Microwave assisted synthesis & properties of nano HA-TCP biphasic calcium phosphate

Biphasic calcium phosphate （BCP） nanopowders were synthesized by using microwave and non-microwave irradiation assisted processes. The synthesized powders were pressed under a pressure of 90 MPa, and then were sintered at 1000-1200℃ for 1 h. The mechanical properties of the samples were investigated. The formed phases and microstructures were characterized by X-ray diffraction （XRD） and scanning electron microscopy （SEM）. The results showed that the synthesis time was shorter, along with a more homogeneous microstructure, when the microwave irradiation assisted method was applied. The compression strength and the Young＇s modulus of the samples synthesized with microwave irradiation were about 60 MPa and 3 GPa, but those of the samples synthesized without microwave irradiation were about 30 MPa and 2 GPa, respectively. XRD patterns of the microwave irradiation assisted and non-microwave irradiation assisted nanopowders showed the coexistence of hydroxyapatite （HA） and lricalcium phosphate （TCP） phases in the system.

Biodegradation of Tricalcium Phosphate Ceramics by Osteoclasts

Biodegradation of tricalcium phosphate (TCP) ceramics was observed through mixed culture of osteoclasts and TCP discs in vitro in this study. Osteo-clasts were isolated from newborn SD rat's marrow of long bone and cultured on TCP discs. The culture terminated at the 48th h and 96th h respectively. Under an inverted microscope, the osteoclasts imparted round or oval body with multinu-clear and many thin processes. These cells were positively stained for tartrate-re-sistance acid phosphatase (TRAP). Scanning electron microscope showed that many resorption lacunae on TCP disc surface and their diameters were smaller than 20 μm. Osteoclasts were located in the lacunae. At the 96th h, the resorption lacunae become larger and osteoclasts showed degeneration. It is suggested that osteoclasts possess ability to re-absorb TCP ceramics under in vitro culturing condition.

Characterization of tricalcium phosphate solubilization by Stenotrophomonas maltophilia YC isolated from phosphate mines

The phosphate solubilizing characteristics of a strain YC, which was isolated from phosphate mines (Hubei, China), were studied in National Botanical Research Institute’s phosphate (NBRIP) growth medium containing tricalcium phosphate (TCP) as sole phosphorus (P) source. The strain YC is identified as Stenotrophomonas maltophilia (S. maltophilia) based upon the results of morphologic, physiological and biochemical characteristics and 16S rRNA sequences analysis. The results show that the strain S. maltophilia YC can solubilize TCP and release soluble P in NBRIP growth medium. A positive correlation between concentration of soluble P and population of the isolate and a negative correlation between concentration of soluble P and pH in the culture medium are observed from statistical analysis results. Moreover, gluconic acid is detected in the culture medium by HPLC analysis. It indicates that the isolate can release gluconic acid during the solubilizing experiment, which causes acidification of the culture medium and then TCP solubilization. S. maltophilia YC has a maximal TCP solubilizing capability when using maltose as carbon source and ammonium nitrate as nitrogen source, respectively, in NBRIP growth medium.

Osteogenesis process of tricalcium phosphate ceramics in vivo

To investigate the osteogenesis of calcium phosphate ceramics, β-TCP ceramics were implanted into the condyle femur of rabbits, and tetracycline was injected termly. Specimens were host at 1, 2, 3, 4, 5, 6 months after implanted. The new bone formation and osteogenesis process were observed by the histomorphology, fluorescent microscope, SEM and EPMA. The results demonstrate that, osteogenesis is active, there are abundant osteoblasts on the surface of osteoid, mesenchymal cell hyperplasia and incursion is found in materials after 1 month. After 2 months, there is blood vessel formation and macrophage soakage within materials. Bone-island appears and connects by bone-bridge after 3 months. β-TCP ceramics degrade and are dispersed by new formation bone. Woven bone turns into bone lamella by rebuilding and calcification. The materials entirely change their original shape and combines with bone tissue as a whole after 6 months. The typical structure of spongy bone forms. It is confirmed that β-TCP is a degradable biocompatible artificial bone material which can incorporating in life.

