Monoclonal antibodies OC 125, OV632, OV-TL 3, MOv18 and OV-TL 23, directed against distinct ovari... more Monoclonal antibodies OC 125, OV632, OV-TL 3, MOv18 and OV-TL 23, directed against distinct ovarian carcinoma-associated antigens, were examined for their value in cytopathologic diagnosis. Their sensitivity and specificity in staining ovarian carcinoma cells in serous effusions we determined using the indirect immunoperoxidase technique. Smears prepared from 140 serous effusions (73 benign, 67 malignant) were immunostained with the five antibodies. OC 125 and MOv18 reacted positively with 96% and 81% of the smears of effusions from ovarian carcinoma patients, respectively, while OV-TL 3, OV632 and OV-TL 23 stained a lower percentage of the samples (73%, 65% and 62%, respectively). In discriminating ovarian carcinoma cells from benign (mesothelial or inflammatory) cells in serous effusions, MOv18 demonstrated the highest specificity (100%) since none of the 73 samples from benign effusions were stained upon incubation with this antibody. OC 125 cannot be used for this purpose due to its reactivity with mesothelial cells in benign samples. Staining cytologic preparations of malignant effusions from cancer patients with carcinomas not originating in the ovary revealed that OV632 and OV-TL 23 may be useful adjuncts to determine the origin of the carcinoma cells found in serous effusions. The reactivity of these antibodies was highly selective for ovarian carcinoma cells, staining only 6% and 0% of the samples from the non-ovarian carcinoma samples, respectively. It is concluded that MOv18 is the most suitable antibody for distinguishing ovarian carcinoma cells from mesothelial cells in serous effusions, while OV632 and OV-TL 23 especially may help to assess whether carcinoma cells found in effusions originate in the ovary.
Purpose: To investigate the optimal silver nitrate concentration and tooth immersion period for a... more Purpose: To investigate the optimal silver nitrate concentration and tooth immersion period for assessing marginal leakage at the sealant-enamel interface, using micro-CT. Methods: Two experiments were conducted, using eight teeth sealed with a resin composite without etching, in dried but not desiccated, pits and fissures. Five teeth were immersed in 50% silver nitrate for 0, 1, 2, 3, and 4 hours and scanned, using micro-CT, and two evaluators using a newly developed index assessed marginal leakage at the sealant-enamel interface (Experiment A). Experiment B was carried out in the same way as Experiment A, with the difference that teeth were immersed in 50% and 60% silver nitrate for 2 and 3 hours. Reliability of scores was estimated, using weighted kappa coefficient. ANOVA and t-test were applied to test for period and concentration effects. Results: Reliability of collected data was substantial. A statistically significant difference was observed between 50% silver nitrate penetration scores at 0-1 and 2-4 hours of immersion (P < 0.0001) but not between 50% silver nitrate penetration scores at 2-, 3-, and 4-hours of immersion (P > 0.05) (experiment A). Analysis in experiment B showed a period effect (P = 0.03) but no concentration (P = 0.07) and interaction effect (P = 0.64). Combining all data from 50% silver nitrate immersion, analysis showed no statistically significant difference in penetration scores between 2, 3 and 4 hours of immersion (P = 0.33). The newly developed index showed substantial reliability.
The aim of this study was to validate the micro-CT and related software against the section metho... more The aim of this study was to validate the micro-CT and related software against the section method using the stereomicroscope for marginal leakage assessment along the sealant-enamel interface. Pits and fissures of the occlusal surface of 10 teeth were sealed with a resin-fissure sealant material without acid etching, thermocycled for 5000 cycles, immersed in 50% silver nitrate for three hours and scanned using micro-CT. Teeth were embedded in epoxy resin and cut in three sections. The middle section was subjected to micro-CT and stereomicroscopy. Images were taken from the left and right sides of the sealant-enamel interface at both the left and the right site of the section. Two experienced evaluators assessed marginal leakage. Both assessment instruments observed no leakage in 37 out of the 40 images evaluated. Leakage at the sealant-enamel interface was observed in three stereomicroscopy images only. A fracture line in the sealant was seen on eight stereomicroscopy images and observed in only two micro-CT images. The quality of the micro-CT and related software used in the present study does not qualify it to replace the section method as the gold standard for marginal leakage assessment at the sealant-enamel interface of permanent teeth.
