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OBJECTIVE@#To study the preparation and properties of the hyaluronic acid (HA)/α-calcium sulfate hemihydrate (α-CSH)/β-tricalcium phosphate (β-TCP) material (hereinafter referred to as composite material).@*METHODS@#Firstly, the α-CSH was prepared from calcium sulfate dihydrate by hydrothermal method, and the β-TCP was prepared by wet reaction of soluble calcium salt and phosphate. Secondly, the α-CSH and β-TCP were mixed in different proportions (10∶0, 9∶1, 8∶2, 7∶3, 5∶5, and 3∶7), and then mixed with HA solutions with concentrations of 0.1%, 0.25%, 0.5%, 1.0%, and 2.0%, respectively, at a liquid-solid ratio of 0.30 and 0.35 respectively to prepare HA/α-CSH/ β-TCP composite material. The α-CSH/β-TCP composite material prepared with α-CSH, β-TCP, and deionized water was used as the control. The composite material was analyzed by scanning electron microscope, X-ray diffraction analysis, initial/final setting time, degradation, compressive strength, dispersion, injectability, and cytotoxicity.@*RESULTS@#The HA/α-CSH/β-TCP composite material was prepared successfully. The composite material has rough surface, densely packed irregular block particles and strip particles, and microporous structures, with the pore size mainly between 5 and 15 μm. When the content of β-TCP increased, the initial/final setting time of composite material increased, the degradation rate decreased, and the compressive strength showed a trend of first increasing and then weakening; there were significant differences between the composite materials with different α-CSH/β-TCP proportion ( P<0.05). Adding HA improved the injectable property of the composite material, and it showed an increasing trend with the increase of concentration ( P<0.05), but it has no obvious effect on the setting time of composite material ( P>0.05). The cytotoxicity level of HA/α-CSH/β-TCP composite material ranged from 0 to 1, without cytotoxicity.@*CONCLUSION@#The HA/α-CSH/β-TCP composite materials have good biocompatibility. Theoretically, it can meet the clinical needs of bone defect repairing, and may be a new artificial bone material with potential clinical application prospect.
Subject(s)
Calcium Phosphates , Bone and Bones , PhosphatesABSTRACT
Bone defects caused by trauma, infection, tumor and other factors is a thorny problem in orthopedic clinic, and promoting bone repair and regeneration is the key and difficult point of treatment. In addition to autologous bone grafting, artificial bone materials are often used for large bone defects. β-tricalcium phosphate has good biocompatibility, bone conduction and bone induction properties, and has been studied deeply because of its excellent drug delivery performance and has shown broad application prospects. In this paper, the author will summarize the research progress of β-tricalcium phosphate composites loaded with different drugs in the treatment of bone defects caused by trauma, infection and tumor.
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Objective: This study aimed to examine differences in platelet-rich plasma (PRP) soak-loaded volumes of β-tricalcium phosphate (β-TCP) with or without a unidirectional porous structure.Materials and Methods: Leukocyte-rich PRP was extracted from 15 healthy volunteers by centrifugation. Two types of artificial bones were soaked for either ten seconds or ten minutes. The volume ratios of PRP soak-loaded onto the artificial bone and soaked area ratios were evaluated. Statistical analyses were performed using the Tukey-Kramer HSD test and the Games-Howell method. A P-value of <0.05 was considered statistically significant.Results: Regardless of the soaking time, the PRP soak-loaded volume ratio and soaked area ratio were significantly higher in the unidirectional porous β-TCP (UDPTCP) group than in the spherical porous β-TCP (SPTCP) group.Conclusion: PRP can be soak-loaded faster and in larger amounts onto UDPTCP compared to SPTCP. Understanding the basic biology of β-TCP soak-loaded with PRP can help develop more novel and effective β-TCP treatments for orthopedic surgery.
