This study endeavors to investigate the effects of Bifidobacterium breve CCFM1078 on bone formation and resorption balance in growing BALB/c mice.Newborn BALB/c mice were assigned to the control group(administration s...This study endeavors to investigate the effects of Bifidobacterium breve CCFM1078 on bone formation and resorption balance in growing BALB/c mice.Newborn BALB/c mice were assigned to the control group(administration saline)and the CCFM1078 group(administration B.breve CCFM1078,3×10^(9) CFU/day)in 3-,4-,and 5-week tests.All the groups have male and female distinctions.Our findings demonstrate that B.breve CCFM1078 exerts on the dynamic equilibrium between bone formation and resorption during the critical period of growth in mice by modulating the composition of gut microbiota and metabolites(hexadecanamide,linoleoyl ethanolamide,and palmitoyl ethanolamide),the genes and proteins expression related to the growth hormone(GH)/insulin-like growth factors-1(IGF-1)axis and Gs/PKA/CREB signaling pathways,as well as downstream osteogenic and osteoclastic differentiation factors.The effects of B.breve CCFM1078 were different with age and gender dependent.This finding suggests B.breve CCFM1078 may have potential applications in regulating bone metabolism in the growth period population.展开更多
How osteoblast cells are induced is a central question for understanding skeletal formation. Abnormal osteoblast differentiation leads to a broad range of devastating craniofacial diseases. Here we have investigated i...How osteoblast cells are induced is a central question for understanding skeletal formation. Abnormal osteoblast differentiation leads to a broad range of devastating craniofacial diseases. Here we have investigated intramembranous ossification during cranial bone development in mouse models of skeletal genetic diseases that exhibit craniofacial bone defects. The GNAS gene encodes Gαs that transduces GPCR signaling. GNAS activation or loss-of-function mutations in humans cause fibrous dysplasia(FD) or progressive osseous heteroplasia(POH) that shows craniofacial hyperostosis or craniosynostosis, respectively. We find here that, while Hh ligand-dependent Hh signaling is essential for endochondral ossification, it is dispensable for intramembranous ossification, where Gαsregulates Hh signaling in a ligand-independent manner. We further show that Gαscontrols intramembranous ossification by regulating both Hh and Wnt/β-catenin signaling. In addition, Gαsactivation in the developing cranial bone leads to reduced ossification but increased cartilage presence due to reduced cartilage dissolution, not cell fate switch. Small molecule inhibitors of Hh and Wnt signaling can effectively ameliorate cranial bone phenotypes in mice caused by loss or gain of Gnas function mutations, respectively. Our work shows that studies of genetic diseases provide invaluable insights in both pathological bone defects and normal bone development, understanding both leads to better diagnosis and therapeutic treatment of bone diseases.展开更多
This experiment was conducted to investigate the effect of dietary calcium(Ca)or phosphorus(P)deficiency on bone development and related Ca or P metabolic utilization parameters of broilers from 22 to 42 days of age b...This experiment was conducted to investigate the effect of dietary calcium(Ca)or phosphorus(P)deficiency on bone development and related Ca or P metabolic utilization parameters of broilers from 22 to 42 days of age based on our previous study,which indicated that dietary Ca or P deficiency impaired the bone development by regulating related Ca or P metabolic utilization parameters of broilers from 1 to 21 days of age.A total of 504 one-day-old Arbor Acres male broilers were randomly assigned to 1 of 4 treatments with 7 replicates in a completely randomized design,and fed the normal control and Ca-or P-deficient diets from 1 to 21 days of age.At 22 days of age,the broilers were further fed the normal control diet(0.90%Ca+0.35%non-phytate P(NPP)),the P-deficient diet(0.90%Ca+0.18%NPP),the Ca-deficient diet(0.30%Ca+0.35%NPP)or the Ca and P-deficient diet(0.30%Ca+0.18%NPP),respectively.The results showed that dietary Ca or P deficiency decreased(P<0.05)tibia bone mineral density(BMD),bone breaking strength(BBS),ash content,tibia ash Ca content and serum P content on days 28 and 42,but increased(P<0.05)tibia alkaline phosphatase(ALP)activity of broilers on day 42 compared with the control group.Furthermore,the broilers fed the P-deficient diet had the lowest(P<0.05)tibia BMD,BBS,ash content,serum P content and the highest(P<0.05)serum Ca content on day 28 compared with those fed the Ca-deficient or Ca and P-deficient diets.The results from the present study indicated that the bone development and related Ca or P metabolic utilization parameters of broilers were the most sensitive to dietary P deficiency,followed by dietary Ca deficiency or Ca and P-deficiency;dietary Ca or P deficiency impaired the bone development possibly by regulating serum Ca and P contents as well as tibia Ca content and ALP activity of broilers from 22 to 42 days of age.展开更多
The bone morphogenetic protein(BMP)and mitogen-activated protein kinase(MAPK)signaling pathways play an important role in regulation of bone formation and development,however,it remains unclear that the effect of diet...The bone morphogenetic protein(BMP)and mitogen-activated protein kinase(MAPK)signaling pathways play an important role in regulation of bone formation and development,however,it remains unclear that the effect of dietary different levels of non-phytate phosphorus(NPP)on these signaling pathways and their correlations with bone phosphorus(P)retention and bone development in broilers.Therefore,this experiment was conducted to investigate the effect of dietary P supplementation on BMP and MAPK signaling pathways and their correlations with bone P retention and bone development in broilers.A total of 800 one-day-old Arbor Acres male broilers were randomly allotted to 1 of 5 treatments with 8 replicates in a completely randomized design.The 5 treatments of dietary NPP levels were 0.15,0.25,0.35,0.45 and 0.55%or 0.15,0.22,0.29,0.36 and 0.43%for broilers from 1 to 21 days of age or 22 to 42 days of age,respectively.The results