The aim of the present real time in vivo micro-computed tomography （pCT） and histologic experiment was to assess the efficacy of guided bone regeneration （GBR） around standardized calvarial critical size defects ...The aim of the present real time in vivo micro-computed tomography （pCT） and histologic experiment was to assess the efficacy of guided bone regeneration （GBR） around standardized calvarial critical size defects （CSD） using bone marrow-derived mesenchymal stem cells （BMSCs）, and collagen membrane （CM） with and without tricalcium phosphate （TCP） graft material. In the calvaria of nine female Sprague-Dawley rats, full-thickness CSD （diameter 4.6 mm） were created under general anesthesia. Treatment-wise, rats were divided into three groups. In group 1, CSD was covered with a resorbable CM; in group 2, BMSCs were filled in CSD and covered with CM; and in group 3, TCP soaked in BMSCs was placed in CSD and covered with CM. All defects were closed using resorbable sutures. Bone volume and bone mineral density of newly formed bone （NFB） and remaining TCP particles and rate of new bone formation was determined at baseline, 2, 4, 6, and 10 weeks using in vivo pCT. At the lOth week, the rats were killed and calvarial segments were assessed histologically. The results showed that the hardness of NFB was similar to that of the native bone in groups I and 2 as compared to the NFB in group 3. Likewise, values for the modulus of elasticity were also significantly higher in group 3 compared to groups 1 and 2. This suggests that TCP when used in combination with BMSCs and without CM was unable to form bone of significant strength that could possibly provide mechanical ＂lock＂ between the natural bone and NFB. The use of BMSCs as adjuncts to conventional GBR initiated new bone formation as early as 2 weeks of treatment compared to when GBR is attempted without adiunct BMSC therapy.展开更多
Natural diatomite （DT） is the ancient deposit of diatom skeleton with many regular pores of 50-200 nm and also an abundant source of biogenic silica. Although silica is considered biologically safe and there is an i...Natural diatomite （DT） is the ancient deposit of diatom skeleton with many regular pores of 50-200 nm and also an abundant source of biogenic silica. Although silica is considered biologically safe and there is an increasing interest of using natural diatomite for biomedical applications, the toxicity information about natural diatomite is still missing. Here, cytotoxicity of natural diatomite on osteoblasts and fibroblasts were compared to hydroxyapatite and the relationships between cytotoxicity and diatomite sizes, dose, geometry or impurity were systematically investigated. Cell adhesion and interaction with diatomite particles were also fluorescently observed, The results clearly suggested a size-, dose- and shape-dependent cytotoxicity of natural diatomite. Disk-shaped diatomite particles with average size of 30μm in diameter revealed the least toxicity, while the diatomite particles with irregular shapes and sizes less than 10 μm were remarkably toxic. Diatomite particles with proper sizes were then selected to investigate the reinforcing effect on injectable calcium phosphate bone cement. Results showed that diatomite significantly improved the compressive strength of bone cement but did not alter the injectability of the cement, This work provided important biocompatibility information of natural diatomite and demonstrated the feasibility of using selected diatomite as bone implant material.展开更多
Experiments on maxillofacial bone tissue engineering showed the promising result;however, its healing mechanisms and effectiveness had not been fully understood. The aim of this study is to compare the bone healing me...Experiments on maxillofacial bone tissue engineering showed the promising result;however, its healing mechanisms and effectiveness had not been fully understood. The aim of this study is to compare the bone healing mechanism and osteogenic capacity between bovine bone mineral loaded with hAMSC and autogenous bone graft in the reconstruction of critical size mandibular bone defect. Critical size defects were made at the mandible of 45 New Zealand white rabbits reconstructed with BBM-hAMSC, BBM alone, and ABG, respectively. At the end of first, second, and twelfth weeks, five