Hydroxyapatite/β-tricalcium Phosphate Composite for Guiding Bone Tissue Growth into a Titanium Tube in 8 mm Dog Tibia Cavity Defects

We developed a fixation method and evaluate bone regrowth in the cavities of a Ф4 mm× 8 mm titanium（Ti）tube through porous hydroxyapatite（HAP）/β-tricalcium phosphate（β-TCP）composite filling（group A）,chitosan/calcium phosphate composite filling（group B）,and HAP particle modification（group C）.After 2 and 5 months of implantation in dog tibia defects,new bone formation in the three groups was studied by histology and histomorphometry.Group A displayed the most bone regenerated area in both 2 and 5 months post-operation.The chitosan/calcium phosphate composite in group B mostly degraded 2 months after implantation,leading to fibrous tissue invasion after 5 months.By contrast,less bone formation was observed in group C.These results indicated that filling the cavities of metalprostheses with a porous HAP/β-TCP composite can be used for stable long-term fixation in clinicalsettings.

Osteoinduction and Osteoconduction with Porous Beta-Tricalcium Phosphate Implanted after Fibular Resection in Humans

Osteoinductive properties of β-TCP remain unknown in humans. It is important to improve the bone grafts which have been the standard treatment for bone defect due to their biocompatibility and bone-healing properties. The purpose of this study was to radiologically clarify the bone forming property of β-TCP by evaluating the replacement of β-TCP by newly formed bone in the defect after fibular resection and to examine the histological features of a β-TCP specimen three months after grafting. Radiographs of 17 patients who underwent β-TCP grafting were evaluated. Osteoinductive and osteoconductive properties were assessed by examining bone formation from the remnant fibula, periosteum, and β-TCP alone. In one case, β-TCP was removed later because of postoperative complications and was evaluated histologically. Twenty two of 34 sites between the remnant fibula and β-TCP had achieved good bone regeneration. Five of 14 sites between the periosteum and β-TCP had achieved good bone regeneration. We found immature but evident bone formation in three cases with no osseous and periosteal sites. Histological analysis revealed bone formation on the outer macropore surface of β-TCP. Some blood vessels formed in the macropores expressed CD31 and CD34, while a few lymphatic vessels expressed CD34 and podoplanin. Thus, the osteoinductive ability of β-TCP alone was demonstrated in humans radiographically for the first time. The histological morphology of β-TCP was demonstrated at an early stage after grafting in humans.

The Role of Microstructure of Highly Purified Beta-Tricalcium Phosphate for Osteoinduction in Canine Dorsal Muscles

Porous β-tricalcium phosphate (TCP) displays osteoinductivity in certain animals in the absence of osteoinductive agents. We evaluated whether the microstructure may be an important determinant of osteoinduction, and also investigated how bone formation was promoted using β-TCP combined with bone marrow aspirates. We prepared two types of β-TCP, namely, β-TCP A, which possessed interconnected macropores and micropores, and β-TCP B, which possessed macropores but had less detectable micropores. These were implanted with or without marrow in canine muscles. Bone formation and the resorption of each β-TCP implant were evaluated histologically. Newly formed bone began to appear at day 42 in the implants of β-TCP A alone, but the implants of β-TCP B alone did not show any bone formation by day 42. Meanwhile, bone formation was already evident on day 14 by loading with bone marrow aspirates with or without micropores. By immunohistochemistry, the number of cathepsin K-positive cells (osteoclasts) increased as time passed in the implants of β-TCP A alone, while the number of the osteoclasts did not change obviously in the implants of β-TCP B alone from day 14 to 56. Reticular fibrils were evident within the β-TCP A, and were barely observed in the β-TCP B in the silver impregnation. The present result would bring about the possible role to enhance the importance of the surface microstructure for the better osteoinductivity. Our findings suggest that the combination of porous β-TCP and bone marrow facilitates bone formation.

