Tissue engineering of cartilage using poly-epsilon-caprolactone nanofiber scaffolds seeded in vivo with periosteal cells.

OBJECTIVE: To determine the potential of periosteal cells to infiltrate poly-epsilon-caprolactone (PCL) nanofiber scaffolds in vivo and subsequently produce cartilage in vitro. DESIGN: PCL nanofiber scaffolds, with or without chitosan-coating were implanted under periosteum in 6-month-old rabbits. Transforming growth factor-beta1 (TGF-beta1) or vehicle was injected into each implant site. After 1, 3, 5 or 7 days, scaffolds were removed, separated from the periosteum, and the scaffolds and periosteum were cultured separately for 6 weeks under chondrogenic conditions. Sulfated glycosaminoglycan (GAG), type II collagen, DNA content, cartilage yield, and calcium deposition were then analyzed. RESULTS: Cell infiltration was observed in all scaffolds. Cartilage formation in the uncoated scaffolds increased with duration of implantation (maximum at 7 days). Cells in the uncoated scaffolds implanted for 7 days produced significantly higher levels of both GAG [560 (95% confidence interval (CI), 107-1013) vs 228 (95% CI, 177-278) microg GAG/microg DNA] and cartilage yield [9% (95% CI, 3-14%) vs 0.02% (95% CI, 0-0.22%)] compared to chitosan-coated scaffolds (P=0.006 or less). There was no significant difference in GAG content or cartilage yield between the TGF-beta1-injected and vehicle-injected scaffolds. However, significantly more mineral deposition was detected in TGF-beta1-injected scaffolds compared to vehicle-injected scaffolds (P<0.0001). Cartilage yield from the periosteum, moreover, was significantly increased by subperiosteal TGF-beta1 injections (P<0.001). However, this response was reduced when chitosan-coated scaffolds were implanted. CONCLUSIONS: This study demonstrates that it is possible to seed PCL nanofiber scaffolds with periosteal cells in vivo and subsequently produce engineered cartilage in vitro.

Pretreatment of periosteum with TGF-beta1 in situ enhances the quality of osteochondral tissue regenerated from transplanted periosteal grafts in adult rabbits.

OBJECTIVE: To compare the efficacy of in situ transforming growth factor-beta1 (TGF-beta1)-pretreated periosteum to untreated periosteum for regeneration of osteochondral tissue in rabbits. METHODS: In the pretreatment group, 12 month-old New Zealand white rabbits received subperiosteal injections of 200 ng of TGF-beta1 percutaneously in the medial side of the proximal tibia, 7 days prior to surgery. Control rabbits received no treatment prior surgery. Osteochondral transverse defects measuring 5mm proximal to distal and spanning the entire width of the patellar groove were created and repaired with untreated or TGF-beta1-pretreated periosteal grafts. Post-operatively the rabbits resumed normal cage activity for 6 weeks. RESULTS: Complete filling of the defects with regenerated tissue was observed in both the TGF-beta1-pretreated and control groups with reformation of the original contours of the patellar groove. The total histological score (modified O'Driscoll) in the TGF-beta1-pretreated group, 20 (95% Confidence Interval (CI), 19-21), was significantly higher (P=0.0001) than the control group, 18 (16-19). The most notable improvements were in structural integrity and subchondral bone regeneration. No significant differences in glycosaminoglycan or type II collagen content, or equilibrium modulus were found between the surgical groups. The cambium of the periosteum regenerated at the graft harvest site was significantly thicker (P=0.0065) in the TGF-beta1-pretreated rabbits, 121 microm (94-149), compared to controls, 74 microm (52-96), after 6 weeks. CONCLUSIONS: This study demonstrates that in situ pretreatment of periosteum with TGF-beta1 improves osteochondral tissue regeneration at 6-weeks post-op compared to untreated periosteum in 12 month-old rabbits.

Rejuvenation of periosteal chondrogenesis using local growth factor injection.