Purpose: To evaluate the reliability of marginal leakage assessment of self-etch adhesive Class I... more Purpose: To evaluate the reliability of marginal leakage assessment of self-etch adhesive Class II resin composite restorations in primary molars prepared in vivo using the micro-CT. Methods: Fourteen primary molars, divided over seven ART and seven traditional resin composite Class II restorations, were subjected to marginal leakage test using a 50% (w/v) silver nitrate solution for 4 hours by a micro-CT. Two evaluators selected the image with the deepest marginal dye penetration for each restoration from the occlusal and approximal surfaces on consensus. The deepest dye penetration depth and total length of the axial cavity wall from both the occlusal and from the approximl surface were measured manually using morphometric quantification software by three evaluators independently. ANOVA was applied to test for differences in marginal leakage scores between the three evaluators. Results: There was no marginal leakage found in five traditional and one ART restoration. The percentage of silver nitrate penetration depth from the cervical-approximal site ranged from 10.2-92.6%. There was only a statistically significant difference observed in measuring total axial wall length from the cervical-approximal site between the three evaluators (P = 0.02). This explanatory investigation showed that the micro-CT was a very useful device for developing a standardized method for measuring marginal leakage from samples obtained in vivo. In combination with 50% (w/v) silver nitrate and 4-hour immersion, marginal leakage along the restoration-tooth tissue interface was accurately and reliably measured. (Am J Dent 2008;21:393-397).
Magnetic Resonance Materials in Physics Biology and Medicine, Nov 1, 2008
Histopathology of prostate needle biopsies (PNBs) is an important part in the diagnosis, prognosi... more Histopathology of prostate needle biopsies (PNBs) is an important part in the diagnosis, prognosis and treatment evaluation of prostate cancer. The determination of metabolite levels in the same biopsies may have additional clinical value. Here, we demonstrate the use of non-destructive high resolution magic angle spinning (HRMAS) proton NMR Spectroscopy for the assessment of metabolic profiles of prostate tissue in PNBs as commonly obtained in standard clinical practice. PNBs that were taken routinely from 48 patients suspected of having prostate cancer were subjected to HRMAS proton NMR spectroscopy. Subsequent histopathology of the same biopsies classified the tissue either as cancer (n = 10) or benign (n = 30). Some practical aspects of this assessment were evaluated, such as typical spectral contamination caused by the PNB procedure. Significant metabolic differences were found between malignant and benign tissue using a small set of ratio&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s involving signals of choline compounds, citrate and lactate. Moreover, significant correlations were observed between choline, total choline, and citrate over creatine signal ratios and the Gleason scores of tumor in PNBs and of tumor in the whole prostate. This preliminary study indicates that HRMAS NMR of routinely obtained PNBs can provide detailed metabolic information of intact prostate tissue with clinical relevance.
Abstract This chapter focuses on the application of high-resolution X-ray imaging modalities curr... more Abstract This chapter focuses on the application of high-resolution X-ray imaging modalities currently available for the assessment of biomaterials and (bone) tissue-engineered constructs, with a specific focus on micro-computed tomography (CT) and CT-derived techniques. It also discusses the development, applications, and limitations of both in vivo and ex vivo micro-CT imaging methods. Moreover, it describes state-of-the-art X-ray imaging techniques, like X-ray phase contrast, scatter contrast, fluorescence contrast, and hybrid X-ray imaging. Finally, it presents challenging nano-resolution multimodal in vivo imaging. Such techniques are providing a simultaneous view into associated molecular, functional, and anatomical changes.
In tissue engineering research, various three-dimensional (3D) techniques are available to study ... more In tissue engineering research, various three-dimensional (3D) techniques are available to study cell morphology, biomaterials, and their relations. To overcome disadvantages of frequently used imaging techniques, in the current study stereoimaging scanning electron microscopy (SEM) is proposed. First, the 3D SEM application was validated using a series of standardized microspheres. Thereafter, MC-3T3 cell morphology was visualized and cell parameters as cell height were quantified on titanium and calcium-phosphate materials using 3D reconstruction software. Besides 3D visualization of the cells, quantitative assessment showed significant substrate dependency of cell spreading in time. Such quantification of cell spreading kinetics can be used for optimization of tissue engineering scaffold surface properties. However, further standardization of SEM image acquisition and 3D SEM software settings are still essential for 3D cell analysis.
