Emerging infectious diseases are a major threat to biodiversity and an important public health issue.Flaviviruses are the cause of several emerging vector-borne zoonotic arboviruses whose distribution is currently inc...Emerging infectious diseases are a major threat to biodiversity and an important public health issue.Flaviviruses are the cause of several emerging vector-borne zoonotic arboviruses whose distribution is currently increasing in Europe.The evidence that West Nile virus(WNV)circulates in resident and migratory species has implications for both animal and public health and should therefore be studied in depth.USUTU(USUV),Bagaza(BAGV)and tick-borne encephalitis virus(TBEV)are other viruses that are beginning to spread more widely.An integrated surveillance program,namely in birds,is essential for reducing the risk of infection in human populations within the One Health principles.In the present study,wild birds admitted to wildlife rehabilitation centers in Portugal were sampled.Two hundred eight blood samples were assayed serologically for antibodies to flaviviruses by using a commercial ELISA kit.An over-all seroprevalence of 19.6%(95%confidence interval[CI]:13.7-26.7%)was observed.Antibodies against flaviviruses were detected in 13(35.1%)different species of wild birds.Accipitriformes(26.7%;95%CI:18.5-36.2%)and Strigi-formes(26.7%;95%CI:14.6-42.0%)were the orders with the highest seroprevalence rates recorded.There were no statistically significant differences(p=0.725)between the geographical regions(NUTSⅡ)studied,but a statistically significant difference(p=0.017)was found between sex(male:34.4%;female:4.8%).A higher seroprevalence was detected in adults(32.1%)than in juvenile birds(9.3%)(p=0.014),and age was considered a risk factor for flavivirus infection in wild birds(odds ratio 1.4;95%CI:0.5-4.0).More epidemiological studies are needed in Portugal since the actual spread of the genus Flavivirus throughout the country is unknown.展开更多
This work aimed to evaluate the physical-chemical characteristics of pork from a local breed （Portuguese black pork called Preto Alentejano） and a commercial breed. Sixteen animals were used, 4 females and 4 males f...This work aimed to evaluate the physical-chemical characteristics of pork from a local breed （Portuguese black pork called Preto Alentejano） and a commercial breed. Sixteen animals were used, 4 females and 4 males from each breed. Animals had 80-100 kg of live weight. The longissimus muscle between the 5th thoracic vertebra and the 10th lumbar vertebra was used in the analysis. Samples were analysed for protein, fat, pigments, ashes, dry mater, water-holding capacity, and texture. Results of fat and pigments contents indicate signiifcant （P〈0.05 and P〈0.01） differences for all treatments. For protein, ashes, dry mater, water-holding capacity and texture, no signiifcant differences were found for sex. In the analysis of fatty acids composition, ten were detected, being the main ones C16：0, C18：0, C16：1, C18：1, C18：2. There was a predominance of monounsaturated fatty acids （MUFA）, followed by saturated （SFA） and polyunsaturated （PUFA）. Differences were signiifcant for sex （P〈0.01） and breed （P〈0.01）. Preto Alentejano breed and females presented the higher percentages of SFA （P〈0.01） and MUFA （P〈0.001） fatty acids. Differences between breeds might be due to breed production system and feed differences.展开更多
This work had an objective to evaluate the sensory quality of two categories of pork meat from a commercial pork meat and a selected meat from the Portuguese black pork (Preto Alentejano breed). Sixteen animals were u...This work had an objective to evaluate the sensory quality of two categories of pork meat from a commercial pork meat and a selected meat from the Portuguese black pork (Preto Alentejano breed). Sixteen animals were used, 8 females and 8 males from each breed. Animals had 80 - 100 kg of live weight. The longissimus muscle between the 5th thoracic vertebra and the 10th lumbar vertebra was used in the analysis. Sensory analysis was performed by a trained taste panel of 10 elements, in 5 sessions. All evaluation conditions were standardized, and the attributes studied were odor intensity, toughness, juiciness and flavor intensity. The taste panel found differences mainly between breeds. The panellists scored Preto Alentejano meat as being juicier, tenderer, and with richer taste than Commercial meat. The higher juiciness score of Preto Alentejano meat was probably attributable to the higher intramuscular fat content compared with Commercial meat. The Commercial pork was characterized mainly by high toughness.展开更多
