with computed tomography (SPECT/CT) devices owing to differences of imaging acquisition and analysis methods. Therefore, the purpose of this study was to compare the SUV obtained from different SPECT/CT devices using the ratio to normal bone, and to analyze the correlation between them. SUBJECTS AND METHODS: A total of 27 prostate cancer patients who underwent bone scintigraphy either using Symbia T16 or Symbia Intevo (Siemens Medical Solutions, Erlangen, Germany) were retrospectively analyzed. In each patient, spherical voxels of interest were placed on the following 5 reference points: humeral head (humerus), femoral neck (femur), lower part of the ilium (ilium), first lumbar vertebra (L1), fifth lumbar vertebra (L5), and the maximum SUV (SUVmax) and average SUV (SUVave) of these regions were measured. RESULTS: The mean and variance of SUVave (humerus) was the smallest on both SPECT/CT. To compare the SUV obtained from the 2 devices, the SUVave ratio and SUVmax ratio of each region were calculated as the SUVave and SUVmax of each region divided by the SUVave of the humeral head in each patient. Median values of the SUVmax ratio and SUVave ratio of each region showed similar tendencies in both devices, with correlation coefficients between 0.93 and 1.19. CONCLUSION: Our results demonstrated that by expressing the quantitative value of SUVave of each region as a ratio to the SUVave of the humeral head, accumulation in the targeted bone can be compared even when the imaging acquisition and analysis methodsare different.

Usefulness of texture analysis in 123I-FP-CIT for the differentiation of AD and DLB subtypes.

OBJECTIVE: I-2ß-carbomethoxy-3ß-(4-iodophenyl)-N-(3-fluoropropyl) nortropane (123I-FP-CIT) is well known to be a useful tracer for differentiating dementia with Lewy bodies (DLB) and Alzheimer disease (AD). However, clinically, there are some cases in which these diseases cannot be differentiated by ordinary quantitative methods. Therefore, in this study, we established an index that reflects not only the total count but also the distribution and heterogeneity of tracer uptake. We investigated whether assessment of the heterogeneous depletion of 123I-FP-CIT is useful for the differentiation of various types of dementia, i.e., probable DLB, possible DLB, and AD, using texture analysis. MATERIALS AND METHODS: A total of 122 patients with either probable DLB (n=35), possible DLB (n=23), AD (n=44), and normal controls (n=20) were analyzed. Summated single photon emission computed tomography (SPECT) images (7 to 10 slices) of the patients, including the bilateral striatum, were analyzed using the gray-level histogram method (GLHM) of texture analysis. Mean, variance, skewness, and kurtosis of GLHM were compared with the specific binding ratio by Livia Tossici-Bolt's method (SBR). RESULTS: The sensitivity and specificity for differentiating probable DLB from possible DLB, AD, and normal controls were 97.1% and 77.0%, respectively, for skewness, using a cut-off point of 6.8%, and 97.1% and 81.6%, respectively, for kurtosis, using a cut-off point of 53.4%. The sensitivity and specificity for differentiating probable and possible DLB from AD and normal controls was 65.5% and 98.4%, respectively, for skewness, using a cut-off point of 6.4%, and 79.3% and 93.8%, respectively, for kurtosis, using a cut-off point of 53.4%. CONCLUSION: In the assessment of the efficacy of 123I-FP-CIT to differentiate AD and DLB subtypes, mean, variance, skewness, and kurtosis by GLHM was as useful as the SBR method. Moreover, possible DLB and probable DLB could be differentiated by skewness and kurtosis. Our results demonstrate that texture analysis is more useful than conventional quantitative methods for obtaining valuable information of the brain. Textural features as such may have considerable potential as imaging biomarkers of DLB progression.

Reproducibility and Variability of Quantitative Cerebral Blood Flow Measured by Multi-delay 3D Arterial Spin Labeling According to Sex and Menstrual Cycle.