showed that extracellular signal-regulated kinase 1(ERK1)mRNA expression in the tibia of broilers on days 14 and 28,phosphorylated-ERK1(p-ERK1)on day 14,and BMP2 protein expression on days 28 and42 decreased linearly(P<0.04),while c-Jun N-terminal kinase 1(JNK1)mRNA expression on day 42 increased linearly(P<0.02)with the increase of dietary NPP level.At 14 days of age,total P accumulation in tibia ash(TP),bone mineral concentration(BMC),bone mineral density(BMD),bone breaking strength(BBS)and tibia ash were negatively correlated(r=-0.726 to-0.359,P<0.05)with ERK1 and JNK1 mRNA as well as p-ERK1;tibia alkaline phosphatase(ALP)and bone gal protein(BGP)were positively correlated(r=0.405 to 0.665,P<0.01)with ERK1 mRNA and p-ERK1.At 28 days of age,TP,BMC,BMD,BBS and tibia ash were negatively correlated(r=-0.518 to-0.370,P<0.05)with ERK1 mRNA and BMP2 protein,while tibia ALP was positively correlated(r=0.382 to 0.648,P<0.05)with them.The results indicated that TP,BMC,BMD,BBS or tibia ash had negative correlations,while tibia ALP and BGP had positive correlations with ERK1 and JNK1 mRNAs,BMP2 protein and p-ERK1,suggesting that bone P retention and bone development might be regulated by BMP and MAPK signaling pathways in broiler chickens.展开更多
Accumulating data suggest that FGFs/FGFR1 plays essential roles in the bone development and human skeletal diseases. Conditional inactivation of fgfrl caused different phenotypes displaying in different cells or speci...Accumulating data suggest that FGFs/FGFR1 plays essential roles in the bone development and human skeletal diseases. Conditional inactivation of fgfrl caused different phenotypes displaying in different cells or specific organs and revealed some novel functions of FGFR1 in bone development. Fgfrl mutation mainly induced 2 types of human skeletal diseases, craniosynostosis syndrome and dysplasias. Similar mutation of fgfrl in mouse model just mimicked the phenotype that happened in human. These fa- cilitate the investigation on the underlying mechanism of the diseases. Here we mainly focused on the ad- vance of FGFR1 function in the bone development and its mutation caused skeletal diseases.展开更多
We assessed genetic and environmental effectson bone development of the hand and wrist, andon key anthropometric measures in Chinese youngtwins. In total, 139 monozygotic and 95 dizygotictwin pairs aged from 5 to 18 y...We assessed genetic and environmental effectson bone development of the hand and wrist, andon key anthropometric measures in Chinese youngtwins. In total, 139 monozygotic and 95 dizygotictwin pairs aged from 5 to 18 years were recruited.The twin correlations of total hand and wrist scoresfor monozygotic （MZ） and dizygotic （DZ） twins were0.71 and 0.36, respectively. Bivariate modelanalysis showed moderate genetic correlations onlyfor total skeletal maturity vs. weight and totalskeletal maturity vs. waist circumference （r, 0.51and 0.46, respectively）. Our findings demonstratedthat genetic factors played important roles in bonedevelopment of the hand and wrist in Chineseyoung twins, and that these genetic effects mightbe distinct from those influencing anthropometricmeasures.展开更多
The growth plate plays a crucial role in long bone development and elongation,housing stem cells that contribute to bone formation.This study investigates the role of primary cilia,specialized organelles that regulate...The growth plate plays a crucial role in long bone development and elongation,housing stem cells that contribute to bone formation.This study investigates the role of primary cilia,specialized organelles that regulate stem cell fate,in the development and repair of long bones.Here,we report the presence of primary cilia in all zones of the growth plate,particularly during embryonic development.Spatial transcriptomics and analysis of conditional knockout(CKO)mice identified that primary cilia mediate critical developmental signaling pathways within the growth plate.Disruption of primary cilia in growth plate chondroblasts or osteoblasts impaired long bone development by activating the Wnt signaling pathway and disrupting the cellular stemness.This resulted in elevated Mmp13 secretion,abnormal mineralization,and structural defects,ultimately hindering bone elongation.Time-lapse imaging showed an increased frequency of abnormal mitosis and a reduced rate of asymmetric division in skeletal stem cells(SSCs)from CKO mice.In conclusion,our findings indicated that primary cilia are critical for long bone development,regulating stem cell fate through key signaling pathways.Loss of primary cilia leads to excessive Wnt signaling and disruption of SSC stemness,impairing bone elongation.This study highlights the essential role of primary cilia in bone development and suggests potential therapeutic targets for skeletal disorders.展开更多
Tissue-resident stem cells are essential for development and repair,and in the skeleton,this function is fulfilled by recently identified skeletal stem cells(SSCs).However,recent work has identified that SSCs are not ...Tissue-resident stem cells are essential for development and repair,and in the skeleton,this function is fulfilled by recently identified skeletal stem cells(SSCs).However,recent work has identified that SSCs are not monolithic,with long bones,craniofacial sites,and the spine being formed by distinct stem cells.Recent studies have utilized techniques such as fluorescence-activated cell sorting,lineage tracing,and single-cell sequencing to investigate the involvement of ssCs in bone development,homeostasis,and disease.These investigations have allowed researchers to map the lineage commitment trajectory of ssCs in different parts of the body and at different time points.Furthermore,recent studies have shed light on the characteristics of ssCs in both physiological and pathological conditions.This review focuses on discussing the spatiotemporal distribution of ssCs and enhancing our understanding of the diversity and plasticity of ssCs by summarizing recent discoveries.展开更多
