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Inflammation is the defense response of the living tissues possessing vascular system to stimulations of various injury factors ,which plays a vital role in the initiation and progression of many major diseases .Drugs used to treat inflammation in the clinical mainly include non‐steroidal anti‐inflammatory drugs (NSAIDs) ,steroidal anti‐inflammatory drugs (SAIDs) and traditional Chinese medicine .As synthetic anti‐inflammatory drugs used in clinical currently have obvious adverse reactions , more and more attention were paid to seek anti‐inflammatory drugs from natural medicines .Reviews reported before mainly fo‐cus on anti‐inflammation mechanism of natural medicine ,however ,there are few reports on the summary of anti‐inflammatory natural products .Active natural products which were reported to possess anti‐inflammatory effects in recent years were summa‐rized in order to provide information for further study of anti‐inflammatory drugs research .
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To investigate the pharmacokinetic and bioavailability of polydatin [PD] in rats after oral and intravenous administration, a simple, rapid and sensitive liquid chromatography-tandem mass spectroscopy [LC-MS/MS] method was developed and validated for the determination of polydation. After precipitating the plasma proteins with methanol, the analytes were separated on a C[18] column [3.5 microm, 2.1×100 mm] with an isocratic mobile phase consisting of methanol-acetonitrile-0.1% formic acid [18: 15: 67, v/v/v] at a flow rate of 0.3 mL/min. The Agilent G6410A triple quadrupole LC/MS system was operated under the multiple reaction monitoring [MRM] mode and the electrospray ionization technique was in negative mode. Linear responses were obtained for PD ranging from 1.0-5000.0 ng/mL [r=0.9984] and the LLOQ was 1.0 ng/ml and was sufficient for the pharmacokinetic studies. The intra-day and inter-day accuracy and precision of the assay were less than 8.0%. The method is capable of quantifying PD. The pharmacokinetic parameters of polydatin after intragastric administration of PD with different doses [50, 100 and 300 mg/kg] and intravenous administration at the dose of 20 mg/kg, were obtained, with t[1/2] of 200.30 min, 210.30 min, 272.26 min, and 112.5 min, and AUC[0- infinity] of 125626.41 microg/L.min, 250433.47 microg/L.min, 693722.60 microg/L.min and 1723509.57 microg/L.min, respectively. The absolute bioavailability of PD was somewhat low to 2.9%. The results were firsly reported, as far as we know, about bioavailability of PD and seem important for linking PD and other phenolic glycosides-related drugs administration to their medicinal effects
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Animais de Laboratório , Estilbenos/farmacocinética , Farmacocinética , Disponibilidade Biológica , Ratos Sprague-Dawley , Plasma , Cromatografia Líquida , Espectrometria de Massas em TandemRESUMO
Linalool is an important monoterpene, and widely used in food, pharmaceutical and cosmetic industry. The low concentration in plants and the difficulties in extraction restrict its large scale production. Saccharomyces cerevisiae can provide the monoterpene precursor, geranyl diphosphate (GPP) through its endogenous isoprenoid pathway. Therefore, it could be used as the host for monoterpene production. However, the weak metabolic flux through the isoprenoid pathway leads to the insufficient supply of GPP, and results in low monoterpene productivity. In order to increase the metabolic flux, we constructed the integrated expression plasmid pRS305-tHMG1 and free expression plasmid pYLIS-IDI1 to enhance the expression levels of isopentenyl diphosphate isomerase (IDI1) and a truncated 3-hydroxyl-3-methylglutaryl-CoA reductase gene (tHMG1). The two plasmids were separately transformed into S. cerevisiae CEN.PK2-1C, resulting in strains LS01 and LS02. The plasmid pYLIS-IDI1 was further transformed into strain LS01, resulting in strain LS03. GC-MS analysis showed that the linalool concentration was increased by 1.3 times and reached (127.71 +/- 7.68) microg/L. In conclusion, enhancement of the supply of GPP precursors through the regulation of isoprenoid pathway could increase the linalool production in S. cerevisiae.