OBJECTIVE: To examine the potential for rejuvenation of aged periosteum by local injection of transforming growth factor-beta1 (TGF-beta1) and insulin-like growth factor-1 (IGF-1) alone or in combination to induce cambium cell proliferation and enhance in vitro periosteal cartilage formation. METHODS: A total of 367 New Zealand white rabbits (6, 12, and 24+ month-old) received subperiosteal injections of TGF-beta1 and/or IGF-1 percutaneously. After 1, 3, 5, or 7 days, the rabbits were sacrificed and cambium cellularity or in vitro cartilage forming capacity was determined. RESULTS: A significant increase in cambium cellularity and thickness, and in vitro cartilage formation was observed after injection of TGF-beta1 alone or in combination with IGF-1. In 12 month-old rabbits, mean cambium cellularity increased 5-fold from 49 to 237 cells/mm and in vitro cartilage production increased 12-fold from 0.8 to 9.7 mg 7 days after TGF-beta1 (200 ng) injection compared to vehicle controls (P<0.0001). A correlation was observed between cambium cellularity and in vitro cartilage production (R2=0.98). An added benefit of IGF-1 plus TGF-beta1 on in vitro cartilage production compared to TGF-beta1 alone was observed in the 2 year-old rabbits. IGF-1 alone generally had no effect on either cambium cellularity or in vitro cartilage production in any of the age groups. CONCLUSIONS: These results clearly demonstrate that it is possible to increase cambium cellularity and in vitro cartilage production in aged rabbit periosteum, to levels comparable to younger rabbits, using local injection of TGF-beta1 alone or in combination with IGF-1, thereby rejuvenating aged periosteum.

Diffusion tractography of the fornix in schizophrenia.

BACKGROUND: White matter fiber tracts, especially those interconnecting the frontal and temporal lobes, are likely implicated in pathophysiology of schizophrenia. Very few studies, however, have focused on the fornix, a compact bundle of white matter fibers, projecting from the hippocampus to the septum, anterior nucleus of the thalamus and the mamillary bodies. Diffusion Tensor Imaging (DTI), and a new post-processing method, fiber tractography, provides a unique opportunity to visualize and to quantify entire trajectories of fiber bundles, such as the fornix, in vivo. We applied these techniques to quantify fornix diffusion anisotropy in schizophrenia. METHODS: DTI images were used to evaluate the left and the right fornix in 36 male patients diagnosed with chronic schizophrenia and 35 male healthy individuals, group matched on age, parental socioeconomic status, and handedness. Regions of interest were drawn manually, blind to group membership, to guide tractography, and fractional anisotropy (FA), a measure of fiber integrity, was calculated and averaged over the entire tract for each subject. The Doors and People test (DPT) was used to evaluate visual and verbal memory, combined recall and combined recognition. RESULTS: Analysis of variance was performed and findings demonstrated a difference between patients with schizophrenia and controls for fornix FA (p=0.006). Protected post-hoc independent sample t-tests demonstrated a bilateral FA decrease in schizophrenia, compared with control subjects (left side: p=0.048; right side p=0.006). Higher fornix FA was statistically significantly correlated with DPT and measures of combined visual memory (r=0.554, p=0.026), combined verbal memory (r=0.647, p=0.007), combined recall (r=0.516, p=0.041), and combined recognition (r=0.710, p=0.002) for the control group. No such statistically significant correlations were found in the patient group. CONCLUSIONS: Our findings show the utility of applying DTI and tractography to study white matter fiber tracts in vivo in schizophrenia. Specifically, we observed a bilateral disruption in fornix integrity in schizophrenia, thus broadening our understanding of the pathophysiology of this disease.

Abnormalities in thalamo-cortical connections in patients with first-episode schizophrenia: a two-tensor tractography study.

