A decellularized extracellular matrix(dECM)constitutes a pivotal biomaterial created by decellularizing the natural extracellular matrix(ECM).This material serves as a supportive medium for intricate cellular interact...A decellularized extracellular matrix(dECM)constitutes a pivotal biomaterial created by decellularizing the natural extracellular matrix(ECM).This material serves as a supportive medium for intricate cellular interactions,fostering cell growth,differentiation,and organization.However,challenges persist in decellularization,necessitating a balance between preserving the ECM structural integrity and achieving effective cellular removal.An approach to enhancing decellularization involves pre-eliminating unnecessary tissues and effectively reducing final DNA levels to lower than 50 ng/mg ECM on preprocessed tissues.Although this strategic step augments decellularization efficiency,the current manual execution method depends on the operator’s skill.To address this limitation,this study proposed an automated raw tissue slicing system that does not require tissue preparation for slicing.Through carefully controlled tissue applanation pressure and oscillatory incisions with optimized parameters,the system achieved a precision within±10µm in obtaining submillimeter-scale tissue slices of the porcine cornea while avoiding significant microscopic complications in the tissue structure,as observed by tissue histology.These findings suggested the system’s capability to streamline and automate preliminary tissue slicing operations.The efficacy of this approach for decellularization was validated by processing porcine corneas using the proposed system and subsequently decellularizing the processed tissues.DNA level analysis revealed that sliced,subdivided tissues created by this system could expedite DNA reduction even at the initial steps of decellularization,enhancing the overall decellularization procedure.展开更多
Background: Although a large number of studies have confirmed that the different levels of reactive oxygen species (ROS) in cytoplasm and nucleus have effects on cell growth, proliferation, differentiation and apoptos...Background: Although a large number of studies have confirmed that the different levels of reactive oxygen species (ROS) in cytoplasm and nucleus have effects on cell growth, proliferation, differentiation and apoptosis, the exact mechanism of ROS action is unclear. An important reason is that the production and degradation time of ROS in cells is very short, and therefore it’s difficult to understand the mechanism of action based on the traditional molecular action process through the ROS diffusion and target binding. Methods: The fresh liver tissue slices were prepared and the nuclei of hepatocytes were separated from Kunming mice according to the reported method. Liver tissue slices and hepatocyte nuclei were perfused with extracellular or intracellular fluids containing different concentrations of hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), and real-time imaging monitoring of biophotonic emission was carried out using an ultra-weak biophoton imaging system. Results: The results showed that the continuous perfusion with different concentrations of H<sub>2</sub>O<sub>2</sub> (300, 400 and 500 μM, respectively) resulted in significant increase of biophotonic emissions, presenting a concentration-dependent effect in liver tissue slices and achieving the maximum effect at 400 μM, while the significant enhancement was found after 500 μM treatment on the hepatocyte nuclei. Conclusion: This study suggests that ROS generated in cells may achieve its physiological and pathological effects via biophotonic emissions, which provides a new quantum biological mechanism of ROS, while the detailed clarification requires further research.展开更多
Imaging mass spectrometry (IMS) methodology for biological tissue component distribution map using MALDI-TOF MS and MALDI-TOF/TOF MS was established.Peak density distribution was probed to be quite useful for MS image...Imaging mass spectrometry (IMS) methodology for biological tissue component distribution map using MALDI-TOF MS and MALDI-TOF/TOF MS was established.Peak density distribution was probed to be quite useful for MS image classification.More than 40 000 spectra from 200 tissue sections were acquired and reproducibility between various of species groups was great than 80%.Tens of differentiately expressed components were detected by t-test (P<0.01).Classification modeling was created based on the differentiate components,blind species were analyzed for model validation,accuracy was above 90%.展开更多
基金supported by the Alchemist Project 1415180884(No.20012378,Development of Meta Soft Organ Module Manufacturing Technology without Immunity Rejection and Module Assembly Robot System)funded by the Ministry of Trade,Industry&Energy(MOTIE,Republic of Korea)the Technology Development Program(No.S3318933)funded by the Ministry of SMEs and Startups(MSS,Republic of Korea).
文摘A decellularized extracellular matrix(dECM)constitutes a pivotal biomaterial created by decellularizing the natural extracellular matrix(ECM).This material serves as a supportive medium for intricate cellular interactions,fostering cell growth,differentiation,and organization.However,challenges persist in decellularization,necessitating a balance between preserving the ECM structural integrity and achieving effective cellular removal.An approach to enhancing decellularization involves pre-eliminating unnecessary tissues and effectively reducing final DNA levels to lower than 50 ng/mg ECM on preprocessed tissues.Although this strategic step augments decellularization efficiency,the current manual execution method depends on the operator’s skill.To address this limitation,this study proposed an automated raw tissue slicing system that does not require tissue preparation for slicing.Through carefully controlled tissue applanation pressure and oscillatory incisions with optimized parameters,the system achieved a precision within±10µm in obtaining submillimeter-scale tissue slices of the porcine cornea while avoiding significant microscopic complications in the tissue structure,as observed by tissue histology.These findings suggested the system’s capability to streamline and automate preliminary tissue slicing operations.The efficacy of this approach for decellularization was validated by processing porcine corneas using the proposed system and subsequently decellularizing the processed tissues.DNA level analysis revealed that sliced,subdivided tissues created by this system could expedite DNA reduction even at the initial steps of decellularization,enhancing the overall decellularization procedure.
文摘Background: Although a large number of studies have confirmed that the different levels of reactive oxygen species (ROS) in cytoplasm and nucleus have effects on cell growth, proliferation, differentiation and apoptosis, the exact mechanism of ROS action is unclear. An important reason is that the production and degradation time of ROS in cells is very short, and therefore it’s difficult to understand the mechanism of action based on the traditional molecular action process through the ROS diffusion and target binding. Methods: The fresh liver tissue slices were prepared and the nuclei of hepatocytes were separated from Kunming mice according to the reported method. Liver tissue slices and hepatocyte nuclei were perfused with extracellular or intracellular fluids containing different concentrations of hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), and real-time imaging monitoring of biophotonic emission was carried out using an ultra-weak biophoton imaging system. Results: The results showed that the continuous perfusion with different concentrations of H<sub>2</sub>O<sub>2</sub> (300, 400 and 500 μM, respectively) resulted in significant increase of biophotonic emissions, presenting a concentration-dependent effect in liver tissue slices and achieving the maximum effect at 400 μM, while the significant enhancement was found after 500 μM treatment on the hepatocyte nuclei. Conclusion: This study suggests that ROS generated in cells may achieve its physiological and pathological effects via biophotonic emissions, which provides a new quantum biological mechanism of ROS, while the detailed clarification requires further research.
文摘Imaging mass spectrometry (IMS) methodology for biological tissue component distribution map using MALDI-TOF MS and MALDI-TOF/TOF MS was established.Peak density distribution was probed to be quite useful for MS image classification.More than 40 000 spectra from 200 tissue sections were acquired and reproducibility between various of species groups was great than 80%.Tens of differentiately expressed components were detected by t-test (P<0.01).Classification modeling was created based on the differentiate components,blind species were analyzed for model validation,accuracy was above 90%.
