Rabbit Elastase Aneurysm Model Mimics the Recurrence Rate of Human Intracranial Aneurysms following Platinum Coil Embolization.

BACKGROUND AND PURPOSE: Intracranial aneurysms treated with coils have been associated with incomplete occlusion, particularly in large or wide-neck aneurysms. This study aimed to validate the accuracy of the rabbit elastase model in predicting aneurysm recurrence in humans treated with platinum coils. MATERIALS AND METHODS: Elastase-induced saccular aneurysms were induced in rabbits and embolized with conventional platinum coils. The recurrence rates of aneurysms were retrospectively analyzed. Morphologic characteristics of aneurysms, angiographic outcomes, and histologic healing were evaluated. RESULTS: A total of 28 (15.3%) of 183 aneurysms recurred. The aneurysm recurrence rate observed in this study (15.3%) is similar to those reported in multiple analyses of aneurysm recurrence rates in humans (7%-27%). The rate of recurrence was higher in aneurysms treated without balloon assistance (19/66, 28.8%) compared with those treated with balloon assistance (9/117, 7.7%). Aneurysms treated with balloon-assisted coiling had a lower recurrence rate (OR = 0.17; 95% CI, 0.05-0.47; P = .001) and higher occlusion rate (OR = 6.88; 95% CI, 2.58-20.37; P < .001) compared with those treated without balloon-assisted coiling. In this rabbit elastase-induced aneurysm model, packing density and aneurysm volume were weak predictors of aneurysm recurrence; however, the packing density was a good predictor of the occlusion rate (OR = 1.05; 95% CI, 1.02-1.10; P = .008). CONCLUSIONS: The rabbit elastase aneurysm model may mimic aneurysm recurrence rates observed in humans after platinum coil embolization. Moreover, balloon assistance and high packing densities were significant predictors of aneurysm recurrence and occlusion.

Evaluation of Outcome Prediction of Flow Diversion for Intracranial Aneurysms.

BACKGROUND AND PURPOSE: Identifying and predicting which aneurysms are likely to quickly occlude and which ones are likely to remain open following treatment with flow-diverting devices is important to develop optimal patient management strategies. The purpose of this study was to evaluate predictions based on computational fluid dynamics models using the elastase rabbit aneurysm model. MATERIALS AND METHODS: A series of 13 aneurysms created in rabbits were treated with flow diverters, and outcomes were angiographically assessed at 8 weeks' follow-up. Computational fluid dynamics models were constructed from pretreatment 3D rotational angiograms and Doppler ultrasound flow velocity measurements. Postimplantation mean aneurysm inflow rate and flow velocity were used to prospectively predict aneurysm occlusion blinded to the actual outcomes. Specifically, if both variables were below their corresponding thresholds, fast occlusion was predicted, while if one of them was above the threshold, slow or incomplete occlusion was predicted. RESULTS: Of the 13 aneurysms included, 8 were incompletely occluded 8 weeks after treatment, and 5 were completely occluded. A total of 10 computational fluid dynamics-based predictions agreed with the angiographic outcome, reaching 77% accuracy, 80% sensitivity, and 75% specificity. Posttreatment mean velocity alone was able to achieve the same predictive power as the combination of inflow rate and velocity. CONCLUSIONS: Subject-specific computational fluid dynamics models of the hemodynamic conditions created immediately after implantation of flow-diverting devices in experimental aneurysms created in rabbits are capable of prospectively predicting, with a reasonable accuracy, which aneurysms will completely occlude and which ones will remain incompletely occluded.

Adaptive Remodeling in the Elastase-induced Rabbit Aneurysms.