Purpose : To determine the reproducibility of corrected quantitative cerebral blood flow (qCBF) through measurement of transit flow time using multi-delay three-dimensional pseudo-continuous arterial spin labeling (pCASL) in healthy men and women and to evaluate the differences in qCBF between not only men and women, but also the follicular and luteal phases of the women's menstrual cycle. Methods : The participants were 16 healthy volunteers (8 men and 8 women ; mean age, 25.3 years). Two MRI were conducted for all participants ; female participants were conducted in the follicular and luteal phases. The reproducibility of qCBF values was evaluated by the intraclass correlation coefficient (ICC) and differences between the two groups were estimated by voxel-based morphometry (VBM) analysis. Results : The qCBF values were lower in men than in women, and those in females were significantly different between the follicular and luteal phases (P < 0.05). In VBM analysis, the qCBF values of the lower frontal lobes were significantly higher in women than in men (P < 0.05). The qCBF values of the frontal pole were significantly higher in the follicular phase than in the luteal phase (P < 0.01). Conclusion : Multi-delay pCASL can reveal physiological and sex differences in cerebral perfusion. J. Med. Invest. 67 : 321-327, August, 2020.

Combination of BMP-2-releasing gelatin/ß-TCP sponges with autologous bone marrow for bone regeneration of X-ray-irradiated rabbit ulnar defects.

The objective of this study is to evaluate the feasibility of gelatin sponges incorporating ß-tricalcium phosphate (ß-TCP) granules (gelatin/ß-TCP sponges) to enhance bone regeneration at a segmental ulnar defect of rabbits with X-ray irradiation. After X-ray irradiation of the ulnar bone, segmental critical-sized defects of 20-mm length were created, and bone morphogenetic protein-2 (BMP-2)-releasing gelatin/ß-TCP sponges with or without autologous bone marrow were applied to the defects to evaluate bone regeneration. Both gelatin/ß-TCP sponges containing autologous bone marrow and BMP-2-releasing sponges enhanced bone regeneration at the ulna defect to a significantly greater extent than the empty sponges (control). However, in the X-ray-irradiated bone, the bone regeneration either by autologous bone marrow or BMP-2 was inhibited. When combined with autologous bone marrow, the BMP-2 exhibited significantly high osteoinductivity, irrespective of the X-ray irradiation. The bone mineral content at the ulna defect was similar to that of the intact bone. It is concluded that the combination of bone marrow with the BMP-2-releasing gelatin/ß-TCP sponge is a promising technique to induce bone regeneration at segmental bone defects after X-ray irradiation.

Statistical evaluation of single-photon emission computed tomography image using smoothed bootstrap method.

Many of the neurodegenerative diseases associated with a decrease in regional cerebral blood flow (rCBF) are untreatable, and the appropriate therapeutic strategy is to slow the progression of the disease. Therefore, it is important that a definitive diagnosis is made as soon as possible when such diseases are suspected. Diagnostic imaging methods, such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT), play an important role in such a definitive diagnosis. Since several problems arise when evaluating these images visually, a procedure to evaluate them objectively is necessary, and studies of image analyses using statistical evaluations have been suggested. However, the assumed data distribution in a statistical procedure may occasionally be inappropriate. Therefore, to evaluate the decrease of rCBF, it is important to use a statistical procedure without assumptions about the data distribution. In this study, we propose a new procedure that uses nonparametric or smoothed bootstrap methods to calculate a standardized distribution of the Z-score without assumptions about the data distribution. To test whether the judgment of the proposed procedure is equivalent to that of an evaluation based on the Z-score with a fixed threshold, the procedure was applied to a sample data set whose size was large enough to be appropriate for the assumption of the Z-score. As a result, the evaluations of the proposed procedure were equivalent to that of an evaluation based on the Z-score.

Quantitative assessment of cerebral hemodynamics using single photon emission computed tomography with venous blood sampling.