This study was performed conducting surveys to assess the Knowledge Level (KL) and Self-Reported Attitudes (SRA) of Food Handlers (FH) in order to evaluate their food safety perception. Food handlers working in 5 cafe...This study was performed conducting surveys to assess the Knowledge Level (KL) and Self-Reported Attitudes (SRA) of Food Handlers (FH) in order to evaluate their food safety perception. Food handlers working in 5 cafes and 6 canteens in a university campus responded to a questionnaire about food hygiene. The knowledge level about food hygiene was obtained by answering five question groups (G): Agents involved in food borne diseases (G1), Food handling hygiene (G2), Cross contamination (G3), Heat treatment/cooling techniques (G4), Reduced temperatures (G5). The SRA level was obtained through seven questions with multiple choice options on behaviors of health and safety applied to the work routine, which were considered as percentage of hits. The hygiene’s knowledge average was 75%, ranging from 63.3% (G4) to 94.5% (G3). Significant associations between establishment type and G1 (p = 0.027), professional experience and G5 (p = 0.020), training and G5 (p = 0.037) were found. Food handlers knowledge did not have effect in FH attitude (p = 0.371). From 25 FH (46.0%) who had high KL, 17 had reported incorrect attitudes. The level of hits is, in general, more than 75%, except for matters relating to the use of different cutting tables (44.4%) and knives (51.9%). Significant differences of values and odds for handlers’ knowledge were observed between cafes’ and canteens’ FH. No difference (p > 0.05) was observed in FH SRA scores according to the type of establishment. The results reveal a reduced application of knowledge acquired by food handlers, evidenced by the low level of attitudes considered correct. This clearly justifies the implementation of additional measures, including on job training as part of an effective strategy to control establishment’s food safety.展开更多
AIM: To study the therapeutic effect of three tubeguides with electrical conductivity associated to mesenchymal stem cells(MSCs) on neuro-muscular regeneration after neurotmesis.METHODS: Rats with 10-mm gap nerve inju...AIM: To study the therapeutic effect of three tubeguides with electrical conductivity associated to mesenchymal stem cells(MSCs) on neuro-muscular regeneration after neurotmesis.METHODS: Rats with 10-mm gap nerve injury were tested using polyvinyl alcohol(PVA), PVA-carbon nanotubes(CNTs) and MSCs, and PVA-polypyrrole(PPy). The regenerated nerves and tibialis anterior muscles were processed for stereological studies after 20 wk. The functional recovery was assessed serially for gait biomechanical analysis, by extensor postural thrust, sciatic functional index and static sciatic functionalindex(SSI), and by withdrawal reflex latency(WRL). In vitro studies included cytocompatibility, flow cytometry, reverse transcriptase polymerase chain reaction and karyotype analysis of the MSCs. Histopathology of lung, liver, kidneys, and regional lymph nodes ensured the biomaterials biocompatibility. RESULTS: SSI remained negative throughout and independently from treatment. Differences between treted groups in the severity of changes in WRL existed, showing a faster regeneration for PVA-CNTs-MSCs(P < 0.05). At toe-off, less acute ankle joint angles were seen for PVA-CNTs-MSCs group(P = 0.051) suggesting improved ankle muscles function during the push off phase of the gait cycle. In PVA-PPy and PVA-CNTs groups, there was a 25% and 42% increase of average fiber area and a 13% and 21% increase of the "minimal Feret's diameter" respectively. Stereological analysis disclosed a significantly(P < 0.05) increased myelin thickness(M), ratio myelin thickness/axon diameter(M/d) and ratio axon diameter/fiber diameter(d/D; g-ratio) in PVA-CNT-MSCs group(P < 0.05). CONCLUSION: Results revealed that treatment with MSCs and PVA-CNTs tube-guides induced better nerve fiber regeneration. Functional and kinematics analysis revealed positive synergistic effects brought by MSCs and PVA-CNTs. The PVA-CNTs and PVA-PPy are promising scaffolds with electric conductive properties, bio- and cytocompatible that might prevent the secondary neurogenic muscular atrophy by improving the reestablishment of the neuro-muscular junction.展开更多
A significant number of stallions produce low quality ejaculates with high sensibility to chilling. Single Layer Centrifugation (SLC) with Andro-coll-ETM has been presented as an efficient method of selecting good qua...A significant number of stallions produce low quality ejaculates with high sensibility to chilling. Single Layer Centrifugation (SLC) with Andro-coll-ETM has been presented as an efficient method of selecting good quality spermatozoa. In the current study, changes in sperm quality (motility, viability, acrosome integrity and DNA damage) occurring during storage at 5?C for a maximum of 72 h, were investigated. For that, one ejaculate from 12 stallions was split in two aliquots: control and SLC-selected. Both aliquots were chilled and stored at 5?C and spermatozoa were evaluated for motility, viability and acrosome integrity at 24, 48 and 72 h post collection. DNA damage was evaluated at 48 h post collection using the comet assay. In the SLC-selected aliquots, there was a significant improvement in terms of progressive motility (0 h: P = 0.005;24 h: P 0.05). SLC with Androcoll-ETM improved semen quality prolonging sperm longevity of chilled semen (P = 0.012). This positive effect was more evident in ejaculates most sensitive to chilling that had a sharp decrease in motility in the first 24 h of refrigeration and for all ejaculates at 72 h post-chilling. Therefore, this method reveals to be a useful technique for selecting spermatozoa and maintain sperm quality during storage.展开更多