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Vias Biossintéticas , Genética , Butadienos , Metabolismo , Hemiterpenos , Metabolismo , Monoterpenos , Metabolismo , Pentanos , Metabolismo , Saccharomyces cerevisiae , Genética , MetabolismoRESUMO
Objective To explore the effect of hepatocyte growth factor on peripheral nerve regeneration. Methods Sciatic nerve contusion injury was made by a custom-made clamp in Wistar rats,in which human hepatocyte growth factor expressed by adenoviral vector(Ad-HGF)was injected into the muscle around the injured nerve.The results of nerve regeneration were evaluated by sciatic nerve function index(SFI),muscle wet weight,neural electrophysiology and image analysis. Results Four weeks after sciatic nerve injury,the results of sciatic nerve function index(SFI),muscle wet weight,neural electrophysiology and image analysis showed better nerve regeneration in group injected with HGF than control group(P<0.05). Conclusion Hepatocyte growth factor can promote axon regeneration and functional recovery and is an effective neurotrophic factor for peripheral nerve regeneration after injury.
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BACKGROUND:Conventjonally,the osteoinduction of bone substitute materials is detected in vivo,which is unsatisfactory regarding the reliability,chronergy and precision,especially for a large amount of substitute materials.OBJECTIVE:To search an in vitro assay for determining in vitro osteogenic activities of bone graft substitutes.DESIGN:Controlled cytological trials.TIME AND SETTING:The experiments were carried out in the Institute of Orthopaedics.Chinese PLA General Hospital(Beijing,China)from August 2006 to May 2007.MATERIALS:C2C 12 cells was offered by the Cell Center of Peking Union Medical College;Human decalcified bone matrix and bone protein were offered by the Institute of Orthopaedics in Chinese PLA General Hospital;Type Ⅰ collagen extracted from bovine tendon was purchased from Beijing Yierkang Bioengineering Development Center;Recombinant human bone morphogenetic protein 2 was purchased from Hangzhou Gene Technology of Huadong Medicine Group.METHODS:By means of dialysis,a composite material was prepared with the bone protein extracted from human,human decalcified bone matrix and type Ⅰ collagen of bovine tendon.The samples of decalcified bone matrix.composite material and recombinant human bone morphogenetic protein 2 were respectively co-incubated with C2C12cells for 72 hours.Negative control group comprised pure cells without materials.Then C2C 1 2 cells were lysed and the lysate were assayed for the absorbance of alkaline phosphatase(ALP)and total protein by chromatometry.ALP is (the specific mark of osteoblastic phenotype.The relative ratio of absorbance value between ALP and total protein could represent ALP activity in the unit quantitative C2C12 cells.MAIN OUTCOME MEASURES:The ratio of absorbance value between ALP and total protein.RESULTS:The ALP activity was the highest in the recombinant human bone morphogenetic protein 2 group,then in the decalcified bone matrix group and composite material group,and the lowest in the negative control group.There were significant differences in the ALP activity between the three trial groups and the negative control group(P<0.05).CONCLUSION:The assay in vitro is effective to detect the ALP activity and it can be used to determine the osteoinduction of bone graft substitutes.
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@#: The management of peripheral nerve injury is a tough problem clinically.Intensive studies in the past were focused on the bridging of nerve defects and the improvement of regeneration rate.But actually the clinical results of functional recovery after peripheral nerve lesion is mainly decided by the accurate regeneration of axons to their original target tissues and structures.Therefore,better clinical results could be obtained by a greater understanding of the cellular and molecular biology of selective nerve regeneration and the application of this theory clinically.This paper summarized recent studies on the cellular and molecular biology mechanisms of peripheral nerve selective regeneration.
