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
The healing of critical-sized bone defects(CSD)remains a challenge in orthopedic medicine.In recent years,scaffolds with sophisticated microstructures fabricated by the emerging three-dimensional(3D)printing technolog...The healing of critical-sized bone defects(CSD)remains a challenge in orthopedic medicine.In recent years,scaffolds with sophisticated microstructures fabricated by the emerging three-dimensional(3D)printing technology have lighted up the treatment of the CSD due to the elaborate microenvironments and support they may build.Here,we established a magnesium oxide-reinforced 3D-printed biocompos-ite scaffold to investigate the effect of magnesium-enriched 3D microenvironment on CSD repairing.The composite was prepared using a biodegradable polymer matrix,polycaprolactone(PCL),and the disper-sion phase,magnesium oxide(MgO).With the appropriate surface treatment by saline coupling agent,the MgO dispersed homogeneously in the polymer matrix,leading to enhanced mechanical performance and steady release of magnesium ion(Mg^(2+))for superior cytocompatibility,higher cell viability,advanced osteogenic differentiation,and cell mineralization capabilities in comparison with the pure PCL.The in-vivo femoral implantation and critical-sized cranial bone defect studies demonstrated the importance of the 3D magnesium microenvironment,as a scaffold that released appropriate Mg^(2+) exhibited remarkably increased bone volume,enhanced angiogenesis,and almost recovered CSD after 8-week implantation.Overall,this study suggests that the magnesium-enriched 3D scaffold is a potential candidate for the treatment of CSD in a cell-free therapeutic approach.展开更多
Introduction Mandibular segmental defects result in significant cosmetic and functional deficiencies.Meanwhile,the reconstruction of both the contour and function of the mandible is a challenging task.At present,autol...Introduction Mandibular segmental defects result in significant cosmetic and functional deficiencies.Meanwhile,the reconstruction of both the contour and function of the mandible is a challenging task.At present,autologous vascularized fibula transplantation is the most common method to reconstruct a mandible with long-span defects[1].展开更多
Objective To study feasibility and value of repair of oral mandibular defects with rib composite flap pedicled with internal thoracic vessels in basic level hospitals. Methods The clinical materials in 13 cases uith m...Objective To study feasibility and value of repair of oral mandibular defects with rib composite flap pedicled with internal thoracic vessels in basic level hospitals. Methods The clinical materials in 13 cases uith mandibular defects which were repaired with rib compos-展开更多
Mandibular defect occurs more frequently in recent years,and clinical repair operations via bone transplantation are difficult to be further improved due to some intrinsic flaws.Tissue engineering,which is a hot resea...Mandibular defect occurs more frequently in recent years,and clinical repair operations via bone transplantation are difficult to be further improved due to some intrinsic flaws.Tissue engineering,which is a hot research field of biomedical engineering,provides a new direction for mandibular defect repair.As the basis and key part of tissue engineering,scaffolds have been widely and deeply studied in regards to the basic theory,as well as the principle of biomaterial,structure,design,and fabrication method.However,little research is targeted at tissue regeneration for clinic repair operations.Since mandibular bone has a special structure,rather than uniform and regular structure in existing studies,a methodology based on tissue engineering is proposed for mandibular defect repair in this paper.Key steps regarding scaffold digital design,such as external shape design and internal microstructure design directly based on triangular meshes are discussed in detail.By analyzing the theoretical model and the measured data from the test parts fabricated by rapid prototyping,the feasibility and effectiveness of the proposed methodology are properly verified.More works about mechanical and biological improvements need to be done to promote its clinical application in future.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
Additive manufacturing has received attention for the fabrication of medical implants that have customized and complicated structures.Biodegradable Zn metals are revolutionary materials for orthopedic implants.In this...Additive manufacturing has received attention for the fabrication of medical implants that have customized and complicated structures.Biodegradable Zn metals are revolutionary materials for orthopedic implants.In this study,pure Zn porous scaffolds with diamond structures were fabricated using customized laser powder bed fusion(L-PBF)technology.First,the mechanical properties,corrosion behavior,and biocompatibility of the pure Zn porous scaffolds were characterized in vitro.The scaffolds were then implanted into the rabbit femur critical-size bone defect model for 24 weeks.The results showed that the pure Zn porous scaffolds had compressive strength and rigidity comparable to those of cancellous bone,as well as relatively suitable degradation rates for bone regeneration.A benign host response was observed using hematoxylin and eosin(HE)staining of the heart,liver,spleen,lungs,and kidneys.Moreover,the pure Zn porous scaffold showed good biocompatibility and osteogenic promotion ability in vivo.This study showed that pure Zn porous scaffolds with customized structures fabricated using L-PBF represent a promising biodegradable solution for treating large bone defects.展开更多
