Engineered polycaprolactone – magnesium hybrid biodegradable porous scaffold for bone tissue engineering

In this paper, we describe the fabrication of a new biodegradable porous scaffold composed of polycaprolactone(PCL) and magnesium(Mg)micro-particles. The compressive modulus of PCL porous scaffold was increased to at least 150% by incorporating 29% Mg particles with the porosity of 74% using Micro-CT analysis. Surprisingly, the compressive modulus of this scaffold was further increased to at least 236% when the silane-coupled Mg particles were added. In terms of cell viability, the scaffold modified with Mg particles significantly convinced the attachment and growth of osteoblasts as compared with the pure PCL scaffold. In addition, the hybrid scaffold was able to attract the formation of apatite layer over its surface after 7 days of immersion in normal culture medium, whereas it was not observed on the pure PCL scaffold. This in vitro result indicated the enhanced bioactivity of the modified scaffold. Moreover, enhanced bone forming ability was also observed in the rat model after 3 months of implantation. Though bony in-growth was found in all the implanted scaffolds. High volume of new bone formation could be found in the Mg/PCL hybrid scaffolds when compared to the pure PCL scaffold. Both pure PCL and Mg/PCL hybrid scaffolds were degraded after 3 months. However, no tissue inflammation was observed. In conclusion, these promising results suggested that the incorporation of Mg micro-particles into PCL porous scaffold could significantly enhance its mechanical and biological properties. This modified porous bio-scaffold may potentially apply in the surgical management of large bone defect fixation.     

A new bone repair scaffold combined with chitosan/ hydroxyapatite and sustained releasing icariin

Icariin, a plant-derived flavonol glycoside, has been proved as an osteoinductive agent for bone tissue engineering. A new bone repair scaffold was generated by thorough mixing of icariin and chitosan/ hydroxyapatite （icariin-CS/HA） using freeze-drying technigue. Characteristics of morphology, mechanical properties, biocompatibility, drug release behavior and bone repair abilities in vivo were evaluated. The results show that drug loading process of icariin did not affect physical structure of CS/HA composite significantly but decreased mechanical properies of CS/HA composite, which happened with a high dosage; icariin-CS/HA had favorable cell compatibility and promoted osteogenic differentiation of hBMSCs; the controlled release of icariin was satisfactory and the release retained after 90 d in vitro. In addition, icariin-CS/HA scaffolds had favorable osteoconduction and osteoinduction in vivo, and could fill bone defect sites and stimulate newborn bone tissues formation at early stage. On the basis of these data, icariin-CS/HA is believed to be an optical bone repair scaffold for tissue engineering.     

Fabrication and characterization of a PAM modified PHBV/BG scaffold

In order to improve the bioactivity and mechanical strength of the scaffold used in bone repair simultaneously, a novel porous PAM-poly （β-hydroxybutyrate-co-β-hydroxyvalerate） （PHBV）/bioactive glass （BG） scaffold was prepared by photo-initiated polymerization. PAM was used to improve the hydrophilicity of PHBV matrix while the BG particles were added to increase the bioactivity and strength of the matrix synchronously. The grafted amide group and Si-O moieties from acrylamide and the added BG were confirmed by Fourier Transform Infrared Spectrometry （FTIR）. The micromorphology of the scaffolds before and after grafting was observed by scanning electron microscopy （SEM）. The resulting images demonstrate that the PAM-PHBV/BG scaffold has a well connected pore structure and appropriate pore size which may be convenient for cells to grow and discharge metabolites. The specific gravity method was used to evaluate the pore property of the scaffold and the result shows that the scaffold has an average porosity up to 82.0%. Mercury intrusion porosimetry （MIP） indicated that the pores of PAM-PHBV/BG scaffold were mainly distributed between 75 and 150 μm. The compressive strength test was adopted to evaluate the mechanical property of the scaffold. The result shows that the PAM-PHBV/BG scaffold has a relatively high compressive strength （0.91 MPa） when compared with the pure PHBV scaffold. Besides, the properties of the pure PHBV scaffold, PHBV/BG scaffold were also evaluated. The newly prepared PAM-PHBV/BG scaffold may be worthy of further studying as a bone repair material.     

