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Introducción. El objetivo de este estudio consistió en investigar el efecto aditivo de las células madre mesenquimales (MSC, por sus siglas en inglés) y del defibrótido (DFT) en un modelo de trombosis arterial femoral en ratas. Métodos. Se incluyeron 30 ratas Sprague Dawley. Se desarrolló un modelo de trombosis arterial mediante cloruro de hierro (FeCl3) en la arteria femoral izquierda. Las ratas se asignaron equitativamente en cinco grupos: grupo 1, intervención quirúrgica simulada (sin lesión arterial); grupo 2, inyección de solución salina tamponada con fosfato (PBS); grupo 3, MSC; grupo 4, DFT; grupo 5, MSC + DFT. Todas las ratas recibieron dos inyecciones intraperitoneales de 0,5 ml: la primera se administró 4 horas después del procedimiento y la segunda 48 horas después de la primera. Se sacrificó a las ratas siete días después de la segunda inyección. Resultados. Aunque el uso por separado de MSC derivadas de médula ósea humana (hBM-MSC) o de DFT permitió una resolución parcial del trombo, la combinación de ambos tuvo como resultado la resolución casi completa. La neovascularización fue doblemente mejor en las ratas tratadas con hBM-MSC + DFT (11,6 ± 2,4 canales) en comparación con los grupos asignados por separado a hBM-MSC (3,8 ± 2,7 canales) y DFT (5,5 ± 1,8 canales) (P < 0,0001 y P= 0,002, respectivamente). Conclusión. El uso combinado de hBM-MSC y DFT en un modelo de trombosis arterial en ratas mostró que el efecto aditivo tuvo como resultado la resolución casi completa del trombo.

Background/aim. In this study, we aimed to investigate the additive effect of mesenchymal stem cells (MSC) and defibrotide (DFT) in a rat model of femoral arterial thrombosis. Methods. Thirty Sprague Dawley rats were included. An arterial thrombosis model by ferric chloride (FeCl3) was developed in the left femoral artery. The rats were equally assigned to 5 groups: Group 1-Sham-operated (without arterial injury); Group 2-Phosphate buffered saline (PBS) injected; Group 3-MSC; Group 4-DFT; Group 5-MSC + DFT. All had two intraperitoneal injections of 0.5 ml: the 1st injection was 4 h after the procedure and the 2nd one 48 h after the 1st injection. The rats were sacrificed 7 days after the 2nd injection. Results. Although the use of human bone marrow-derived (hBM) hBM-MSC or DFT alone enabled partial resolution of the thrombus, combining them resulted in near-complete resolution. Neovascularization was two-fold better in hBM-MSC + DFT treated rats (11.6 ± 2.4 channels) compared with the hBM-MSC (3.8 ± 2.7 channels) and DFT groups (5.5 ± 1.8 channels) (P < 0.0001 and P= 0.002, respectively). Conclusion. The combined use of hBM-MSC and DFT in a rat model of arterial thrombosis showed additive effect resulting in near-complete resolution of the thrombus.

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BACKGROUND/AIM: In this study, we aimed to investigate the additive effect of mesenchymal stem cells (MSC) and defibrotide (DFT) in a rat model of femoral arterial thrombosis. METHODS: Thirty Sprague Dawley rats were included. An arterial thrombosis model by ferric chloride (FeCl3) was developed in the left femoral artery. The rats were equally assigned to 5 groups: Group 1-Sham-operated (without arterial injury); Group 2-Phosphate buffered saline (PBS) injected; Group 3-MSC; Group 4-DFT; Group 5-MSC + DFT. All had two intraperitoneal injections of 0.5 ml: the 1st injection was 4 h after the procedure and the 2nd one 48 h after the 1st injection. The rats were sacrificed 7 days after the 2nd injection. RESULTS: Although the use of human bone marrow-derived (hBM) hBM-MSC or DFT alone enabled partial resolution of the thrombus, combining them resulted in near-complete resolution. Neovascularization was two-fold better in hBM-MSC + DFT treated rats (11.6 ± 2.4 channels) compared with the hBM-MSC (3.8 ± 2.7 channels) and DFT groups (5.5 ± 1.8 channels) (P < 0.0001 and P= 0.002, respectively). CONCLUSION: The combined use of hBM-MSC and DFT in a rat model of arterial thrombosis showed additive effect resulting in near-complete resolution of the thrombus.