BACKGROUND CONTEXT Lumbar interbody fusion is an effective treatment for unstable spinal segments... more BACKGROUND CONTEXT Lumbar interbody fusion is an effective treatment for unstable spinal segments. However, the time needed to establish a solid bony interbody fusion between the two vertebrae may be longer than twelve months after surgery. During this time window, the instrumented spinal segment is assumed to be at increased risk for instability related complications such as cage migration or subsidence. It is hypothesized that the design of new interbody cages that enable direct osseointegration of the cage at the vertebral endplates, without requiring full bony fusion between the two vertebral endplates, might shorten the time window that the instrumented spinal segment is susceptible to failure. PURPOSE To quantify the bone ingrowth and resulting segmental stability during consolidation of lumbar interbody fusion using two different cage types. STUDY DESIGN Preclinical ovine model. METHODS Seven skeletally mature sheep underwent bi-segmental lumbar interbody fusion surgery with one conventional polyether ether ketone (PEEK) cage, and one newly developed trussed titanium (TT) cage. After a postoperative time period of 13 weeks, non-destructive range of motion testing and histological analysis were performed. Additionally, sample specific finite element (FE) analysis was performed to predict the stability of the interbody fusion region alone. RESULTS Physiological movement of complete spinal motion segments did not reveal significant differences between the segments operated with PEEK and TT cages. The onset of creeping substitution within the cage seemed to be sooner for PEEK cages, which led to significantly higher bone volume over total volume (BV/TV) compared to the TT cages. TT cages showed significantly more direct bone to implant contact (BIC). Although the mean stability of the interbody fusion region alone was not statistically different between the PEEK and TT cages, the variation within the cage types illustrated an all-or-nothing response for the PEEK cages while a more gradual increase in stability was found for the TT cages. CONCLUSIONS Spinal segments operated with conventional PEEK cages were not different from those operated with newly developed TT cages in terms of segmental stability but did show a different mechanism of bone ingrowth and attachment. Based on the differences in development of bony fusion, we hypothesize that TT cages might facilitate increased early segmental stability by direct osseointegration of the cage at the vertebral endplates without requiring complete bony bridging through the cage. CLINICAL SIGNIFICANCE Interbody cage type affects the consolidation process of spinal interbody fusion. Whether different consolidation processes of spinal interbody fusion result in clinically significant differences requires further investigation.
Monoclonal antibodies OC 125, OV632, OV-TL 3, MOv18 and OV-TL 23, directed against distinct ovari... more Monoclonal antibodies OC 125, OV632, OV-TL 3, MOv18 and OV-TL 23, directed against distinct ovarian carcinoma-associated antigens, were examined for their value in cytopathologic diagnosis. Their sensitivity and specificity in staining ovarian carcinoma cells in serous effusions we determined using the indirect immunoperoxidase technique. Smears prepared from 140 serous effusions (73 benign, 67 malignant) were immunostained with the five antibodies. OC 125 and MOv18 reacted positively with 96% and 81% of the smears of effusions from ovarian carcinoma patients, respectively, while OV-TL 3, OV632 and OV-TL 23 stained a lower percentage of the samples (73%, 65% and 62%, respectively). In discriminating ovarian carcinoma cells from benign (mesothelial or inflammatory) cells in serous effusions, MOv18 demonstrated the highest specificity (100%) since none of the 73 samples from benign effusions were stained upon incubation with this antibody. OC 125 cannot be used for this purpose due to its reactivity with mesothelial cells in benign samples. Staining cytologic preparations of malignant effusions from cancer patients with carcinomas not originating in the ovary revealed that OV632 and OV-TL 23 may be useful adjuncts to determine the origin of the carcinoma cells found in serous effusions. The reactivity of these antibodies was highly selective for ovarian carcinoma cells, staining only 6% and 0% of the samples from the non-ovarian carcinoma samples, respectively. It is concluded that MOv18 is the most suitable antibody for distinguishing ovarian carcinoma cells from mesothelial cells in serous effusions, while OV632 and OV-TL 23 especially may help to assess whether carcinoma cells found in effusions originate in the ovary.