Ameloblastoma is the most frequent odontogenic tumor of the jaw. If not treated, ameloblastoma can gain an enormous size and cause severe facial disfigurement and functional impairment. Here we report two patients aff...Ameloblastoma is the most frequent odontogenic tumor of the jaw. If not treated, ameloblastoma can gain an enormous size and cause severe facial disfigurement and functional impairment. Here we report two patients afflicted with extensive mandibular ameloblastoma (sized in 23 cm × 18 cm × 17 cm and 15 cm × 12 cm × 10 cm, respectively). Both the patients received the same sequential treatment including radical tumor resection, simultaneous reconstruction with fibula free flap graft, vertical distraction osteogenesis on the fibula graft, placement of endosseous dental implants, and final pros-thodontic rehabilitation. It took about 15 months to finish the entire course of treatment. And after the four-year follow-up, neither soft tissue related, nor hard tissue related problems were observed. Satisfactory facial symmetry, chewing and speech functions of the patients were restored. So this sequential treatment for extensive mandibular ameloblastoma can obtain an excellent effect by the shortest time and the lowest economical cost. Furthermore, the series also can be used to reconstruct giant mandibular defects caused by different reasons.展开更多
A novel biodegradable metal system,ZnLiCa ternary alloys,were systematically investigated both in vitro and in vivo.The ultimate tensile strength(UTS)of Zn0.8Li0.1Ca alloy reached 567.60±9.56 MPa,which is compara...A novel biodegradable metal system,ZnLiCa ternary alloys,were systematically investigated both in vitro and in vivo.The ultimate tensile strength(UTS)of Zn0.8Li0.1Ca alloy reached 567.60±9.56 MPa,which is comparable to pure Ti,one of the most common material used in orthopedics.The elongation of Zn0.8Li0.1Ca is 27.82±18.35%,which is the highest among the ZnLiCa alloys.The in vitro degradation rate of Zn0.8Li0.1Ca alloy in simulated body fluid(SBF)showed significant acceleration than that of pure Zn.CCK-8 tests and hemocompatibility tests manifested that ZnLiCa alloys exhibit good biocompatibility.Real-time PCR showed that Zn0.8Li0.1Ca alloy successfully stimulated the expressions of osteogenesis-related genes(ALP,COL-1,OCN and Runx-2),especially the OCN.An in vivo implantation was conducted in the radius of New Zealand rabbits for 24 weeks,aiming to treat the bone defects.The Micro-CT and histological evaluations proved that the regeneration of bone defect was faster within the Zn0.8Li0.1Ca alloy scaffold than the pure Ti scaffold.Zn0.8Li0.1Ca alloy showed great potential to be applied in orthopedics,especially in the load-bearing sites.展开更多
Background:Three-dimensional(3D)printed tissue engineered bone was used to repair the bone tissue defects in the oral and maxillofacial(OMF)region of experimental dogs.Material and methods:Canine bone marrow stromal c...Background:Three-dimensional(3D)printed tissue engineered bone was used to repair the bone tissue defects in the oral and maxillofacial(OMF)region of experimental dogs.Material and methods:Canine bone marrow stromal cells(BMSCs)were obtained from 9 male Beagle dogs and in vitro cultured for osteogenic differentiation.The OMF region was scanned for 3D printed surgical guide plate and mold by ProJet1200 high-precision printer using implant materials followed sintering at 1250℃.The tissue engineered bones was co-cultured with BASCs for 2 or 8 d.The cell scaffold composite was placed in the defects and fixed in 9 dogs in 3 groups.Postoperative CT and/or micro-CT scans were performed to observe the osteogenesis and material degradation.Results:BMSCs were cultured with osteogenic differentiation in the second generation(P2).The nanoporous hydroxyapatite implant was made using the 3D printing mold with the white porous structure and the hard texture.BMSCs with osteogenic induction were densely covered with the surface of the material after co-culture and ECM was secreted to form calcium-like crystal nodules.The effect of the tissue engineered bone on the in vivo osteogenesis ability was no significant difference between 2 d and 8 d of the compositing time.Conclusions:The tissue-engineered bone was constructed by 3D printing mold and hightemperature sintering to produce nanoporous hydroxyapatite scaffolds,which repair in situ bone defects in experimental dogs.The time of compositing for tissue engineered bone was reduced from 8 d to 2 d without the in vivo effect.展开更多
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The authors would like to thank Li LI and H.Z.Xie for the technical support.This work was financially supported by the National Natural Science Foundation of China(Nos.82002303 and 81702171)the Guangdong Basic and Applied Basic Research Foundation(Nos.2022A1515011536,2021A1515220093,2021A1515220086,2019A1515111156,and 2022A1515011815)+7 种基金the Scientific Research Foundation of Peking University Shenzhen hospital(No.KYQD2021064)the Health and Medical Research Fund(No.19180712)the Shenzhen Double Chain Project for Innovation and Development Industry supported by the Bureau of Industry and Information Technology of Shenzhen(No.201806081018272960)the Shenzhen Science and Technology Innovation Committee Projects(Nos.JCYJ20190809182213535 and JSGG20180507183242702)the program from Shanghai Municipal Health Commission(No.201740165)the National Key R&D Program of China(No.2018YFC1105100)the Hong Kong Innovation Technology Fund(Nos.ITS/287/17 and ITS/405/18)the Hong Kong Research Grant Council General Research Fund(No.17214516).