INTRODUCCIÓN: El objetivo de este estudio consistió en investigar el efecto aditivo de las células madre mesenquimales (MSC, por sus siglas en inglés) y del defibrótido (DFT) en un modelo de trombosis arterial femoral en ratas. MÉTODOS: Se incluyeron 30 ratas Sprague Dawley. Se desarrolló un modelo de trombosis arterial mediante cloruro de hierro (FeCl3) en la arteria femoral izquierda. Las ratas se asignaron equitativamente en cinco grupos: grupo 1, intervención quirúrgica simulada (sin lesión arterial); grupo 2, inyección de solución salina tamponada con fosfato (PBS); grupo 3, MSC; grupo 4, DFT; grupo 5, MSC + DFT. Todas las ratas recibieron dos inyecciones intraperitoneales de 0,5 ml: la primera se administró 4 horas después del procedimiento y la segunda 48 horas después de la primera. Se sacrificó a las ratas siete días después de la segunda inyección. RESULTADOS: Aunque el uso por separado de MSC derivadas de médula ósea humana (hBM-MSC) o de DFT permitió una resolución parcial del trombo, la combinación de ambos tuvo como resultado la resolución casi completa. La neovascularización fue doblemente mejor en las ratas tratadas con hBM-MSC + DFT (11,6 ± 2,4 canales) en comparación con los grupos asignados por separado a hBM-MSC (3,8 ± 2,7 canales) y DFT (5,5 ± 1,8 canales) (P < 0,0001 y P= 0,002, respectivamente). CONCLUSIÓN: El uso combinado de hBM-MSC y DFT en un modelo de trombosis arterial en ratas mostró que el efecto aditivo tuvo como resultado la resolución casi completa del trombo.

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UNLABELLED: An increasing body of evidence suggests that the use of probiotic bacteria is a promising intervention approach for the treatment of inflammatory diseases with a polymicrobial etiology. P. gingivalis has been noted to have a different way of interacting with the innate immune response of the host compared to other pathogenic bacteria, which is a recognized feature that inhibits CXCL8 expression. OBJECTIVE: The aim of the study was to determine if P. gingivalis infection modulates the inflammatory response of gingival stromal stem cells (G-MSSCs), including the release of CXCL8, and the expression of TLRs and if immunomodulatory L. rhamnosus ATCC9595 could prevent CXCL8 inhibition in experimental inflammation. MATERIAL AND METHODS: G-MSSCs were pretreated with L. rhamnosus ATCC9595 and then stimulated with P. gingivalis ATCC33277. CXCL8 and IL-10 levels were investigated with ELISA and the TLR-4 and 2 were determined through flow cytometer analysis. RESULTS: CXCL8 was suppressed by P. gingivalis and L. rhamnosus ATCC9595, whereas incubation with both strains did not abolish CXCL8. L. rhamnosus ATCC9595 scaled down the expression of TLR4 and induced TLR2 expression when exposed to P. gingivalis stimulation (p<0.01). CONCLUSIONS: These findings provide evidence that L. rhamnosus ATCC9595 can modulate the inflammatory signals and could introduce P. gingivalis to immune systems by inducing CXCL8 secretion.

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ABSTRACT An increasing body of evidence suggests that the use of probiotic bacteria is a promising intervention approach for the treatment of inflammatory diseases with a polymicrobial etiology. P. gingivalis has been noted to have a different way of interacting with the innate immune response of the host compared to other pathogenic bacteria, which is a recognized feature that inhibits CXCL8 expression. Objective The aim of the study was to determine if P. gingivalis infection modulates the inflammatory response of gingival stromal stem cells (G-MSSCs), including the release of CXCL8, and the expression of TLRs and if immunomodulatory L. rhamnosus ATCC9595 could prevent CXCL8 inhibition in experimental inflammation. Material and Methods G-MSSCs were pretreated with L. rhamnosus ATCC9595 and then stimulated with P. gingivalis ATCC33277. CXCL8 and IL-10 levels were investigated with ELISA and the TLR-4 and 2 were determined through flow cytometer analysis. Results CXCL8 was suppressed by P. gingivalis and L. rhamnosus ATCC9595, whereas incubation with both strains did not abolish CXCL8. L. rhamnosus ATCC9595 scaled down the expression of TLR4 and induced TLR2 expression when exposed to P. gingivalis stimulation (p<0.01). Conclusions These findings provide evidence that L. rhamnosus ATCC9595 can modulate the inflammatory signals and could introduce P. gingivalis to immune systems by inducing CXCL8 secretion.

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