BACKGROUND: Rupture of brain aneurysms is associated with high fatality and morbidity rates. Through remodeling of the collagen matrix, many aneurysms can remain unruptured for decades, despite an enlarging and evolving geometry. OBJECTIVE: Our objective was to explore this adaptive remodeling for the first time in an elastase induced aneurysm model in rabbits. METHODS: Saccular aneurysms were created in 22 New Zealand white rabbits and remodeling was assessed in tissue harvested 2, 4, 8 and 12 weeks after creation. RESULTS: The intramural principal stress ratio doubled after aneurysm creation due to increased longitudinal loads, triggering a remodeling response. A distinct wall layer with multi-directional collagen fibers developed between the media and adventitia as early as 2 weeks, and in all cases by 4 weeks with an average thickness of 50.6 ± 14.3 µm. Collagen fibers in this layer were multi-directional (AI = 0.56 ± 0.15) with low tortuosity (1.08 ± 0.02) compared with adjacent circumferentially aligned medial fibers (AI = 0.78 ± 0.12) and highly tortuous adventitial fibers (1.22 ± 0.03). A second phase of remodeling replaced circumferentially aligned fibers in the inner media with longitudinal fibers. A structurally motivated constitutive model with both remodeling modes was introduced along with methodology for determining material parameters from mechanical testing and multiphoton imaging. CONCLUSIONS: A new mechanism was identified by which aneurysm walls can rapidly adapt to changes in load, ensuring the structural integrity of the aneurysm until a slower process of medial reorganization occurs. The rabbit model can be used to evaluate therapies to increase aneurysm wall stability.

Histologic and Biomolecular Similarities in Healing between Aneurysms and Cutaneous Skin Wounds.

The poorly understood mechanisms of aneurysm healing contribute substantially to the pressing medical problem of coiled aneurysm recanalization. Using an established saccular aneurysm model, we developed an animal model system in rabbits to study aneurysm and skin wound healing concurrently in the same animal. We found treated aneurysm healing to be similar to skin wound healing both histologically and in biomarker gene and protein expression, but in a delayed fashion.

Wall Apposition Is a Key Factor for Aneurysm Occlusion after Flow Diversion: A Histologic Evaluation in 41 Rabbits.

BACKGROUND AND PURPOSE: Robust wall apposition for flow-diverter stents may be important for endothelialization. Using a large series of experimental aneurysms treated with the Pipeline Embolization Device, the objectives of this study were to 1) assess interobserver agreement for the evaluation of wall apposition on posttreatment DSA and evaluate its association with aneurysm occlusion, and 2) measure the relationship between wall apposition assessed with histology and aneurysm occlusion rate after treatment. MATERIALS AND METHODS: Saccular aneurysms were created in 41 rabbits and treated with the Pipeline Embolization Device. DSA was performed just after the deployment of the device and at follow-up. Three investigators independently graded wall apposition on posttreatment DSA as good or poor. A histopathologist blinded to the angiographic results graded the wall apposition on histologic samples. We examined the correlation between angiographic occlusion and wall apposition with histology and angiography. RESULTS: Wall apposition evaluated on histology was strongly associated with saccular aneurysm occlusion. Sensitivity and specificity of wall apposition to predict complete occlusion at follow-up were 76.9% and 84.0%, respectively, with an overall accuracy of 81.6%. In this experimental study, DSA was suboptimal to assess flow-diverter apposition, with moderate interobserver agreement and low accuracy. CONCLUSIONS: Good wall apposition is strongly associated with complete occlusion after flow-diverter therapy. In this study, DSA was suboptimal for assessing wall apposition of flow-diverter stents. These findings suggest that improved tools for assessing flow diverter-stent wall apposition are highly relevant.

Differential Gene Expression in Coiled versus Flow-Diverter-Treated Aneurysms: RNA Sequencing Analysis in a Rabbit Aneurysm Model.