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Introduction: Unidirectional porous β-tricalcium phosphate (UDPTCP) consists of a novel porous artificial bone that is structurally different from conventional artificial bone comprised of spherical porous β-tricalcium phosphate (SPTCP).Case presentation: We present our first four clinical cases of opening-wedge high tibial osteotomy (OWHTO) using UDPTCP and SPTCP together. The patients’ mean age was 54.5 ± 5.9 years, and the mean observation period was 20.8 ± 2.8 months. In OWHTO, two wedge shaped pieces of UDPTCP and SPTCP were cut to fit the gap and implanted parallel to each other in the anterior and posterior parts, respectively. We evaluated the correction loss and bone remodeling for UDPTCP and SPTCP over time using radiography and computed tomography, and evaluated the clinical outcomes.Conclusion: There was no correction loss reported in any case, and early bone remodeling was observed with UDPTCP. All patients achieved satisfactory clinical results with no adverse events.
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BACKGROUND: Chitosan exhibits good physiochemical properties and biocompatibility, but it has poor biological activity of osseointegration. Therefore, it needs to combine with other materials for bone repair. OBJECTIVE: Calcined bone/chitosan composite was prepared and its physiochemical properties and biocompatibility were analyzed. METHODS: Calcined bone/chitosan composite was prepared at a mass ratio of 1:2, 1:1, 2:1 respectively by solution blending method. The physicochemical properties of three composite materials were characterized. Passage 5 mouse L929 fibroblasts were treated with the leaching solution of three composite materials. The cytotoxicity of three composite materials was detected by the CCK-8 test. RESULTS AND CONCLUSION: (1) X-ray diffraction and Fourier Transform Infrared Spectroscopy showed that the essential components of three composite materials were hydroxyapatite and β-tricalcium phosphate. The characteristic diffraction peaks of hydroxyapatite/β-tricalcium phosphate increased with the increase of the proportion of calcined bone. (2) Scanning electron microscopy showed that calcined bone particles were evenly dispersed in chitosan medium. (3) With the increase of the proportion of calcined bone, the compressive strength of the composite decreased gradually. (4) After 7 days of culture, the cells in the leaching solution of three composite materials grew well without obvious change in morphology. By the ninth day, the relative proliferation rate of the cells in the leaching solution of three composite materials was over 90%. Cytotoxicity was grade 1, which meets the safety standard of biomaterials. (5) These results suggest that the calcined bone/chitosan composite has good structural characteristics, physicochemical properties and suitable compressive strength and is safe and non-toxic.
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Objective: To treat vertebral fractures with posterior wall injury in the elderly, vertebral bone grafting is generally performed through a posterior transpedicular approach, combined with pedicle screw fixation. An autologous bone is ideal to treat this disorder. However, harvesting autologous bones from the elderly with osteoporosis is limited by the amount and quality of available autologous bone. Thus, we developed a bone-grafting substitute. The newly developed unidirectional porous β-tricalcium phosphate, with a porosity of 57% (UDPTCP; Affinos®, Kuraray Co., Ltd., Tokyo, Japan), is used in the bone-grafting procedure. This is the first report of UDPTCP used as an artificial bone graft in patients with an acute vertebral burst fracture.Materials and Methods: UDPTCP (mean: 4.2 g) was implanted through the pedicle, and posterior instrumentation was achieved with pedicle screws in five elderly patients. Resorption of UDPTCP and substitution with the autologous bone were evaluated on computed tomography (CT) and plain X-ray performed immediately and at 3, 6, and 12 months after the operation.Results: In case 1, the pedicle screws did not loosen, and UDPTCP was completely resorbed and replaced with the autologous bone at 3 postoperative months. In the other four cases, although the pedicle screws or the caudal part loosened because of osteoporosis, resorption of UDPTCP was observed at 3 postoperative months. At 6 postoperative months, progressive substitution with the autologous bone was confirmed, and at 12 postoperative months, formation of the good autologous bone was confirmed.Conclusion: This preliminary case series demonstrated that the newly developed UDPTCP shows good clinical potential as a bone-graft substitute for acute vertebral burst fractures in the elderly, including patients with osteoporosis.