Aquaculture procedures such as capturing,handling,and trahsporting fish can lead to stress in animals.For this reason,anaesthesia has been recommended to reduce fish stress during these procedures.Synthetic and natura...Aquaculture procedures such as capturing,handling,and trahsporting fish can lead to stress in animals.For this reason,anaesthesia has been recommended to reduce fish stress during these procedures.Synthetic and natural anaesthetics such as tricaine methanesulfonate(MS-222)and eugenol(monoterpene)are commonly used;however,biochemical changes and aversive effects have been reported.Other monoterpene compounds such as thymol and menthol,have shown potential anaesthetic effects in fish species.Thus,further investigation of its anaesthetic properties seems imperative,and the use of the zebrafish in the early stages of its development is an appropriate model due to its advantages.Therefore,the aimed to evaluate the anaesthetic profile of thymol and menthol in zebrafish at 72 hpf(hours post fertisation).Following the determination of the effective anaesthetic concentration,three concentrations(100,200 and 300 mg/L for thymol and 200,400 and 500 mg/L for menthol)were further tested by focusing on mortality rate,heart rate,behaviour features of induction and oxidative effects in comparison to MS-222 and eugenol.Both monoterpenes caused anaesthetic effects in a concentration-dependent manner,with concentrations of 200 mg/L exhibiting a similar profile to MS 222 and eugenol,as evidenced by the increased recovery times.Furthermore,anaesthesia with thymol and menthol caused post-anaesthesia motor pattern abnormalities,which supported some indicators of stress.Slight modifications in physiological parameters and antioxidant changes were observed,but their association with thymol and menthol anaesthesia is unclear.Although sup-porting its anaesthetic use,more extensive research will be required to fully understand the impact of these natural anaesthetics on fish behaviour and stress levels,ultimately leading to safer and more successful anaes-thetic operations in the future.展开更多
The development of injectable bone substitutes(IBS)have obtained great importance in the bone re-generation field,as a strategy to reach hardly accessible defects using minimally invasive techniques and able to fit to...The development of injectable bone substitutes(IBS)have obtained great importance in the bone re-generation field,as a strategy to reach hardly accessible defects using minimally invasive techniques and able to fit to irregular topographies.In this scenario,the association of injectable hydrogels and bone graft granules is emerging as a well-established trend.Particularly,in situ forming hydrogels have arisen as a new IBS generation.An in situ forming and injectable dextrin-based hydrogel(HG)was developed,aiming to act as a carrier of granular bone substitutes and bioactive agents.In this work,the HG was associated to a granular bone substitute(Bonelike)and implanted in goat critical-sized calvarial defects(14mm)for 3,6 and 12weeks.The results showed that HG improved the han-dling properties of the Bonelike granules and did not affect its osteoconductive features,neither impairing the bone regener ation process.Human multipotent mesenchymal stromal cells from the umbilical cord,extracellular matrix hydrolysates and the pro-angiogenic peptide LLKKK18 were also combined with the IBS.These bioactive agents did not enhance the new bone formation significantly under the conditions tested,according to micro-computed tomography and histological analysis.展开更多
基金funded by the Portuguese Foundation for Science and Technology(FCT)sup-ported by the projects UIDB/00772/2020(https://doi.org/10.54499/UIDB/00772/2020)。
文摘Emerging infectious diseases are a major threat to biodiversity and an important public health issue.Flaviviruses are the cause of several emerging vector-borne zoonotic arboviruses whose distribution is currently increasing in Europe.The evidence that West Nile virus(WNV)circulates in resident and migratory species has implications for both animal and public health and should therefore be studied in depth.USUTU(USUV),Bagaza(BAGV)and tick-borne encephalitis virus(TBEV)are other viruses that are beginning to spread more widely.An integrated surveillance program,namely in birds,is essential for reducing the risk of infection in human populations within the One Health principles.In the present study,wild birds admitted to wildlife rehabilitation centers in Portugal