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BACKGROUND:Because human cells for culturing alveolar bone cell line are from alveolar bone, which is in oral cavity,and easily polluted, so laboratory study is often unsuccessful. Because the samples are from adults, so cell division index and the successful rate of culture are low.OBJECTIVE: To compare the biological characteristics of survived cell line established through passage,cryopreservation and revitalization following in vitro culturing the alveolar bone tissue obtained from normal persons and patients with chronic periodontitis accompanied with osteoporosis in aseptic operation; To compare the biological characteristics of two kinds of cells so as to provide theoretical and related experimental evidence for defect, repair and treatment of alveolar bone.DESIGN: Controlled observation.SETTING: Department of Stomatology, Beijing Chaoyang Hospital, Capital Medical University; Institute of Orthopaedics,General Hospital of Chinese PLA.MATERIALS: Alveolar bone tissue obtained from normal persons and patients with chronic periodontitis confirmed in clinic was used in aseptic operation.METHODS: Alveolar bone tissue from normal persons and chronic periodontitis accompanied with osteoporosis were cultured in vitro. In the four cell lines (H-171, H-258, 261, 262) cultured primarily, cell lines H-171 and H-258 were chosen from periodonitis patients group and normal group respectively, and stained with histochemical and immunohistochemical methods. Cell morphology was observed. Doubling time and division index of two kinds of cells were calculated with cytometry. After several circles of passage, cryopreservation and revitalization, growth and aging rule of cells were compared.MAIN OUTCOME MEASURES: Passage and biological characteristics of two groups of cell lines.RESULTS: ①In the abnormal alveolar bone group, there was one successful primary culture and cells presented short-spindle shape. There were 3 times of cryopreservation and 3 times of revitalization. Its doubling time was 53.4 hours. The average division index was about 4‰. Cells well grew after 20 times of passages. ②In the normal alveolar bone group, there were 26 cases of cell lines cultured primarily, but passage was found in only 3 cases of cell lines due to various causes. There were 10 passages and the cells presented long-spindle shape. After two circles of cryopreservation and revitalization, the survival and growth rate of cells were inferior as compared with cell line H-171.Doubling time was 65.9 hours and the average division index was 3.5‰. ③Both two kinds of cells adhered the wall, with the characteristics of osteoblasts: AKP, toluidine blue, PAS, tetracycline-labeled mineralized nodus, type Ⅰ collagen and BMP-2 immunohistochemical staining all presented positive.CONCLUSION: Both two kinds of cultured cells have the characteristics of osteoblasts. The growth speed of cell line H-171 is faster than that of cell line H-258. No obvious mutation is found in 20 passages. In the 8th generation of H-258,aging appears and growth speed becomes slow.
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Objective To study the cooperative method of goat's cruciate ligament reconstruction by bone-tendon autograft with interference fixation. Methods A customized reamer was used to make the bottleneck-like femoral tunnel, and the patellar tendon-tibial tuberosity autograft was harvested. Make sufficient preparation for this operation. Results The operation was successful, the laboratory animals all survived without any postoperative infection. Conclusion Scientific and careful operative-cooperation can guarantee the successful operation.
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0.05).There were better effect of removal of myelin(P2
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[Objective]To evaluate the effect of post-operative rehabilitative nursing of patients repaired with acellular nerve allograft.[Method]From April 2003 to April 2006,39 inpatients with peripheral nerve defect were subjected to receive acellular nerve allograft in order to repair nerve defect.The patients were rehabilitated with special nursing after being operated and discharged.Among of them,21 patients were followed up over 6 months,the effect of treatment was analyzed.[Result]Among 21 patients,16 people had excellent and good effect of treatment and the efficient rate was 71.4%.[Conclusion]Post-operative rehabili tative nursing is important and effective for rehabilitation patients of peripheral nerve injuries repaired with acellular nerve allograft.
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OBJECTIVE To observe the effect of electrolyzed oxidizing water(EOW) on the surface sterilization of indoor environment.METHODS Sterilized the surface of ground,wall,table,mob and thermostat-controlled water-bath in cell culture room and good laboratory practice(GLP) with EOW,then to draw the materials and culture with culture dish for 48 h in a 37℃ incubator,and count the colony number.Sterilization method and grouping: group A treated as a control,group B sterilized with EOW,group C sterilized with ultraviolate ray for 30 min and group D first treated with ultraviolate ray for 30 min,then sterilized with EOW.RESULTS In group A,the bacteria were overgrew and formed flakiness in 10/10 culture dishes;1 colony was formed in group B,the sterilization effective rate was 90%.The bacteria culture of group C found no bacteria growing after sterilization with ultraviolate ray,however,sample from surface and culture after sterilization were seen bacteria,though the number of bacteria was less than group A.The bacteria culture outcome of group D was negative.CONCLUSIONS The EOW has a good sterilization effect,it is safe and untoxic,costly cheap and convenient to use,and fit to claim of environmental protection.