The study evaluated the effects of nutritional strategies on broilers challenged with Eimeria from d 14 to 26.A total of 840 Cobb male broilers were fed five diets in a 2×5 factorial arrangement:1)nutrient adequa...The study evaluated the effects of nutritional strategies on broilers challenged with Eimeria from d 14 to 26.A total of 840 Cobb male broilers were fed five diets in a 2×5 factorial arrangement:1)nutrient adequate diet(PC;0.84%calcium[Ca],0.42%available phosphorus[avP]);2)Ca-P deficient diet(NC;0.64%Ca,0.22%avP);3)NC+1500 FTU/kg phytase of diet(NC+PHY);4)NC+5000 IU/kg 25-hydroxycholecalciferol of diet(NC+25OHD);and 5)NC with both supplements(NC+PHY+25OHD),with and without Eimeria challenge.All treatments had six replicate cages with 14 birds per cage.At 5 days post inoculation(DPI),the challenged birds exhibited higher serum fluorescein isothiocyanated(FITC-d)levels than the unchallenged birds(P<0.001).The NC+PHY and NC+PHY+25OHD groups exhibited lower FITC-d levels compared to the NC+25OHD group(P=0.012).Significant interaction effects between Eimeria challenge and dietary treatments were observed on various parameters.During 0 to 6 and 0 to 12 DPI,Eimeria challenge resulted in decreased the body weight gain(BWG)(P<0.05)but had a negative effect on the feed conversion ratio(FCR)in birds compared to the unchallenged group(P<0.05).Reducing Ca and avP levels in the diet(NC)did not adversely affect BWG,but negatively impacted FCR,bone ash weight,ash concentration,and femur bone microstructure parameters(P<0.05).On 12 DPI,Eimeria challenge led to decreased tibia bone weight,bone volume,fatfree bone weight(FFBW),and ash weight of birds(P<0.05).Supplementation with phytase alone or in combination with 25OHD improved growth performance,gut permeability,bone ash and bone microstructure parameters in birds(P<0.05).However,the group fed 25OHD alone showed enhancements on growth performance,mineral apposition rate(MAR),bone ash concentration and ash percentage of the birds(P<0.05).In conclusion,lowering Ca and avP levels in the diet negatively affected FCR and bone development but did not affect intestinal integrity in broilers.Dietary supplementation of phytase,25OHD,or phytase in combination of 25OHD could enhance the growth performance and bone quality of broilers infected with Eimeria.Notably,the benefits of phytase supplementation were generally more pronounced than those associated with 25OHD supplementation;however,the combination of phytase and 25OHD could induce optimum effects.展开更多
INTRODUCTIONThe transforming growth factor-β （TGF-β） superfamily com- prises TGF-βs, Activin, bone morphogenetic proteins （BMPs） and other related proteins. TGF-β superfamily members act through a heteromeric ...INTRODUCTIONThe transforming growth factor-β （TGF-β） superfamily com- prises TGF-βs, Activin, bone morphogenetic proteins （BMPs） and other related proteins. TGF-β superfamily members act through a heteromeric receptor complex,, comprised of type I and type II receptors at the cell surface that transduce intracellular signals via Smad complex or mitogen-activated protein kinase （MAPK） cascade.展开更多
OBJECTIVE:This study was conducted to evaluate the effect of Amomum villosum on longitudinal bone growth.METHODS:Adolescent female Sprague-Dawley rats were divided into 3 groups and treated for 4 days:control(distille...OBJECTIVE:This study was conducted to evaluate the effect of Amomum villosum on longitudinal bone growth.METHODS:Adolescent female Sprague-Dawley rats were divided into 3 groups and treated for 4 days:control(distilled water,p.o.),recombinant human growth hormone(rhGH;100 μg/kg,s.c.),and A.villosum(500 mg/kg,p.o.) groups.On day 3,tetracycline(20 g/kg,i.p.) was injected for growth plate identification.On days 2,3 and 4,5-bromo-2'-deoxyuridine(BrdU)(50 mg/kg,i.p.) was injected to label proliferating cells.On day 5,tibias were dissected and fixed in 4% paraformaldehyde,dehydrated,and sectioned for immunohistochemistry and histomorphometry.RESULTS:The rate of bone growth in the A.villosum and rhGH groups increased to(410 ± 44) and(389 ± 46) μm/day(P<0.01),respectively,as compared with the control(330.7 ± 34.7) μm/day.The thickness of the growth plates also increased to(591 ± 37) and(598 ± 32) μm,respectively,as compared with the control(524± 89) μm(P<0.001).The number of BrdU-positive cells in the chondrocytes of the A.villosum and rhGH groups was also significantly higher(126±24) and(143±18) cells/mm 2,respectively) than in the control(109 ± 25) mm 2(P<0.05).Insulin-like growth factor-1 and bone morphogenetic protein-2 in the A.villosum and rhGH groups were highly expressed in the growth plate as compared with the control samples,indicating increased bone formation.CONCLUSIONS:A.villosum could be used to treat growth retardation during adolescence.展开更多
Skeletal stem/progenitor cells(SSPCs)are tissue-specific stem/progenitor cells localized within skeletons and contribute to bone development,homeostasis,and regeneration.However,the heterogeneity of SSPC populations i...Skeletal stem/progenitor cells(SSPCs)are tissue-specific stem/progenitor cells localized within skeletons and contribute to bone development,homeostasis,and regeneration.However,the heterogeneity of SSPC populations in mouse long bones and their respective regenerative capacity remain to be further clarified.In this study,we perform integrated analysis using single-cell RNA sequencing(scRNA-seq)datasets of mouse hindlimb buds,postnatal long bones,and fractured long bones.Our analyses reveal the heterogeneity of osteochondrogenic lineage cells and recapitulate the developmental trajectories during mouse long bone growth.In addition,we identify a novel Cd168þSSPC population with highly replicating capacity and osteochondrogenic potential in embryonic and postnatal long bones.Moreover,the Cd168þSSPCs can contribute to newly formed skeletal tissues during fracture healing.Furthermore,the results of multicolor immunofluorescence show that Cd168þSSPCs reside in the superficial zone of articular cartilage as well as in growth plates of postnatal mouse long bones.In summary,we identify a novel Cd168þSSPC population with regenerative potential in mouse long bones,which adds to the knowledge of the tissuespecific stem cells in skeletons.展开更多