The "cognitive dysmetria" hypothesis suggests that impairments in cognition and behavior in patients with schizophrenia can be explained by disruptions in the cortico-cerebellar-thalamic-cortical circuit. In this study we examine thalamo-cortical connections in patients with first-episode schizophrenia (FESZ). White matter pathways are investigated that connect the thalamus with three frontal cortex regions including the anterior cingulate cortex (ACC), ventrolateral prefrontal cortex (VLPFC), and lateral oribitofrontal cortex (LOFC). We use a novel method of two-tensor tractography in 26 patients with FESZ compared to 31 healthy controls (HC), who did not differ on age, sex, or education. Dependent measures were fractional anisotropy (FA), Axial Diffusivity (AD), and Radial Diffusivity (RD). Subjects were also assessed using clinical functioning measures including the Global Assessment of Functioning (GAF) Scale, the Global Social Functioning Scale (GF: Social), and the Global Role Functioning Scale (GF: Role). FESZ patients showed decreased FA in the right thalamus-right ACC and right-thalamus-right LOFC pathways compared to healthy controls (HCs). In the right thalamus-right VLPFC tract, we found decreased FA and increased RD in the FESZ group compared to HCs. After correcting for multiple comparisons, reductions in FA in the right thalamus- right ACC and the right thalamus- right VLPC tracts remained significant. Moreover, reductions in FA were significantly associated with lower global functioning scores as well as lower social and role functioning scores. We report the first diffusion tensor imaging study of white matter pathways connecting the thalamus to three frontal regions. Findings of white matter alterations and clinical associations in the thalamic-cortical component of the cortico-cerebellar-thalamic-cortical circuit in patients with FESZ support the cognitive dysmetria hypothesis and further suggest the possible involvement of myelin sheath pathology and axonal membrane disruption in the pathogenesis of the disorder.

The role of the posterior bundle of the medial collateral ligament in posteromedial rotatory instability of the elbow.

Aims: The aim of this study was to evaluate two hypotheses. First, that disruption of posterior bundle of the medial collateral ligament (PMCL) has to occur for the elbow to subluxate in cases of posteromedial rotatory instability (PMRI) and second, that ulnohumeral contact pressures increase after disruption of the PMCL. Materials and Methods: Six human cadaveric elbows were prepared on a custom-designed apparatus which allowed muscle loading and passive elbow motion under gravitational varus. Joint contact pressures were measured sequentially in the intact elbow (INTACT), followed by an anteromedial subtype two coronoid fracture (COR), a lateral collateral ligament (LCL) tear (COR + LCL), and a PMCL tear (COR + LCL + PMCL). Results: There was no subluxation or joint incongruity in the INTACT, COR, and COR + LCL specimens. All specimens in the COR + LCL + PMCL group subluxated under gravity-varus loads. The mean articular contact pressure of the COR + LCL group was significantly higher than those in the INTACT and the COR groups. The mean articular contact pressure of the COR + LCL + PMCL group was significantly higher than that of the INTACT group, but not higher than that of the COR + LCL group. Conclusion: In the presence of an anteromedial fracture and disruption of the LCL, the posterior bundle of the MCL has to be disrupted for gross subluxation of the elbow to occur. However, elevated joint contact pressures are seen after an anteromedial fracture and LCL disruption even in the absence of such subluxation. Cite this article: Bone Joint J 2018;100-B:1060-5.

Porous tantalum biocomposites for osteochondral defect repair: A follow-up study in a sheep model.