OBJECTIVE: Quantitative cerebral blood flow (CBF) measured by single photon emission computed tomography (SPECT) with arterial blood sampling is one of the most reliable methods to assess the hemodynamics in individual patients. SPECT with venous blood sampling is less invasive. The present study compared the measurement of CBF using N-isopropyl-p-(iodine-123)-iodoamphetamine SPECT with venous blood sampling and with arterial blood sampling in patients with major cerebral artery occlusive disease. METHODS: Two normal subjects and 14 patients with major cerebral artery occlusive disease underwent SPECT with arterial and venous blood sampling. The microsphere method was used for quantitative SPECT imaging. Whole brain radioactivity was corrected when the detectors rotated in the forward direction (F1-F7). Venous sampling was performed 30min after radiotracer injection. Arterial blood radioactivity was estimated by multiple regression analysis from these parameters. The cerebrovascular reactivity to acetazolamide was also measured. RESULTS: Multiple regression analysis established the following formula:(where Ca10 is the arterial blood radioactivity at 10min, F1-F7 are the whole brain radioactivity in the forward direction, Cv30 is the venous blood radioactivity at 30min). Mean CBF values were 32.2±6.6ml/100g/min for measured arterial radioactivity and 42.2±7.8ml/100g/min for calculated arterial radioactivity based on venous radioactivity. CONCLUSIONS: The present modified method of calculating quantitative CBF from whole brain and venous blood radioactivities correlated well with values determined with arterial blood radioactivity.

Estimation of ¹²³I-IMP arterial blood activity using ¹²³I-IMP acquisition data from the lungs and brain without any blood sampling: validation of its usefulness for quantification of regional cerebral blood flow.

OBJECTIVE: The conventional methods for the estimation of regional cerebral blood flow (rCBF) using ¹²³I-labeled N-isopropyl-p-iodoamphetamine (¹²³I IMP) autoradiography (ARG) require continuous or 1-point arterial blood sampling. Patients who need rCBF quantification benefit from the avoidance of arterial puncture. In this study, we attempted to develop a method without any blood sampling to estimate ¹²³I IMP activity in the arterial blood sample at 10 minutes after injection of ¹²³I IMP (Ca10) for the purpose of rCBF quantification. For the evaluation of validity of this method, the mean of rCBFs in various regions of the brain (mean CBF) calculated by ¹²³I IMP ARG method using the estimated Ca10 was compared with that calculated using the Ca10 directly measured with the actual arterial blood sample. Both groups of the mean CBF values were also compared with those measured by O-15 H2O PET ARG method. METHODS: I-123 IMP ARG study was applied to 23 patients, and O-15 H2O PET ARG was applied to 20 patients of them. Dynamic images of the lungs, time series of static images of the brain, and brain SPECT images were acquired after injection of ¹²³I IMP. Arterial blood sampling was done 10 minutes after injection of ¹²³I IMP. Multiple regression analysis was used to estimate Ca10 using 5 parameters from the lung washout counts, time series of brain static counts, and brain SPECT average counts as the explanatory variables and the Ca10 directly measured with the actual arterial blood sample as the objective variable, and the regression equation was calculated. RESULTS: The regression equation was calculated by multiple regression analysis as follows: Estimated Ca10 = (2.09 × 10⁻² · LW3) - (2.29 × 10⁻4 · Cb5) - (9.87 × 10⁻³ · Cbpre-SPECT) + (1.06 · CbSPECTav) + (1.03 × 10⁻² · Cbpost-SPECT) + 165 (counts/s/g), where LW3: lung washout count at 3 minutes after injection, Cb5: brain count at 5 minutes, Cb pre-SPECT: brain count before SPECT, Cb SPECT av: average brain count during SPECT, and Cb post-SPECT: brain count after SPECT. The estimated Ca10 values closely correlated with the directly measured Ca10 values (r = 0.907, P < 0.01). The mean CBF values (mL/min/100 g) calculated by ¹²³I IMP ARG method using the estimated Ca10 also closely correlated with those calculated using the directly measured Ca10 (r = 0.818, P < 0.01). The mean CBF values calculated by the ¹²³I IMP ARG method using either the directly measured or the estimated Ca10 significantly correlated (r = 0.698 and 0.590, respectively; P < 0.01) with those measured by O-15 H2O PET ARG method. CONCLUSIONS: The ¹²³I IMP arterial blood activity can be estimated reliably without any blood sampling using the ¹²³I IMP acquisition data from the lungs and brain. This method can serve for a convenient and noninvasive rCBF quantification technique instead of the conventional methods requiring arterial blood sampling.