were sampled.Two hundred eight blood samples were assayed serologically for antibodies to flaviviruses by using a commercial ELISA kit.An over-all seroprevalence of 19.6%(95%confidence interval[CI]:13.7-26.7%)was observed.Antibodies against flaviviruses were detected in 13(35.1%)different species of wild birds.Accipitriformes(26.7%;95%CI:18.5-36.2%)and Strigi-formes(26.7%;95%CI:14.6-42.0%)were the orders with the highest seroprevalence rates recorded.There were no statistically significant differences(p=0.725)between the geographical regions(NUTSⅡ)studied,but a statistically significant difference(p=0.017)was found between sex(male:34.4%;female:4.8%).A higher seroprevalence was detected in adults(32.1%)than in juvenile birds(9.3%)(p=0.014),and age was considered a risk factor for flavivirus infection in wild birds(odds ratio 1.4;95%CI:0.5-4.0).More epidemiological studies are needed in Portugal since the actual spread of the genus Flavivirus throughout the country is unknown.
文摘This work aimed to evaluate the physical-chemical characteristics of pork from a local breed （Portuguese black pork called Preto Alentejano） and a commercial breed. Sixteen animals were used, 4 females and 4 males from each breed. Animals had 80-100 kg of live weight. The longissimus muscle between the 5th thoracic vertebra and the 10th lumbar vertebra was used in the analysis. Samples were analysed for protein, fat, pigments, ashes, dry mater, water-holding capacity, and texture. Results of fat and pigments contents indicate signiifcant （P〈0.05 and P〈0.01） differences for all treatments. For protein, ashes, dry mater, water-holding capacity and texture, no signiifcant differences were found for sex. In the analysis of fatty acids composition, ten were detected, being the main ones C16：0, C18：0, C16：1, C18：1, C18：2. There was a predominance of monounsaturated fatty acids （MUFA）, followed by saturated （SFA） and polyunsaturated （PUFA）. Differences were signiifcant for sex （P〈0.01） and breed （P〈0.01）. Preto Alentejano breed and females presented the higher percentages of SFA （P〈0.01） and MUFA （P〈0.001） fatty acids. Differences between breeds might be due to breed production system and feed differences.
文摘This work had an objective to evaluate the sensory quality of two categories of pork meat from a commercial pork meat and a selected meat from the Portuguese black pork (Preto Alentejano breed). Sixteen animals were used, 8 females and 8 males from each breed. Animals had 80 - 100 kg of live weight. The longissimus muscle between the 5th thoracic vertebra and the 10th lumbar vertebra was used in the analysis. Sensory analysis was performed by a trained taste panel of 10 elements, in 5 sessions. All evaluation conditions were standardized, and the attributes studied were odor intensity, toughness, juiciness and flavor intensity. The taste panel found differences mainly between breeds. The panellists scored Preto Alentejano meat as being juicier, tenderer, and with richer taste than Commercial meat. The higher juiciness score of Preto Alentejano meat was probably attributable to the higher intramuscular fat content compared with Commercial meat. The Commercial pork was characterized mainly by high toughness.
文摘This study was performed conducting surveys to assess the Knowledge Level (KL) and Self-Reported Attitudes (SRA) of Food Handlers (FH) in order to evaluate their food safety perception. Food handlers working in 5 cafes and 6 canteens in a university campus responded to a questionnaire about food hygiene. The knowledge level about food hygiene was obtained by answering five question groups (G): Agents involved in food borne diseases (G1), Food handling hygiene (G2), Cross contamination (G3), Heat treatment/cooling techniques (G4), Reduced temperatures (G5). The SRA level was obtained through seven questions with multiple choice options on behaviors of health and safety applied to the work routine, which were considered as percentage of hits. The hygiene’s knowledge average was 75%, ranging from 63.3% (G4) to 94.5% (G3). Significant associations between establishment type and G1 (p = 0.027), professional experience and G5 (p = 0.020), training and G5 (p = 0.037) were found. Food handlers knowledge did not have effect in FH attitude (p = 0.371). From 25 FH (46.0%) who had high KL, 17 had reported incorrect attitudes. The level of hits is, in general, more than 75%, except for matters relating to the use of different cutting tables (44.4%) and knives (51.9%). Significant differences of values and odds for handlers’ knowledge were observed between cafes’ and canteens’ FH. No difference (p > 0.05) was observed in FH SRA scores according to the type of establishment. The results reveal a reduced application of knowledge acquired by food handlers, evidenced by the low level of attitudes considered correct. This clearly justifies the implementation of additional measures, including on job training as part of an effective strategy to control establishment’s food safety.