OBJECTIVES: We sought to determine if a durable bilayer implant composed of trabecular metal with autologous periosteum on top would be suitable to reconstitute large osteochondral defects. This design would allow for secure implant fixation, subsequent integration and remodeling. MATERIALS AND METHODS: Adult sheep were randomly assigned to one of three groups (n = 8/group): 1. trabecular metal/periosteal graft (TMPG), 2. trabecular metal (TM), 3. empty defect (ED). Cartilage and bone healing were assessed macroscopically, biochemically (type II collagen, sulfated glycosaminoglycan (sGAG) and double-stranded DNA (dsDNA) content) and histologically. RESULTS: At 16 weeks post-operatively, histological scores amongst treatment groups were not statistically different (TMPG: overall 12.7, cartilage 8.6, bone 4.1; TM: overall 14.2, cartilage 9.5, bone 4.9; ED: overall 13.6, cartilage 9.1, bone 4.5). Metal scaffolds were incorporated into the surrounding bone, both in TM and TMPG. The sGAG yield was lower in the neo-cartilage regions compared with the articular cartilage (AC) controls (TMPG 20.8/AC 39.5, TM 25.6/AC 33.3, ED 32.2/AC 40.2 µg sGAG/1 mg respectively), with statistical significance being achieved for the TMPG group (p < 0.05). Hypercellularity of the neo-cartilage was found in TM and ED, as the dsDNA content was significantly higher (p < 0.05) compared with contralateral AC controls (TM 126.7/AC 71.1, ED 99.3/AC 62.8 ng dsDNA/1 mg). The highest type II collagen content was found in neo-cartilage after TM compared with TMPG and ED (TM 60%/TMPG 40%/ED 39%). Inter-treatment differences were not significant. CONCLUSIONS: TM is a highly suitable material for the reconstitution of osseous defects. TM enables excellent bony ingrowth and fast integration. However, combined with autologous periosteum, such a biocomposite failed to promote satisfactory neo-cartilage formation.Cite this article: E. H. Mrosek, H-W. Chung, J. S. Fitzsimmons, S. W. O'Driscoll, G. G. Reinholz, J. C. Schagemann. Porous tantalum biocomposites for osteochondral defect repair: A follow-up study in a sheep model. Bone Joint J 2016;5:403-411. DOI: 10.1302/2046-3758.59.BJR-2016-0070.R1.

Large scale accelerator production of 225Ac: Effective cross sections for 78-192MeV protons incident on 232Th targets.

Actinium-225 and 213Bi have been used successfully in targeted alpha therapy (TAT) in preclinical and clinical research. This paper is a continuation of research activities aiming to expand the availability of 225Ac. The high-energy proton spallation reaction on natural thorium metal targets has been utilized to produce millicurie quantities of 225Ac. The results of sixteen irradiation experiments of thorium metal at beam energies between 78 and 192MeV are summarized in this work. Irradiations have been conducted at Brookhaven National Laboratory (BNL) and Los Alamos National Laboratory (LANL), while target dissolution and processing was carried out at Oak Ridge National Laboratory (ORNL). Excitation functions for actinium and thorium isotopes, as well as for some of the fission products, are presented. The cross sections for production of 225Ac range from 3.6 to 16.7mb in the incident proton energy range of 78-192MeV. Based on these data, production of curie quantities of 225Ac is possible by irradiating a 5.0gcm-2 232Th target for 10 days in either BNL or LANL proton irradiation facilities.

Diagnosis of posttraumatic stress disorder in Viet Nam veterans: preliminary findings.

Sixty-six Viet Nam veterans were evaluated for posttraumatic stress disorder. Several of the DSM-III criteria for the disorder, but no other clinical features, distinguished patients diagnosed as having the disorder from others. The findings tend to validate the DSM-III construct for this disorder.

Disruption of the blood-brain barrier in experimental optic neuritis: immunocytochemical co-localization of H2O2 and extravasated serum albumin.