Tussah Silk Fibroin Porous Scaffolds Prepared with a Mild Self-assembly Process for Controlled Drug Release

Besides excellent biodegradability and biocompatibility,a useful tissue engineering scaffold should provide favorable surface properties,outstanding mechanical strength and controlled drug release property. In this paper,a mild process to prepare porous tussah silk fibroin（ TSF） scaffolds from aqueous solution was described. The n-butanol was used to control the self-assembly of tussah silk. The scaffolds with different TSF concentrations and the same volume showed differences in pore size and distribution. The maximum porosity of the poprepared porous scaffolds was 80% in this paper. And the pore size of the prepared porous scaffolds with different concentrations was between 10μm and 230 μm. X-ray diffraction（ XRD） analysis revealed that amorphous TSF was crystallized to β-sheet secondary structure upon gelatin. The TSF scaffolds for controlled drug release was studied and the result showed that the time of drug release was significantly longer. The produced TSF scaffolds with sustained drug release have potential application in tissue engineering.     

Preparation of fiber-microsphere scaffolds for loading bioactive substances in gradient amounts

Gradient scaffolds are needed for interface tissue regeneration. In this study, a technique combining electrospinning and electrospraying was developed for preparing poly(L-lactide-co-glycolide) (PLGA) fiber-microsphere scaffolds for loading bioactive substances in gradient amounts. The gradient fiber-microsphere scaffolds contain two sheets of electrospun membranes and a sheet of microspheres loaded with bioactive substances in gradient amounts between the electrospun membranes. The morphologies of the gradient scaffolds were characterized and bovine serum albumin (BSA) was loaded as a model bioactive substance. The amount of BSA-loaded microspheres decreased gradually along the length of the gradient scaffold. The addition of poly (ethylene glycol) significantly improved the hydrophilicity of the gradient scaffold and the release behavior of BSA with respect to the gradient became apparent, with differences in the release amounts along the length of the gradient scaffold being observed. The biocompatibility of the gradient scaffold was verified using MC3T3-E1 pre-osteoblastic cells. The study demonstrated that the combination of electrospinning and electrospraying was a feasible method for the preparation of gradient scaffolds for potential applications in interface tissue engineering.     

Restoration of rat calvarial defects by poly（lactide-co-glycolide）/ hydroxyapatite scaffolds loaded with bone mesenchymal stem cells and DNA complexes

A composite construct comprising of a bone mesenchymal stem cell (BMSC) sheet, plasmid DNA, encoding human bone morphogenic protein-2 (hBMP-2), and poly(lactide-co-glycolide)/hydroxyapatite (PLGA/HA) sponge was designed and employed in the restoration of rat calvarial defects. To improve gene transfection efficiency, a cationic chitosan derivative, N,N,N,-trimethyl chitosan chloride (TMC), was employed as the vector. The TMC/DNA complexes had a transfection efficiency of 13% in rat BMSCs, resulting in heterogeneous hBMP-2 expression in a 10-d culture period in vitro. In vivo culture of the composite constructs was performed by implantation into rat full-thickness calvarial defects, using constructs lacking pDNA-hBMP-2 or BMSC sheets as controls. Significantly higher heterogeneous expression of hBMP-2 was detected in vivo at 2 weeks for the cell sheet/DNA complex/scaffold constructs, compared with the constructs lacking pDNA-hBMP-2 or BMSC sheets. New bone formation was evident as early as 4 weeks in the experimental constructs. At 8 weeks, partial bridging of calvarial defects was observed in the experimental constructs, which was significantly better than the constructs lacking pDNA-hBMP-2 or BMSC sheets. Therefore, the combination of the PLGA/HA scaffold with BMSC sheets and gene therapy vectors is effective at enhancing bone formation.     

