Osteoinduction of hydroxyapatite/β-tricalcium phosphate bioceramics in mice with a fractured fibula

Many studies have shown that calcium phosphate ceramics can induce bone formation in non-osseous sites without the application of any osteoinductive biomolecules, but the mechanisms of this phenomenon (intrinsic osteoinduction of bioceramics) remain unclear. In this study, we compared the intrinsic osteoinduction of porous hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) implanted in mice at different sites. In 30 mice the left fibula was fractured and the right fibula was kept intact. A porous HA/β-TCP cylinder was implanted into both the left (group 1) and right (group 2) leg muscles of each animal. In addition, two HA/β-TCP cylinders were bilaterally implanted into leg subcutaneous pockets (group 3) in each of the remaining 15 mice. New bone formation was studied in the three groups by histology, histomorphometry and immunostaining. In group 1 new bone was observed at week 6 and bone marrow appeared at week 12. In group 2 new bone was observed at week 8 and bone marrow appeared at week 12. The new bone area percentage in group 1 was significantly higher than in group 2 at both weeks 8 and 12. In contrast, group 3 did not show any new bone within the period studied. These differences were explained based on the location of the implants and thus their proximity to the osteogenic environment of fracture healing. The results support the hypothesis that intrinsic osteoinduction by calcium phosphate ceramics is the result of adsorption of osteoinductive substances on the surface.     

3D microenvironment as essential element for osteoinduction by biomaterials

In order to unravel the mechanism of osteoinduction by biomaterials, in this study we investigated the influence of the specific surface area on osteoinductive properties of two types of calcium phosphate ceramics. Different surface areas of the ceramics were obtained by varying their sintering temperatures. Hydroxyapatite (HA) ceramic was sintered at 1150 and 1250 degrees C. Biphasic calcium phosphate (BCP) ceramic, consisting of HA and beta-tricalcium phosphate (beta-TCP), was sintered at 1100, 1150 and 1200 degrees C. Changes in sintering temperature did not influence the chemistry of the ceramics; HA remained pure after sintering at different temperatures and the weight ratio of HA and beta-TCP in the BCP was independent of the temperature as well. Similarly, macroporosity of the ceramics was unaffected by the changes of the sintering temperature. However, microporosity (pore diameter <10 microm) significantly decreased with increasing sintering temperature. In addition to the decrease of the microporosity, the crystal size increased with increasing sintering temperature. These two effects resulted in a significant decrease of the specific surface area of the ceramics with increasing sintering temperatures. Samples of HA1150, HA1250, BCP1100, BCP1150 and BCP1200 were implanted in the back muscles of Dutch milk goats and harvested at 6 and 12 weeks post implantation. After explantation, histomorphometrical analysis was performed on all implants. All implanted materials except HA1250 induced bone. However, large variations in the amounts of induced bone were observed between different materials and between individual animals. Histomorphometrical results showed that the presence of micropores within macropore walls is necessary to make a material osteoinductive. We postulate that introduction of microporosity within macropores, and consequent increase of the specific surface area, affects the interface dynamics of the ceramic in such a way that relevant cells are triggered to differentiate into the osteogenic lineage.     

A comparison of bone formation in biphasic calcium phosphate (BCP) and hydroxyapatite (HA) implanted in muscle and bone of dogs at different time periods

Physicochemical modification could implement synthetic materials into osteoinductive materials, which induce bone formation in nonosseous tissues. We hereby studied the relevance between the osteogenic capacities of osteoinductive materials in nonosseous tissues and in osseous sites. Biphasic calcium phosphate ceramic (BCP) and hydroxyapatite ceramic (HA) were implanted in femoral muscles and femoral cortical bone of dogs for 7, 14, 21, 30, 45, 60, 90, 180, and 360 days, respectively. Two dogs were used in each time point. In each dog, four cylinders (phi5x6 mm) per material were implanted in femoral muscles and 2 cylinders (phi5x6 mm) per material in femoral cortical bone. The harvested samples were processed for both histological and histomorphometric analyses. Bone was observed in BCP implanted in femoral muscles since day 30, while in HA since day 45. Quantitatively, more bone was formed in BCP than in HA at each time point after day 30 (p<0.05). The earlier and more bone formed in BCP than in HA suggests BCP a higher osteoinductive potential than HA in muscle. In femoral cortical bone defects, a bridge of bone in the defect with BCP was observed at day 21, while with HA at day 30. At days 14, 21, and 30, significantly more bone was formed in BCP than in HA (p<0.05). The results herein show that osteogenic capacities of osteoinductive materials in nonosseous tissues and osseous sites are correlated: the higher the osteoinductive potential of the material, the faster the bone repair.     