Estimation of 123I-IMP arterial blood activity from dynamic planar imaging of the chest using a graph plot method for the quantification of regional cerebral blood flow.

PURPOSE: I-123-labeled N-isopropyl-p-iodoamphetamine ((123)I-IMP) is used for the measurement of regional cerebral blood flow (rCBF). A continuous or single arterial blood sampling (ABS) is necessary to estimate an integral of arterial input function (AIF) for the measurement of rCBF by using a microsphere model analysis. Therefore, a method of measuring rCBF without any blood sampling is desired. The aim of this study was to establish a method to estimate the AIF from the time-activity curve of the lungs after an injection of (123)I-IMP, using a regression analysis for the measurement of rCBF without any blood sampling. MATERIALS AND METHODS: Thirty-seven prospective studies in 10 consecutive patients were enrolled. A chest planar dynamic imaging for 3 min and continuous ABS for 5 min after a bolus injection of 167MBq (123)I-IMP were performed in all studies. Data from the chest imaging were analyzed in comparison with ABS data (AIF(5)) in the first 10 studies, and an equation for estimation yielding accurate AIF(5) from the total counts cleared from the lungs, during 5 min after injection of (123)I-IMP (TCL(5)), was derived. The validity of the proposed method was evaluated in the subsequent 27 studies. RESULTS: A good correlation was obtained between the AIF and TCL by regression analysis in the first 10 studies (r = 0.94, P < 0.001). An equation for the estimation of AIF by the regression analysis in the first 10 studies was defined as follows: estimated AIF = 2147 + 4.174 x TCL(5). In the subsequent 27 studies, a good linear correlation was obtained between the measured and the estimated AIF(5) by using the equation (r = 0.79, P < 0.001). CONCLUSION: AIF(5) can be accurately estimated from TCL(5). Therefore, estimated AIF(5) can be used for the measurement of rCBF instead of ABS data.

Skull bone regeneration in nonhuman primates by controlled release of bone morphogenetic protein-2 from a biodegradable hydrogel.

The objective of this study was to investigate the feasibility of biodegradable gelatin hydrogels as the controlled-release carrier of bone morphogenetic protein-2 (BMP-2) to enhance bone regeneration at a skull defect of nonhuman primates. Hydrogels with 3 different water contents were prepared through glutaraldehyde crosslinking of gelatin with an isoelectric point of 9.0 under varied reaction conditions. A critical-sized defect (6 mm in diameter) was prepared at the skull bone of skeletally mature cynomolgus monkeys, and gelatin hydrogels incorporating various doses of BMP-2 were applied to the defects. When the bone regeneration was evaluated by soft radiography and bone mineral density (BMD) examinations, the gelatin hydrogel incorporating BMP-2 exhibited significantly higher osteoinduction activity than did an insoluble bone matrix that incorporated BMP-2 (one of the best osteoinduction systems), although the activity depended on the water content of hydrogels. BMD enhancement was highest for the gelatin hydrogel that had a water content of 97.8 wt% among all types of hydrogels. Moreover, the gelatin hydrogel enabled BMP-2 to induce the bone regeneration in nonhuman primates even at low doses. We conclude that the controlled release of BMP-2 for a certain time period was essential to inducing the osteoinductive potential of BMP-2.