Purpose: To investigate the optimal silver nitrate concentration and tooth immersion period for a... more Purpose: To investigate the optimal silver nitrate concentration and tooth immersion period for assessing marginal leakage at the sealant-enamel interface, using micro-CT. Methods: Two experiments were conducted, using eight teeth sealed with a resin composite without etching, in dried but not desiccated, pits and fissures. Five teeth were immersed in 50% silver nitrate for 0, 1, 2, 3, and 4 hours and scanned, using micro-CT, and two evaluators using a newly developed index assessed marginal leakage at the sealant-enamel interface (Experiment A). Experiment B was carried out in the same way as Experiment A, with the difference that teeth were immersed in 50% and 60% silver nitrate for 2 and 3 hours. Reliability of scores was estimated, using weighted kappa coefficient. ANOVA and t-test were applied to test for period and concentration effects. Results: Reliability of collected data was substantial. A statistically significant difference was observed between 50% silver nitrate penetration scores at 0-1 and 2-4 hours of immersion (P < 0.0001) but not between 50% silver nitrate penetration scores at 2-, 3-, and 4-hours of immersion (P > 0.05) (experiment A). Analysis in experiment B showed a period effect (P = 0.03) but no concentration (P = 0.07) and interaction effect (P = 0.64). Combining all data from 50% silver nitrate immersion, analysis showed no statistically significant difference in penetration scores between 2, 3 and 4 hours of immersion (P = 0.33). The newly developed index showed substantial reliability.
The aim of this study was to validate the micro-CT and related software against the section metho... more The aim of this study was to validate the micro-CT and related software against the section method using the stereomicroscope for marginal leakage assessment along the sealant-enamel interface. Pits and fissures of the occlusal surface of 10 teeth were sealed with a resin-fissure sealant material without acid etching, thermocycled for 5000 cycles, immersed in 50% silver nitrate for three hours and scanned using micro-CT. Teeth were embedded in epoxy resin and cut in three sections. The middle section was subjected to micro-CT and stereomicroscopy. Images were taken from the left and right sides of the sealant-enamel interface at both the left and the right site of the section. Two experienced evaluators assessed marginal leakage. Both assessment instruments observed no leakage in 37 out of the 40 images evaluated. Leakage at the sealant-enamel interface was observed in three stereomicroscopy images only. A fracture line in the sealant was seen on eight stereomicroscopy images and observed in only two micro-CT images. The quality of the micro-CT and related software used in the present study does not qualify it to replace the section method as the gold standard for marginal leakage assessment at the sealant-enamel interface of permanent teeth.
Purpose: To evaluate the reliability of marginal leakage assessment of self-etch adhesive Class I... more Purpose: To evaluate the reliability of marginal leakage assessment of self-etch adhesive Class II resin composite restorations in primary molars prepared in vivo using the micro-CT. Methods: Fourteen primary molars, divided over seven ART and seven traditional resin composite Class II restorations, were subjected to marginal leakage test using a 50% (w/v) silver nitrate solution for 4 hours by a micro-CT. Two evaluators selected the image with the deepest marginal dye penetration for each restoration from the occlusal and approximal surfaces on consensus. The deepest dye penetration depth and total length of the axial cavity wall from both the occlusal and from the approximl surface were measured manually using morphometric quantification software by three evaluators independently. ANOVA was applied to test for differences in marginal leakage scores between the three evaluators. Results: There was no marginal leakage found in five traditional and one ART restoration. The percentage of silver nitrate penetration depth from the cervical-approximal site ranged from 10.2-92.6%. There was only a statistically significant difference observed in measuring total axial wall length from the cervical-approximal site between the three evaluators (P = 0.02). This explanatory investigation showed that the micro-CT was a very useful device for developing a standardized method for measuring marginal leakage from samples obtained in vivo. In combination with 50% (w/v) silver nitrate and 4-hour immersion, marginal leakage along the restoration-tooth tissue interface was accurately and reliably measured. (Am J Dent 2008;21:393-397).
Magnetic Resonance Materials in Physics Biology and Medicine, Nov 1, 2008
Histopathology of prostate needle biopsies (PNBs) is an important part in the diagnosis, prognosi... more Histopathology of prostate needle biopsies (PNBs) is an important part in the diagnosis, prognosis and treatment evaluation of prostate cancer. The determination of metabolite levels in the same biopsies may have additional clinical value. Here, we demonstrate the use of non-destructive high resolution magic angle spinning (HRMAS) proton NMR Spectroscopy for the assessment of metabolic profiles of prostate tissue in PNBs as commonly obtained in standard clinical practice. PNBs that were taken routinely from 48 patients suspected of having prostate cancer were subjected to HRMAS proton NMR spectroscopy. Subsequent histopathology of the same biopsies classified the tissue either as cancer (n = 10) or benign (n = 30). Some practical aspects of this assessment were evaluated, such as typical spectral contamination caused by the PNB procedure. Significant metabolic differences were found between malignant and benign tissue using a small set of ratio&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;s involving signals of choline compounds, citrate and lactate. Moreover, significant correlations were observed between choline, total choline, and citrate over creatine signal ratios and the Gleason scores of tumor in PNBs and of tumor in the whole prostate. This preliminary study indicates that HRMAS NMR of routinely obtained PNBs can provide detailed metabolic information of intact prostate tissue with clinical relevance.
