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
Anti-perovskites A3SnO(A=Ca,Sr,and Ba)are an important class of materials due to the emergence of Dirac cones and tiny mass gaps in their band structures originating from an intricate interplay of crystal symmetry,spi...Anti-perovskites A3SnO(A=Ca,Sr,and Ba)are an important class of materials due to the emergence of Dirac cones and tiny mass gaps in their band structures originating from an intricate interplay of crystal symmetry,spin–orbit coupling,and band overlap.This provides an exciting playground for modulating their electronic properties in the two-dimensional(2D)limit.Herein,we employ first-principles density functional theory(DFT)calculations by combining dispersion-corrected SCAN+rVV10 and mBJ functionals for a comprehensive side-by-side comparison of the structural,thermodynamic,dynamical,mechanical,electronic,and thermoelectric properties of bulk and monolayer(one unit cell thick)A3SnO anti-perovskites.Our results show that 2D monolayers derived from bulk A3SnO anti-perovskites are structurally and energetically stable.Moreover,Rashba-type splitting in the electronic structure of Ca3SnO and Sr3SnO monolayers is observed owing to strong spin–orbit coupling and inversion asymmetry.On the other hand,monolayer Ba3SnO exhibits Dirac cone at the high-symmetryΓpoint due to the domination of band overlap.Based on the predicted electronic transport properties,it is shown that inversion asymmetry plays an essential character such that the monolayers Ca3SnO and Sr3SnO outperform thermoelectric performance of their bulk counterparts.展开更多
The key to high performance for GPU architecture lies in its massive threading capability to drive a large number of cores and enable execution overlapping among threads. However, in reality, the number of threads tha...The key to high performance for GPU architecture lies in its massive threading capability to drive a large number of cores and enable execution overlapping among threads. However, in reality, the number of threads that can simultaneously execute is often limited by the size of the register file on GPUs. The traditional SRAM-based register file takes up so large amount of chip area that it cannot scale to meet the increasing demand of GPU applications. Racetrack memory (RM) is a promising technology for designing large capacity register file on GPUs due to its high data storage density. However, without careful deployment of RM-based register file, the lengthy shift operations of RM may hurt the performance. In this paper, we explore RM for designing high-performance register file for GPU architecture. High storage density RM helps to improve the thread level parallelism (TLP), but if the bits of the registers are not aligned to the ports, shift operations are required to move the bits to the access ports before they are accessed, and thus the read/write operations are delayed. We develop an optimization framework for RM-based register file on GPUs, which employs three different optimization techniques at the application, compilation, and architecture level, respectively. More clearly, we optimize the TLP at the application level, design a register mapping algorithm at the compilation level, and design a preshifting mechanism at the architecture level. Collectively, these optimizations help to determine the TLP without causing cache and register file resource contention and reduce the shift operation overhead. Experimental results using a variety of representative workloads demonstrate that our optimization framework achieves up to 29% (21% on average) performance improvement.展开更多
Molecularly imprinted polymer(MIP)beads imprinted with vitamin E(V_(E))were synthesized via a two-step swelling polymerization method,using methacrylic acid(MAA)and other functional monomers together with polystyrene(...Molecularly imprinted polymer(MIP)beads imprinted with vitamin E(V_(E))were synthesized via a two-step swelling polymerization method,using methacrylic acid(MAA)and other functional monomers together with polystyrene(PS)microspheres as the substrate.In initial trials,different functional monomers were assessed,along with a variety of solvents and imprinting ratios.The MIP_(MAA) sample exhibited the highest performance and was characterized and also assessed with regard to V_(E) adsorption under various experimental conditions.This material exhibited reasonably good sphericity,porosity and stability,along with an excellent adsorption capacity of 46 mg/g.This was much higher than the value for the corresponding non-imprinted polymer made with the same monomer,and 20%higher than that for an acrylamide-based MIP synthesized using a precipitation polymerization method.The core-shell structured MIP_(MAA) also demonstrated fast adsorption kinetics,a low pressure drop and good reusability in fixed bed experiments.The V_(E) adsorption equilibrium,selectivity,kinetics,thermodynamics and regeneration capacity of this material were also examined and are reported herein.展开更多
Development of synthetic bone substitutes has arisen as a major research interest in the need to find an alternative to autologous bone grafts.Using an ovine model,the present pre-clinical study presents a synthetic b...Development of synthetic bone substitutes has arisen as a major research interest in the need to find an alternative to autologous bone grafts.Using an ovine model,the present pre-clinical study presents a synthetic bone graft(BonelikeVR)in combination with a cellular system as an alternative for the regeneration of non-critical defects.The association of biomaterials and cell-based therapies is a promising strategy for bone tissue engineering.Mesenchymal stem cells(MSCs)from human dental pulp have demonstrated both in vitro and in vivo to interact with diverse biomaterial systems and promote mineral deposition,aiming at the reconstruction of osseous defects.Moreover,these cells can be found and isolated from many species.Non-critical bone defects were treated with BonelikeVR with or without MSCs obtained from the human dental pulp.Results showed that BonelikeVR and MSCs treated defects showed improved bone regeneration compared with the defects treated with BonelikeVR alone.Also,it was observed that the biomaterial matrix was reabsorbed and gradually replaced by new bone during the healing process.We therefore propose this combination as an efficient binomial strategy that promotes bone growth and vascularization in non-critical bone defects.展开更多