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Objective: To explore the osteogenesis effect of advanced-platelet-rich fibrin (A-PRF) and β-tricalcium phosphate (β-TCP) composite. Methods: Thirty-two healthy female New Zealand rabbits were randomly selected. A-PRF was prepared by collecting blood from middle auricular artery. Rabbits were randomly divided into 6 groups: groups A, B, C, D, and E (6 rabbits in each group) and group F (2 rabbits). Bone defects (6 mm in diameter, 8 mm in depth) were drilled into femur condyle of each rabbit's both back legs. Then A-PRF and β-TCP composites manufactured by different proportion were planted into bone defects of group A (1∶1), group B (2∶1), group C (4∶1), group D (1∶2) and group E (1∶4) ( V/ V). The bone defect was not repaired in group F. The specimens were collected at 8 and at 12 weeks after operation. Then gross observation, X-ray examination, Micro-CT examination, and biomechanical test were performed. The bone volume/total volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular spacing (Tb.Sp), compressive strength, and modulus of elasticity were calculated. Results: The gross observation and X-ray examination showed that the osteogenesis effect at 12 weeks was better than that at 8 weeks. At the same time point, the repair of bone defect and the formation of new bone in group B were better than those in other groups. Micro-CT examination showed that the trabeculae of new bone in group B were the most and the trabeculae arranged closely at 8 and 12 weeks. Besides there were significant differences in BV/TV, Tb.N, and Tb.Sp between group B and the other groups ( P<0.05). There were significant differences in Tb.N and Tb.Th in group B, BV/TV and Tb.Sp in group C, Tb.Sp in group D between 8 weeks and 12 weeks ( P<0.05). Biomechanical tests showed that the compression strength and elastic modulus of group B were the highest, and the compression strength and elastic modulus of group C were the lowest at 8 and at 12 weeks, showing significant differences ( P<0.05). There were significant differences in compression strength and elastic modulus of each group between 8 weeks and 12 weeks ( P<0.05). Conclusion: The A-PRF and β-TCP composite can repair femoral condylar defects in rabbits, and the osteogenesis is better in proportion of 2∶1.
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Objective To study the mechanical properties and biological characteristics of 3D-printed porous β-tricalcium phosphate ［β-Ca3(PO4)2, β-TCP］ scaffolds, so as to provide guidance for the design of composite scaffolds in animal experimentation. Methods Poly 1,8-octanediol citrate (POC), a kind of novel biodegradable materials, was used as the adhesive. The 3D-printed porous β-TCP scaffolds were fabricated by fused deposition modeling (FDM) technology, and Gly-Arg-Gly-Asp-Ser (GRGDS), a kind of polypeptides, was added into the scaffolds to improve the adhesive property of cells. The optical microscope and scanning electron microscope (SEM) were used to observe the micro-pore architectures of those scaffolds. The material testing machine was used to conduct compressive test on the scaffolds, and the water contact angles of the scaffolds were measured. The cell adhesion rate and proliferation rate of the scaffolds were also tested by in vitro cell experiment. The model of SD rat skull defects was repaired by the scaffolds, and the osteogenic ability in vivo was further studied. Results The GRGDS, remaining active, was evenly distributed in the composite scaffolds. The micro-pore architectures of the polypeptide modified scaffolds changed, with improvement in cell adhesion rate, while the compressive modulus, water contact angle and osteogenic ability in vivo of the scaffolds were not obviously affected. Conclusions The cell adhesion capacity of β-TCP composite scaffolds modified by polypeptide improved significantly, while the mechanical properties and hydrophilicity, osteogenic ability in vivo of the scaffolds were not affected very much. These research results provide new ideas for reconstruction of scaffolds for repairing bone defects in clinic, and a laboratory basis for further clinical application of this scaffold.
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Objective To investigate the effects of human leptin ( hLEP) gene transfection on rat bone marrow stro-mal cells ( rBMSCs) .Methods rBMSCs were cultured and transfected with adenoviruses encoding hLEP ( Ad5-hLEP-EGFP) in vitro as experimental group while rBMSCs transfected with Ad 5-EGFP and non-transfected were control groups.The proliferation was detected by MTT and the expression of collagen type Ⅰ(Col-Ⅰ) and alkaline phosphatase ( ALP) were assessed by real-time PCR.The ability of mineralized nodule forming was also examined by Alizarin red staining .The combination of transfected rBMSCs and β-tricalcium phosphate (β-TCP ) was con-structed and the osteogenic ability of the construction was evaluated in nude mice .Results hLEP could be trans-fected into rBMSCs successfully by adenovirous .After transfection , the proliferation was not affected while Col-Ⅰand ALP expressions were more pronounced in rBMSCs transfected with Ad 5-hLEP-EGFP ( P<0.05 ) .Alizarin red staining showed the ability of mineralized nodule forming was also up-regulated in Ad5-hLEP-EGFP group (P<0.05).In addition, the transfected rBMSCs adhered to β-TCP and survived well and the combination showed more new bone like tissue formation in nude mice compared to control groups .Conclusions rBMSCs transfected with hLEP might be potently used in bone or periodontal tissue regeneration .