文摘The healing of critical-sized bone defects(CSD)remains a challenge in orthopedic medicine.In recent years,scaffolds with sophisticated microstructures fabricated by the emerging three-dimensional(3D)printing technology have lighted up the treatment of the CSD due to the elaborate microenvironments and support they may build.Here,we established a magnesium oxide-reinforced 3D-printed biocompos-ite scaffold to investigate the effect of magnesium-enriched 3D microenvironment on CSD repairing.The composite was prepared using a biodegradable polymer matrix,polycaprolactone(PCL),and the disper-sion phase,magnesium oxide(MgO).With the appropriate surface treatment by saline coupling agent,the MgO dispersed homogeneously in the polymer matrix,leading to enhanced mechanical performance and steady release of magnesium ion(Mg^(2+))for superior cytocompatibility,higher cell viability,advanced osteogenic differentiation,and cell mineralization capabilities in comparison with the pure PCL.The in-vivo femoral implantation and critical-sized cranial bone defect studies demonstrated the importance of the 3D magnesium microenvironment,as a scaffold that released appropriate Mg^(2+) exhibited remarkably increased bone volume,enhanced angiogenesis,and almost recovered CSD after 8-week implantation.Overall,this study suggests that the magnesium-enriched 3D scaffold is a potential candidate for the treatment of CSD in a cell-free therapeutic approach.
基金supported in part by grants from the Clinical Research for Crossing Item of Shandong University(No.2020SDUCRCB001)the Shandong Provincial Natural Science Foundation(No.ZR2020MH180)+1 种基金the Rongxiang Regeneration Medicine Fund of Shandong University(No.2019SDRX-11)the Science and Technology Development Plan of Jinan City(No.201805041)。
文摘Introduction Mandibular segmental defects result in significant cosmetic and functional deficiencies.Meanwhile,the reconstruction of both the contour and function of the mandible is a challenging task.At present,autologous vascularized fibula transplantation is the most common method to reconstruct a mandible with long-span defects[1].
文摘Objective To study feasibility and value of repair of oral mandibular defects with rib composite flap pedicled with internal thoracic vessels in basic level hospitals. Methods The clinical materials in 13 cases uith mandibular defects which were repaired with rib compos-
基金Project supported by the National Natural Science Foundation of China (No. 50905164)the Zhejiang Provincial Natural Science Foundation of China (No. Y2090835)
文摘Mandibular defect occurs more frequently in recent years,and clinical repair operations via bone transplantation are difficult to be further improved due to some intrinsic flaws.Tissue engineering,which is a hot research field of biomedical engineering,provides a new direction for mandibular defect repair.As the basis and key part of tissue engineering,scaffolds have been widely and deeply studied in regards to the basic theory,as well as the principle of biomaterial,structure,design,and fabrication method.However,little research is targeted at tissue regeneration for clinic repair operations.Since mandibular bone has a special structure,rather than uniform and regular structure in existing studies,a methodology based on tissue engineering is proposed for mandibular defect repair in this paper.Key steps regarding scaffold digital design,such as external shape design and internal microstructure design directly based on triangular meshes are discussed in detail.By analyzing the theoretical model and the measured data from the test parts fabricated by rapid prototyping,the feasibility and effectiveness of the proposed methodology are properly verified.More works about mechanical and biological improvements need to be done to promote its clinical application in future.
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.
基金supported by the National Key R&D Program of China[grant number 2018YFE0104200]the National Natural Science Foundation of China[grant numbers 51901003,51931001,52171233,51875310]+1 种基金the Beijing Natural Science Foundation[grant number L212014]the Open Project of NMPA Key Laboratory for Dental Materials[grant number PKUSS20200401].