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[Objective]To study the effect of()~(60)Co-irridiated and ethylene oxide sterilization on acellular Achilles tendon-bone biomechanical properties.[Method]Twelve fresh Achilles tendon-bone were harvested from New-Zealand white rabbits.Those tendons were soaked in 1%TnBP[tri(n-butyl)phosphate] for 48h,then they were rinsed with deionized water and ethyl alcohol.They were conducted mechanics testing after lyophilization and received()~(60)Co-irridiated or ethylene oxide sterilization.Fresh Achilles tendon-bones were as positive control.[Result]There were no significant difference between fresh and sterilized by ethylene oxide Achilles tendon-bone on biomechanical properties(P
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[Objective]The purpose of this paper is to demonstrate whether the nerve length could affect the quality of acellular nerve and investigate the properly repairable distance of nerve defects with acellular nerve allografts.[Method]Fresh sciatic nerves were obtained from adult dogs and divided into 12 cm long segments.The nerve segments were decellularized via an improved chemical decelluarization treatment as following: Nerve segments were rinsed with cold sterile Ringer's solution and submergedin 5% Triton-100 solution 12h,and then soaked the nerve segments into 5% sodium deoxycholate for 12h.The treated nerve segments were washed in distilled water for 3h.This procedures were repeated once again.In vitro,the degrees of decellularization,demyelination and integrity of nerve fiber tubal of chemically extracted acellular nerves were observed with microscope and assessed by a score system.In vivo,the sciatic nerve of dogs on the right was exposed.In 8 cm grafted group(n=6),a 7 cm segment of sciatic nerve was removed from the midthigh level.In 10 cm grafted group(n=6),a 9 cm segment of sciatic nerve was removed at the same level.The gaps were bridged with acellular nerve allografts by 8 cm and 10 cm long segments respectively.The follow-up period was 12 month postoperatively.Motor functional recovery of the right hind following allografting was examined by neurobehavioral,electrophysiological,histological and immunohistochemical assessment.[Result]There was no difference on the degrees of decellularization,demyelination and integrity of nerve fiber tubal among every fraction of the acellular nerve from the two ends to the central portion.In 8 cm grafted group,all survival dogs(n=5) were held upright with the affected hindlimb extended so that the body's weight was supported by the distal metatarsus and toes.In 10 cm grafted group,animals were failed to held upright with the affected hindlimb.Electrophysiological studies showed that elctromyographic activity was observed in both groups.After 12 month the conduction velocity was 32.1+5.1 m/s in 8 cm grafted animals and 18.3+6.0m/s in 10 cm grafted group.In normal animals, the conduction velocity was 106.6+16.4 m/s.The conduction velocity in 10 cm grafted group was lower than 8 cm grafted(P
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0.05).[Conclusion]The values of three-dimensional parameters in old femoral neck fracture patient are different with that of normal persons in the loading region of the femoral head although there is no difference in BMD values between them.This may provide new evidence to predict osteoporotic femoral neck fracture.
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BACKGROUND: Elimination of antigenic substances from natural extracellular matrix with the integrity of the tissue structure retained renders the matrix to possess better biocompatibility and provides a cell culture environment close to conditions of the internal environment. Such materials are the primary choice for cell culture scaffold in tissue engineering.OBJECTIVE: To prepare human articular cartilage acellular matrix so as to provide a methodological basis for further study of articular cartilage acellular matrix as cell scaffold materials.DESIGN: A single sample study of bone tissues.SETTING: The experiment was performed in Institute of Orthopedics, General Hospital of PLA, between January and May in 2004. The specimens were obtained from patients requiring joint replacement for femoral neck fracture.MATERIAIS: The experiment was conducted in the Department of Orthopedics, General Hospital of PLA from January to May in 2004. Human articular cartilage specimens were obtained from the femoral head of patients with total hip arthroplasty for femoral neck fracture.METHODS: Totally 10 specimens of fresh articular cartilage(3.5 mm × 4. 5 mm × 2.0 mm) were obtained and freeze-dried for 12 hours. Cartilage acellular matrix was prepared using Triton X-100, Dnase and Rnase and identified by means of hematoxylin-eosin(HE) and safranine O staining and immunohistochemical staining for cartilage proteoglycan.MAIN OUTCOME MEASURES: Histological observation of the articular cartilage acellular matrix and immunohistochemical staining of cartilage proteoglycan.RESULTS: HE and safranine O staining both showed no cellular structure in the matrix with only recesses left by the removed cells. Immunohistochemical staining for cartilage proteoglycan yielded positive results, suggesting the presence of cartilage proteoglycan in the acellular matrix.CONCLUSION: Human articular cartilage acellular matrix can be prepared using the modified four-step procedures with detergent and enzymatic extraction with lyophilization, and the preserved cartilage proteoglycan in the material may retain good pressure resistance.