BACKGROUND The epiphyseal growth plate is an important anatomical segment localized on the ends of a long bone.Despite the abovementioned atractive reasons for alendronate’s use,few data on the effect of alendronate ...BACKGROUND The epiphyseal growth plate is an important anatomical segment localized on the ends of a long bone.Despite the abovementioned atractive reasons for alendronate’s use,few data on the effect of alendronate during epiphyseal growth exist.AIM Verify the effect of alendronate on the growth epiphyseal plate,and compare its effect with the size of the femur during the double-staining of the immunolocalization of transforming growth factor-β1(TGF-β1)and bone morphogenetic protein-2(BMP2)in endochondral ossifing in specimens that have received alendronate.METHODS Forty newborn rats were randomly divided into two groups:a control group(were given applications of 1 mg/kg physiologic saline)and a group that received Alendronate(a dose of 2.5 mg/kg).These groups were then divided into two subgroups for euthanasia in two and 12 d of life.After euthanasia,the femurs were removed,and the femoral bones were measured linearly between the apex of the greater trochanter until the lower intercondylar midlle face to verify the probable bone growth between 3 and 12 d in control and alednroanto treated rats.Posteriorly,the surgical pieces were also sent to the histopathology laboratory to produce histological slides.The obtained slides were stained with hematoxylin and eosin to measure each of the cartilage zones in endochondral development.and other slides were immunohistochemically tested for anti-TGF-β1 and BMP-2 antibodies to investigate the immunolocalization of these proteins in the epiphyseal plaque area.RESULTS On the third day,some diferences between the control group and specimens treated with alendronate were verified.Macroscopiccaly,we found similarities in size between the femoral bones when we compared the control group with the specimens that received alendronate.On the 12^th day,the bone size of the mice receiving the drug was significantly smaller than those of the control group.These results coincide with changes in the TGF-β1 and BMP-2 expression.In the specimens that received alendronate,the TGF-β1 was expressed in some sites of trabecular bone that was neoformed,peripherally to the bone marrow area.The BMP-2 was also positive in proliferative chondrocytes and hypertrofic chondrocytes.On the 12^th day,all layers of chondrocytes exhibited positivity for BMP-2 in the specimens that received alendronate.In the interface between the trabecular bone and cartilage,an area of disorganized bone deposition was evident.Neoformed bone also appeared to be different at 12 d.In the control group,BMP-2 was positive in an intense area of bone trabeculae,whereas the alendronate-treated group showed TGF-β1 positive trabeculae and a greater bone area.CONCLUSION Alendronate alters the immunolocalization of TGF-β1 and BMP-2 simultaneously,a condition that changes the usual histological aspects of the cartilage zone and impairs epiphysis growth and femur growth.展开更多
BMP2 plays crucial roles in vertebrate developmental process and acts as a bone inducer during osteogenesis. We present here the molecular cloning of bmp2 cDNA from the marine flatfish Cynoglossus semilaevis, and the ...BMP2 plays crucial roles in vertebrate developmental process and acts as a bone inducer during osteogenesis. We present here the molecular cloning of bmp2 cDNA from the marine flatfish Cynoglossus semilaevis, and the analysis of bmp2 expression profiling and promoter function. The full length of bmp2 cDNA sequence is 2 048 bp,which encodes a protein of 422 amino acids. Tissue expression distribution of bmp2 was examined in 14 tissues of mature individuals by quantitative real time PCR（qRT-PCR）. The results revealed that bmp2 was expressed ubiquitously, and the highest expression level was detected in the spinal cord. Moreover, bmp2 expression levels were detected at 15 sampling time points of early developmental stages（egg, larva, juvenile and fingerling stages）.The highest expression level of bmp2 was observed at the gastrula stage, which was about ten times higher than those at the other three embryo stages. Whole-mount in situ hybridization showed that the bmp2 signal was strongly detected at the location of the crown-like larval fin, heart and liver, and slightly expressed in the notochord at one day post hatch（dph）; then the expression of bmp2 started to be concentrated in notochord at three dph. Subsequently, we characterized the 5′-flanking region of bmp2 by testing the promoter activity by Luciferase reporter assays. Positive regulatory region was detected at the location of –179 to ＋109. The predicted transcription factor binding sites（E-box binding factors, zinc finger transcription factor, etc.） in this region might participate in the transcriptional regulation of the bmp2 gene.展开更多
Scaffold-free techniques in the developmental tissue engineering area are designed to mimic in vivo embryonic processes with the aim of biofabricating,in vitro,tissues with more authentic properties.Cell clusters call...Scaffold-free techniques in the developmental tissue engineering area are designed to mimic in vivo embryonic processes with the aim of biofabricating,in vitro,tissues with more authentic properties.Cell clusters called spheroids are the basis for scaffold-free tissue engineering.In this review,we explore the use of spheroids from adult mesenchymal stem/stromal cells as a model in the developmental engineering area in order to mimic the developmental stages of cartilage and bone tissues.Spheroids from adult mesenchymal stromal/stem cells lineages recapitulate crucial events in bone and cartilage formation during embryogenesis,and are capable of spontaneously fusing to other spheroids,making them ideal building blocks for bone and cartilage tissue engineering.Here,we discuss data from ours and other labs on the use of adipose stromal/stem cell spheroids in chondrogenesis and osteogenesis in vitro.Overall,recent studies support the notion that spheroids are ideal"building blocks"for tissue engineering by“bottom-up”approaches,which are based on tissue assembly by advanced techniques such as three-dimensional bioprinting.Further studies on the cellular and molecular mechanisms that orchestrate spheroid fusion are now crucial to support continued development of bottom-up tissue engineering approaches such as three-dimensional bioprinting.展开更多