Viable osteogenic cells are obligatory for tissue-engineered ectopic bone formation in goats

In this study we investigated the bone-forming capacity of tissue-engineered (TE) constructs implanted ectopically in goats. As cell survival is questionable in large animal models, we investigated the significance of vitality, and thus whether living cells instead of only the potentially osteoinductive extracellular matrix are required to achieve bone formation. Vital TE constructs of porous hydroxyapatite (HA) covered with differentiated bone marrow stromal cells (BMSCs) within an extracellular matrix (ECM) were compared with identical constructs that were devitalized before implantation. The devitalized implants did contain the potentially osteoinductive ECM. Furthermore, we evaluated HA impregnated with fresh bone marrow and HA only. Two different types of HA granules with a volume of approximately 40 microm were investigated: HA70/800, a microporous HA with 70% interconnected macroporosity and an average pore size of 800 microm, and HA60/400, a smooth HA with 60% interconnected macropores and an average size of 400 microm. Two granules of each type were combined and then treated as a single unit for cell seeding, implantation, and histology. The tissue-engineered samples were obtained by seeding culture-expanded goat BMSCs on the HA and subsequently culturing these constructs for 6 days to allow cell differentiation and ECM formation. To devitalize, TE constructs were frozen in liquid nitrogen according to a validated protocol. Fresh bone marrow impregnation was performed perioperatively (4 mL per implant unit). All study groups were implanted in bilateral paraspinal muscles. Fluorochromes were administered at three time points to monitor bone mineralization. After 12 weeks the units were explanted and analyzed by histology of nondecalcified sections. Bone formation was present in all vital tissue-engineered implants. None of the other groups showed any bone formation. Histomorphometry indicated that microporous HA70/800 yielded more bone than did HA60/400. Within the newly formed bone, the fluorescent labels showed that mineralization had occurred before 5 weeks of implantation and was directed from the HA surface toward the center of the pores. In conclusion, tissue-engineered bone formation in goats can be achieved only with viable constructs of an appropriate scaffold and sufficient BMSCs.     

Material-dependent bone induction by calcium phosphate ceramics: a 2.5-year study in dog

Bone induction by different calcium phosphate biomaterials has been reported previously. With regard to (1) whether the induced bone would disappear with time due to the absence of mechanical stresses and (2) whether this heterotopically formed bone would give rise to uncontrolled growth, a long-time investigation of porous hydroxyapatite ceramic (HA), porous biphasic calcium phosphate ceramic (TCP/HA, BCP), porous alpha-tricalcium phosphate ceramic (alpha-TCP) and porous beta-tricalcium phosphate ceramic (beta-TCP) was performed in dorsal muscles of dog, for 2.5 years. Histological observation, backscattered scanning electron microscopy observation and histomorphometric analysis were made on thin un-decalcified sections of retrieved samples. Normal compact bone with bone marrow was found in all HA implants (n = 4) and in all BCP implants (n = 4), 48 +/- 4% pore area was filled with bone in HA implants and 41 +/- 2% in BCP implants. Bone-like tissue, which was a mineralised bone matrix with osteocytes but lacked osteoblasts and bone marrow, was found in all beta-TCP implants (n = 4) and in one of the four alpha-TCP implants. Both normal bone and bone-like tissues were confined inside the pores of the implants. The results show that calcium phosphate ceramics are osteoinductive in muscles of dogs. Although the quality and quantity varied among different ceramics, the induced bone in both HA and BCP ceramics did neither disappear nor grow uncontrollably during the period as long as 2.5 years.     

Implanted octacalcium phosphate is more resorbable than beta-tricalcium phosphate and hydroxyapatite.