Osteogenesis and Degradation Behavior of rhBMP-2／β-Tricalcium Phosphate Porous Composite Materials

Ultrafine-tricalcium phosphate(β-TCP)powders with good crystalline structure were produced by a new process through bone tissue engineering approach rorous β-TCPcermic was combined with recombined human bone morphogenetic proteins-2(rhBMP-2)to develop a novel composite material ,osteogenesis capacity of the composite was investigated intramuscularly in rat with histological analyses and SEM examination pureβ-TCP porous carmic wsa investigated as the control results show that the compostie materials possess good bilcompatibility biodegradation and strong osteogenesis capacity through inductive process after implantation material degradation began from 2 weeks post-implantation accompanying with the changing o pore structure with the enwrapping and separation fo materials by hyperplatic mesenchymal cells and fibroblast and with the phagocytose reaction of multinucleated giant cells early in 72h immature cartilage could be found within novel composite mature lamellar bone was induced to generate after 3 weeks with strong osteoinduction capacity and controlable bildegradation the novel rhBMP-2\β-TCP porous ceramic is expected to be a promising bone grafting substitute for bone tissue engineering

The Bacterial Inhibitory Ability and In Vivo Drug Release Pattern of a New Drug Delivery System:Ciprofloxacine/tricalcium Phosphate Delivery Capsule

The bacterial inhibitory ability of a new drug delivery system (DDS):Ciprofloxacine/tricalcium phosphate delivery capsule (CTDC), its in vivo drug release pattern, and the influence of ultrasonic irradiation on its drug release were investigated. It was found that CTDC had a strong and sustained inhibitory ability to some common pathogens of bone and joint infections, such as staphylococcus aureus, escherichia coli and pseudomonas aeruginosa. In vivo drug-release study in animals demonstrated a high concentration of ciprofloxacine in the bone tissue surrounding CTDC which was placed in the greater trochanter of the rabbit and continued to release ciprofloxacine for at least 5 weeks and the blood level of ciprofloxacine was low. In vivo study also showed ultrasonic irradiation could increase the amount of ciprofloxacine released from CTDC, which may be an economical, effecient and safe new method to achieve the control of drug release from DDS.

The Experimental Study on Mixed Culture of Osteoblasts and Tricalcium Phosphate Ceramics In Vitro

To study the effects of tricalcium phosphate (TCP) ceramics on osteoblasts, the rat osteoblasts were cultured with the TCP ceramics in vitro . Scanning electron microscopy and the colorimetric methyl thiazol tetrazolium assay showed that the osteoblasts could adhere well to the surface of the ceramics and the culture dish, and the proliferation of the cells was not inhibited. The results demonstrated that TCP ceramics possessed an excellent cytocompatibility with the osteoblasts, and had some promoting effects on proliferation of osteoblasts.

Expression of TGF-β in Region of Bone Defect Repaired by Collagen/Nano-beta-Tricalcium Phosphate Composite Artificial Bone

The distribution and function of transforming growth factor-beta (TGF-β) in the region of bone defect repaired by collagen/nano-beta-tricalcium phosphate composite artificial bone (Co/N-TCP) and the ability of Co/N-TCP recruiting osteoblasts to precipitate the repair of bone defect were investigated Twenty-four domestic rabbits were operated on bilateral cranial bone to create an experimental bone defect of 8 0 mm in diameter through the whole bone On the left, Co/N-TCP was implanted as experimental group, but on the right, Co/TCP was implanted as control group At 2nd, 4th, 8th, 12th week after operation, all animals were sacrificed and the implanted materials with surrounding bone were taken out Immunohistochemical staining was performed for TGF-β assay by avidin-biotin complex method (SABC) Simultaneously, TGF-β was quantitatively analyzed by HPIAS-1000 imaging analysis system The inmmunohistochemical staining for TGF-β revealed that osteoblasts and immature osteocytes highly expressed TGF-β Diffused TGF-β positive staining particles appeared in the mesenchymal and fibrous-tissue There was no significant difference in the TGF-β positive staining between two groups in the medial region to original osseous beds at different time points ( P >0 05) However, in distal original osseous bed of the defected region, the positive expression of TGF-β in the Co/N-TCP group was significantly stronger than in the control group ( P <0 05 or 0 01) The Co/N-TCP has good bioactivities and ability of stimulating and conducting TGF-β to aggregate and precipitate the healing of bone defect