目的:分析和总结自体股骨头结构植骨重建髋臼辅助全髋关节置换术(total hip arthroplasty,THA)治疗改良Crowe TypeⅣB型成人髋关节发育不良性脱位(developmental dysplasia of the hip,DDH)患者的临床疗效。方法:按照改良Crowe分型,选...目的:分析和总结自体股骨头结构植骨重建髋臼辅助全髋关节置换术(total hip arthroplasty,THA)治疗改良Crowe TypeⅣB型成人髋关节发育不良性脱位(developmental dysplasia of the hip,DDH)患者的临床疗效。方法:按照改良Crowe分型,选取山东大学齐鲁医院德州医院关节外科2015年8月至2023年3月收治的TypeⅣB型DDH患者26例,其中男25例,女1例,采用自体股骨头结构植骨重建髋臼辅助THA,记录患者手术时间、术中失血量、术中术后输血量、术后血红蛋白、手术相关并发症和骨愈合时间等,并行骨盆正位X线片了解假体位置、假体骨长入、假体松动以及骨愈合情况等,采用视觉模拟评分表(visual analogue scale,VAS)评价髋关节的疼痛不适,采用髋关节Harris评分和Western Ontario and McMaster Universities Osteoarthritis Index(WOMAC)来评价髋关节功能及临床疗效。结果:所有患者的平均随访时间(9.73±8.35)个月,术中出血平均为(715.38±143.37) mL,术中平均输血(415.38±282.41) mL,手术时间平均为(118.62±18.27) min,术后平均输血为(192.31±236.51) mL。所有患者转子下骨端、自体股骨头和假臼之间均骨愈合良好。髋关节VAS评分从术前6.73±0.45,至术后末次随访时VAS评分1.73±0.53,差异有统计学意义(P=0.000),髋关节活动度均较术前明显改善,髋关节Harris评分从术前24.27±1.66,至术后末次随访时Harris评分74.77±2.89,差异有统计学意义(P=0.000),WOMAC术前术后评分分别为130.08±5.72和67.85±3.23,差异均有统计学意义(P=0.000)。结论:自体股骨头结构植骨重建髋臼辅助THA治疗改良Crowe TypeⅣB型DDH,具有操作相对简单、固定牢固、手术相对安全和疗效确切的优点。展开更多
基金supported by the National Key R&D Program of China(2021YFD2100700)National Natural Science Foundation of China(32021005)+1 种基金111 project(BP0719028)Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province。
文摘This study endeavors to investigate the effects of Bifidobacterium breve CCFM1078 on bone formation and resorption balance in growing BALB/c mice.Newborn BALB/c mice were assigned to the control group(administration saline)and the CCFM1078 group(administration B.breve CCFM1078,3×10^(9) CFU/day)in 3-,4-,and 5-week tests.All the groups have male and female distinctions.Our findings demonstrate that B.breve CCFM1078 exerts on the dynamic equilibrium between bone formation and resorption during the critical period of growth in mice by modulating the composition of gut microbiota and metabolites(hexadecanamide,linoleoyl ethanolamide,and palmitoyl ethanolamide),the genes and proteins expression related to the growth hormone(GH)/insulin-like growth factors-1(IGF-1)axis and Gs/PKA/CREB signaling pathways,as well as downstream osteogenic and osteoclastic differentiation factors.The effects of B.breve CCFM1078 were different with age and gender dependent.This finding suggests B.breve CCFM1078 may have potential applications in regulating bone metabolism in the growth period population.
基金supported by the NIH grants R01DE025866 from NIDCRR01AR070877 from NIAMSsupported by the 111 Project, MOE (B14038), China
文摘How osteoblast cells are induced is a central question for understanding skeletal formation. Abnormal osteoblast differentiation leads to a broad range of devastating craniofacial diseases. Here we have investigated intramembranous ossification during cranial bone development in mouse models of skeletal genetic diseases that exhibit craniofacial bone defects. The GNAS gene encodes Gαs that transduces GPCR signaling. GNAS activation or loss-of-function mutations in humans cause fibrous dysplasia(FD) or progressive osseous heteroplasia(POH) that shows craniofacial hyperostosis or craniosynostosis, respectively. We find here that, while Hh ligand-dependent Hh signaling is essential for endochondral ossification, it is dispensable for intramembranous ossification, where Gαsregulates Hh signaling in a ligand-independent manner. We further show that Gαscontrols intramembranous ossification by regulating both Hh and Wnt/β-catenin signaling. In addition, Gαsactivation in the developing cranial bone leads to reduced ossification but increased cartilage presence due to reduced cartilage dissolution, not cell fate switch. Small molecule inhibitors of Hh and Wnt signaling can effectively ameliorate cranial bone phenotypes in mice caused by loss or gain of Gnas function mutations, respectively. Our work shows that studies of genetic diseases provide invaluable insights in both pathological bone defects and normal bone development, understanding both leads to better diagnosis and therapeutic treatment of bone diseases.
基金The present study was financially supported by the National Key R&D Program of China(2017YF0502200)the Key Program of the National Natural Science Foundation of China(31630073)+2 种基金the earmarked fund for China Agriculture Research System(CARS-41)the Agricultural Science and Technology Innovation Program,China(ASTIP-IAS09)the earmarked fund for Hebei Chicken Innovation Team of Modern Agro-Industry Technology Research System,China(HBCT2018150203 and HBCT2018150206).
文摘This experiment was conducted to investigate the effect of dietary calcium(Ca)or phosphorus(P)deficiency on bone development and related Ca or P metabolic utilization parameters of broilers from 22 to 42 days of age based on our previous study,which indicated that dietary Ca or P deficiency impaired the bone development by regulating related Ca or P metabolic utilization parameters of broilers from 1 to 21 days of age.A total of 504 one-day-old Arbor Acres male broilers were randomly assigned to 1 of 4 treatments with 7 replicates in a completely randomized design,and fed the normal control and Ca-or P-deficient diets from 1 to 21 days of age.At 22 days of age,the broilers were further fed the normal control diet(0.90%Ca+0.35%non-phytate P(NPP)),the P-deficient diet(0.90%Ca+0.18%NPP),the Ca-deficient diet(0.30%Ca+0.35%NPP)or the Ca and P-deficient diet(0.30%Ca+0.18%NPP),respectively.The results showed that dietary Ca or P deficiency decreased(P<0.05)tibia bone mineral density(BMD),bone breaking strength(BBS),ash content,tibia ash Ca content and serum P content on days 28 and 42,but increased(P<0.05)tibia alkaline phosphatase(ALP)activity of broilers on day 42 compared with the control group.Furthermore,the broilers fed the P-deficient diet had the lowest(P<0.05)tibia BMD,BBS,ash content,serum P content and the highest(P<0.05)serum Ca content on day 28 compared with those fed the Ca-deficient or Ca and P-deficient diets.The results from the present study indicated that the bone development and related Ca or P metabolic utilization parameters of broilers were the most sensitive to dietary P deficiency,followed by dietary Ca deficiency or Ca and P-deficiency;dietary Ca or P deficiency impaired the bone development possibly by regulating serum Ca and P contents as well as tibia Ca content and ALP activity of broilers from 22 to 42 days of age.
