Shock Wave-Induced Damage and Poration in Eukaryotic Cell Membranes.

Shock waves are known to permeabilize eukaryotic cell membranes, which may be a powerful tool for a variety of drug delivery applications. However, the mechanisms involved in shock wave-mediated membrane permeabilization are still poorly understood. In this study, the effects on both the permeability and the ultrastructural features of two human cell lineages were investigated after the application of underwater shock waves in vitro. Scanning Electron Microscopy of cells derived from a human embryo kidney (HEK)-293 and Michigan Cancer Foundation (MCF)-7 cells, an immortalized culture derived from human breast adenocarcinoma, showed a small amount of microvilli (as compared to control cells), the presence of hole-like structures, and a decrease in cell size after shock wave exposure. Interestingly, these effects were accompanied by the permeabilization of acid and macromolecular dyes and gene transfection. Trypan blue exclusion assays indicated that cell membranes were porated during shock wave treatment but resealed after a few seconds. Deformations of the cell membrane lasted for at least 5 min, allowing their observation in fixed cells. For each cell line, different shock wave parameters were needed to achieve cell membrane poration. This difference was correlated to successful gene transfection by shock waves. Our results demonstrate, for the first time, that shock waves induce transient micro- and submicrosized deformations at the cell membrane, leading to cell transfection and cell survival. They also indicate that ultrastructural analyses of cell surfaces may constitute a useful way to match the use of shock waves to different cells and settings.

Is quantitative diffusion-weighted MRI a valuable technique for the detection of changes in kidneys after extracorporeal shock wave lithotripsy?

OBJECTIVE: The aim of this study was to evaluate the capability and the reliability of diffusion-weighted imaging (DWI) in the changes of kidneys occurring after extracorporeal shock wave lithotripsy (ESWL) treatment for renal stones. MATERIALS AND METHODS: A total of 32 patients who underwent ESWL treatment for renal stone disease between June and December 2011 were enrolled in this prospective study. Color Doppler ultrasonography (CDUS) and DWI were performed before and within 24 hours after ESWL. DWI was obtained with b factors of 0, 500 and 1000 s/ mm2 at 1.5 T MRI. Each of Resistive index (RI) and ADC values were calculated from the three regions of renal upper, middle and lower zones for both of the affected and contralateral kidneys. Paired sample t test was used for statistical analyses. RESULTS: After ESWL, the treated kidneys had statistically significant lower ADC values in all different regions compared with previous renal images. The best discriminative parameter was signal intensity with a b value of 1000 s/mm2. The changes of DWI after ESWL were noteworthy in the middle of the treated kidney (p < 0.01). There were no significant difference between RI values in all regions of treated and contralateral kidneys before and after treatment with ESWL (p>0.05). CONCLUSION: DWI is a valuable technique enables the detection of changes in DWI after ESWL treatment that may provide useful information in prediction of renal damage by shock waves, even CDUS is normal.

Is quantitative diffusion-weighted MRI a valuable technique for the detection of changes in kidneys after extracorporeal shock wave lithotripsy?

Objective The aim of this study was to evaluate the capability and the reliability of diffusion-weighted imaging (DWI) in the changes of kidneys occurring after extracorporeal shock wave lithotripsy (ESWL) treatment for renal stones. Materials and Methods A total of 32 patients who underwent ESWL treatment for renal stone disease between June and December 2011 were enrolled in this prospective study. Color Doppler ultrasonography (CDUS) and DWI were performed before and within 24 hours after ESWL. DWI was obtained with b factors of 0, 500 and 1000 s/mm2 at 1.5 T MRI. Each of Resistive index (RI) and ADC values were calculated from the three regions of renal upper, middle and lower zones for both of the affected and contralateral kidneys. Paired sample t test was used for statistical analyses. Results After ESWL, the treated kidneys had statistically significant lower ADC values in all different regions compared with previous renal images. The best discriminative parameter was signal intensity with a b value of 1000 s/mm2. The changes of DWI after ESWL were noteworthy in the middle of the treated kidney (p<0.01). There were no significant difference between RI values in all regions of treated and contralateral kidneys before and after treatment with ESWL (p>0.05). Conclusion DWI is a valuable technique enables the detection of changes in DWI after ESWL treatment that may provide useful information in prediction of renal damage by shock waves, even CDUS is normal. .

