Cell sheet(CS)-based approaches hold significant potential for tissue regeneration,relying on the extracellular matrix(ECM)for success.Like in native tissues,the ECM provides structural and biochemical support for cel...Cell sheet(CS)-based approaches hold significant potential for tissue regeneration,relying on the extracellular matrix(ECM)for success.Like in native tissues,the ECM provides structural and biochemical support for cellular homeostasis and function.Effective preservation strategies that maintain ECM integrity are critical to enhance the therapeutic potential of CS-based approaches.While cryogenic and hypothermic preservation methods offer potential solutions,their impact on CS ECM structure is not fully understood.Therefore,a comprehensive analysis of the ECM of hASCs CS following cryogenic and hypothermic preservation for 3 and 7 days,was conducted.Although proteomic analysis indicated that cryopreservation had no significant effect on the overall composition of the ECM,it induced significant ECM structural alterations,particularly disrupting collagen organization,which was not observed following hypothermic preservation.These structural changes were accompanied by alterations in mechanical properties,including a reduction in elastic modulus.In contrast,hypothermic preservation maintained ECM integrity and mechanical properties similar to the control.The notable ECM structural changes following cryogenic preservation can potentially impact cellular behavior,including adhesion,proliferation,and differentiation,thereby affecting the efficacy of CS therapies in vivo.This suggests that hypothermia may offer a promising alternative to cryopreservation for preserving CS integrity and functionality.展开更多
Background Current diagnostic criteria for hypoxic–ischemic encephalopathy in the early hours lack objective measurement tools.Therefore,this systematic review aims to identify putative molecules that can be used in ...Background Current diagnostic criteria for hypoxic–ischemic encephalopathy in the early hours lack objective measurement tools.Therefore,this systematic review aims to identify putative molecules that can be used in diagnosis in daily clinical practice(PROSPERO ID:CRD42021272610).Data sources Searches were performed in PubMed,Web of Science,and Science Direct databases until November 2020.English original papers analyzing samples from newborns>36 weeks that met at least two American College of Obstetricians and Gynecologists diagnostic criteria and/or imaging evidence of cerebral damage were included.Bias was assessed by the Newcastle–Ottawa Scale.The search and data extraction were verified by two authors separately.Results From 373 papers,30 met the inclusion criteria.Data from samples collected in the first 72 hours were extracted,and increased serum levels of neuron-specific enolase and S100-calcium-binding protein-B were associated with a worse prognosis in newborns that suffered an episode of perinatal asphyxia.In addition,the levels of glial fibrillary acidic protein,ubiquitin carboxyl terminal hydrolase isozyme-L1,glutamic pyruvic transaminase-2,lactate,and glucose were elevated in newborns diagnosed with hypoxic–ischemic encephalopathy.Moreover,pathway analysis revealed insulin-like growth factor signaling and alanine,aspartate and glutamate metabolism to be involved in the early molecular response to insult.Conclusions Neuron-specific enolase and S100-calcium-binding protein-B are potential biomarkers,since they are correlated with an unfavorable outcome of hypoxic-ischemic encephalopathy newborns.However,more studies are required to determine the sensitivity and specificity of this approach to be validated for clinical practice.展开更多
Background The identification of circulating biomarkers that closely correlate with Parkinson’s Disease(PD)has failed several times in the past.Nevertheless,in this pilot study,a translational approach was conducted,...Background The identification of circulating biomarkers that closely correlate with Parkinson’s Disease(PD)has failed several times in the past.Nevertheless,in this pilot study,a translational approach was conducted,allowing the evaluation of the plasma levels of two mitochondrial-related proteins,whose combination leads to a robust model with potential diagnostic value to discriminate the PD patients from matched controls.Methods The proposed translational approach was initiated by the analysis of secretomes from cells cultured under control or well-defined oxidative stress conditions,followed by the identification of proteins related to PD pathologic mechanisms that were altered between the two states.This pipeline was further translated into the analysis of undepleted plasma samples from 28 control and 31 PD patients.Results From the secretome