Our previous studies have suggested that synthetic octacalcium phosphate (OCP) could be resorbed and replaced by newly formed bone if implanted in rat skull defects. We hypothesized that the implanted OCP is more resorbable than other commonly used bone graft substitutes of calcium phosphate compounds, such as hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP). To test the hypothesis, the present study was designed to compare histomorphometrically resorption of the implanted OCP, HA, and beta-TCP, which were kept in the experimental cranial defect of rats for a long term. A full thickness of standardized trephine defect was made in the rat parietal bone, and the same volume of granules of OCP, HA, and beta-TCP were implanted into the defect. Five specimens of each group were fixed 6 months after implantation. The percentage of remaining implants (r-Imp%) and newly formed bone (n-Bone%) in the defect was analyzed histomorphometrically. The statistical analysis showed that the r-Imp% of OCP was significantly lower than that of HA and beta-TCP. In contrast, the n-Bone% of OCP was significantly higher than that of HA and beta-TCP. The present study has shown that the implanted OCP in the rat cranial defect is more resorbable than the implanted beta-TCP and HA, whereas the implanted OCP enhances bone formation more than the implanted beta-TCP and HA.     

Osteointegration of femoral stem prostheses with a bilayered calcium phosphate coating

Our purpose was to evaluate the osteointegration of bilayered calcium phosphate (CaP)-coated femoral hip stems in a canine model. A first layer of hydroxyapatite (HA) 20 microm thick and a superficial layer of Biphasic Calcium Phosphate (BCP) 30 microm thick were plasma-sprayed on to the proximal region of sandblasted Ti6Al4V prostheses. Bilayered CaP-coated and non-coated canine femoral stems were implanted bilaterally under general anesthesia in 6 adult female Beagle dogs. After 6 and 12 months, a significant degradation of the bilayered coating occurred with a remainder of 33.1+/-12.4 and 23.6+/-9.2 microm in thickness, respectively. Lamellar bone apposition was observed on bilayered coated implants while fibrous tissue encapsulation was observed on non-coated femoral stems. The bone-implant contacts (BIC) were 91+/-3% and 81+/-8% for coated and 7+/-8% and 8+/-12% for non-coated implants, at 6 and 12 months, respectively. Our study supports the concept of a direct relationship between the biodegradation of CaP coating and the enhanced osteointegration of titanium prostheses. A bilayered CaP coating might therefore enhance bone apposition in the early stages because of the superior bioactivity of the BCP layer while the more stable HA layer might sustain bone bonding over long periods.     

Efficacité et performance des substituts osseux pour remplacer les allogreffes et autogreffes

Although bone tissue possesses the capacity for regenerative growth, the bone repair process is impaired in many clinical and pathological situations. For example massive bone loss caused by trauma and tumor resection as well as deformities require reconstructive surgery. In this context, there was a critical need to develop implant technologies to promote bone healing. Cortical and cancellous bone grafts are the materials of choice for bone filling or reconstruction, but their clinical use involves some difficulties. Septic complications, viral transmission and unavailability of native bone have therefore led to the development of synthetic bone substitutes. Allograft bone, or tissue harvested from a cadaver, while more readily available, may carry with it the risk of disease transmission and is also difficult to shape [1–3]. A significant additional limitation of allograft bone is the delayed remodeling by the host. In the case of very large defects, the allograft may remain in the implant site throughout the patient's life, creating an area more prone to fracture or infection. The development of calcium phosphate ceramics and other related biomaterials for bone graft involved a better control of the process of biomaterials resorption and bone substitution. Synthetic bone graft materials available as alternatives to autogeneous bone for repair, substitution or augmentation, in particular synthetic biomaterials include, special glass ceramics described as bioactive glasses; calcium phosphates (calcium hydroxyapatite, HA; tricalcium phosphate, TCP; and biphasic calcium phosphate, BCP). These materials differ in composition and physical properties from each other and from bone; and must be take in consideration for more efficient bone ingrowth at the expense of the biomaterials and to adapt to new development of dedicated biomaterials. In the last decade synthetic calcium phosphate materials, principally calcium hydroxyapatite (HA) ceramics, was commercially used. However the concept of bioactivity (release of ions of biological interest) well described for glass ceramic was not particularly take in account for HA and other related biomaterials(ACP Amorphous Calcium Phosphate, CdA Calcium Phosphate deficient Apatite). HA until recently was considered to be non able to be resorbed. Calcium phosphate biomaterials differ in their solubility or extent of dissolution: ACP > > α-TCP > > β-TCP > CdA > > ACP. These ceramics are osteoconductive (act as a support for new bone formation requiring the presence of porosity) and able to be resorbed (degradable through chemical and cellular processes). They are also biocompatible (do not induce adverse local tissue reaction, immunogenicity or systematically toxicity); and more recently, some papers report osteoinductive properties associated to the chemical nature (biphasic Ca P) and the microstruture. Past decade, these bioceramics have been marketed and approved for use in humans as bone substitutes. Various presentations are currently used in orthopaedic and maxillo-facial surgery such as wedges, blocks or granules. Owing to their bone substitution properties, CaP ceramics have naturally been considered as a potential matrix for tissue engineering and the development of a bioactive drug delivery system (DDS) in bone sites. The paper presents the current knowledge on Calcium phosphate bioceramics, Bone tissue engineering and Calcium Phosphate Drug Delivery.     