rabbits from each experimental week were sacrificed for histology and immunohistochemistry staining. Expressions of vascular endothelial growth factor (VEGF), bone mor-phogenic proteins-2 (BMP2), Runx2 and the amount of angiogenesis were analyzed in the first and second week groups, while expressions of Runx2, osteocalcin, collagen type-I fibres, trabecular area and bone incorporation were analyzed in the twelfth week groups. The result showed that expressions of VEGF, BMP2 and Runx2 as well as the amount of angiogenesis were higher in ABG compared with BBM-hAMSC group in the first and second weeks of healing. The result of twelfth week of healing showed that expressions of Runx2 and osteocalcin as well as the thickness of collagen type-I fibres were significantly higher in BBM-hAMSC compared to ABG group, while there was no statistically difference in trabecular area and bone incorporation between BBM-hAMSC and ABG group. This study concluded that early healing activities were higher in auto-genous bone graft than in BBM-hAMSC, while osteogenic activities in the late stage of healing were higher in BBM-hAMSC compared to autogenous bone graft. It was also concluded that the osteo-genic capacity of BBM-hAMSC was comparable to autogenous bone graft in the reconstruction of critical size defect in the mandible.展开更多
BACKGROUND Critically sized bone defects represent a significant challenge to orthopaedic surgeons worldwide.These defects generally result from severe trauma or resection of a whole large tumour.Autologous bone graft...BACKGROUND Critically sized bone defects represent a significant challenge to orthopaedic surgeons worldwide.These defects generally result from severe trauma or resection of a whole large tumour.Autologous bone grafts are the current gold standard for the reconstruction of such defects.However,due to increased patient morbidity and the need for a second operative site,other lines of treatment should be introduced.To find alternative unconventional therapies to manage such defects,bone tissue engineering using a combination of suitable bioactive factors,cells,and biocompatible scaffolds offers a promising new approach for bone regeneration.AIM To evaluate the healing capacity of platelet-rich fibrin(PRF)membranes seeded with allogeneic mesenchymal bone marrow-derived stem cells(BMSCs)on critically sized mandibular defects in a rat model.METHODS Sixty-three Sprague Dawley rats were subjected to bilateral bone defects of critical size in the mandibles created by a 5-mm diameter trephine bur.Rats were allocated to three equal groups of 21 rats each.Group I bone defects were irrigated with normal saline and designed as negative controls.Defects of group II were grafted with PRF membranes and served as positive controls,while defects of group III were grafted with PRF membranes seeded with allogeneic BMSCs.Seven rats from each group were killed at 1,2 and 4 wk.The mandibles were dissected and prepared for routine haematoxylin and eosin(HE)staining,Masson's trichrome staining and CD68 immunohistochemical staining.RESULTS Four weeks postoperatively,the percentage area of newly formed bone was significantly higher in group III(0.88±0.02)than in groups I(0.02±0.00)and II(0.60±0.02).The amount of granulation tissue formation was lower in group III(0.12±0.02)than in groups I(0.20±0.02)and II(0.40±0.02).The number of inflammatory cells was lower in group III(0.29±0.03)than in groups I(4.82±0.08)and II(3.09±0.07).CONCLUSION Bone regenerative quality of critically sized mandibular bone defects in rats was better promoted by PRF membranes seeded with BMSCs than with PRF membranes alone.展开更多
AIM To investigate the effects of heme oxygenase-1(HO-1)-modified bone marrow mesenchymal stem cells(BMMSCs)on the microcirculation and energy metabolism of hepatic sinusoids following reduced-size liver transplantati...AIM To investigate the effects of heme oxygenase-1(HO-1)-modified bone marrow mesenchymal stem cells(BMMSCs)on the microcirculation and energy metabolism of hepatic sinusoids following reduced-size liver transplantation(RLT)in a rat model.METHODS BMMSCs were isolated and cultured in vitro using an adherent method,and then transduced with HO-1-bearing recombinant adenovirus to construct HO-1/BMMSCs.A rat acute rejection model following 50%RLT was established using a