[Study on the accuracy of regional cerebral blood flow measurement using N-isopropyl-p-[123I]iodoamphetamine: effects of differences in venous sampling site and static imaging direction in one-point venous sampling method based on the causality model].

PURPOSE: We have previously reported the method of regional cerebral blood flow measurement using N-isopropyl-p-[123I]iodoamphetamine, in which the input function into brain was estimated from one-point venous blood sampling value based on the method of causality analysis between input and output functions. In the present study, we examined the effects of differences in blood sampling site and direction of static image collection on the accuracy of estimating input function using this method. METHODS: The subjects consisted of 50 patients of right forearm venous sampling and 50 patients of left forearm venous sampling. As the static imaging directions, the following four combinations were compared 10 all four directions, 2) anterior and posterior directions, 3) right and left directions, and 4) an anterior direction. The accuracy of measurement was evaluated by comparing the estimated and directly measured value of input/output function (Caoct/Cvoct), and by the error indices and the correlation coefficients between the estimated and directly measured value. RESULT: In both groups of venous sampling, there was no significant difference between the estimated and directly measured value of Caoct/Cvoct. The error indices and correlation coefficient showed no significant difference between the right and left venous sampling groups. Similarly, no significant influence on Caoct/Cvoct value was observed by the difference of static imaging direction. Finally, the rCBF values calculated using these estimations were not significantly different from those by continuous arterial sampling method. CONCLUSIONS: These results indicate that both the difference in venous sampling site and the static imaging direction have little effect on the accuracy in our new method of rCBF measurement, and suggest its clinical versatility.

Enhanced bone regeneration at a segmental bone defect by controlled release of bone morphogenetic protein-2 from a biodegradable hydrogel.

The objective of this study is to investigate the feasibility of a biodegradable hydrogel of gelatin as the controlled release carrier of bone morphogenetic protein-2 (BMP-2) suitable for enhancement of bone regeneration at a segmental bone defect. Hydrogels with three different water contents were prepared through glutaraldehyde crosslinking of gelatin with an isoelectric point of 9.0 under varied reaction conditions. Segmental critical-sized defects (20 mm) were created at the ulnar bone of skeletally mature New Zealand white rabbits, and gelatin hydrogels incorporating BMP-2 (17 microg/hydrogel) were implanted into the defects. When bone regeneration was evaluated by soft x-ray observation and bone mineral density (BMD) measurement, the gelatin hydrogels incorporating BMP- 2 exhibited significantly high osteoinduction activity compared with that of free BMP-2, although the activity depended on the water content of the hydrogels. Significantly higher BMD enhancement was observed in the gelatin hydrogel with a water content of 97.8 wt% than that with the lower or higher water content. We concluded that the biodegradable gelatin hydrogel is a promising controlled release carrier of BMP-2 for bone regeneration at the segmental bone defect.

Measurement of regional cerebral blood flow with 123I-IMP using one-point venous blood sampling and causality analysis: evaluation by comparison with conventional continuous arterial blood sampling.