Electrospun composites of PHBV/pearl powder for bone repairing

Electrospun fi ber has highly structural similarity with natural bone extracelluar matrix(ECM). Many researches about fabricating organic–inorganic composite materials have been carried out in order to mimic the natural composition of bone and enhance the biocompatibility of materials. In this work, pearl powder was added to the poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(PHBV) and the composite nano fi ber scaffold was prepared by electrospinning. Mineralization ability of the composite scaffolds can be evaluated by analyzing hydroxyapatite(HA)formation on the surface of nano fi ber scaffolds. The obtained composite nano fi ber scaffolds showed an enhanced mineralization capacity due to incorporation of pearl powder. The HA formed amount of the composite scaffolds was raised as the increase of pearl powder in composite scaffolds. Therefore, the prepared PHBV/pearl composite nano fi ber scaffolds would be a promising candidate as an osteoconductive composite material for bone repairing.     

Preparation and bioactivity of sol-gel macroporous bioactive glass

Bioactive glass is well known for its ability of bone regeneration, and sol-gel bioactive glass has many advantages compared with melt-derived bioactive glass. 3-D scaffold prepared by the sol-gel method is a promising substrate material for bone tissue engineering and large-scale bone repair. Porous sol-gel glass in the CaO-SiO₂-P₂0₅ system with macropores larger than 100 μm was prepared by the addition of stearic acid as a pore former. The diameter of the pore created by the pore former varied from 100 to 300 μm. The formation of a hydroxyapatite layer on the glass was analyzed by studying the surface of the porous glass by scanning electron microscopy, energy dispersive spectroscopy, X-ray diffraction, and Raman spectra after they had been immersed in simulated body fluid (SBF) for some time, and the porous glass shows good bioactivity.     

Effects of the rare earth ions on bone resorbing function of rabbit mature osteoclasts<Emphasis Type="Italic">in vitro</Emphasis>

The osteoclast plays initiating and vanguard roles in the course of bone remodeling. The resorption and formation of bone is the basic clues of bone remodeling, which results from the mutual dependent functions of osteoclasts and osteoblasts. When the osteoclasts are activated, the bone resorption function is increased. The excess bone resorption leads to osteoporosis, characterized by formation of many hollows, lacunae and tunnels on the bone surface or in depth[1,2]. The differentiation, pro…     

On Modeling Bio-Scaffolds:Structural and Fluid Transport Characterization Based on 3-D Imaging Data

Bio-scaffolds which are most commonly open celled porous structures are increasingly used for tissue engineering and regenerative medicine. A number of studies have shown that the bulk properties of such irregular structures are poorly modeled using idealized unit cell approaches. The paper therefore uses novel image based meshing techniques to explore both fluid flow and bulk structural properties of a bone scaffold, as accurate modeling of bio-scaffolds with non-uniform cellular structures is very important for the development of optimal scaffolds for tissue engineering application. In this study, a porous hydroxyapatite/tricalcium phosphate (HA/TCP) bone scaffold has been scanned in a Micro-CT scanner, and converted into a volumetric mesh using image processing software developed by the authors. The resulting mesh was then exported to commercial FEA and CFD solvers for analysis. Initial FEA and CFD studies have shown promising results and have highlighted the importance of accurate modeling to understand how microstructures influence the mechanical property of the scaffold, and to analyze flow regimes through the sample. The work highlights the potential use of image based meshing for the ad hoc characterization of scaffolds as well as for assisting in the design of scaffolds with tailored strength, stiffness, and transport properties.     

Preparation and evaluation of biomimetric nano-hydroxyapatite-based composite scaffolds for bone-tissue engineering

In the present study,novel biomimetic composite scaffolds with a composition similar to that of natural bone were prepared,using nano-hydroxyapatite,collagen,and phosphatidylserine.The scaffolds possess an interconnected porous structure with a porosity of 84%.The pore size ranges from several micrometers up to about 400 m.In-vitro studies in simulated body fluids showed that the morphologies of the products derived from mineralization can be regulated by the extracellular matrix components of the scaffolds;this in turn leads to creation of a large number of hydroxyapatite crystals on the scaffold surface.The regulatory properties of collagen and phosphatidylserine also influenced the cell response to the composite scaffolds.MC3T3-E1 cells attached and spread on the surfaces of the materials and interacted with the substrates;this may be the result of charged groups on the composite materials.Radiological analysis suggested that calluses and bone bridges formed in defects within 12 weeks.These composite scaffolds may therefore be a suitable replacement in bone-tissue engineering.     