Effect on Hydration and Hardening of Tricalcium Phosphate Bone Cement

The bioactive α-Ca3（ PO4 ）2 bone cement was studied by XRD , SEM and isothermal calorimetric measurements. The results showed that a mixed pattern of TCP and hydroxylapatite were obtained after hydration and hardening. The mechanism of hydration and hardening of the α-Ca3 （ PO4）2 was dissolution-precipitation, （ NH4 ） 112 PO4 was the best set accelerator to the α-Ca3 （ PO4）2 cement, and the HAP powers and the （ NH4）H2 PO4 concentration had a great effect on the hydration rate of a-Ca3（ PO4）2.

Preparation and properties of β-tricalcium phosphate porous bioceramic

Porous β-tricalcium phosphate bioceramic （PTCP） has important roles in surgical implants because of good biocompatibility. But the low compressive strength of the ceramic limits its application. The preparation of PTCP was improved with the adjustment of the constituents and the sintering-process. A new type of PTCP material with high compressive strength was made. The compositions, microstructure and properties of PTCP were analyzed by TG-DSC, XRD, TEM, SEM and so on. The result indicates that stearic acid burns sufficiently and gives out carbon dioxide and water vapor when slowly heated between 200 ℃ and 400 ℃ so that the porous structure like coral in β-TCP bioceramic is formed. Through crystallization at 470 ℃ and 570 ℃, more CaO-P2O5 glass-cement is converted into crystallite-glass, which is beneficial for improving the compressive strength of β-TCP bioceramic. PTCP can form a support action in bone imperfect section with good solubility.

PLLA/β-TCP支架表面多孔结构的构建及其对细胞黏附的影响

目的通过NaOH溶液的简单处理构建3D打印PLLA/β-TCP骨组织工程支架表面多孔结构,增加支架的粗糙度和亲水性,促进支架表面的细胞黏附。方法通过3D打印熔融沉积成型技术制备PLLA/β-TCP网状支架,并通过NaOH蚀刻的方法进行支架的粗糙化改性,依据支架表面微观形貌、能谱、接触角、力学、细胞黏附等观察NaOH浓度、时间两项反应参数对支架的影响。结果通过熔融沉积成型技术制备的PLLA/β-TCP复合支架存在预先设置的网状结构;经NaOH蚀刻构建了兼有宏观和微观空隙的多孔形态。NaOH浓度、时间中任意一种参数的增加都会导致支架表面微观孔隙的孔径、孔密度增加。NaOH处理参数为0.1 mol/L(9 h)时,可显著减小支架表面水接触角,且对支架的压缩强度无显著影响。体外细胞检测显示,经NaOH蚀刻后的表面多孔复合支架在骨髓间充质干细胞(BMSCs)的黏附增殖上更具优势。结论用NaOH处理3D打印PLLA/β-TCP骨组织工程支架可有效改善支架表面形态,优化该类支架的亲水性及细胞黏附。

Experimental Study of Porous Beta-tricalcium Phosphate and Bone Morphogenetic Protein/porous Beta-tricalcium Phosphate Complex in Rabbit Mandibular Reconstruction