two-cuff technique.Recipients were divided into three groups based on the treatment received:normal saline(NS),BMMSCs and HO-1/BMMSCs.Liver function was examined at six time points.The levels of endothelin-1(ET-1),endothelial nitric-oxide synthase(e NOS),inducible nitric-oxide synthase(i NOS),nitric oxide(NO),and hyaluronic acid(HA)were detected using an enzyme-linked immunosorbent assay.The portal vein pressure(PVP)was detected by Power Lab ML880.The expressions of ET-1,i NOS,e NOS,and von Willebrand factor(v WF)protein in the transplanted liver were detected using immunohistochemistry and Western blotting.ATPase in the transplanted liver was detected by chemical colorimetry,and the ultrastructural changes were observed under a transmission electron microscope.RESULTS HO-1/BMMSCs could alleviate the pathological changes and rejection activity index of the transplanted liver,and improve the liver function of rats following 50%RLT,with statistically significant differences compared with those of the NS group and BMMSCs group(P<0.05).In term of the microcirculation of hepatic sinusoids:The PVP on POD7 decreased significantly in the HO-1/BMMSCs and BMMSCs groups compared with that of the NS group(P<0.01);HO-1/BMMSCs could inhibit the expressions of ET-1 and i NOS,increase the expressions of e NOS and inhibit amounts of NO production,and maintain the equilibrium of ET-1/NO(P<0.05);and HO-1/BMMSCs increased the expression of v WF in hepatic sinusoidal endothelial cells(SECs),and promoted the degradation of HA,compared with those of the NS group and BMMSCs group(P<0.05).In term of the energy metabolism of the transplanted liver,HO-1/BMMSCs repaired the damaged mitochondria,and improved the activity of mitochondrial aspartate aminotransferase(ASTm)and ATPase,compared with the other two groups(P<0.05).CONCLUSION HO-1/BMMSCs can improve the microcirculation of hepatic sinusoids significantly,and recover the energy metabolism of damaged hepatocytes in rats following RLT,thus protecting the transplanted liver.展开更多
Recently, porous titanium granules （PTGs） have been indicated for the preservation of the dimensions of post-extraction sockets, as a filler in sinus lift procedures and for the treatment of peri-implant and periodo...Recently, porous titanium granules （PTGs） have been indicated for the preservation of the dimensions of post-extraction sockets, as a filler in sinus lift procedures and for the treatment of peri-implant and periodontal defects, based on the osteoconductivity and dimensional stability of the titanium granules. However, there is a lack of information regarding the use of this material in larger defects and in conjunction with membranes. The objective of this study is to test the behavior of PTGs used to fill critical size defects in rabbit tibiae, with and without membranes. Critical defects were created in both tibiae of rabbits, divided randomly into three groups： Group A （defect filled with PTG）, Group B （defect filled with PTG＋collagen membrane） and a control group （empty defect）. After six weeks, histomorphometric analysis was performed. The results showed more defect closures at the cortical area （87.37%±2.2%） and more bone formation at the marrow area （57.6%± 1.3%） in Group B, in comparison with the other groups （P〈0.05）; the use of membranes improved the material stability expressed as more percentages of the original material when membranes were used （P〈0.05）. Finally, inflammatory reactions were observed when the granules were not protected by membranes. In spite of the limitations of this animal study, it may be concluded that PTG particles are osteoconductive and allow bone growth. The PTG particles must be covered by a membrane, especially when grafting larger defects, in order to control particle migration, promote clot stabilization and separate the PTG graft from undesired soft tissue cells.展开更多