analysis,several mitochondria-related proteins were found to be differentially released between control and stress conditions and to be able to distinguish the two secretomes.Similarly,two mitochondrial-related proteins were found to be significantly changed in a PD cohort compared to matched controls.Moreover,a linear discriminant model with potential diagnostic value to discriminate PD patients was obtained using the combination of these two proteins.Both proteins are associated with apoptotic mitochondrial changes,which may correspond to potential indicators of cell death.Moreover,one of these proteins,the VPS35 protein,was reported in plasma for the first time,and its quantification was only possible due to its previous identification in the secretome analysis.Conclusions In this work,an adaptation of a translational pipeline for biomarker selection was presented and transposed to neurological diseases,in the present case Parkinson’s Disease.The novelty and success of this pilot study may arise from the combination of:i)a translational research pipeline,where plasma samples are interrogated using knowledge previously obtained from the evaluation of cells’secretome under oxidative stress;ii)the combined used of statistical analysis and an informed selection of candidates based on their link with relevant disease mechanisms,and iii)the use of SWATH-MS,an untargeted MS method that allows a complete record of the analyzed samples and a targeted data extraction of the quantitative values of proteins previously identified.展开更多
The number of patients with Alzheimer’s disease(AD)is increasing worldwide due to extended life expectancy,with AD being the most common cause of dementia.AD pathological hallmarks consist of brain depositions of agg...The number of patients with Alzheimer’s disease(AD)is increasing worldwide due to extended life expectancy,with AD being the most common cause of dementia.AD pathological hallmarks consist of brain depositions of aggregated amyloid beta(Aβ)into neuritic plaques and neurofibrillary tangles of hyperphosphorylated tau,lead-ing to synaptic dysfunction and neuronal loss[1].Prot-eomic studies of cerebrospinal fluid(CSF)have shown that several biological processes are dysregulated in AD,such as the innate immune system,inflammatory response,hemostasis,lipid processing,oxidative stress response and synaptic functioning[2].展开更多
基金funded by EU Horizon 2020 research and innovation program under the European Research Council(ERC)grant CapBed(805411)the Portuguese Science and Technology Foundation(FCT)UIDB/04539/2020,UIDP/04539/2020+1 种基金The National Mass Spectrometry Network(POCI-01–0145-FEDER-402–022125 Ref.ROTEIRO/0028/2013)SFR acknowledges doctoral fellowship PD/BD/135252/2017 and RAP acknowledges individual grant CEECIND/05623/2022.
文摘Cell sheet(CS)-based approaches hold significant potential for tissue regeneration,relying on the extracellular matrix(ECM)for success.Like in native tissues,the ECM provides structural and biochemical support for cellular homeostasis and function.Effective preservation strategies that maintain ECM integrity are critical to enhance the therapeutic potential of CS-based approaches.While cryogenic and hypothermic preservation methods offer potential solutions,their impact on CS ECM structure is not fully understood.Therefore,a comprehensive analysis of the ECM of hASCs CS following cryogenic and hypothermic preservation for 3 and 7 days,was conducted.Although proteomic analysis indicated that cryopreservation had no significant effect on the overall composition of the ECM,it induced significant ECM structural alterations,particularly disrupting collagen organization,which was not observed following hypothermic preservation.These structural changes were accompanied by alterations in mechanical properties,including a reduction in elastic modulus.In contrast,hypothermic preservation maintained ECM integrity and mechanical properties similar to the control.The notable ECM structural changes following cryogenic preservation can potentially impact cellular behavior,including adhesion,proliferation,and differentiation,thereby affecting the efficacy of CS therapies in vivo.This suggests that hypothermia may offer a promising alternative to cryopreservation for preserving CS integrity and functionality.
基金funding provided by FCT|FCCN(b-on)financed by the European Regional Development Fund(ERDF),through the COMPETE 2020—Operational Programme for Competitiveness and Internationalization and Portuguese national funds via FCT-Fundcao para a Ciencia e a Tecnologia,under projects POCI-01-0145-FEDER-029311,POCI-01-0247-FEDER-045311,UIDB/04539/2020 and UIDP/04539/2020individual Ph.D.fellowships PD/BD/135178/2017(Margarida Coelho),SFRH/BD/143442/2019(Ines Caramelo),and 2020.07749.BD(Miguel Rosado).