基金Supported by System of Incentives for Research and Technological development of QREN in the scope of project n°38853/2013-DEXGELERATION:"Solucoes avancadas de regeneracao ossea com base em hidrogeis de dextrin"the European Community FEDER fund through ON2-O Novo Norte-North Portugal Regional Operational Program 2007-2013+4 种基金Project n°34128-BEPIM II:"Microdispositivos biomédicos com capacidade osteointegrativa porμPIM"funded by Ad Ithe program COMPETE-Programa Operacional Factores de Competitividade,projects Pest-OE/AGR/UI0211/2011PTDC/CVT/103081/2008CDRsp’s Strategic Project-UI-4044-2011-2012(Pest-OE/EME/UI4044/2011)funding from FCT
文摘AIM: To study the therapeutic effect of three tubeguides with electrical conductivity associated to mesenchymal stem cells(MSCs) on neuro-muscular regeneration after neurotmesis.METHODS: Rats with 10-mm gap nerve injury were tested using polyvinyl alcohol(PVA), PVA-carbon nanotubes(CNTs) and MSCs, and PVA-polypyrrole(PPy). The regenerated nerves and tibialis anterior muscles were processed for stereological studies after 20 wk. The functional recovery was assessed serially for gait biomechanical analysis, by extensor postural thrust, sciatic functional index and static sciatic functionalindex(SSI), and by withdrawal reflex latency(WRL). In vitro studies included cytocompatibility, flow cytometry, reverse transcriptase polymerase chain reaction and karyotype analysis of the MSCs. Histopathology of lung, liver, kidneys, and regional lymph nodes ensured the biomaterials biocompatibility. RESULTS: SSI remained negative throughout and independently from treatment. Differences between treted groups in the severity of changes in WRL existed, showing a faster regeneration for PVA-CNTs-MSCs(P < 0.05). At toe-off, less acute ankle joint angles were seen for PVA-CNTs-MSCs group(P = 0.051) suggesting improved ankle muscles function during the push off phase of the gait cycle. In PVA-PPy and PVA-CNTs groups, there was a 25% and 42% increase of average fiber area and a 13% and 21% increase of the "minimal Feret's diameter" respectively. Stereological analysis disclosed a significantly(P < 0.05) increased myelin thickness(M), ratio myelin thickness/axon diameter(M/d) and ratio axon diameter/fiber diameter(d/D; g-ratio) in PVA-CNT-MSCs group(P < 0.05). CONCLUSION: Results revealed that treatment with MSCs and PVA-CNTs tube-guides induced better nerve fiber regeneration. Functional and kinematics analysis revealed positive synergistic effects brought by MSCs and PVA-CNTs. The PVA-CNTs and PVA-PPy are promising scaffolds with electric conductive properties, bio- and cytocompatible that might prevent the secondary neurogenic muscular atrophy by improving the reestablishment of the neuro-muscular junction.
基金partially financed by Projects PTDC/CVT/108456/2008(FCT)and COMPETE:FCOMP-01-0124-FEDER-009565(FEDER),“Development of methods to increase the fertilizing ability of chilled and frozen stallion semen:a multidisciplinary ap-proach”.
文摘A significant number of stallions produce low quality ejaculates with high sensibility to chilling. Single Layer Centrifugation (SLC) with Andro-coll-ETM has been presented as an efficient method of selecting good quality spermatozoa. In the current study, changes in sperm quality (motility, viability, acrosome integrity and DNA damage) occurring during storage at 5?C for a maximum of 72 h, were investigated. For that, one ejaculate from 12 stallions was split in two aliquots: control and SLC-selected. Both aliquots were chilled and stored at 5?C and spermatozoa were evaluated for motility, viability and acrosome integrity at 24, 48 and 72 h post collection. DNA damage was evaluated at 48 h post collection using the comet assay. In the SLC-selected aliquots, there was a significant improvement in terms of progressive motility (0 h: P = 0.005;24 h: P 0.05). SLC with Androcoll-ETM improved semen quality prolonging sperm longevity of chilled semen (P = 0.012). This positive effect was more evident in ejaculates most sensitive to chilling that had a sharp decrease in motility in the first 24 h of refrigeration and for all ejaculates at 72 h post-chilling. Therefore, this method reveals to be a useful technique for selecting spermatozoa and maintain sperm quality during storage.