基金supported by the Key Program of the National Natural Science Foundation of China(31630073)the China Agriculture Research System of MOF and MARA(CARS-41)the Agricultural Science and Technology Innovation Program,China(ASTIPIAS09)。
文摘The bone morphogenetic protein(BMP)and mitogen-activated protein kinase(MAPK)signaling pathways play an important role in regulation of bone formation and development,however,it remains unclear that the effect of dietary different levels of non-phytate phosphorus(NPP)on these signaling pathways and their correlations with bone phosphorus(P)retention and bone development in broilers.Therefore,this experiment was conducted to investigate the effect of dietary P supplementation on BMP and MAPK signaling pathways and their correlations with bone P retention and bone development in broilers.A total of 800 one-day-old Arbor Acres male broilers were randomly allotted to 1 of 5 treatments with 8 replicates in a completely randomized design.The 5 treatments of dietary NPP levels were 0.15,0.25,0.35,0.45 and 0.55%or 0.15,0.22,0.29,0.36 and 0.43%for broilers from 1 to 21 days of age or 22 to 42 days of age,respectively.The results showed that extracellular signal-regulated kinase 1(ERK1)mRNA expression in the tibia of broilers on days 14 and 28,phosphorylated-ERK1(p-ERK1)on day 14,and BMP2 protein expression on days 28 and42 decreased linearly(P<0.04),while c-Jun N-terminal kinase 1(JNK1)mRNA expression on day 42 increased linearly(P<0.02)with the increase of dietary NPP level.At 14 days of age,total P accumulation in tibia ash(TP),bone mineral concentration(BMC),bone mineral density(BMD),bone breaking strength(BBS)and tibia ash were negatively correlated(r=-0.726 to-0.359,P<0.05)with ERK1 and JNK1 mRNA as well as p-ERK1;tibia alkaline phosphatase(ALP)and bone gal protein(BGP)were positively correlated(r=0.405 to 0.665,P<0.01)with ERK1 mRNA and p-ERK1.At 28 days of age,TP,BMC,BMD,BBS and tibia ash were negatively correlated(r=-0.518 to-0.370,P<0.05)with ERK1 mRNA and BMP2 protein,while tibia ALP was positively correlated(r=0.382 to 0.648,P<0.05)with them.The results indicated that TP,BMC,BMD,BBS or tibia ash had negative correlations,while tibia ALP and BGP had positive correlations with ERK1 and JNK1 mRNAs,BMP2 protein and p-ERK1,suggesting that bone P retention and bone development might be regulated by BMP and MAPK signaling pathways in broiler chickens.
基金the National Key Basic Research and Devel opment Plan of China(973 Projects,2005CB522604)the Research Project of Scientific Committee of Chongqing(2004BA5016)
文摘Accumulating data suggest that FGFs/FGFR1 plays essential roles in the bone development and human skeletal diseases. Conditional inactivation of fgfrl caused different phenotypes displaying in different cells or specific organs and revealed some novel functions of FGFR1 in bone development. Fgfrl mutation mainly induced 2 types of human skeletal diseases, craniosynostosis syndrome and dysplasias. Similar mutation of fgfrl in mouse model just mimicked the phenotype that happened in human. These fa- cilitate the investigation on the underlying mechanism of the diseases. Here we mainly focused on the ad- vance of FGFR1 function in the bone development and its mutation caused skeletal diseases.
文摘We assessed genetic and environmental effectson bone development of the hand and wrist, andon key anthropometric measures in Chinese youngtwins. In total, 139 monozygotic and 95 dizygotictwin pairs aged from 5 to 18 years were recruited.The twin correlations of total hand and wrist scoresfor monozygotic （MZ） and dizygotic （DZ） twins were0.71 and 0.36, respectively. Bivariate modelanalysis showed moderate genetic correlations onlyfor total skeletal maturity vs. weight and totalskeletal maturity vs. waist circumference （r, 0.51and 0.46, respectively）. Our findings demonstratedthat genetic factors played important roles in bonedevelopment of the hand and wrist in Chineseyoung twins, and that these genetic effects mightbe distinct from those influencing anthropometricmeasures.
基金supported by National Key R&D Program of China(2022YFA1103200)National,Natural Science Foundation Projects of China(U23A20444,82270963,82425014,82301048)+2 种基金Shanghai Municipal Education Commission Scientific Research Innovation Plan Project(2023ZKZD29)China Postdoctoral Science Foundation General Project(2022M722406)Shanghai Science and Technology Innovation Action Plan:Qimingxing Project(Sailing Special Program,23YF1450500).
文摘The growth plate plays a crucial role in long bone development and elongation,housing stem cells that contribute to bone formation.This study investigates the role of primary cilia,specialized organelles that regulate stem cell fate,in the development and repair of long bones.Here,we report the presence of primary cilia in all zones of the growth plate,particularly during embryonic development.Spatial transcriptomics and analysis of conditional knockout(CKO)mice identified that primary cilia mediate critical developmental signaling pathways within the growth plate.Disruption of primary cilia in growth plate chondroblasts or osteoblasts impaired long bone development by activating the Wnt signaling pathway and disrupting the cellular stemness.This resulted in elevated Mmp13 secretion,abnormal mineralization,and structural defects,ultimately hindering bone elongation.Time-lapse imaging showed an increased frequency of abnormal mitosis and a reduced rate of asymmetric division in skeletal stem cells(SSCs)from CKO mice.In conclusion,our findings indicated that primary cilia are critical for long bone development,regulating stem cell fate through key signaling pathways.Loss of primary cilia leads to excessive Wnt signaling and disruption of SSC stemness,impairing bone elongation.This study highlights the essential role of primary cilia in bone development and suggests potential therapeutic targets for skeletal disorders.