We investigated the osteogenic characteristic and biodegradation behavior of porous beta-tricalcitum phosphate （ β- TCP ） and bone morphogenetic protein/beta-tricalcium phosphate （ BMP/ β- TCP ） complex in mandibular reconstruction and evaluated the advantages of BMP / β- TCP complex in repairing bone defects. Bone defects created in the lower margin of bilateral mandible bodies in 12 rabbits were repaired with β- TCP （ control group ） and BMP/ β- TCP complex （ experimental group ） respectively. The rabbits were euthanized after 2, 4, 8 and 12 weeks and examined by macroscopy, radiography, histology, histomorphometry and image analysis. 2 weeks after surgery, ossftcation of newly-generated tissue in BMP/ β- TCP complex group appeared and after 12 weeks, massive new bone and ossification maturation were seen. However, in β- TCP group without BMP , ossification was not found until 12 weeks after operation. Image analysis showed that bone regeneration rate of BMP/ β- TCP was 30%-40% higher and the degradation rate was 20%- 30% higher than that of β- TCP. Therefore, as a reconstructive material for bone defects, BMP/ β-TCP complex is superior to β-TCP and can be used in oral and maxillofacial surgery.

Local Application of Alendronate on <i>β</i>-Tricalcium Phosphate Accelerated Induction of Osteogenesis with Formation of Giant Osteoclast-Like Cell

Intrinsic osteoinductivity—the ability to induce bone formation in ectopic sites without addition of osteogenic factors has been reported in various porous materials. Tartrate-resistant acid phosphatase (TRAP)-positive osteoclast-like cells are thought to play an important role in material-induced osteoinduction. To investigate the influence of osteoclastic activity on intrinsic osteoinduction, we loaded alendronate (10–2 , 10–4 , and 10–6 M) onto porous β-tricalcium phosphate (β-TCP) blocks to inhibit osteoclastic activity, and evaluated osteoinductivity by implantation of the blocks into the dorsal muscles of adult beagle dogs. Alendronate-loaded porous β-TCP blocks increased both speed and amount of osteoinduction, as measured 4 weeks after implantation, with the 10–4 M alendronate-loaded β-TCP being especially active. This finding indicates that β-TCP loaded with 10–4 M alendronate might prove crucial in providing the desirable balance between the degradation rate of bone scaffolds and their osteoinductive replacement. Thus, material-induced osteoinduction may be controlled by local application of alendronate, establishing alendronate loading as a promising therapeutic approach.

Raman Spectroscopy Analysis of Wollastonite/Tricalcium Phosphate Glass-Ceramics after Implantation in Critical Bone Defect in Rats

In this work wollastonite/tricalcium phosphate (W/TCP) glass-ceramics with three W/TCP weight ratios (20/80;60/40 and 80/20) were implanted in rat calvaria and the modifications taking place during implantation were studied by Raman spectroscopy. The experimental glass-ceramics were composed of different contents of βW, αW, βTCP, αTCP, and glassy phases. Materials were implanted for 7-, 15-, 45- and 120-day periods after which the implanted materials were recovered and analyzed by FT-Raman spectroscopy. The results suggested that the αW phase reabsorbs fast during implantation in the glass-ceramics 60/40 and 80/20, whereas βTCP and αTCP glass-ceramic are gradually attenuated and replaced by biological apatite-like bands. In the glass-ceramic 20/80, the bands related to the βTCP phase remained unvaried in all analyzed periods. New bands associated with the deposition of collagenous material appeared during implantation for all 60/40 and 80/20 glass-ceramics experimental groups, but important differences in intensities between both groups. The spectra corresponding to implants of 60/40 glass-ceramic at the 120-day period were very similar to those of the control group (normal cortical bone), with regards to Raman shifts and intensities, as well as in the FWHM value of the 962 cm-1 apatite band (ν1 PO4 in hydroxyapatite), evidencing that apatite deposited at the implant site has the same crystallinity than biological apatite in normal bone mineral. The glass-ceramic 20/80 behaved just as an osteoconductive filling material, while glass-ceramics 60/40 and 80/20 were able to induce deposition of organic matrix mineralized new tissue. The 60/40 glass-ceramic showed the best performance and the most similar Raman spectrum to normal cortical bone.