Effects of repeated extracorporeal shock wave on kidney apoptosis of normal and diabetic rat.

OBJECTIVE: To assess the effect of repeated extracorporeal shock waves (ESW) on the apoptosis of renal parenchyma in normal and diabetic rats by the caspase-3 expression. MATERIALS AND METHODS: 80 rats were assigned for ESW (Direx Tripter X1(R) - 14 KVA) to one of three groups with 50% rats with diabetes (A) and 50% normal (B) in each: G1 (n = 40): no ESW; G2 (n = 20): one ESW; G3 (n = 20): two ESWin an interval of 14 days. The animals were sacrificed 3 days after the ESW and samples of the renal parenchyma were histologically prepared, stained by the caspase-3 cleavage immunohistochemical method, and the apoptotic index (ApIn) was calculated by computer program Image Pro-Plus V 4,5,1 Media Cybernetics ( ApIn = number of apoptotic cells in the microscopic section / total cells in the microscopic section). RESULTS: There was no statistical difference in ApIn of normal rats with one ESW (8.4 +/- 0.2) or two ESW (9.1 +/- 0.3) and both were similar to the control group with no ESW (7.9 +/- 0.2). The ApIn of diabetic rats with one ESW (7.4 +/- 0.3) was similar to the normal rats with one or two ESW. On the other hand, the ApIn was statistically higher in diabetic rats with two ESW (11.9 +/- 0.4) than in diabetic rats with one ESW (7.4 +/- 0.3). CONCLUSIONS: The ApIn of normal rats with a single (2,000 SW) or repeated ESW (4,000 SW) was similar to the control animals without any ESW. In diabetic rats, the repeated ESW (4,000 SW) may result in an additional risk of tissue damage expressed by the higher ApIn.

Effects of repeated extracorporeal shock wave on kidney apoptosis of normal and diabetic rat

OBJECTIVE: To assess the effect of repeated extracorporeal shock waves (ESW) on the apoptosis of renal parenchyma in normal and diabetic rats by the caspase-3 expression. MATERIALS AND METHODS: 80 rats were assigned for ESW (Direx Tripter X1® - 14 KVA) to one of three groups with 50 percent rats with diabetes (A) and 50 percent normal (B) in each: G1 (n = 40): no ESW; G2 (n = 20): one ESW; G3 (n = 20): two ESWin an interval of 14 days. The animals were sacrificed 3 days after the ESW and samples of the renal parenchyma were histologically prepared, stained by the caspase-3 cleavage immunohistochemical method, and the apoptotic index (ApIn) was calculated by computer program Image Pro-Plus V 4,5,1 Media Cybernetics® ( ApIn = number of apoptotic cells in the microscopic section / total cells in the microscopic section). RESULTS: There was no statistical difference in ApIn of normal rats with one ESW (8.4 + 0.2) or two ESW (9.1 + 0.3) and both were similar to the control group with no ESW (7.9 + 0.2). The ApIn of diabetic rats with one ESW (7.4 + 0.3) was similar to the normal rats with one or two ESW. On the other hand, the ApIn was statistically higher in diabetic rats with two ESW (11.9 + 0.4) than in diabetic rats with one ESW (7.4 + 0.3). CONCLUSIONS: The ApIn of normal rats with a single (2,000 SW) or repeated ESW (4,000 SW) was similar to the control animals without any ESW. In diabetic rats, the repeated ESW (4,000 SW) may result in an additional risk of tissue damage expressed by the higher ApIn.

The repeated extracorporeal shock waves and the renal parenchyma injury on normal and diabetic rats