骨诱导性磷酸钙陶瓷材料修复牙槽突裂

BACKGROUND: In our previous studies, we had prepared calcium phosphate ceramics with better ectopic osteogenesis. OBJECTIVE: To explore the effect of novel osteoinductive calcium phosphate ceramics in the repairing of alveolar cleft. METHODS: Bilaterl alveolar defects were created in nine immature beagles. Three months later, osteoinductive calcium phosphate ceramics with high modular surface (experimental group) and smooth surface (control group) were randomly implanted in each side of the defect. Meanwhile, the corresponding material was implanted into the thigh muscle. New bone formation in the implanted region, osteogenesis in the implanted region and muscle, and respair results were respectively observed by fluorescence microscope, light microscope and CT at 4, 8 and 12 weeks after implantation. RESULTS AND CONCLUSION: (1) Fluorescence microscope observation: A circular permutation of red, yellow and green fluorescent strip could be observed in both two groups. (2) Light microscope observation: At 12 weeks after implantation, in the experimental group, the bone reconstruction was obvious, the implant material was decomposed gradually, the gap was filled with a large number of mature bone that combined with the rest material closely, and numerous Haversian canals appeared; the control group was similar but slightly inferior to the experimental group in the quality of new bone. The experimental group material successfully induced heterotopic osteogenesis in muscle, while the control did not. (3) CT examination: The two group materials restored the appearance and continuity of the alveolar ridge, and made no effect on the eruption of permanent teeth in both sides of the defect. To conclude, our findings suggest that the novel osteoinductive calcium phosphate ceramic exhibits advantages in alveolar cleft repair with earlier osteogenesis activation, faster osteogenesis rate and more bone formation than those traditional materials.     

Differential osteogenic activity of osteoprogenitor cells on HA and TCP/HA scaffold of tissue engineered bone

Biomaterial, an essential component of tissue engineering, serves as a scaffold for cell attachment, proliferation, and differentiation; provides the three dimensional (3D) structure and, in some applications, the mechanical strength required for the engineered tissue. Both synthetic and naturally occurring calcium phosphate based biomaterial have been used as bone fillers or bone extenders in orthopedic and reconstructive surgeries. This study aims to evaluate two popular calcium phosphate based biomaterial i.e., hydroxyapatite (HA) and tricalcium phosphate/hydroxyapatite (TCP/HA) granules as scaffold materials in bone tissue engineering. In our strategy for constructing tissue engineered bone, human osteoprogenitor cells derived from periosteum were incorporated with human plasma-derived fibrin and seeded onto HA or TCP/HA forming 3D tissue constructs and further maintained in osteogenic medium for 4 weeks to induce osteogenic differentiation. Constructs were subsequently implanted intramuscularly in nude mice for 8 weeks after which mice were euthanized and constructs harvested for evaluation. The differential cell response to the biomaterial (HA or TCP/HA) adopted as scaffold was illustrated by the histology of undecalcified constructs and evaluation using SEM and TEM. Both HA and TCP/HA constructs showed evidence of cell proliferation, calcium deposition, and collagen bundle formation albeit lesser in the former. Our findings demonstrated that TCP/HA is superior between the two in early bone formation and hence is the scaffold material of choice in bone tissue engineering.     