OBJECTIVE: Arterial input function represents the delivery of intravascular tracer to the brain. The optimal setting of this function is essential for measuring regional cerebral blood flow (rCBF) based on the microsphere model using N-isopropyl-4-[123I]iodoamphetamine (123I-IMP), in which the arterial 123I-IMP concentration (integral value) during the initial 5 min is usually applied. We developed a novel method in which the arterial 123I-IMP concentration is estimated from that in venous blood samples. METHODS: Brain perfusion SPECT with 123I-IMP was performed in 110 patients with disorders of the central nervous system. A causality analysis determined the relationship between various SPECT parameters and the ratio of the octanol-extracted arterial radioactivity concentration during the first 5 min (Caoct) to the octanol-extracted venous radioactivity concentration at 27 min after an intravenous injection of 123I-IMP (Cvoct). The Caoct/Cvoct value was estimated using various SPECT parameters and compared with the directly measured value. RESULTS: The measured and estimated values of Caoct/ Cvoct (r = 0.856, n = 50) closely correlated when the following 7 parameters were included in the regression formula: radioactivity concentration in venous blood sampled at 27 min (Cv), Cvoct, Cvoct/Cv, and 4 parameters related to cerebral tissue accumulation that were measured using a four-head gamma camera 5 and 28 min after 123I-IMP injection. Furthermore, the rCBF values obtained using the input function estimated by this method also closely correlated with the rCBF values measured using the continuous arterial blood sampling (r = 0.912, n = 180). CONCLUSION: These results suggest that this method would serve as a convenient and less invasive method of rCBF measurement in the clinical setting.

Viability and function of autologous and allogeneic fibroblasts seeded in dermal substitutes after implantation.

BACKGROUND: Fibroblast-seeded collagen sponges have been used for the treatment of skin defects and skin ulcers. However, the viability of the fibroblasts after implantation is still unknown. The objective of this study was to investigate the viability and distribution of autologous and allogeneic fibroblasts after implantation and to clarify which type is more effective for wound healing. MATERIALS AND METHODS: Skin samples of Hartley guinea pigs were retrieved and autologous fibroblasts were isolated and cultured. Fibroblasts isolated from the skin of a Strain2 guinea pig were used as allogeneic fibroblasts. Three full-thickness wounds were created on the backs of guinea pigs and an acellular collagen sponge, a collagen sponge seeded with autologous fibroblasts, and a collagen sponge seeded with allogeneic fibroblasts were transplanted. Before implantation, fibroblasts were labeled with PKH26. The guinea pigs were sacrificed 1, 2, and 3 weeks after implantation. The epithelization and contraction of the wounds were assessed, and the viability and distribution of the seeded fibroblasts were observed in cross sections. RESULTS: Three weeks after implantation, the PKH26-labeled autologous and allogeneic fibroblasts remained viable. In the wounds covered with the autologous fibroblast-seeded collagen sponge, the epithelization was fastest, and the percent wound contraction was smallest. In contrast, in the wounds covered with allogeneic fibroblasts, the epithelization was slowest and the percent contraction was largest. CONCLUSION: The allogeneic fibroblasts seeded in the collagen sponge survived and remained viable on the grafted area, but did not accelerate wound healing.

Enhanced osteoinduction by controlled release of bone morphogenetic protein-2 from biodegradable sponge composed of gelatin and beta-tricalcium phosphate.

Biodegradable gelatin sponges at different contents of beta-tricalcium phosphate (beta-TCP) were fabricated to allow bone morphogenetic protein (BMP)-2 to incorporate into them. The in vivo osteoinduction activity of the sponges incorporating BMP-2 was investigated, while their in vivo profile of BMP-2 release was evaluated. The sponges prepared had an interconnected pore structure with an average pore size of 200 microm, irrespective of the beta-TCP content. The in vivo release test revealed that BMP-2 was released in vivo at a similar time profile, irrespective of the beta-TCP content. The in vivo time period of BMP-2 retention was longer than 28 days. When the osteoinduction activity of gelatin or gelatin-beta-TCP sponges incorporating BMP-2 was studied following the implantation into the back subcutis of rats in terms of histological and biochemical examinations, homogeneous bone formation was histologically observed throughout the sponges, although the extent of bone formation was higher in the sponges with the lower contents of beta-TCP. On the other hand, the level of alkaline phosphatase activity and osteocalcin content at the implanted sites of sponges decreased with an increase in the content of beta-TCP. The gelatin sponge exhibited significantly higher osteoinduction activity than that of any gelatin-beta-TCP sponge, although every sponge with or without beta-TCP showed a similar in vivo profile of BMP-2 release. In addition, the in vitro collagenase digestion experiments revealed that the gelatin-beta-TCP sponge collapsed easier than the gelatin sponge without beta-TCP incorporation. These results suggest that the maintenance of the intrasponge space necessary for the osteoinduction is one factor contributing to the osteoinduction extent of BMP-2-incorporating sponges.