文摘Background Current diagnostic criteria for hypoxic–ischemic encephalopathy in the early hours lack objective measurement tools.Therefore,this systematic review aims to identify putative molecules that can be used in diagnosis in daily clinical practice(PROSPERO ID:CRD42021272610).Data sources Searches were performed in PubMed,Web of Science,and Science Direct databases until November 2020.English original papers analyzing samples from newborns>36 weeks that met at least two American College of Obstetricians and Gynecologists diagnostic criteria and/or imaging evidence of cerebral damage were included.Bias was assessed by the Newcastle–Ottawa Scale.The search and data extraction were verified by two authors separately.Results From 373 papers,30 met the inclusion criteria.Data from samples collected in the first 72 hours were extracted,and increased serum levels of neuron-specific enolase and S100-calcium-binding protein-B were associated with a worse prognosis in newborns that suffered an episode of perinatal asphyxia.In addition,the levels of glial fibrillary acidic protein,ubiquitin carboxyl terminal hydrolase isozyme-L1,glutamic pyruvic transaminase-2,lactate,and glucose were elevated in newborns diagnosed with hypoxic–ischemic encephalopathy.Moreover,pathway analysis revealed insulin-like growth factor signaling and alanine,aspartate and glutamate metabolism to be involved in the early molecular response to insult.Conclusions Neuron-specific enolase and S100-calcium-binding protein-B are potential biomarkers,since they are correlated with an unfavorable outcome of hypoxic-ischemic encephalopathy newborns.However,more studies are required to determine the sensitivity and specificity of this approach to be validated for clinical practice.
基金This work was financed by the European Regional Development Fund(ERDF)through the COMPETE 2020-Operational Programme for Competitiveness and Internationalisation and Portuguese national funds via FCT–Fundação para a Ciência e a Tecnologia,I.P.,under projects:POCI-01-0145-FEDER-029311(ref.:PTDC/BTM-TEC/29311/2017),PTDC/NEU-NMC/0205/2012,UIDB/04539/2020,POCI-01-0145-FEDER-016428(ref.:SAICTPAC/0010/2015),POCI-01-0145-FEDER-016795(ref.:PTDC/NEU-SCC/7051/2014),and POCI-01-0145-FEDER-029516(PTDC/MED-NEU/29516/2017).
文摘Background The identification of circulating biomarkers that closely correlate with Parkinson’s Disease(PD)has failed several times in the past.Nevertheless,in this pilot study,a translational approach was conducted,allowing the evaluation of the plasma levels of two mitochondrial-related proteins,whose combination leads to a robust model with potential diagnostic value to discriminate the PD patients from matched controls.Methods The proposed translational approach was initiated by the analysis of secretomes from cells cultured under control or well-defined oxidative stress conditions,followed by the identification of proteins related to PD pathologic mechanisms that were altered between the two states.This pipeline was further translated into the analysis of undepleted plasma samples from 28 control and 31 PD patients.Results From the secretome analysis,several mitochondria-related proteins were found to be differentially released between control and stress conditions and to be able to distinguish the two secretomes.Similarly,two mitochondrial-related proteins were found to be significantly changed in a PD cohort compared to matched controls.Moreover,a linear discriminant model with potential diagnostic value to discriminate PD patients was obtained using the combination of these two proteins.Both proteins are associated with apoptotic mitochondrial changes,which may correspond to potential indicators of cell death.Moreover,one of these proteins,the VPS35 protein,was reported in plasma for the first time,and its quantification was only possible due to its previous identification in the secretome analysis.Conclusions In this work,an adaptation of a translational pipeline for biomarker selection was presented and transposed to neurological diseases,in the present case Parkinson’s Disease.The novelty and success of this pilot study may arise from the combination of:i)a translational research pipeline,where plasma samples are interrogated using knowledge previously obtained from the evaluation of cells’secretome under oxidative stress;ii)the combined used of statistical analysis and an informed selection of candidates based on their link with relevant disease mechanisms,and iii)the use of SWATH-MS,an untargeted MS method that allows a complete record of the analyzed samples and a targeted data extraction of the quantitative values of proteins previously identified.
基金funded by the Portuguese Science and Technology Foundation(FCT)Beyond Beta Amyloid:Deciphering Early Pathogenic Changes in Alzheimer’s disease project PTDC/MED-NEU/27946/2017,by The National Mass Spectrometry Network(POCI-01–0145-FEDER-402–022125 Ref.ROTEIRO/0028/2013),UIDB/04539/2020,UIDP/04539/2020,through funding of LASIGE Research Unit(UIDB/00408/2020 and UIDP/00408/2020).
文摘The number of patients with Alzheimer’s disease(AD)is increasing worldwide due to extended life expectancy,with AD being the most common cause of dementia.AD pathological hallmarks consist of brain depositions of aggregated amyloid beta(Aβ)into neuritic plaques and neurofibrillary tangles of hyperphosphorylated tau,lead-ing to synaptic dysfunction and neuronal loss[1].Prot-eomic studies of cerebrospinal fluid(CSF)have shown that several biological processes are dysregulated in AD,such as the innate immune system,inflammatory response,hemostasis,lipid processing,oxidative stress response and synaptic functioning[2].