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We investigated the use of Polycaprolactone (PCL)/β-tricalcium phosphate (β-TCP) composites with applied mechanical stimulation as scaffold for bone tissue engineering. PCL-based three-dimensional (3D) structures were fabricated in a solvent-free process using a 3D-printing technique. The mass fraction of β-TCP was varied in the range 0–30%, and the structure and compressive modulus of the specimens was characterized. The shape and interconnectivity of the pores was found to be satisfactory, and the compressive modulus of the specimens was comparable with that of human trabecular bone. Human mesenchymal stem cells were seeded on the composites, and various biological evaluations were performed over 9 days. With a mass fraction of β-TCP of 30%, differentiation began earlier; however, the cell proliferation rate was lower. Through the use of mechanical stimulation, however, the proliferation rate recovered, and was comparable with that of the other groups. This stimulation effect was also observed in ECM generation and other biological assays. With mechanical stimulation, expression of osteogenic markers was lower on samples with a β-TCP content of 10 wt% than without β-TCP; however, with mechanical stimulation, the sample with a β-TCP content of 30 wt% exhibited significantly greater expression of those markers than the other samples. We found that mechanical stimulation and the addition of β-TCP interacted closely, and that a mass fraction of β-TCP of 30% was particularly useful as a bone tissue scaffold when accompanied by mechanical stimulation.
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Humans , Biological Assay , Bone and Bones , Cell Proliferation , Mesenchymal Stem CellsABSTRACT
PURPOSE: The purpose of this study was to analyze patient factors including smoking, body mass index, correction angle, graft material, presence of lateral cortex fracture, and age for the effect on bone union after open-wedge high tibial osteotomy and the effect of graft material used for lateral cortex fractures. MATERIALS AND METHODS: This retrospective study was conducted on 54 patients and 58 cases with osteoarthritic change Kallgren-Lawrence grade 2 or less from May 2012 to June 2014. Average follow-up period was 22 months (14–38 months). The patients were divided into two groups according to patient related factors and graft materials (allograft, n=6; beta-tricalcium phosphate [β-TCP], n=6) used for lateral cortex fractures and were analyzed for the relationship with bone union after open-wedge high tibial osteotomy. Radiographic and clinic analyses were performed, and van Hemert grading was used for grading bone union at 6 weeks, 3 months, 6 months, and 1 year postoperatively. RESULTS: The non-smoking group and the group without lateral cortex fracture showed significantly higher bone union rates than the control group. No significant clinical or radiological difference was observed between the two groups in 12 cases and the allograft group showed significantly higher rates of union at 6 months and 1 year postoperatively according to the van Hemert grading. CONCLUSION: Smoking and the presence of a lateral cortex fracture is a risk factor for nonunion in medial open-wedge high tibial osteotomy. The use of allograft material rather than β-TCP for lateral cortex fractures is thought to result in better bone union.
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Humans , Allografts , Body Mass Index , Follow-Up Studies , Knee , Osteoarthritis , Osteotomy , Retrospective Studies , Risk Factors , Smoke , Smoking , TransplantsABSTRACT
Secreted protein, acidic, cysteine-rich (SPARC)-related modular calcium binding 1 (SMOC1) has been implicated in the regulation of osteogenic differentiation of human bone marrow mesenchymal stem cells (BMSCs). In this study, we found that a peptide (16 amino acids in length), which is located in the extracellular calcium (EC) binding domain of SMOC1, stimulated osteogenic differentiation of human BMSCs in vitro and calvarial bone regeneration in vivo. Treatment of BMSCs with SMOC1-EC peptide significantly stimulated their mineralization in a dose-dependent manner without changing their rate of proliferation. The expression of osteogenic differentiation marker genes, including type 1 collagen and osteocalcin, also increased in a dose-dependent manner. To examine the effect of the SMOC1-EC peptide on bone formation in vivo, the peptide was covalently immobilized onto hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) particles. X-ray photoelectron spectroscopy analysis showed that the peptide was successfully immobilized onto the surface of HA/β-TCP. Implantation of the SMOC1-EC peptide-immobilized HA/β-TCP particles into mouse calvarial defects and subsequent analyses using microcomputed tomography and histology showed significant bone regeneration compared with that of calvarial defects implanted with unmodified HA/β-TCP particles. Collectively, our data suggest that a peptide derived from the EC domain of SMOC1 induces osteogenic differentiation of human BMSCs in vitro and efficiently enhances bone regeneration in vivo.