Ceramic drug delivery: a perspective

Different ceramic substances are offered in the market as bone substitute materials. These include monophasic calcium phosphate ceramics of tricalciumphosphate (TCP) or hydroxyapatite (HA), biphasic calcium phosphate ceramics and multiphasic bio-glasses synthetic calcium phosphate cements. Ceramics with appropriate three-dimensional geometry are able to bind and concentrate bone morphogenetic proteins in circulation and may become osteoinductive (capable of osteogenesis) and can be effective carriers of bioactive peptide or bone cell seeds and are therefore potentially useful in tissue engineering and drug delivery. An attempt has been made to review various drug delivery applications of ceramics.     

Ectopic bone formation in rats: the importance of vascularity of the acceptor site

Bone graft substitutes (BGS) can be fabricated by the combination of three key ingredients: (1) competent bone-forming cells, (2) a suitable framework or scaffold, and (3) the presence of biological stimulants. Although much research has been done to develop the ideal BGS, still the results are not very consistent. In view of this, the cellularity and vascularity of the recipient site are supposed to be important for the osteoinductive capacity of BGS. Therefore, we hypothesized that a muscle recipient site could favor bone formation in a cell-based BGS compared to a subcutaneous recipient site due to the higher vascularity of muscle tissue. To prove this hypothesis, 48 titanium fiber mesh implants were seeded with rat bone marrow stromal cells (RBM) and implanted subcutaneously and intramuscularly in the adductor thigh muscle of rats. The amount of bone formation after 1, 3 and 6 weeks was evaluated by histology and histomorphometry as well as by calcium content. Analysis revealed that the bone formation increased during implantation. However, bone formation did not exceed 12% of the implant surface, both for the intramuscular and subcutaneous recipient site. Also, no significant differences in bone amount between these two sites existed. Consequently, our hypothesis could not be confirmed.     

Osteogenicity of biphasic calcium phosphate ceramics and bone autograft in a goat model

The aim of this work was to compare the osteogenicity of calcium phosphate ceramic granules with autologous bone graft in ectopic and orthotopic sites. Biphasic calcium phosphate (BCP) granules composed of hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP) in a 60/40 ratio were sintered at 1050, 1125 and 1200 degrees C, producing different microporosities. Either BCP ceramic granules or autologous bone chips (n=7) were implanted into paraspinal muscles. Osteoinduction was not observed in either the BCP implants or autologous bone chips after 6 or 12 weeks in the ectopic sites. Hollow and bored polytetrafluoroethylene (PTFE) cylinders were filled with autologous bone, BCP granules or left empty, then implanted into critical-sized defects in femoral epiphyses. The PTFE cylinders left empty contained marrow and blood vessels but not mineralized bone, indicating that this model prevented bone ingrowth (0.56+/-0.43% at 12 weeks). Bone formation was observed in contact with the BCP1050 and BCP1125 granules in the femoral sites after 6 weeks. The amount of bone after 12 weeks was 5.6+/-7.3 and 9.6+/-6.6% for BCP1050 and BCP1125, respectively. Very little bone formation was observed with the BCP1200 implants (1.5+/-1.3% at 12 weeks). In both the ectopic and orthotopic sites, autologous bone chips were drastically resorbed (from 19.4+/-3.7% initially to 1.7+/-1.2% at 12 weeks). This study shows that synthetic bone substitutes may have superior stability and osteogenic properties than autologous bone grafts in critical-sized bone defects.     

On the feasibility of phosphate glass and hydroxyapatite engineered coating on titanium

In this report, bioactive calcium phosphate (CaP) coatings were produced on titanium (Ti) by using phosphate-based glass (P-glass) and hydroxyapatite (HA), and their feasibility for hard tissue applications was addressed in vitro. P-glass and HA composite slurries were coated on Ti under mild heat treatment conditions to form a porous thick layer, and then the micropores were filled in with an HA sol-gel precursor to produce a dense layer. The resultant coating product was composed of HA and calcium phosphate glass ceramics, such as tricalcium phosphate (TCP) and calcium pyrophosphate (CPP). The coating layer had a thickness of approximately 30-40 microm and adhered to the Ti substrate tightly. The adhesion strength of the coating layer on Ti was as high as 30-33 MPa. The human osteoblastic cells cultured on the coatings produced by the combined method attached and proliferated favorably. Moreover, the cells on the coatings expressed significantly higher alkaline phosphatase activity than those on pure Ti, suggesting the stimulation of the osteoblastic activity on the coatings. On the basis of these observations, the engineered CaP coating layer is considered to be potentially applicable as a hard tissue-coating system on Ti-based implants.     