Peroxisome proliferator-activated receptorγ(PPARγ)is a transcriptional coactivator that binds to a diverse range of transcription factors.PPARγcoactivator 1(PGC-1)coactivators possess an extensive range of biologic...Peroxisome proliferator-activated receptorγ(PPARγ)is a transcriptional coactivator that binds to a diverse range of transcription factors.PPARγcoactivator 1(PGC-1)coactivators possess an extensive range of biological effects in different tissues,and play a key part in the regulation of the oxidative metabolism,consequently modulating the production of reactive oxygen species,autophagy,and mitochondrial biogenesis.Owing to these findings,a large body of studies,aiming to establish the role of PGC-1 in the neuromuscular system,has shown that PGC-1 could be a promising target for therapies targeting neuromuscular diseases.Among these,some evidence has shown that various signaling pathways linked to PGC-1αare deregulated in muscular dystrophy,leading to a reduced capacity for mitochondrial oxidative phosphorylation and increased reactive oxygen species(ROS)production.In the light of these results,any intervention aimed at activating PGC-1 could contribute towards ameliorating the progression of muscular dystrophies.PGC-1αis influenced by different patho-physiological/pharmacological stimuli.Natural products have been reported to display modulatory effects on PPARγactivation with fewer side effects in comparison to synthetic drugs.Taken together,this review summarizes the current knowledge on Duchenne muscular dystrophy,focusing on the potential effects of natural compounds,acting as regulators of PGC-1α.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The computations were enabled by resources provided by the Swedish National Infrastructure for Computing(SNIC)at HPC2N and NSC partially funded by the Swedish Research Council through grant agreement no.2018-05973.
文摘Anti-perovskites A3SnO(A=Ca,Sr,and Ba)are an important class of materials due to the emergence of Dirac cones and tiny mass gaps in their band structures originating from an intricate interplay of crystal symmetry,spin–orbit coupling,and band overlap.This provides an exciting playground for modulating their electronic properties in the two-dimensional(2D)limit.Herein,we employ first-principles density functional theory(DFT)calculations by combining dispersion-corrected SCAN+rVV10 and mBJ functionals for a comprehensive side-by-side comparison of the structural,thermodynamic,dynamical,mechanical,electronic,and thermoelectric properties of bulk and monolayer(one unit cell thick)A3SnO anti-perovskites.Our results show that 2D monolayers derived from bulk A3SnO anti-perovskites are structurally and energetically stable.Moreover,Rashba-type splitting in the electronic structure of Ca3SnO and Sr3SnO monolayers is observed owing to strong spin–orbit coupling and inversion asymmetry.On the other hand,monolayer Ba3SnO exhibits Dirac cone at the high-symmetryΓpoint due to the domination of band overlap.Based on the predicted electronic transport properties,it is shown that inversion asymmetry plays an essential character such that the monolayers Ca3SnO and Sr3SnO outperform thermoelectric performance of their bulk counterparts.
基金This work was supported by the National Natural Science Foundation of China under Grant No. 61300005.
文摘The key to high performance for GPU architecture lies in its massive threading capability to drive a large number of cores and enable execution overlapping among threads. However, in reality, the number of threads that can simultaneously execute is often limited by the size of the register file on GPUs. The traditional SRAM-based register file takes up so large amount of chip area that it cannot scale to meet the increasing demand of GPU applications. Racetrack memory (RM) is a promising technology for designing large capacity register file on GPUs due to its high data storage density. However, without careful deployment of RM-based register file, the lengthy shift operations of RM may hurt the performance. In this paper, we explore RM for designing high-performance register file for GPU architecture. High storage density RM helps to improve the thread level parallelism (TLP), but if the bits of the registers are not aligned to the ports, shift operations are required to move the bits to the access ports before they are accessed, and thus the read/write operations are delayed. We develop an optimization framework for RM-based register file on GPUs, which employs three different optimization techniques at the application, compilation, and architecture level, respectively. More clearly, we optimize the TLP at the application level, design a register mapping algorithm at the compilation level, and design a preshifting mechanism at the architecture level. Collectively, these optimizations help to determine the TLP without causing cache and register file resource contention and reduce the shift operation overhead. Experimental results using a variety of representative workloads demonstrate that our optimization framework achieves up to 29% (21% on average) performance improvement.
基金This work was supported by the Jiang Su Yu Xin Vitamin Co.,Ltd.,China.