Osteogenic differentiation of mesenchymal stem cells in biodegradable sponges composed of gelatin and beta-tricalcium phosphate.

Biodegradable gelatin sponges incorporating various amounts of beta-tricalcium phosphate (betaTCP) (gelatin-betaTCP) were fabricated and the in vitro osteogenic differentiation of mesenchymal stem cells (MSC) isolated from the rat bone marrow in the sponges was investigated. The gelatin sponges incorporating betaTCP have an interconnected pore structure with the average size of 180-200 microm, irrespective of the betaTCP amount. The stiffness of the sponges became higher with an increase in the amount of betaTCP. When seeded into the sponges by an agitated method, MSC were homogeneously distributed throughout the sponge. The morphology of cells attached got more spreaded with the increased betaTCP amount. The rate of MSC proliferation depended on the betaTCP amount and culture method: the higher the betaTCP amount in the stirring culture, the higher the proliferation rate. The deformed extent of gelatin-betaTCP sponges was suppressed with the increased amount of betaTCP. When measured to evaluate the osteogenic differentiation of MSC, the alkaline phosphatase activity and osteocalcin content became maximum for the sponge with a betaTCP amount of 50 wt%, although both the values were significantly high in the stirring culture compared with those in the static culture. We concluded that the attachment, proliferation, and osteogenic differentiation of MSC were influenced by sponge composition of gelatin and betaTCP as the cell scaffold.

Prefabrication of vascularized bone graft using guided bone regeneration.

This article describes the prefabrication of a vascularized bone graft composed of autologous particulate cancellous bone and marrow (PCBM), a vessel bundle, and a biodegradable membrane. The PCBM was placed around the saphenous vessel bundle of rats and rolled with a biodegradable membrane of L-lactide-epsilon-caprolactone copolymer to prepare the prefabricated vascularized bone graft (group A). As controls, combinations of PCBM and membrane (group B), vessel bundle and membrane (group C), and PCBM and vessel bundle (group D) were prepared. A radiographic study revealed radio-opacity in the implantation site of group A 1 week later, in contrast to the other groups. Newly formed bone in the membrane roll was histologically confirmed, and neomicrovasculature circulating from the vessel bundle through the newly formed bone tissue was observed. The increase in alkaline phosphatase activity and osteocalcin content was significant for the group A preparation compared with the other groups. We concluded that the combination of autologous PCBM, a vessel bundle, and a biodegradable membrane was promising in the prefabrication of vascularized bone with good blood circulation.

Effect of the fiber diameter and porosity of non-woven PET fabrics on the osteogenic differentiation of mesenchymal stem cells.

The proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) was investigated in three-dimensional non-woven fabrics prepared from polyethylene terephthalate (PET) fiber with different diameters. When seeded into the fabrics of cell scaffold, more MSC attached in the fabric of thicker PET fibers than that of thinner ones, irrespective of the fabric porosity. The morphology of cells attached became more spreaded with an increase in the fiber diameter of fabrics. The rate of MSC proliferation depended on the PET fiber diameter and porosity of fabrics: the bigger the fiber diameter of fabrics with higher porosity, the higher their proliferation rate. When the alkaline phosphatase (ALP) activity and osteocalcin content of MSC cultured in different types of fabrics was measured to evaluate the ostegenic differentiation, they became maximum for the non-woven fabrics with a fiber diameter of 9.0 microm, although the values of low-porous fabrics were significantly high compared with those of high porous fabrics. We concluded that the attachment, proliferation and bone differentiation of MSC was influenced by the fiber diameter and porosity of non-woven fabrics as the scaffold.