PURPOSE: To probe the role of endogenous hydrogen peroxide (H2O2) in the pathogenesis of disruption of the blood-brain barrier (BBB) associated with experimental allergic encephalomyelitis (EAE), an animal model for primary central nervous system demyelination. METHODS: Strain-13 guinea pigs were sensitized for EAE with central myelin in complete Freund's adjuvant. Magnetic resonance imaging with Gd-DTPA was performed twice a week for 2 weeks to assess disruption of the BBB, in vivo, by the enhancement of the optic nerves. Two weeks after antigenic sensitization, ultracytochemical localization of endogenous H2O2 was performed using the cerium perhydroxide method, with co-localization of endogenous serum albumin extravasation using gold-labeled antibodies against serum albumin. Examination of blood vessels for perivascular immunogold-labeled serum albumin and H2O2 derived reaction product began in the optic nerve head and proceeded toward the retrobulbar optic nerve until a total of 20 vessels were evaluated per animal. RESULTS: Magnetic resonance imaging revealed Gd-DTPA enhancement of the optic nerves in all animals sensitized for EAE. Optic nerve ultrastructure revealed colloidal gold-labeled antibodies against serum albumin in the perivascular and adjacent interstitial spaces of capillaries and small venules in which H2O2 derived cerium perhydroxide reaction product was also simultaneously evident. Immunogold-labeled serum albumin was predominantly confined to the intravascular compartment of the optic nerve in the absence of perivascular H2O2 and/or perivascular foci of inflammatory cells. The difference between the mean percentage of blood vessels (61.8%) with co-localization of perivascular immunogold-labeled serum albumin and cerium perhydroxide reaction product, to the mean percentage of blood vessels (9.5%) with perivascular immunogold-labeled serum albumin in the absence of cerium perhydroxide, was statistically significant (P = 0.0019). CONCLUSIONS: Endogenous H2O2, found at the foci of BBB disruption, may be one of the mediators involved in the alteration of vascular permeability in experimental optic neuritis.

Reversals of blood-brain barrier disruption by catalase: a serial magnetic resonance imaging study of experimental optic neuritis.

PURPOSE: To investigate serially the role of catalase detoxification of endogenous H2O2 in the disruption of the blood-brain barrier (BBB) and demyelination of experimental optic neuritis. METHODS: Serial contrast-enhanced magnetic resonance imaging (MRI) of the optic nerves (T1 weighted) and T2 weighted MRI without contrast were performed on 18 guinea pigs 3 to 14 days after sensitization with central myelin for experimental allergic encephalomyelitis. Sex and age-matched littermates were paired and sensitized with the identical antigenic emulsion. To detoxify endogenous hydrogen peroxide (H2O2), animals received daily intraperitoneal injections of polyethylene glycol (PEG)-catalase at a dose of 12,000 U/kg per day for 3 days, then 1,200 U/kg daily for the next week, commencing 3 days after antigenic sensitization. Littermates received an equal volume of preservative-free saline. The intensity of gadolinium-DTPA (Gd-DTPA) enhancement was quantitated by obtaining the value for a region of interest (ROI) of the right optic nerve and the left optic nerve. The effect of H2O2 detoxification by catalase was evaluated by differences in the intensity of Gd-DTPA enhancement and T2 weighted signal in the ROI of the right and the left optic nerves at 7, 10, and 14 days after antigenic sensitization, from the pretreatment value obtained at day 3. The effectiveness of catalase detoxification of H2O2 was assessed with quantitative ultracytochemical localization of electron-dense, H2O2-derived cerium perhydroxide in the optic nerves. RESULTS: With PEG-catalase treatment, mean differences for Gd-DTPA enhancement in the ROI at 7, 10, and 14 days after antigenic sensitization were significantly reduced from the pretreatment values obtained 3 days after antigenic sensitization compared with the comparable interval values for untreated littermates. For T2 weighted signal intensity, only the 7- and 14-day values were significantly less with PEG-catalase compared with values for littermates obtained at comparable intervals. Quantitative ultracytochemical localization of H2O2-derived cerium perhydroxide reaction product revealed significant reductions in the medium number of cerium particle counts of the optic nerve head, sheath, and myelinated retrobulbar nerve. CONCLUSIONS: PEG-catalase reduced H2O2-derived cerium perhydroxide reaction product in the optic nerve but did not eliminate it, reversed disruption of the BBB as measured by Gd-DTPA enhancement, and reduced demyelination and edema as measured by T2 weighted signal intensity, suggesting detoxification of H2O2 as a new treatment strategy for disorders of primary demyelination of the central nervous system.

Importance of imaging plane for magnetic resonance imaging of the normal left ventricle.