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Animals , Humans , Mice , Amino Acids , Bone Marrow , Bone Regeneration , Calcium , Ceramics , Collagen Type I , In Vitro Techniques , Mesenchymal Stem Cells , Miners , Osteocalcin , Osteogenesis , Photoelectron Spectroscopy , Regeneration , X-Ray MicrotomographyABSTRACT
Objective To explore the changes of PH values of N,O-CMC/β-TCP compositive materials with different mass fractions in simulated body fluids (SBF)and their influence in the growth of MG63 cells, and to illustrate their mechanisms, and to provide reference for the further research on the bone repair materials. Methods The N,O-CMC/β-TCP with mass fractions of 2/1,1/1 and 1/2 were used as experimental groups,and the collagen nano calcium phosphate bone repair material as control group. The materials with different mass fractions were immersed in SBF and the pH values were measured by pH meter after soaking for 7,14,21 and 28d,respectively.The MG63 cells with the concentration of 1 × 105 mL-1 were inoculated and co-cultured in experimental and control groups,the adhesion and morphological changes of MG63 cells in each group were observed by scanning electron microscope and the cell proliferation was detected by MTT method after co-culturing for 2,4 and 6 d.Results The pH values were 6.70-7.25 in N,O-CMC/β-TCP (1/2)group and N,O-CMC/β-TCP (2/1)groups and the pH value in N,O-CMC/β-TCP (1/1)group was basically 7.15. The cells in N,O-CMC/β-TCP (2/1)group formed owe full,spreading face small and less secretion,but the cells in N,O-CMC/β-TCP 1/2 and 1/1 groups formed in full, pseudopodia interconnection, widely spreading and more secretions under electron microscope. The proliferation rate of the cells in N,O-CMC/β-TCP with (1/1 ) and N,O-CMC/β-TCP (2/1)groups had no statistical difference compared with control group (P>0.05),but there was significant difference between control group and N,O-CMC/β-TCP (1/2)group (P<0.05).Conclusion The changes of pH values of N,O-CMC/β-TCP materials with different mass fractions in SBF are small and the pH values are neutral;the order of the mass fraction of N,O-CMC/β-TCP to promote the growth of MG63 cells is 1/1,2/1,and 1/2.
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BACKGROUND:As a bone scaffold material,β-tricalcium phosphate has good biocompatibility, osteoinductive, and biomechanical properties. OBJECTIVE:To study the effect of al ogeneic osteoblasts compounded withβ-tricalcium phosphate in repairing rabbit radial defects. METHODS:A total of 45 rabbit radial defect models were made and divided into three groups in random. Experimental group was repaired with the compound of al ogeneic osteoblasts andβ-tricalcium phosphate; control group withβ-tricalcium phosphate;and blank control group with nothing. The new bone formation of each group was observed and assessed by X-ray and histopathological analysis at weeks 4, 8, 16 after implantation for evaluation of the bone repairing effect. RESULTS AND CONCLUSION:With the repair time, the experimental group appeared to complete bone defect repair gradual y. By the end of 16 weeks, the X-ray showed that the bone cal us between the scaffold and the host was completely ossified, and bone defects were completely repaired in the experimental group. Histopathological observation showed continuous cortical bone formed in the defect area, and canal recanalization realized in the experimental group. Additional y, the repair effect in the experimental group was better than that in the control and blank control group at different time points (P<0.01). It is suggested that the al ogenic osteoblasts/β-tricalcium phosphate compound has the better effects on guiding bone regeneration and preventing from nonunion.