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<p><b>OBJECTIVE</b>To investigate the biological characteristics of cell lines of healthy and diseased human dental alveoli.</p><p><b>METHODS</b>Primary cell lines from either healthy or diseased human dental alveoli were obtained. Two cell lines, H-258 and H-171 derived from healthy and diseased human tissues respectively, were selected for morphological study and research on their growth and aging, using cell counting, and histochemical and immunohistochemical staining.</p><p><b>RESULTS</b>Primary cell lines were successfully established from innormal dental alveoli. After freezing and thawing for three times, cell growth was continued and no morphological alterations were observed. The doubling time was 53.4 hours and mean division index (MDI) was 4 per thousand. Cells were kept normal after twenty generations with no obvious reduction of doubling time and MDI. Of twenty-six primary cell lines derived from healthy human dental alveoli, only three cell lines achieved generation. After freezing and thawing for twice, cultured cells were still alive at a decreased growth speed, with doubling time of 85.9 hours and MDI of 3 per thousand. Both cell lines, H-171 and H-258, shared the characteristics of osteoblast.</p><p><b>CONCLUSIONS</b>Primary cell lines of diseased human dental alveoli show greater growth potential. All cell lines of dental alveoli share characteristics of osteoblast. The technique we developed may be put into practice for the treatment of abnormal dental alveoli.</p>
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Humanos , Divisão Celular , Fisiologia , Linhagem Celular , Alvéolo Dental , Biologia CelularRESUMO
Objective The tissue-engineered composite graft was formed with induced marrow-derived stromal cells (MSCs)and PLGA double-layer scaffold. The effectiveness of this graft for the repair of osteochondral defects in the knee of rabbits was investigated. Methods MSCs were isolated from 20 adult rabbits with density gradient centrifugation and was divided into two groups. In group A, the MSCs were cultivated with regular medium. In group B they were cultivated with chondrogenic differentiation medium. The mRNA of MSCs and articular cartilage cells were extracted, and the expression of mRNA for type Ⅰ and Ⅱ collagen was tested by RT-PCR. The distribution and compound of MSCs with PLGA double-layer scaffold was examined with scanning electron microscopy. 28 adult rabbits were divided into 3 groups, osteochondral defect of 3.5 mm in diameter and 3 to 4 mm in depth were created in the patellar groove. Group A (10 rabbits), the MSCs cultivated with regular medium was grafted into the defects. In group B (10 rabbits), the MSCs cultivated with chondrogenic differentiation medium was grafted into the defects. In group C (8 rabbits), the defects were repaired with autologous osteochondral grafts as control. Specimens were harvested at 4th, 8th, 16th and 24th week post operation respectively, histological examination was performed and graded. Results For the MSCs cultivated with regular medium, the expression of mRNA for type Ⅰ collagen was found with RT-PCR, but no expression for Ⅱ collagen was found. For the induced MSCs, the expression of mRNA both for type Ⅰ and type Ⅱ collagen were found. The adhesion and growth of MSCs on the PLGA double-layer scaffold were well visualized with scanning electron microscopy, and some cells were found in the deep porotic area. For the specimens of group B, no significant difference was found comparing with normal cartilage at 24th week, and the specimens were defined as matured hyaline-like cartilage(4/6)with histological examination, superior to those specimens of group A (1/4). Conclusion The MSCs have osteogenic and chondrogenic potentiality. Combined with PLGA double-layer scaffold, it can be served as seeded cell to form tissue-engineered composite grafts, which can be used to repair osteochondral defects in rabbit models.