Abstract This chapter focuses on the application of high-resolution X-ray imaging modalities curr... more Abstract This chapter focuses on the application of high-resolution X-ray imaging modalities currently available for the assessment of biomaterials and (bone) tissue-engineered constructs, with a specific focus on micro-computed tomography (CT) and CT-derived techniques. It also discusses the development, applications, and limitations of both in vivo and ex vivo micro-CT imaging methods. Moreover, it describes state-of-the-art X-ray imaging techniques, like X-ray phase contrast, scatter contrast, fluorescence contrast, and hybrid X-ray imaging. Finally, it presents challenging nano-resolution multimodal in vivo imaging. Such techniques are providing a simultaneous view into associated molecular, functional, and anatomical changes.
In tissue engineering research, various three-dimensional (3D) techniques are available to study ... more In tissue engineering research, various three-dimensional (3D) techniques are available to study cell morphology, biomaterials, and their relations. To overcome disadvantages of frequently used imaging techniques, in the current study stereoimaging scanning electron microscopy (SEM) is proposed. First, the 3D SEM application was validated using a series of standardized microspheres. Thereafter, MC-3T3 cell morphology was visualized and cell parameters as cell height were quantified on titanium and calcium-phosphate materials using 3D reconstruction software. Besides 3D visualization of the cells, quantitative assessment showed significant substrate dependency of cell spreading in time. Such quantification of cell spreading kinetics can be used for optimization of tissue engineering scaffold surface properties. However, further standardization of SEM image acquisition and 3D SEM software settings are still essential for 3D cell analysis.
BACKGROUND CONTEXT Lumbar interbody fusion is an effective treatment for unstable spinal segments... more BACKGROUND CONTEXT Lumbar interbody fusion is an effective treatment for unstable spinal segments. However, the time needed to establish a solid bony interbody fusion between the two vertebrae may be longer than twelve months after surgery. During this time window, the instrumented spinal segment is assumed to be at increased risk for instability related complications such as cage migration or subsidence. It is hypothesized that the design of new interbody cages that enable direct osseointegration of the cage at the vertebral endplates, without requiring full bony fusion between the two vertebral endplates, might shorten the time window that the instrumented spinal segment is susceptible to failure. PURPOSE To quantify the bone ingrowth and resulting segmental stability during consolidation of lumbar interbody fusion using two different cage types. STUDY DESIGN Preclinical ovine model. METHODS Seven skeletally mature sheep underwent bi-segmental lumbar interbody fusion surgery with one conventional polyether ether ketone (PEEK) cage, and one newly developed trussed titanium (TT) cage. After a postoperative time period of 13 weeks, non-destructive range of motion testing and histological analysis were performed. Additionally, sample specific finite element (FE) analysis was performed to predict the stability of the interbody fusion region alone. RESULTS Physiological movement of complete spinal motion segments did not reveal significant differences between the segments operated with PEEK and TT cages. The onset of creeping substitution within the cage seemed to be sooner for PEEK cages, which led to significantly higher bone volume over total volume (BV/TV) compared to the TT cages. TT cages showed significantly more direct bone to implant contact (BIC). Although the mean stability of the interbody fusion region alone was not statistically different between the PEEK and TT cages, the variation within the cage types illustrated an all-or-nothing response for the PEEK cages while a more gradual increase in stability was found for the TT cages. CONCLUSIONS Spinal segments operated with conventional PEEK cages were not different from those operated with newly developed TT cages in terms of segmental stability but did show a different mechanism of bone ingrowth and attachment. Based on the differences in development of bony fusion, we hypothesize that TT cages might facilitate increased early segmental stability by direct osseointegration of the cage at the vertebral endplates without requiring complete bony bridging through the cage. CLINICAL SIGNIFICANCE Interbody cage type affects the consolidation process of spinal interbody fusion. Whether different consolidation processes of spinal interbody fusion result in clinically significant differences requires further investigation.
Uploads
Papers by Vincent M . J . I . Cuijpers