文摘Additive manufacturing has received attention for the fabrication of medical implants that have customized and complicated structures.Biodegradable Zn metals are revolutionary materials for orthopedic implants.In this study,pure Zn porous scaffolds with diamond structures were fabricated using customized laser powder bed fusion(L-PBF)technology.First,the mechanical properties,corrosion behavior,and biocompatibility of the pure Zn porous scaffolds were characterized in vitro.The scaffolds were then implanted into the rabbit femur critical-size bone defect model for 24 weeks.The results showed that the pure Zn porous scaffolds had compressive strength and rigidity comparable to those of cancellous bone,as well as relatively suitable degradation rates for bone regeneration.A benign host response was observed using hematoxylin and eosin(HE)staining of the heart,liver,spleen,lungs,and kidneys.Moreover,the pure Zn porous scaffold showed good biocompatibility and osteogenic promotion ability in vivo.This study showed that pure Zn porous scaffolds with customized structures fabricated using L-PBF represent a promising biodegradable solution for treating large bone defects.
文摘Ameloblastoma is the most frequent odontogenic tumor of the jaw. If not treated, ameloblastoma can gain an enormous size and cause severe facial disfigurement and functional impairment. Here we report two patients afflicted with extensive mandibular ameloblastoma (sized in 23 cm × 18 cm × 17 cm and 15 cm × 12 cm × 10 cm, respectively). Both the patients received the same sequential treatment including radical tumor resection, simultaneous reconstruction with fibula free flap graft, vertical distraction osteogenesis on the fibula graft, placement of endosseous dental implants, and final pros-thodontic rehabilitation. It took about 15 months to finish the entire course of treatment. And after the four-year follow-up, neither soft tissue related, nor hard tissue related problems were observed. Satisfactory facial symmetry, chewing and speech functions of the patients were restored. So this sequential treatment for extensive mandibular ameloblastoma can obtain an excellent effect by the shortest time and the lowest economical cost. Furthermore, the series also can be used to reconstruct giant mandibular defects caused by different reasons.
基金National Natural Science Foundation of China(Grant No.51931001)International Cooperation and Exchange project between NSFC(China)and CNR(Italy)(NSFC-CNR Grant No.52011530392).
文摘A novel biodegradable metal system,ZnLiCa ternary alloys,were systematically investigated both in vitro and in vivo.The ultimate tensile strength(UTS)of Zn0.8Li0.1Ca alloy reached 567.60±9.56 MPa,which is comparable to pure Ti,one of the most common material used in orthopedics.The elongation of Zn0.8Li0.1Ca is 27.82±18.35%,which is the highest among the ZnLiCa alloys.The in vitro degradation rate of Zn0.8Li0.1Ca alloy in simulated body fluid(SBF)showed significant acceleration than that of pure Zn.CCK-8 tests and hemocompatibility tests manifested that ZnLiCa alloys exhibit good biocompatibility.Real-time PCR showed that Zn0.8Li0.1Ca alloy successfully stimulated the expressions of osteogenesis-related genes(ALP,COL-1,OCN and Runx-2),especially the OCN.An in vivo implantation was conducted in the radius of New Zealand rabbits for 24 weeks,aiming to treat the bone defects.The Micro-CT and histological evaluations proved that the regeneration of bone defect was faster within the Zn0.8Li0.1Ca alloy scaffold than the pure Ti scaffold.Zn0.8Li0.1Ca alloy showed great potential to be applied in orthopedics,especially in the load-bearing sites.
基金The study was approved by the Animal Experimental Enthical Review From Of Southwest Medical University.Informed consent was obtained。
文摘Background:Three-dimensional(3D)printed tissue engineered bone was used to repair the bone tissue defects in the oral and maxillofacial(OMF)region of experimental dogs.Material and methods:Canine bone marrow stromal cells(BMSCs)were obtained from 9 male Beagle dogs and in vitro cultured for osteogenic differentiation.The OMF region was scanned for 3D printed surgical guide plate and mold by ProJet1200 high-precision printer using implant materials followed sintering at 1250℃.The tissue engineered bones was co-cultured with BASCs for 2 or 8 d.The cell scaffold composite was placed in the defects and fixed in 9 dogs in 3 groups.Postoperative CT and/or micro-CT scans were performed to observe the osteogenesis and material degradation.Results:BMSCs were cultured with osteogenic differentiation in the second generation(P2).The nanoporous hydroxyapatite implant was made using the 3D printing mold with the white porous structure and the hard texture.BMSCs with osteogenic induction were densely covered with the surface of the material after co-culture and ECM was secreted to form calcium-like crystal nodules.The effect of the tissue engineered bone on the in vivo osteogenesis ability was no significant difference between 2 d and 8 d of the compositing time.Conclusions:The tissue-engineered bone was constructed by 3D printing mold and hightemperature sintering to produce nanoporous hydroxyapatite scaffolds,which repair in situ bone defects in experimental dogs.The time of compositing for tissue engineered bone was reduced from 8 d to 2 d without the in vivo effect.