Visualization of the left ventricle by magnetic resonance imaging (MRI) is largely a function of imaging planes. The best depiction of the myocardium is possible when the imaging plane is perpendicular to the region of interest. The effect of imaging plane on the visualization of the left ventricle has not been fully described. To assess this effect, 10 normal adults underwent electrocardiogram-gated multiplanar cardiac MRI. The imaging planes being evaluated were standard planes, transverse, sagittal and coronal, and modified planes relating to the left ventricular (LV) long axis: coronal long axis parallel to the ventricular septum (VS), sagittal long axis perpendicular to the VS and transverse short axis perpendicular to the VS and long axis. The myocardium was separated into 9 segments. Using each plane, the LV segments were evaluated for quality of LV depiction. The planes based on the LV long axis were superior to standard planes in depicting the LV segments.

Familial recurrence of achondroplasia.

A family is reported among whom 2 second-cousins-once removed (two half or half third cousins) demonstrate clinical and radiological features of classical achondroplasia.

Subsequent reproductive outcome in couples with repeated pregnancy loss.

While the role of cytogenetic study in couples with repeated pregnancy loss is well-established, little information is available for counseling these couples concerning future reproductive outcome. Couples evaluated by chromosome analysis for recurrent abortion between 1972 and 1979 were contacted by phone in 1981. Of those studied cytogenetically, 195 couples (50.1%) could be located, and information concerning outcome of subsequent pregnancies were obtained. Couples (91) with two consecutive pregnancy losses at the time of initial investigation had a 31.3% subsequent abortion rate, but most (68%) had at least one liveborn child. The rate of infertility following evaluation was slightly increased (18.7%), but that of prematurity (11.2%) and congenital anomalies (2.5%) was not. In contrast, couples with greater than or equal to 3 consecutive losses (84) experienced abortion in 45.7% of subsequent pregnancies, and only 54.8% of them eventually had a liveborn child. Again, the rate of infertility was increased (26.2%), but that of prematurity (10%) and congenital defects (3.2%) was not. Nine couples in which one individual was found to have a chromosome abnormality (two inversions and seven translocations) were considered separately. Of these, seven couples had nine liveborn offspring. Amniocentesis was performed in eight cases, with karyotypes showing a balanced translocation identical to that of the parent in 2. All children were phenotypically normal.

Repeated pregnancy loss.

Debate persists over the value of chromosome analysis of couples with repeated pregnancy loss. Therefore, we studied the records of all patients referred to the Genetics Division at Thomas Jefferson University for repeated pregnancy loss. Couples were divided into three groups according to the reason for evaluation. In group I (two consecutive abortions) significant chromosome abnormalities were found in 1.8% of individuals; in group II (three or more consecutive abortions) 2.3% of individuals had a chromosome abnormality; and in group III (50% fetal loss) 1.8% of persons had abnormal chromosomes. These rates are lower than those reported by others, but are still ten times higher than those expected in the general population and affirm the value of doing a chromosome study in such couples. In addition, we found increased incidence of liveborn offspring with congenital abnormalities in couples evaluated for the above indications, and found a high incidence of a family history of repeated suboptimal pregnancy outcome. The significance of these findings is discussed.

Factors related to subsequent reproductive outcome in couples with repeated pregnancy loss.

Specific factors in a couple's history may influence the recurrence risk following repeated pregnancy loss (RPL). Couples with RPL were contacted several years following evaluation and information concerning subsequent pregnancies was obtained. Linear regression analysis was utilized to determine which factors in the history were significant predictors of pregnancy outcome following evaluation. A family history of RPL or a "genetic defect" was a highly significant predictor of subsequent unsuccessful pregnancies. Surgical, but not medical, treatment for RPL was a significant predictor of eventual successful outcome. The number of abortions prior to evaluation for RPL, presence of a liveborn child, maternal age at evaluation, and intercurrent infertility all failed to be significant predictors of pregnancy outcome after evaluation.

Duplication (6q) syndrome diagnosed in utero.

We report on a case of partial duplication 6q detected ultrasonographically. The clinical picture noted in utero is consistent with the adult phenotype previously reported in the literature.

Eradication of poliomyelitis: progress in the Americas.