Homogeneous seeding of mesenchymal stem cells into nonwoven fabric for tissue engineering.

Mesenchymal stem cells (MSCs) were isolated from the bone marrow of rats and seeded into a nonwoven fabric of polyethylene terephtalate (PET) by agitation and static methods. MSC attachment was investigated in terms of the number of cells attached to the fabric, their distribution inside the fabric, and cell damage. The number of MSCs attached was greater for the agitation seeding method than for the static seeding method. The higher the rotating speed in the agitation seeding method, the greater the number of cells attached. When the cell suspension was seeded into the fabric in culture medium volumes of 50 and 200 microL per well of the culture plate or per culture tube, the best cell attachment was observed for the tube culture group at the larger volume. These cells attached more homogeneously throughout the fabric in greater numbers than was the case for the other culture groups. It is possible that agitation of the cell suspension allows cells to infiltrate uniformly inside the fabric, resulting in a homogeneous distribution of the cells in the fabric. A biochemical study revealed that neither the agitation nor static seeding method damaged cells, irrespective of the medium volume and the type of culture vessel. We conclude that the agitation seeding method is a promising method by which to formulate a homogeneous construct of fabric and MSCs.

Controlled release by biodegradable hydrogels enhances the ectopic bone formation of bone morphogenetic protein.

The objective of this study is to develop a carrier for the controlled release of bone morphogenetic protein-2 (BMP-2) suitable for enhancement of the bone regeneration activity. Hydrogels with different water contents were prepared through glutaraldehyde crosslinking of gelatin with an isoelectric point of 9.0 under varied reaction conditions. Following subcutaneous implantation of the gelatin hydrogels incorporating 125I-labeled BMP-2 into the back of mice, the in vivo retention period of BMP-2 prolonged with a decrease in the water content of hydrogels used, although every time period was much longer than that of BMP-2 solution injection. Ectopic bone formation studies demonstrated that the alkaline phosphatase (ALP) activity and osteocalcin content around the implanted site of BMP-2-incorporated gelatin hydrogels were significantly high compared with those around the injected site of BMP-2 solution. The values became maximum for the gelatin hydrogel incorporating BMP-2 with a middle period of BMP-2 retention, while bone formation was histologically observed around the hydrogel incorporating BMP-2. The ALP activity was significantly higher than that of the collagen sponge incorporating BMP-2. We concluded that the controlled release technology of BMP-2 for a certain time period was essential to induce the potential activity for bone formation.

Cyclooxygenase-2 promotes prostate cancer progression.

BACKGROUND: Cyclooxygenase (COX) -2, an inducible isoform of COX, has been observed to be expressed in prostate cancer. Several studies have reported that COX-2 overexpression is associated with carcinogenesis, cell growth, angiogenesis, apoptosis, and invasiveness in a variety of tumor types. METHODS: To investigate the function of COX-2 in prostate cancer directly, we stably transfected human full-length COX-2 cDNA into LNCaP cells (LNCaP-COX-2), which express low levels of endogenous COX-2. RESULTS: The level of COX-2 mRNA and protein and the COX activity in COX-2 LNCaP-COX-2 cells was significantly increased compared with parent and control-transfected cells. Overexpression of COX-2 increased both proliferation in vitro and tumor growth rate in vivo. However, the pro-tumor effect was neither associated with changes of androgen receptor (AR) expression level nor AR activity. Furthermore, addition of the major metabolites of COX-2-mediated arachidonic acid metabolism did not alter the proliferation of LNCaP-COX-2 cells in vitro. LNCaP-COX-2 cells had increased secretion of vascular endothelial growth factor (VEGF) protein, suggesting that angiogenesis induced by COX-2 stimulates tumor growth in vivo. CONCLUSION: These data demonstrate that COX-2 contributes to prostate cancer progression and suggest that it mediates this effect, in part, through increased VEGF.