基金supported in part by National Natural Science Foundation of China(Grant nos.82372362,81972034,92068104 to Ren Xu and 82002262 to Na Li)National Key R&D Program of China(2020YFA0112900 to Ren Xu)+2 种基金Natural Science Foundation of Fujian Province(2022J06003 to Ren Xu)Project of Xiarmen Cell Therapy Research,Xiamen,Fujian,China(3502Z20214001)supported by a Pershing Square MIND Prize award,an Irma T.Hirschl Career Scientist Award,an NIH award RO1AR075585,a Career Award for Medical Scientists from the Burroughs Welcome Foundation,a William Rhodes and Louise Tilzer-Rhodes Center for Glioblastoma research award,and a Weill Comell Medicine Prostate Cancer SPORE Developmental Research Program Award。
文摘Tissue-resident stem cells are essential for development and repair,and in the skeleton,this function is fulfilled by recently identified skeletal stem cells(SSCs).However,recent work has identified that SSCs are not monolithic,with long bones,craniofacial sites,and the spine being formed by distinct stem cells.Recent studies have utilized techniques such as fluorescence-activated cell sorting,lineage tracing,and single-cell sequencing to investigate the involvement of ssCs in bone development,homeostasis,and disease.These investigations have allowed researchers to map the lineage commitment trajectory of ssCs in different parts of the body and at different time points.Furthermore,recent studies have shed light on the characteristics of ssCs in both physiological and pathological conditions.This review focuses on discussing the spatiotemporal distribution of ssCs and enhancing our understanding of the diversity and plasticity of ssCs by summarizing recent discoveries.
文摘The study evaluated the effects of nutritional strategies on broilers challenged with Eimeria from d 14 to 26.A total of 840 Cobb male broilers were fed five diets in a 2×5 factorial arrangement:1)nutrient adequate diet(PC;0.84%calcium[Ca],0.42%available phosphorus[avP]);2)Ca-P deficient diet(NC;0.64%Ca,0.22%avP);3)NC+1500 FTU/kg phytase of diet(NC+PHY);4)NC+5000 IU/kg 25-hydroxycholecalciferol of diet(NC+25OHD);and 5)NC with both supplements(NC+PHY+25OHD),with and without Eimeria challenge.All treatments had six replicate cages with 14 birds per cage.At 5 days post inoculation(DPI),the challenged birds exhibited higher serum fluorescein isothiocyanated(FITC-d)levels than the unchallenged birds(P<0.001).The NC+PHY and NC+PHY+25OHD groups exhibited lower FITC-d levels compared to the NC+25OHD group(P=0.012).Significant interaction effects between Eimeria challenge and dietary treatments were observed on various parameters.During 0 to 6 and 0 to 12 DPI,Eimeria challenge resulted in decreased the body weight gain(BWG)(P<0.05)but had a negative effect on the feed conversion ratio(FCR)in birds compared to the unchallenged group(P<0.05).Reducing Ca and avP levels in the diet(NC)did not adversely affect BWG,but negatively impacted FCR,bone ash weight,ash concentration,and femur bone microstructure parameters(P<0.05).On 12 DPI,Eimeria challenge led to decreased tibia bone weight,bone volume,fatfree bone weight(FFBW),and ash weight of birds(P<0.05).Supplementation with phytase alone or in combination with 25OHD improved growth performance,gut permeability,bone ash and bone microstructure parameters in birds(P<0.05).However,the group fed 25OHD alone showed enhancements on growth performance,mineral apposition rate(MAR),bone ash concentration and ash percentage of the birds(P<0.05).In conclusion,lowering Ca and avP levels in the diet negatively affected FCR and bone development but did not affect intestinal integrity in broilers.Dietary supplementation of phytase,25OHD,or phytase in combination of 25OHD could enhance the growth performance and bone quality of broilers infected with Eimeria.Notably,the benefits of phytase supplementation were generally more pronounced than those associated with 25OHD supplementation;however,the combination of phytase and 25OHD could induce optimum effects.
基金supported by grants by NIH grant AR-044741(Y-PL) and R01DE023813 (Y-PL)
文摘INTRODUCTIONThe transforming growth factor-β （TGF-β） superfamily com- prises TGF-βs, Activin, bone morphogenetic proteins （BMPs） and other related proteins. TGF-β superfamily members act through a heteromeric receptor complex,, comprised of type I and type II receptors at the cell surface that transduce intracellular signals via Smad complex or mitogen-activated protein kinase （MAPK） cascade.
基金Supported by the Fund of Amomum villosum in bone growth
文摘OBJECTIVE:This study was conducted to evaluate the effect of Amomum villosum on longitudinal bone growth.METHODS:Adolescent female Sprague-Dawley rats were divided into 3 groups and treated for 4 days:control(distilled water,p.o.),recombinant human growth hormone(rhGH;100 μg/kg,s.c.),and A.villosum(500 mg/kg,p.o.) groups.On day 3,tetracycline(20 g/kg,i.p.) was injected for growth plate identification.On days 2,3 and 4,5-bromo-2'-deoxyuridine(BrdU)(50 mg/kg,i.p.) was injected to label proliferating cells.On day 5,tibias were dissected and fixed in 4% paraformaldehyde,dehydrated,and sectioned for immunohistochemistry and histomorphometry.RESULTS:The rate of bone growth in the A.villosum and rhGH groups increased to(410 ± 44) and(389 ± 46) μm/day(P<0.01),respectively,as compared with the control(330.7 ± 34.7) μm/day.The thickness of the growth plates also increased to(591 ± 37) and(598 ± 32) μm,respectively,as compared with the control(524± 89) μm(P<0.001).The number of BrdU-positive cells in the chondrocytes of the A.villosum and rhGH groups was also significantly higher(126±24) and(143±18) cells/mm 2,respectively) than in the control(109 ± 25) mm 2(P<0.05).Insulin-like growth factor-1 and bone morphogenetic protein-2 in the A.villosum and rhGH groups were highly expressed in the growth plate as compared with the control samples,indicating increased bone formation.CONCLUSIONS:A.villosum could be used to treat growth retardation during adolescence.
基金supported by the National Key R&D Program of China(2022YFA1104100,2022YFA1103500)the National Natural Sciences Grants China(82172388,82372373,81871771)the Beijing Natural Sciences Foundation(7222123,L212065).