BACKGROUND AND PURPOSE: The biologic mechanisms leading to aneurysm healing or rare complications such as delayed aneurysm ruptures after flow-diverter placement remain poorly understood. We used RNA sequencing following implantation of coils or flow diverters in elastase aneurysms in rabbits to identify genes and pathways of potential interest. MATERIALS AND METHODS: Aneurysms were treated with coils (n = 5) or flow diverters (n = 4) or were left untreated for controls (n = 6). Messenger RNA was isolated from the aneurysms at 4 weeks following treatment. RNA samples were processed by using RNA-sequencing technology and were analyzed by using the Ingenuity Pathway Analysis tool. RESULTS: With RNA sequencing for coiled versus untreated aneurysms, 464/9990 genes (4.6%) were differentially expressed (58 down-regulated, 406 up-regulated). When we compared flow-diverter versus untreated aneurysms, 177/10,041 (1.8%) genes were differentially expressed (8 down-regulated, 169 up-regulated). When we compared flow-diverter versus coiled aneurysms, 13/9982 (0.13%) genes were differentially expressed (8 down-regulated, 5 up-regulated). Keratin 8 was overexpressed in flow diverters versus coils. This molecule may potentially play a critical role in delayed ruptures due to plasmin production. We identified overregulation of apelin in flow diverters, supporting the preponderance of endothelialization, whereas we found overexpression of molecules implicated in wound healing (dectin 1 and hedgehog interacting protein) for coiled aneurysms. Furthermore, we identified metallopeptidases 1, 12, and 13 as overexpressed in coiled versus untreated aneurysms. CONCLUSIONS: We observed different physiopathologic responses after endovascular treatment with various devices. Flow diverters promote endothelialization but express molecules that could potentially explain the rare delayed ruptures. Coils promote wound healing and express genes potentially implicated in the recurrence of coiled aneurysms.

Evaluation of the Angiographic Grading Scale in Aneurysms Treated with the WEB Device in 80 Rabbits: Correlation with Histologic Evaluation.

BACKGROUND AND PURPOSE: The WEB Occlusion Score has been proposed to assess angiographic outcomes for intracranial aneurysms treated with the Woven EndoBridge (WEB) device. Using a large series of experimental aneurysms treated with the WEB, we had the following objectives: 1) to compare angiographic outcomes as measured by the WEB Occlusion Scale with histologic results, and 2) to assess interobserver and intraobserver agreement of the WEB Occlusion Scale. MATERIALS AND METHODS: Intracranial aneurysms were created in 80 rabbits and treated with WEB devices. Animals were sacrificed at last follow-up for histologic evaluation. DSA was performed just after the deployment of the device and at follow-up. Four investigators independently and retrospectively graded the DSA twice according to the WEB Occlusion Scale. One histopathologist blinded to the angiographic results graded the occlusion according to a 4-point scale patterned on the WEB Occlusion Scale. Intra- and interobserver agreement were evaluated for DSA. Follow-up angiographic grading and histologic reference were compared to determine the WEB Occlusion Scale accuracy for complete (with or without recess filling) versus incomplete occlusion and adequate (complete occlusion or neck remnant) versus inadequate occlusion. RESULTS: Inter- and intraobserver weighted κ for the angiographic WEB Occlusion Scale were, respectively, 0.76 and 0.76, indicating substantial agreement. The sensitivity and specificity of the WEB Occlusion Scale for complete occlusion at follow-up compared with the histologic reference standard were, respectively, 75% and 83.3%, with an overall accuracy of 80%. Similarly, for adequate occlusion at follow-up, sensitivity was 97.7%, specificity was 64.9%, and overall accuracy was 82.5%. CONCLUSIONS: The WEB Occlusion Scale appears to be consistent, reliable, and accurate compared with a histologic reference standard.

RNA-Sequencing Analysis of Messenger RNA/MicroRNA in a Rabbit Aneurysm Model Identifies Pathways and Genes of Interest.

BACKGROUND AND PURPOSE: Rabbit aneurysm models are used for the testing of embolization devices and elucidating the mechanisms of human intracranial aneurysm growth and healing. We used RNA-sequencing technology to identify genes relevant to induced rabbit aneurysm biology and to identify genes and pathways of potential clinical interest. This process included sequencing microRNAs, which are important regulatory noncoding RNAs. MATERIALS AND METHODS: Elastase-induced saccular aneurysms were created at the origin of the right common carotid artery in 6 rabbits. Messenger RNA and microRNA were isolated from the aneurysm and from the control left common carotid artery at 12 weeks and processed by using RNA-sequencing technology. The results from RNA sequencing were analyzed by using the Ingenuity Pathway Analysis tool. RESULTS: A total of 9396 genes were analyzed by using RNA sequencing, 648 (6.9%) of which were found to be significantly differentially expressed between the aneurysms and control tissues (P < .05; false-discovery rate, <0.01; fold change, >2 or <.5). Of these genes, 614 were mapped successfully, 143 were down-regulated, and 471 were up-regulated in the aneurysms as compared with controls. Using the same criteria for significance, 3 microRNAs were identified as down-regulated and 5 were identified as up-regulated. Pathway analysis associated these genes with inflammatory response, cellular migration, and coagulation, among other functions and pathologies. CONCLUSIONS: RNA-sequencing analysis of rabbit aneurysms revealed differential regulation of some key pathways, including inflammation and antigen presentation. ANKRD1 and TACR1 were identified as genes of interest in the regulation of matrix metalloproteinases.