Bone formation in calvarial defects of Sprague-Dawley rats by transplantation of calcium phosphate glass

The purpose of this study was to investigate the bone-regenerative effect of calcium phosphate glass in vivo. We prepared amorphous calcium phosphate glass powder having a mean particle size of 400 microm in the system CaO-CaF2-P2O5-MgO-ZnO. Calvarial critical-sized defects (8 mm) were created in 60 male Sprague-Dawley rats. The animals were divided into an experimental group and control group of 30 animals each. Each defect was filled with a constant weight of 0.5 g calcium phosphate glass powder mixed with saline. As a control, the defect was left empty. The rats were sacrificed 2, 4, or 8 weeks postsurgery, and the results evaluated using radiodensitometric and histological studies; they were also examined histomorphometrically. When the calcium phosphate glass powders with 400-microm particles were grafted, the defects were nearly completely filled with new-formed bone in a clean healing condition after 8 weeks. It was observed that the prepared calcium phosphate glass enhanced new bone formation in the calvarial defect of Sprague-Dawley rats and could be expected to have potential for use as a hard tissue regeneration material.     

Intramembranous bone tissue response to biodegradable octacalcium phosphate implant

Previous studies showed that synthetic octacalcium phosphate (OCP) enhances bone formation coupled with its own osteoclastic biodegradation more than non-biodegradable hydroxyapatite (HA), including sintered HA ceramic, when implanted in animal bone defects. The present study was designed to investigate whether synthetic OCP in granule form has biodegradable characteristics when implanted in the subperiosteal area of mouse calvaria in comparison with non-sintered stoichiometric HA, especially in relatively short periods after implantation. OCP crystals exhibited plate-like morphology, whereas HA crystals had a sphere-like structure. Both crystals had large pore volumes >75% in total, with micropores within the granules. Direct bonding of newly formed bone was discernible in HA until 35 days after implantation by element analysis for calcium and phosphorus. However, histomorphometric analysis demonstrated that bone formation was facilitated on OCP surfaces with greater alkaline phosphatase activity than on HA up to 21 days. The surfaces attacked by tartrate-resistant acid phosphatase positive osteoclast-like cells were significantly greater than those of HA. OCP became encapsulated and replaced with new bone with prolonged implantation periods up to 180 days. The results suggest that the enhanced bone formation in mouse calvaria could be associated with the biodegradable nature of OCP, and that OCP could be used in augmenting intramembranous bone volume.     

Hydroxyapatite granules interposed at bone-cement interface in total hip replacements: histological study of retrieved specimens

The effect of hydroxyapatite (HA) granules interposed between bone and polymethylmethacrylate (PMMA) bone cement in total hip replacement was histologically evaluated. The technique consisted of smearing 2-5 g of HA granules (straight phi = 100-300 microm) onto the bone surface just before cementing. Four specimens containing well-fixed bone-cement interface were retrieved at 1, 2, 6, and 10 years postoperatively and examined with back-scattered electron microscopy and light microscopy. The majority of HA granules were incorporated into remodeled trabeculae, and highly convoluted bone-cement interface was maintained up to 10 years. The presence of active remodeling in the adjacent bone was observed. There were no significant inflammatory or foreign body reactions against interposed HA granules. In one specimen retrieved from a patient with rheumatoid arthritis, bone formation around HA granules was limited after 1 year. These results have provided histological evidence for the significantly reduced incidence of radiolucent lines in total hip replacement with this cementing technique, reported elsewhere.     