In order to formulate feasible conservation measures and develop and utilize Muchuan black-bone chicken rationally,two generations of Muchuan black-bone chicken were bred,and their production performance and slaughter...In order to formulate feasible conservation measures and develop and utilize Muchuan black-bone chicken rationally,two generations of Muchuan black-bone chicken were bred,and their production performance and slaughter traits( at the age of 300 d) were compared. The results showed that the reproductive performance,slaughter performance and body size of the Muchuan black-bone chicken of Generation 1( G_1)were all improved compared with those of the Generation 0( G_0)( P <0. 05). At the age of 300 d,the live weight,carcass weight,and leg muscle weight of the G_1 were significantly higher than those of the G_0( P <0. 05); and the chest width of the cocks of the G_1 was significantly higher than that of the G_0( P <0. 05). It suggests that after rigorous selection and utilization of modern poultry breeding techniques,the reproductive performance,slaughter performance and survival rate of Muchuan black-bone chicken can be improved significantly.展开更多
基金King Saud University,through Vice Deanship of Research Chairs
文摘The aim of the present real time in vivo micro-computed tomography （pCT） and histologic experiment was to assess the efficacy of guided bone regeneration （GBR） around standardized calvarial critical size defects （CSD） using bone marrow-derived mesenchymal stem cells （BMSCs）, and collagen membrane （CM） with and without tricalcium phosphate （TCP） graft material. In the calvaria of nine female Sprague-Dawley rats, full-thickness CSD （diameter 4.6 mm） were created under general anesthesia. Treatment-wise, rats were divided into three groups. In group 1, CSD was covered with a resorbable CM; in group 2, BMSCs were filled in CSD and covered with CM; and in group 3, TCP soaked in BMSCs was placed in CSD and covered with CM. All defects were closed using resorbable sutures. Bone volume and bone mineral density of newly formed bone （NFB） and remaining TCP particles and rate of new bone formation was determined at baseline, 2, 4, 6, and 10 weeks using in vivo pCT. At the lOth week, the rats were killed and calvarial segments were assessed histologically. The results showed that the hardness of NFB was similar to that of the native bone in groups I and 2 as compared to the NFB in group 3. Likewise, values for the modulus of elasticity were also significantly higher in group 3 compared to groups 1 and 2. This suggests that TCP when used in combination with BMSCs and without CM was unable to form bone of significant strength that could possibly provide mechanical ＂lock＂ between the natural bone and NFB. The use of BMSCs as adjuncts to conventional GBR initiated new bone formation as early as 2 weeks of treatment compared to when GBR is attempted without adiunct BMSC therapy.
基金financially supported by the National Natural Science Foundation of China(Nos.81622032,51672184,51525101and 51472279)the Priority Academic Program Development of Jiangsu High Education Institutions(PAPD)+2 种基金the Jiangsu Innovation and Entrepreneurship Program,the National Basic Research Program of China(973 Program,No.2014CB748600)the Jiangsu Provincial Special Program of Medical Science(No.BL2012004)the Jiangsu Six Peak of Talents Program(No.2013-WSW-056)
文摘Natural diatomite （DT） is the ancient deposit of diatom skeleton with many regular pores of 50-200 nm and also an abundant source of biogenic silica. Although silica is considered biologically safe and there is an increasing interest of using natural diatomite for biomedical applications, the toxicity information about natural diatomite is still missing. Here, cytotoxicity of natural diatomite on osteoblasts and fibroblasts were compared to hydroxyapatite and the relationships between cytotoxicity and diatomite sizes, dose, geometry or impurity were systematically investigated. Cell adhesion and interaction with diatomite particles were also fluorescently observed, The results clearly suggested a size-, dose- and shape-dependent cytotoxicity of natural diatomite. Disk-shaped diatomite particles with average size of 30μm in diameter revealed the least toxicity, while the diatomite particles with irregular shapes and sizes less than 10 μm were remarkably toxic. Diatomite particles with proper sizes were then selected to investigate the reinforcing effect on injectable calcium phosphate bone cement. Results showed that diatomite significantly improved the compressive strength of bone cement but did not alter the injectability of the cement, This work provided important biocompatibility information of natural diatomite and demonstrated the feasibility of using selected diatomite as bone implant material.