文摘Skeletal stem/progenitor cells(SSPCs)are tissue-specific stem/progenitor cells localized within skeletons and contribute to bone development,homeostasis,and regeneration.However,the heterogeneity of SSPC populations in mouse long bones and their respective regenerative capacity remain to be further clarified.In this study,we perform integrated analysis using single-cell RNA sequencing(scRNA-seq)datasets of mouse hindlimb buds,postnatal long bones,and fractured long bones.Our analyses reveal the heterogeneity of osteochondrogenic lineage cells and recapitulate the developmental trajectories during mouse long bone growth.In addition,we identify a novel Cd168þSSPC population with highly replicating capacity and osteochondrogenic potential in embryonic and postnatal long bones.Moreover,the Cd168þSSPCs can contribute to newly formed skeletal tissues during fracture healing.Furthermore,the results of multicolor immunofluorescence show that Cd168þSSPCs reside in the superficial zone of articular cartilage as well as in growth plates of postnatal mouse long bones.In summary,we identify a novel Cd168þSSPC population with regenerative potential in mouse long bones,which adds to the knowledge of the tissuespecific stem cells in skeletons.
文摘BACKGROUND The epiphyseal growth plate is an important anatomical segment localized on the ends of a long bone.Despite the abovementioned atractive reasons for alendronate’s use,few data on the effect of alendronate during epiphyseal growth exist.AIM Verify the effect of alendronate on the growth epiphyseal plate,and compare its effect with the size of the femur during the double-staining of the immunolocalization of transforming growth factor-β1(TGF-β1)and bone morphogenetic protein-2(BMP2)in endochondral ossifing in specimens that have received alendronate.METHODS Forty newborn rats were randomly divided into two groups:a control group(were given applications of 1 mg/kg physiologic saline)and a group that received Alendronate(a dose of 2.5 mg/kg).These groups were then divided into two subgroups for euthanasia in two and 12 d of life.After euthanasia,the femurs were removed,and the femoral bones were measured linearly between the apex of the greater trochanter until the lower intercondylar midlle face to verify the probable bone growth between 3 and 12 d in control and alednroanto treated rats.Posteriorly,the surgical pieces were also sent to the histopathology laboratory to produce histological slides.The obtained slides were stained with hematoxylin and eosin to measure each of the cartilage zones in endochondral development.and other slides were immunohistochemically tested for anti-TGF-β1 and BMP-2 antibodies to investigate the immunolocalization of these proteins in the epiphyseal plaque area.RESULTS On the third day,some diferences between the control group and specimens treated with alendronate were verified.Macroscopiccaly,we found similarities in size between the femoral bones when we compared the control group with the specimens that received alendronate.On the 12^th day,the bone size of the mice receiving the drug was significantly smaller than those of the control group.These results coincide with changes in the TGF-β1 and BMP-2 expression.In the specimens that received alendronate,the TGF-β1 was expressed in some sites of trabecular bone that was neoformed,peripherally to the bone marrow area.The BMP-2 was also positive in proliferative chondrocytes and hypertrofic chondrocytes.On the 12^th day,all layers of chondrocytes exhibited positivity for BMP-2 in the specimens that received alendronate.In the interface between the trabecular bone and cartilage,an area of disorganized bone deposition was evident.Neoformed bone also appeared to be different at 12 d.In the control group,BMP-2 was positive in an intense area of bone trabeculae,whereas the alendronate-treated group showed TGF-β1 positive trabeculae and a greater bone area.CONCLUSION Alendronate alters the immunolocalization of TGF-β1 and BMP-2 simultaneously,a condition that changes the usual histological aspects of the cartilage zone and impairs epiphysis growth and femur growth.
基金The Special Scientific Research Funds for Central Non-profit Institutes,Chinese Academy of Fishery Sciences under contract No.2016RC-LX02the National Natural Science Foundation of China under contract No.31201981
文摘BMP2 plays crucial roles in vertebrate developmental process and acts as a bone inducer during osteogenesis. We present here the molecular cloning of bmp2 cDNA from the marine flatfish Cynoglossus semilaevis, and the analysis of bmp2 expression profiling and promoter function. The full length of bmp2 cDNA sequence is 2 048 bp,which encodes a protein of 422 amino acids. Tissue expression distribution of bmp2 was examined in 14 tissues of mature individuals by quantitative real time PCR（qRT-PCR）. The results revealed that bmp2 was expressed ubiquitously, and the highest expression level was detected in the spinal cord. Moreover, bmp2 expression levels were detected at 15 sampling time points of early developmental stages（egg, larva, juvenile and fingerling stages）.The highest expression level of bmp2 was observed at the gastrula stage, which was about ten times higher than those at the other three embryo stages. Whole-mount in situ hybridization showed that the bmp2 signal was strongly detected at the location of the crown-like larval fin, heart and liver, and slightly expressed in the notochord at one day post hatch（dph）; then the expression of bmp2 started to be concentrated in notochord at three dph. Subsequently, we characterized the 5′-flanking region of bmp2 by testing the promoter activity by Luciferase reporter assays. Positive regulatory region was detected at the location of –179 to ＋109. The predicted transcription factor binding sites（E-box binding factors, zinc finger transcription factor, etc.） in this region might participate in the transcriptional regulation of the bmp2 gene.
基金the Coordination for the Improvement of Higher Education Personnel(CAPES),No.88882.366181/2019-01the Carlos Chagas Filho Foundation for Research Support of the State of Rio de Janeiro(FAPERJ),No.E-26/202.682/2018National Council for Scientific and Technological Development(CNPq),No.467513/2014-7
文摘Scaffold-free techniques in the developmental tissue engineering area are designed to mimic in vivo embryonic processes with the aim of biofabricating,in vitro,tissues with more authentic properties.Cell clusters called spheroids are the basis for scaffold-free tissue engineering.In this review,we explore the use of spheroids from adult mesenchymal stem/stromal cells as a model in the developmental engineering area in order to mimic the developmental stages of cartilage and bone tissues.Spheroids from adult mesenchymal stromal/stem cells lineages recapitulate crucial events in bone and cartilage formation during embryogenesis,and are capable of spontaneously fusing to other spheroids,making them ideal building blocks for bone and cartilage tissue engineering.Here,we discuss data from ours and other labs on the use of adipose stromal/stem cell spheroids in chondrogenesis and osteogenesis in vitro.Overall,recent studies support the notion that spheroids are ideal"building blocks"for tissue engineering by“bottom-up”approaches,which are based on tissue assembly by advanced techniques such as three-dimensional bioprinting.Further studies on the cellular and molecular mechanisms that orchestrate spheroid fusion are now crucial to support continued development of bottom-up tissue engineering approaches such as three-dimensional bioprinting.