PURPOSE: To assess the effect of repeated extracorporeal shock waves (ESW) on renal parenchyma of normal and diabetic rats. METHODS: 40 normal rats (A) and 40 diabetic rats (B) were assigned for ESW (Direx Tripter X1® - 14 KVA) as follow: A1/B1 and A3/B3 no ESW; A2/B2 one ESW (2,000 SW); A4/B4 two ESW (4,000 SW) in an elapsed 14 days. All the animals were sacrificed 3 days after the ESW and samples of renal parenchyma were histological prepared, stained by H&E. For each animal the frequency of hemorrhage focus (HF) in the subcapasular, interstitial and glomerulus area was calculated (porcentage) on 20 randomly histological sections. RESULTS: No one HF was identified in all normal or diabetic animals without ESW (A1, A3 and B1, B3). In the normal rats the HF frequency was similar to one ESW (subcapsular =15 percent; interstitial =20 percent and glomerular =10 percent) or repetead ESW (subcapsular =25 percent; interstitial =20 percent; glomerular=10 percent). In diabetic rats the occurence of HF with repetead ESW was more frequent (subcapsular =40 percent; interstitial =30 percent and glomerular =10 percent) than with a single ESW (subcapsular =25 percent; interstitial =15 percent and glomerular =15 percent). CONCLUSION: A single ESW or a repeated ESW caused a mild and similar damage on renal cortex of normal rats. In diabetic rats the repetead ESW may result in an accumulated damage, especially with focus of hemorrhage in subcapsular and interstitial tissue and glomerulus edema.

RESUMO OBJETIVO: Avaliar o efeito de repetidas ondas de choque extracorpóreas (OCE) sobre o parênquima renal de ratos normais e diabéticos. MÉTODOS: 40 ratos normais e 40 ratos diabéticos foram distribuídos para aplicação de OCE (Direx Tripter X1® - 14 KVA) como segue: A1/B1 e A3/B3 sem OCE; A2/B2 uma sessão de OCE (2000 OC); A4/B4 duas sessões de OC (4000 OC) num intervalo de 14 dias. Todos os animais foram sacrificados no 3°. dia após a aplicação da OCE e amostras de parênquima renal foram histologicamente preparados e corados em H&E. Para cada animal foi calculado, em 20 campos aleatórios, a freqüência (em porcentagem) de focos hemorrágicos (FH) nas áreas subcapsular, intersticial e glomerular. RESULTADOS: Nenhum foco hemorrágico foi identificado nos animais normais ou diabéticos que não receberam nenhuma OCE (A1, A3 e B1, B3). Nos ratos normais a freqüência de FH foi similar com uma sessão de OCE (subcapsular =15 por cento; intersticial =20 por cento e glomerular =10 por cento) ou duas sessões de OCE (subcapsular =25 por cento; intersticial =20 por cento; glomerular =10 por cento). Nos ratos diabéticos a ocorrência de FH com duas sessões de OCE foi mais freqüente (subcapsular =40 por cento; intersticial =30 por cento e glomerular =10 por cento) do que com uma simples sessão de OCE (subcapsular =25 por cento; intersticial =15 por cento e glomerular =15 por cento). CONCLUSÃO: Uma única ou duas sessões de OCE causa um discreto e semelhante dano no parênquima renal de ratos normais. Nos ratos diabéticos a repetição da OCE pode resultar em acúmulo de danos, especialmente com FH nas áreas subcapsular e intersticial e no edema do glomérulo.

The repeated extracorporeal shock waves and the renal parenchyma injury on normal and diabetic rats.

PURPOSE: To assess the effect of repeated extracorporeal shock waves (ESW) on renal parenchyma of normal and diabetic rats. METHODS: 40 normal rats (A) and 40 diabetic rats (B) were assigned for ESW (Direx Tripter X1 - 14 KVA) as follow: A1/B1 and A3/B3 no ESW; A2/B2 one ESW (2,000 SW); A4/B4 two ESW (4,000 SW) in an elapsed 14 days. All the animals were sacrificed 3 days after the ESW and samples of renal parenchyma were histological prepared, stained by H&E. For each animal the frequency of hemorrhage focus (HF) in the subcapsular, interstitial and glomerulus area was calculated (percentage) on 20 randomly histological sections. RESULTS: No one HF was identified in all normal or diabetic animals without ESW (A1, A3 and B1, B3). In the normal rats the HF frequency was similar to one ESW (subcapsular =15%; interstitial =20% and glomerular =10%) or repeated ESW (subcapsular =25%; interstitial =20%; glomerular=10%). In diabetic rats the occurrence of HF with repeated ESW was more frequent (subcapsular =40%; interstitial =30% and glomerular =10%) than with a single ESW (subcapsular =25%; interstitial =15% and glomerular =15%). CONCLUSION: A single ESW or a repeated ESW caused a mild and similar damage on renal cortex of normal rats. In diabetic rats the repeated ESW may result in an accumulated damage, especially with focus of hemorrhage in subcapsular and interstitial tissue and glomerulus edema.