Analysis of hemodynamics and aneurysm occlusion after flow-diverting treatment in rabbit models.

BACKGROUND AND PURPOSE: Predicting the outcome of flow diversion treatment of cerebral aneurysms remains challenging. Our aim was to investigate the relationship between hemodynamic conditions created immediately after flow diversion and subsequent occlusion of experimental aneurysms in rabbits. MATERIALS AND METHODS: The hemodynamic environment before and after flow-diversion treatment of elastase-induced aneurysms in 20 rabbits was modeled by using image-based computational fluid dynamics. Local aneurysm occlusion was quantified by using a voxelization technique on 3D images acquired 8 weeks after treatment. Global and local voxel-by-voxel hemodynamic variables were used to statistically compare aneurysm regions that later thrombosed to regions that remained patent. RESULTS: Six aneurysms remained patent at 8 weeks, while 14 were completely or nearly completely occluded. Patent aneurysms had statistically larger neck sizes (P = .0015) and smaller mean transit times (P = .02). The velocity, vorticity, and shear rate were approximately 2.8 times (P < .0001) larger in patent regions-that is, they had larger "flow activity" than regions that progressed to occlusion. Statistical models based on local hemodynamic variables were capable of predicting local occlusion with good precision (84% accuracy), especially away from the neck (92%-94%). Predictions near the neck were poorer (73% accuracy). CONCLUSIONS: These results suggests that the dominant healing mechanism of occlusion within the aneurysm dome is related to slow-flow-induced thrombosis, while near the neck, other processes could be at play simultaneously.

Co-precipitation synthesis and electrochemical properties of graphene supported LiMn1/3Ni1/3Co1/3O2 cathode materials for lithium-ion batteries.

Nanostructured graphene supported LiMn1/3Ni1/3Co1/3O2 has been prepared by the co-precipitation method and used as the cathode material for lithium-ion batteries. The structure and morphology of the composite were examined using atomic force microscopy, scanning electron microscopy, high-resolution transmission electron microscopy, and x-ray diffraction. The composite cathodes made of graphene nanosheet supported LiMn1/3Ni1/3Co1/3O2 showed superior electrochemical performance and increased capacity compared to raw LiMn1/3Ni1/3Co1/3O2 electrodes. The outstanding electrochemical behavior of graphene supported LiMn1/3Ni1/3Co1/3O2 was ascribed to the increased electrical conductivity and high structural stability resulting from the addition of graphene.

Relationship between Serum Aminotransferase Levels and Metabolic Disorders in Northern China.

BACKGROUND: Increasing evidence suggests an association between elevated serum aminotransferase levels and metabolic disorders (metabolic syndrome, hyperlipemia and diabetes mellitus). However, the significance of relatively low levels of aminotransferases in relation to metabolic disorders has not been fully investigated in the general population. We investigated the association between serum amiontransferase levels and metabolic disorders using data from a survey in Jilin province, China. METHODS: In 2007, a survey was conducted throughout Jilin, China, covering both urban and rural areas. A total of 3835 people, 18 to 79 years old including 1761 men and 2074 women, underwent real-time ultrasonography, blood tests including aspartate aminotransferase and alanine aminotransferase, and had interviews with a structured questionnaire. RESULTS: Serum aminotransferase levels within the normal range were associated with metabolic syndrome independent of age, occupation, cultural and educational level, income, body mass index, waist circumference, smoking, and alcohol intake. Compared with the lowest level (<20 IU/L), the adjusted odds ratios for ALT levels of 20-29, 30-39, 40-49 and >50 IU/L were 1.92, 2.50, 2.97, and 3.52 in men, and 1.38, 1.54, 3.06, and 2.62 in women, respectively. Near-normal serum aminotransferase levels associated with hyperlipemia, NAFLD, DM were also found in the study. CONCLUSIONS: Normal to near-normal serum aminotransferase levels are associated with metabolic disorders. Serum ALT levels of 21-25 IU/L for men, and 17-22 IU/L for women are suggested as cutoff levels that detect metabolic disorders affecting the liver.