文摘Molecularly imprinted polymer(MIP)beads imprinted with vitamin E(V_(E))were synthesized via a two-step swelling polymerization method,using methacrylic acid(MAA)and other functional monomers together with polystyrene(PS)microspheres as the substrate.In initial trials,different functional monomers were assessed,along with a variety of solvents and imprinting ratios.The MIP_(MAA) sample exhibited the highest performance and was characterized and also assessed with regard to V_(E) adsorption under various experimental conditions.This material exhibited reasonably good sphericity,porosity and stability,along with an excellent adsorption capacity of 46 mg/g.This was much higher than the value for the corresponding non-imprinted polymer made with the same monomer,and 20%higher than that for an acrylamide-based MIP synthesized using a precipitation polymerization method.The core-shell structured MIP_(MAA) also demonstrated fast adsorption kinetics,a low pressure drop and good reusability in fixed bed experiments.The V_(E) adsorption equilibrium,selectivity,kinetics,thermodynamics and regeneration capacity of this material were also examined and are reported herein.
基金This research was supported by Programa Operacional Regional do Norte(ON.2–O Novo Norte),QREN,FEDER with the project‘iBone Therapies:Terapias inovadoras para a regenerac¸~ao o´ssea’,ref.NORTE-01-0247-FEDER-003262,and by the programme COMPETE–Programa Operacional Factores de Competitividade,Projects PEst-OE/AGR/UI0211/2011 and PEst-C/EME/UI0285/2013 funding from FCT.This research was also supported by Programa Operacional Competitividade e Internacionalizac¸~ao(P2020),Fundos Europeus Estruturais e de Investimento(FEEI)and FCT with the project‘BioMate—A novel bio-manufacturing system to produce bioactive scaffolds for tissue engineering’with reference PTDC/EMS-SIS/7032/2014 and by COMPETE 2020,from ANI—Projectos ID&T Empresas em Copromoc¸~ao,Programas Operacionais POCI,by the project‘insitu.Biomas-Reinvent biomanufacturing systems by using an usability approach for in situ clinic temporary implants fabrication’with the reference POCI-01-0247-FEDER-017771.The research was also supported by the research project‘BEPIM III–Microdispositivos me´dicos com capacidades osteintegradoras por micoPIM’,with the reference POCI-01-0247-FEDER-017935,from Fundo Europeu de Desenvolvimento Regional(FEDER),by the Programa Operacional da Competitividade&Internacionalizac¸~ao.Ana Rita Caseiro(SFRH/BD/101174/2014)acknowledges FCT,for financial support.
文摘Development of synthetic bone substitutes has arisen as a major research interest in the need to find an alternative to autologous bone grafts.Using an ovine model,the present pre-clinical study presents a synthetic bone graft(BonelikeVR)in combination with a cellular system as an alternative for the regeneration of non-critical defects.The association of biomaterials and cell-based therapies is a promising strategy for bone tissue engineering.Mesenchymal stem cells(MSCs)from human dental pulp have demonstrated both in vitro and in vivo to interact with diverse biomaterial systems and promote mineral deposition,aiming at the reconstruction of osseous defects.Moreover,these cells can be found and isolated from many species.Non-critical bone defects were treated with BonelikeVR with or without MSCs obtained from the human dental pulp.Results showed that BonelikeVR and MSCs treated defects showed improved bone regeneration compared with the defects treated with BonelikeVR alone.Also,it was observed that the biomaterial matrix was reabsorbed and gradually replaced by new bone during the healing process.We therefore propose this combination as an efficient binomial strategy that promotes bone growth and vascularization in non-critical bone defects.
基金supported by the crowd funding#Sport4Therapy to Giuseppe D’Antona(Italy)supported by Instituto de Salud CarlosⅢ,Grant Number:CIBEROBN CB12/03/30038
文摘Peroxisome proliferator-activated receptorγ(PPARγ)is a transcriptional coactivator that binds to a diverse range of transcription factors.PPARγcoactivator 1(PGC-1)coactivators possess an extensive range of biological effects in different tissues,and play a key part in the regulation of the oxidative metabolism,consequently modulating the production of reactive oxygen species,autophagy,and mitochondrial biogenesis.Owing to these findings,a large body of studies,aiming to establish the role of PGC-1 in the neuromuscular system,has shown that PGC-1 could be a promising target for therapies targeting neuromuscular diseases.Among these,some evidence has shown that various signaling pathways linked to PGC-1αare deregulated in muscular dystrophy,leading to a reduced capacity for mitochondrial oxidative phosphorylation and increased reactive oxygen species(ROS)production.In the light of these results,any intervention aimed at activating PGC-1 could contribute towards ameliorating the progression of muscular dystrophies.PGC-1αis influenced by different patho-physiological/pharmacological stimuli.Natural products have been reported to display modulatory effects on PPARγactivation with fewer side effects in comparison to synthetic drugs.Taken together,this review summarizes the current knowledge on Duchenne muscular dystrophy,focusing on the potential effects of natural compounds,acting as regulators of PGC-1α.
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.