In the span of 5 years since the eradication initiative was launched and only 3 years since external funds were made available, PAHO has been able to develop and implement a comprehensive program strategy for polio eradication that includes the following components: achievement and maintenance of high immunization levels (which include the supplemental strategies of national immunization days and mop-up operations); effective surveillance to detect all new cases; and a rapid response to the occurrence of new cases. Despite yearly increases in the number of cases of acute flaccid paralysis reported to the surveillance system, a decline in reported confirmed cases of polio has occurred since 1986 to record low levels in 1989. Cases in 1989 were reported from only 0.7% of the counties in the Americas. The occurrence of 24 wild-type virus isolates in 1989 were limited to only three geographic areas: northwestern Mexico; the northern Andean Region; and northeastern Brazil. At this writing the clock is ticking with only 3 months left to achieve the goal of interrupting transmission by the end of 1990. If the current level of effort is sustained and special efforts are directed at the remaining foci of infection, the eradication of the transmission of wild-type poliovirus from the Americas can be achieved. Continued external financial support will be critical if the effort is to succeed. The prospect of poliomyelitis eradication in the Americas led the 41st World Health Assembly of WHO to adopt a resolution in May, 1988, to eradicate the indigenous transmission of wild-type poliovirus from the world by the year 2000.(ABSTRACT TRUNCATED AT 250 WORDS)

Phantom evaluation of imaging modalities for silicone breast implants.

RATIONALE AND OBJECTIVES: Recent concern regarding possible adverse effects from silicone breast implants has increased the role of radiologists in assessing augmented breasts. The authors compare the commonly available imaging modalities in evaluating the intact silicone implant as well as free silicone in the adjacent tissue. METHODS: A contrast resolution phantom and breast of veal phantom were tested. Fat was used as a reference material. The phantoms were imaged with xeromammography, film-screen mammography, ultrasound, computed tomography (CT), and magnetic resonance imaging (MRI). Proton MRI spectroscopy also was performed on fat, silicone, water, and water/gelatin samples. The consensus of two radiologists determined whether free silicone was present. RESULTS: CT and MRI provided the best images of the implant and the free silicone. Several features of MRI were useful: spin-density scans and the fast low-angle shot (FLASH) and fast imaging with steady-state precision (FISP) techniques provided excellent resolution, a consistent chemical shift artifact appeared around the silicone, and frequency selective pre-saturation techniques resulted in marked suppression of the silicone. CONCLUSION: Additional testing in a more realistic setting, breast coil design, and improvement of various MRI techniques, particularly the frequency selective pre-saturation techniques, all appear promising in evaluating breast implants, the presence of free silicone, and the adjacent tissues.

Method for automated cartilage histomorphometry.

We have developed and tested a color-based method for automated computerized histomorphometric analysis of cartilage. Histological sections stained with safranin O from 29 rabbit periosteal agarose-cultured explants were selected with various amounts of cartilage (0-100%). Color photomicrographs of these sections were visually assessed by five expert observers who estimated the percent area occupied by cartilage and outlined (in pen) the areas they considered to be cartilage. Manual histomorphometry was performed by cutting out and weighing the outlined areas. The average area for each of the five observers ranged from 31% to 43% (intraclass correlation coefficient = 0.70). The average of these values was used as a "gold standard" against which to compare the computer measurements. When point counting histomorphometry was performed on the 29 sections, the data agreed with the measurements made by the other five cartilage experts (r2 = 0.96; p < 0.0001). The analysis of cartilage is based on safranin O stain, using a custom-designed Vidas 2.1 Image Analysis Program (Zeiss). The computer-based results correlated very closely with those obtained by manual (p = 0.0001; r2 = 0.92) and point counting (r2 = 0.92; p < 0.0001) histomorphometry. The mean percentage of the sections occupied by cartilage measured in the automated mode was only 6% higher than that using the gold standard. Histological complexity had only a minor effect on the computerized values. The automated computerized image analysis system has the advantages of objectivity, accuracy, repeatability, and ease of use.