文摘Experiments on maxillofacial bone tissue engineering showed the promising result;however, its healing mechanisms and effectiveness had not been fully understood. The aim of this study is to compare the bone healing mechanism and osteogenic capacity between bovine bone mineral loaded with hAMSC and autogenous bone graft in the reconstruction of critical size mandibular bone defect. Critical size defects were made at the mandible of 45 New Zealand white rabbits reconstructed with BBM-hAMSC, BBM alone, and ABG, respectively. At the end of first, second, and twelfth weeks, five rabbits from each experimental week were sacrificed for histology and immunohistochemistry staining. Expressions of vascular endothelial growth factor (VEGF), bone mor-phogenic proteins-2 (BMP2), Runx2 and the amount of angiogenesis were analyzed in the first and second week groups, while expressions of Runx2, osteocalcin, collagen type-I fibres, trabecular area and bone incorporation were analyzed in the twelfth week groups. The result showed that expressions of VEGF, BMP2 and Runx2 as well as the amount of angiogenesis were higher in ABG compared with BBM-hAMSC group in the first and second weeks of healing. The result of twelfth week of healing showed that expressions of Runx2 and osteocalcin as well as the thickness of collagen type-I fibres were significantly higher in BBM-hAMSC compared to ABG group, while there was no statistically difference in trabecular area and bone incorporation between BBM-hAMSC and ABG group. This study concluded that early healing activities were higher in auto-genous bone graft than in BBM-hAMSC, while osteogenic activities in the late stage of healing were higher in BBM-hAMSC compared to autogenous bone graft. It was also concluded that the osteo-genic capacity of BBM-hAMSC was comparable to autogenous bone graft in the reconstruction of critical size defect in the mandible.
文摘BACKGROUND Critically sized bone defects represent a significant challenge to orthopaedic surgeons worldwide.These defects generally result from severe trauma or resection of a whole large tumour.Autologous bone grafts are the current gold standard for the reconstruction of such defects.However,due to increased patient morbidity and the need for a second operative site,other lines of treatment should be introduced.To find alternative unconventional therapies to manage such defects,bone tissue engineering using a combination of suitable bioactive factors,cells,and biocompatible scaffolds offers a promising new approach for bone regeneration.AIM To evaluate the healing capacity of platelet-rich fibrin(PRF)membranes seeded with allogeneic mesenchymal bone marrow-derived stem cells(BMSCs)on critically sized mandibular defects in a rat model.METHODS Sixty-three Sprague Dawley rats were subjected to bilateral bone defects of critical size in the mandibles created by a 5-mm diameter trephine bur.Rats were allocated to three equal groups of 21 rats each.Group I bone defects were irrigated with normal saline and designed as negative controls.Defects of group II were grafted with PRF membranes and served as positive controls,while defects of group III were grafted with PRF membranes seeded with allogeneic BMSCs.Seven rats from each group were killed at 1,2 and 4 wk.The mandibles were dissected and prepared for routine haematoxylin and eosin(HE)staining,Masson's trichrome staining and CD68 immunohistochemical staining.RESULTS Four weeks postoperatively,the percentage area of newly formed bone was significantly higher in group III(0.88±0.02)than in groups I(0.02±0.00)and II(0.60±0.02).The amount of granulation tissue formation was lower in group III(0.12±0.02)than in groups I(0.20±0.02)and II(0.40±0.02).The number of inflammatory cells was lower in group III(0.29±0.03)than in groups I(4.82±0.08)and II(3.09±0.07).CONCLUSION Bone regenerative quality of critically sized mandibular bone defects in rats was better promoted by PRF membranes seeded with BMSCs than with PRF membranes alone.