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Objective To research the immunologic reaction and the potential of chemically extracted acellular nerve allograft(CEANA) to repair peripheral nerve defects in primates. Methods Adult SD rats were used as nerve donors and adult male Wistar rats used as nerve recipient hosts. 25 mm long nerve segments were excised from SD rats' sciatic nerves. The nerve segments were decellularized via an improved chemical decelluarization treatment as follows: 1) nerve segments were rinsed with cold sterile Ringer's solution; 2)stabilized by pinning the ends to a thin plastic support, and submerged in 4% Triton-100 solution 12 h; 3)soaked into 72 mM sodium deoxycholate for 12 h; 4)washed in distilled water for 6 h. The procedures were repeated once again. Median nerve segments were obtained from macaques and decellularized according to above procedures. The CEANA from SD rats were implanted into Wistar rats subcutanously. The control group was implantation of fresh nerve allografts from SD rats. The immunogenicity of acellular nerve allograft was tested by immunohistochemical examination of the intensity of CD3+, CD4+ and CD8+ cells that infiltrated the allografts. Median nerve defects for 5 cm were created in three macaques. CEANA were interposed across the gap. The CEANA were anastomosed microsurgically to the epineurium of proximal and distal stumps. Results The number of CD3, CD4 and CD8 positive lymphocytes infiltration in CEANA was far lower than that in the control group of fresh nerve allografts at 2 weeks and 4 weeks after implantation. There was no significant evidence of inflammatory in the CEANA grafted group. In the experiment of nerve regeneration of macaques, electromyographic activity was recorded across the allografts. The conduction velocity of regenerated nerve was (40.5?6.8) m/s. Regenerated axons sprouted from the proximal portion reached the distal portion of the grafts, and Schwann cells were also present in the central portion of the CEANA. Motor end-plates were observed in reinnervated muscles. Conclusion The immunogenicity that would have initiated cell-mediated immunological rejection of CEANA are removed. The implantation of CEANA could repair the defect of median nerve 5 cm long in the arm 5 months postoperatively. The CEANA as a type of substitute of nerve autografts has the potential to repair peripheral nerve defects in primates.
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Objective To observe the effect of based biomimetic biphasic calcium phosphate(BCP)tissue engineered bone masses to repair the bone defects and prevent the collapse of femoral head in dogs.Methods Biomimetic BCP ceramic scaffolds was fabricated using three-dimensional(3D)gel-lamination technology according the micro-computed tomography(Micro-CT)information of cancellous bone of dogs' femoral heads.Tissue engineered bone masses were constructed by the combination of biomimetic BCP scaffolds and induced osteogenic bone marrow stromal cells(BMSCs)in vitro.In 10 dogs,tissue engineered bone was implanted into the bone defect on the weight-bearing area of femoral head.In the control group of another 10 dogs,autogenous cancellous bone chips were impressed into the bone defect on the weight-bearing area of femoral head.The gross appearance and regeneration of new bone were observed in 30 weeks postoperatively.Results The fabricated biomimetic BCP scaffolds had better 3D structure.The trabecular system of the scaffolds had some orientation along the weight-bearing line of femoral head and represented plate-like model.BMSCs grow flourishing on the surface of the scaffolds.After 30 weeks operatively,in the tissue engineered bone implanted dogs,the gross appearances of the femoral head were integrity and a great lot of new bone tissue wraps the trabecula of the scaffolds under microscope.In the control group,the mostly femoral heads were collapse gross appearance.Conclusion Tissue engineered biomimetic BCP had better biocompatibility and could prevent the collapse of femoral head after it is implanted into the bone defects of femoral head.
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0.05), whereas the value of new bone grafted with pDBM was significantly lower than that of the normal group (P0.05); but the CUS of the pDBM grafted group was significantly lower than that of normal radius (P0.05). Histological analysis exhibited that most of the DBM was absorbed and substituted by matured new cortical bone in the treated defects of both groups 6 weeks postoperatively, whereas in the untreated group, the defects were only filled with fibrous connective tissue in their mid-portion. Conclusion The sDBM and pDBM are both effective in repairing segmental bone defects. The properties of new bone induced by grafts with sDBM are superior to that of pDBM in biomechanics. These materials can be used in clinical practice as bone graft extenders or enhancers.