Angiographic and histologic comparison of experimental aneurysms embolized with hydrogel filaments.

BACKGROUND AND PURPOSE: The embolization of aneurysms with hydrogel filaments allow postprocedural CT and MR imaging studies without artifacts. We compared the performance of 3 hydrogel filament formulations in rabbit experimental aneurysms by using angiography and histologic samples. MATERIALS AND METHODS: Embolization of 35 rabbit elastase or bifurcation aneurysms was performed with 3 different formulations of detachable hydrogel filaments, including 1) polyethylene glycol opacified with aromatic iodine (PEG-I; n = 12), 2) polyethylene glycol opacified with barium sulfate (PEG-B; n = 12), or 3) polypropylene glycol opacified with barium sulfate (PPG-B; n = 11). Follow-up angiography was performed before the rabbits were killed at 2 (n = 7), 6 (n = 9), 10 (n = 8), or 26 (n = 11) weeks. Angiographic occlusion was scored according to the Raymond scale, and interval changes were assessed. The harvested aneurysms were evaluated on histologic examination. From the sections, we determined the percentage of the sac excluded from the vasculature and occupied by embolic devices by using image analysis. We compared results using the analysis of variance/t test or chi(2) test. RESULTS: The mean number of devices used to treat aneurysms in the PPG-B group was significantly greater than that used for the other 2 groups, though aneurysm volumes were similar among groups. Compared with immediate posttreatment occlusion scores, mean angiographic occlusion at follow-up was increased for all 3 hydrogel filament groups. On histologic examination, thrombus organization, neointima formation, and inflammation were similar to that observed in rabbit experimental aneurysms with other embolic devices containing platinum coils. CONCLUSIONS: The embolization of experimental aneurysms with hydrogel filaments resulted in durable angiographic and histologic occlusion from 2 to 26 weeks. With improvements, hydrogel filaments free from metallic coils show promise for endovascular use.

Gene expression profiling of experimental saccular aneurysms using deoxyribonucleic acid microarrays.

BACKGROUND AND PURPOSE: The molecular characteristics of the pathophysiology of saccular aneurysms remain poorly understood. The purpose of the current study was to investigate the expression of various groups of genes at different stages of aneurysm age in elastase-induced saccular aneurysms in rabbits through the use of deoxyribonucleic acid (DNA) microarrays. MATERIALS AND METHODS: A microarray consisting of genes related to cell adhesion, apoptosis, cell signaling, growth, inflammation, vascular remodeling, and oxidative stress was constructed by using rabbit nucleotide sequences. Elastase-induced saccular aneurysms were created at the origin of the right common carotid artery (CCA) in 12 rabbits. Two weeks (n=6) and 12 weeks (n=6) after aneurysm creation, ribonucleic acid (RNA) was isolated from the aneurysm and the control unoperated left CCA and was used for microarray experiments. Real-time polymerase chain reaction (RT-PCR) was performed for validation of microarray results. RESULTS: Of 209 genes, 157 (75%) at 2 weeks and 88 (42%) at 12 weeks demonstrated statistically significant differential expression between aneurysm tissue and the control left CCA tissue (P < .05). Multiple genes implicated in vessel wall remodeling were found to be elevated at 2 weeks and at 12 weeks. Expression of cell adhesion molecules and antioxidant enzymes was down-regulated at 2 weeks but was not significantly different from that of controls at 12 weeks. Most transcription factors, inflammatory genes, and structural genes showed underexpression at both time points. The expression profiles of selected genes were confirmed by RT-PCR. CONCLUSION: Multiple genes in diverse pathways have been differentially expressed in the rabbit aneurysm model.