基金Supported by The National Natural Science Foundation of China,No.81670574,No.81441022 and No.81270528The Natural Science Foundation of Tianjin,China,No.08JCYBJC08400,No.11JCZDJC27800,and No.12JCZDJC25200the Technology Foundation of the Health Bureau in Tianjin,China,No.2011KY11
文摘AIM To investigate the effects of heme oxygenase-1(HO-1)-modified bone marrow mesenchymal stem cells(BMMSCs)on the microcirculation and energy metabolism of hepatic sinusoids following reduced-size liver transplantation(RLT)in a rat model.METHODS BMMSCs were isolated and cultured in vitro using an adherent method,and then transduced with HO-1-bearing recombinant adenovirus to construct HO-1/BMMSCs.A rat acute rejection model following 50%RLT was established using a two-cuff technique.Recipients were divided into three groups based on the treatment received:normal saline(NS),BMMSCs and HO-1/BMMSCs.Liver function was examined at six time points.The levels of endothelin-1(ET-1),endothelial nitric-oxide synthase(e NOS),inducible nitric-oxide synthase(i NOS),nitric oxide(NO),and hyaluronic acid(HA)were detected using an enzyme-linked immunosorbent assay.The portal vein pressure(PVP)was detected by Power Lab ML880.The expressions of ET-1,i NOS,e NOS,and von Willebrand factor(v WF)protein in the transplanted liver were detected using immunohistochemistry and Western blotting.ATPase in the transplanted liver was detected by chemical colorimetry,and the ultrastructural changes were observed under a transmission electron microscope.RESULTS HO-1/BMMSCs could alleviate the pathological changes and rejection activity index of the transplanted liver,and improve the liver function of rats following 50%RLT,with statistically significant differences compared with those of the NS group and BMMSCs group(P<0.05).In term of the microcirculation of hepatic sinusoids:The PVP on POD7 decreased significantly in the HO-1/BMMSCs and BMMSCs groups compared with that of the NS group(P<0.01);HO-1/BMMSCs could inhibit the expressions of ET-1 and i NOS,increase the expressions of e NOS and inhibit amounts of NO production,and maintain the equilibrium of ET-1/NO(P<0.05);and HO-1/BMMSCs increased the expression of v WF in hepatic sinusoidal endothelial cells(SECs),and promoted the degradation of HA,compared with those of the NS group and BMMSCs group(P<0.05).In term of the energy metabolism of the transplanted liver,HO-1/BMMSCs repaired the damaged mitochondria,and improved the activity of mitochondrial aspartate aminotransferase(ASTm)and ATPase,compared with the other two groups(P<0.05).CONCLUSION HO-1/BMMSCs can improve the microcirculation of hepatic sinusoids significantly,and recover the energy metabolism of damaged hepatocytes in rats following RLT,thus protecting the transplanted liver.
文摘Recently, porous titanium granules （PTGs） have been indicated for the preservation of the dimensions of post-extraction sockets, as a filler in sinus lift procedures and for the treatment of peri-implant and periodontal defects, based on the osteoconductivity and dimensional stability of the titanium granules. However, there is a lack of information regarding the use of this material in larger defects and in conjunction with membranes. The objective of this study is to test the behavior of PTGs used to fill critical size defects in rabbit tibiae, with and without membranes. Critical defects were created in both tibiae of rabbits, divided randomly into three groups： Group A （defect filled with PTG）, Group B （defect filled with PTG＋collagen membrane） and a control group （empty defect）. After six weeks, histomorphometric analysis was performed. The results showed more defect closures at the cortical area （87.37%±2.2%） and more bone formation at the marrow area （57.6%± 1.3%） in Group B, in comparison with the other groups （P〈0.05）; the use of membranes improved the material stability expressed as more percentages of the original material when membranes were used （P〈0.05）. Finally, inflammatory reactions were observed when the granules were not protected by membranes. In spite of the limitations of this animal study, it may be concluded that PTG particles are osteoconductive and allow bone growth. The PTG particles must be covered by a membrane, especially when grafting larger defects, in order to control particle migration, promote clot stabilization and separate the PTG graft from undesired soft tissue cells.
基金Supported by National Key Technology Research and Development Program(2015BAD03B03)Breeding Program of Sichuan Province during the 13th Five-Year Plan Period(2016NYZ0043)Science and Technology Support Program of Sichuan Province(2016NZ0034)
文摘In order to formulate feasible conservation measures and develop and utilize Muchuan black-bone chicken rationally,two generations of Muchuan black-bone chicken were bred,and their production performance and slaughter traits( at the age of 300 d) were compared. The results showed that the reproductive performance,slaughter performance and body size of the Muchuan black-bone chicken of Generation 1( G_1)were all improved compared with those of the Generation 0( G_0)( P <0. 05). At the age of 300 d,the live weight,carcass weight,and leg muscle weight of the G_1 were significantly higher than those of the G_0( P <0. 05); and the chest width of the cocks of the G_1 was significantly higher than that of the G_0( P <0. 05). It suggests that after rigorous selection and utilization of modern poultry breeding techniques,the reproductive performance,slaughter performance and survival rate of Muchuan black-bone chicken can be improved significantly.