基金supported by Portuguese funds by FCT/MCTES-Portuguese Foundation for Science and Technology/Ministerio da Ciencia,Tecnologia e Ensino Superior,under the project 10.54499/UIDB/04033/2020(CITAB—Centre for Research and Technology of Agro-Environment and Biological Sciences),SFRH/BD/144904/2019 and 10.54499/2021.00458.CEECIND/CP1690/CT0001.
文摘Aquaculture procedures such as capturing,handling,and trahsporting fish can lead to stress in animals.For this reason,anaesthesia has been recommended to reduce fish stress during these procedures.Synthetic and natural anaesthetics such as tricaine methanesulfonate(MS-222)and eugenol(monoterpene)are commonly used;however,biochemical changes and aversive effects have been reported.Other monoterpene compounds such as thymol and menthol,have shown potential anaesthetic effects in fish species.Thus,further investigation of its anaesthetic properties seems imperative,and the use of the zebrafish in the early stages of its development is an appropriate model due to its advantages.Therefore,the aimed to evaluate the anaesthetic profile of thymol and menthol in zebrafish at 72 hpf(hours post fertisation).Following the determination of the effective anaesthetic concentration,three concentrations(100,200 and 300 mg/L for thymol and 200,400 and 500 mg/L for menthol)were further tested by focusing on mortality rate,heart rate,behaviour features of induction and oxidative effects in comparison to MS-222 and eugenol.Both monoterpenes caused anaesthetic effects in a concentration-dependent manner,with concentrations of 200 mg/L exhibiting a similar profile to MS 222 and eugenol,as evidenced by the increased recovery times.Furthermore,anaesthesia with thymol and menthol caused post-anaesthesia motor pattern abnormalities,which supported some indicators of stress.Slight modifications in physiological parameters and antioxidant changes were observed,but their association with thymol and menthol anaesthesia is unclear.Although sup-porting its anaesthetic use,more extensive research will be required to fully understand the impact of these natural anaesthetics on fish behaviour and stress levels,ultimately leading to safer and more successful anaes-thetic operations in the future.
基金This work was funded by the project‘DEXGELERATION-Advanced solu-tions for bone regeneration based on dextrin hydrogels’(Norte-07-0202-FEDER-038853).It was also funded by FCT under the scope of the strategic funding of UID/BIO/04469/2013 and UID/BIM/04293/2013 units and COMPETE 2020(POCI-01-0145-FEDER-006684),BioTecNorte operation(NORTE-01-0145-FEDER-000004)and NORTE-01-0145-FEDER-000012 funded by FEDER under the scope of Norte2020-Programa Operacional Regional do Norte.Isabel Pereira and Ana Rita Caseiro were supported by the grants SFRH/BD90066/2012 and SFRH/BD/101174/2014,respectively,from FCT,Portugal.
文摘The development of injectable bone substitutes(IBS)have obtained great importance in the bone re-generation field,as a strategy to reach hardly accessible defects using minimally invasive techniques and able to fit to irregular topographies.In this scenario,the association of injectable hydrogels and bone graft granules is emerging as a well-established trend.Particularly,in situ forming hydrogels have arisen as a new IBS generation.An in situ forming and injectable dextrin-based hydrogel(HG)was developed,aiming to act as a carrier of granular bone substitutes and bioactive agents.In this work,the HG was associated to a granular bone substitute(Bonelike)and implanted in goat critical-sized calvarial defects(14mm)for 3,6 and 12weeks.The results showed that HG improved the han-dling properties of the Bonelike granules and did not affect its osteoconductive features,neither impairing the bone regener ation process.Human multipotent mesenchymal stromal cells from the umbilical cord,extracellular matrix hydrolysates and the pro-angiogenic peptide LLKKK18 were also combined with the IBS.These bioactive agents did not enhance the new bone formation significantly under the conditions tested,according to micro-computed tomography and histological analysis.