In vivo bone response to 3D periodic hydroxyapatite scaffolds assembled by direct ink writing

The in vivo bone response of 3D periodic hydroxyapatite (HA) scaffolds is investigated. Two groups of HA scaffolds (11 mm diameter x 3.5 mm thick) are fabricated by direct-write assembly of a concentrated HA ink. The scaffolds consist of cylindrical rods periodically arranged into four quadrants with varying separation distances between rods. In the first group, HA rods (250 microm in diameter) are patterned to create pore channels, whose areal dimensions are 250 x 250 microm(2) in quadrant 1, 250 x 500 microm(2) in quadrants 2 and 4, and 500 x 500 microm(2) in quadrant 3. In the second group, HA rods (400 microm in diameter) are patterned to create pore channels, whose areal dimensions of 500 x 500 microm(2) in quadrant 1, 500 x 750 microm(2) in quadrants 2 and 4, and 750 x 750 microm(2) in quadrant 3. Each group of scaffolds is partially densified by sintering at 1200 degrees C prior to being implanted bilaterally in trephine defects of skeletally mature New Zealand White rabbits. Their tissue response is evaluated at 8 and 16 weeks using micro-computed tomography, histology, and scanning electron microscopy. New trabecular bone is conducted rapidly and efficiently across substantial distances within these patterned 3D HA scaffolds. Our observations suggest that HA rods are first coated with a layer of new bone followed by subsequent scaffold infilling via outward and inward radial growth of the coated regions. Direct-write assembly of 3D periodic scaffolds composed of micro-porous HA rods arrayed to produce macro-pores that are size-matched to trabecular bone may represent an optimal strategy for bone repair and replacement structures.     

Stimulatory effect of hydrothermally synthesized biodegradable hydroxyapatite granules on osteogenesis and direct association with osteoclasts

Calcium-deficient hydroxyapatite (HA) granules with a unique spherical shape were prepared using an applied hydrothermal method. Spherical stoichiometric HA granules were also prepared by normal sintering and both granules were used for implantation into rat tibiae to compare the biological responses to each implant. Twelve and 24 weeks after implantation, the volume of calcium-deficient HA granules was significantly less than that of stoichiometric HA granules, and the biodegradability of calcium-deficient HA granules was confirmed. The larger number of osteoclasts, larger osteoblast surface and larger bone volume in the implanted area of calcium-deficient HA than those of stoichiometric HA suggested that osteoclastic resorption of calcium-deficient HA affected osteogenesis in that area. To analyze the direct contribution of osteoclasts to osteogenesis, C2C12 multipotent myoblastic cells, which have the potential to differentiate into osteoblasts in the presence of bone morphogenetic protein 2, were cultured with supernatants of osteoclasts cultured on calcium-deficient HA, stoichiometric HA, β-tricalcium phosphate disks or plastic dishes, or bone marrow macrophages cultured on plastic dishes. Supernatants of osteoclasts but not bone marrow macrophages stimulated the expression of Runx2 and osteocalcin in C2C12 cells in concert with bone morphogenetic protein 2. The expression of alkaline phosphatase was stimulated with supernatants of osteoclasts cultured on ceramic disks. These results suggested that osteoclasts produced certain soluble factors which stimulated osteoblastic differentiation and they were thought to be associated with the induction of a larger osteoblast surface and bone volume in the animals implanted with calcium-deficient HA granules.     

钛合金表面新型生物玻璃/羟基磷灰石涂层的体内动物实验

目的为促进钛合金植入体与骨的结合,在其表面制备了生物玻璃/羟基磷灰石复合涂层,并植入兔子股骨内进行动物试验,采用等离子喷涂羟基磷灰石涂层和未涂层的Ti6Al4V合金作为对照。方法种植到期的植入体取出后进行组织学切片,采用品红-苦味酸染色后进行组织学观察,采用SEM高倍观察种植体与骨的结合界面,并对骨接触率和凹槽内骨长入量进行了统计分析和比较。结果三种植入体都具有良好的生物相容性。Ti6Al4V合金与骨之间是一种形态固定,而生物玻璃/羟基磷灰石涂层、等离子喷涂羟基磷灰石涂层可与骨形成骨键合。生物玻璃/羟基磷灰石涂层在植入期间与基体没有脱落,同时其与骨的接触率和凹槽内骨长入量要明显高于其余两个植入体,显示出促进骨生长的作用。结论由于具有良好的生物相容性和促进新骨生长的能力,生物玻璃/羟基磷灰石涂层可加快植入体与骨的愈合速度,在骨替代修复方面显示出优势和广阔的应用前景。