Acute toxicity and oxidative damage induced by silica nanorattle in vivo

Hollow mesoporous nanomaterial is a kind of promising new drug delivery system due to their unique hollow structures.In order to evaluate the toxicity of silica nanorattle (SN) particles in vivo,40 female mice were used in this study to investigate the acute toxicity and oxidative damage.Mice were intravenously injected with SN suspension in sterile 5% glucose at 40,80 and 240 mg/kg,respectively.The control group was administrated with equal-volume 5% glucose.Weight,feed intake,hematology analysis,blood biochemical assay and histopathology diagnosis were examined.The activities of SOD,GSH and CAT were measured as well.The results demonstrated that the levels of ALT and AST in the mice treated with 240 mg/kg SN increased significantly as compared with the control group (P<0.05),whereas the contents of BUN and CREA changed unremarkably.The activity of SOD induced by SN in the liver decreased significantly (P<0.05).In summary,this study revealed that liver was the target organ of the SN.It also can be concluded that activity of SOD played an important role in liver injury caused by SN.     

Processing, microstructure and mechanical properties of biomedical magnesium with a specific two-layer structure

A novel magnesium based scaffold with a two-layer structure was synthesized by powder metallurgical process using salt particles as space holder. The outer layer of the scaffold shows an interconnected porous structure and the inner layer presents a compact structure reinforced by the salt particles. Such a specific structure is introduced primarily for the purpose of a better combination of biocompatibility and mechanical compatibility. Experimental results demonstrate that the structural features and mechanical properties of the magnesium based scaffold with a salt content of 30 wt% prepared by the current method are quite compatible with the cancellous bone. Such a novel Mg-based scaffold has the potential to act as degradable implants for bone substitute application.     

Osseointegration of hollow porous titanium prostheses loaded with cancellous bone matrix in rabbits

This study aimed to observe the osseointegration of hollow porous titanium prostheses (HPTP) loaded with cancellous bone matrix (CBM) in rabbits using histological and biomechanical perspectives.Experimental samples were implanted into the lateral femoral condyles of 66 New Zealand rabbits,allocated into the following groups:non-porous prosthesis group (Group A,n=22);HPTP group (Group B,n=22);HPTP+CBM group (Group C,n=22).The rabbits were sacrificed at 3,8 and 12 weeks,postoperatively.X-ray analyses,microscopy techniques and morphological measurement software and mechanical tests were used for evaluation.At each time point,the tissues surrounding the implants were similar in all of the groups,with bony in-growth into the 2-mm round holes observed for the defects in Groups B and C.However,the internal bone formation was significantly better in Group C than in Group B at different time points (P<0.01).Biomechanically,the pull-out forces were significantly greater in Groups B and C than in Group A (P<0.01),with no difference between Groups B and C (P>0.05).These results suggest that bone can grow into the cavities of HPTP to achieve more stable locking fixation,and those osteogenic materials,such as CBM,can enhance osteogenesis to achieve better osseointegration between the implant and the host bone.     

Effects of different types of palatal lateral excisions on growth and development of maxilla and dental arch

Objective： This study aimed to explore the effects of different types of palatal lateral excisions on the growth and development of the maxilla and dental arch, and to investigate the underlying mechanisms. Methods： A total of 112 3-week-old Sprague-Dawley （SD） male rats were randomly divided into a control and 3 experimental groups： the mucoperiosteal denudation group, the mucosal flap excision group, and the periosteum excision group. In the experimental groups, bilateral mucoperiosteal, mucosal flap and periosteum were excised respectively in the lateral one half of the palate. Four rats in each group were randomly chosen for sacrifice every two weeks. The maxilla was dissected following the excision. The widths of the maxilla and dental arch were measured and the histological phenomena were investigated at different phases. At the same time, 12 animals in each group were sequentially injected with calcein every two weeks. Three animals in each group, whose fluorescent labeling was used, were sacrificed for investigating bone formation at Week 8 following injection. Results： （1） Each experimental group presented the constriction of the maxilla and dental arch. The upper first molars in the experimental groups inclined medially. The mucoperiosteal denudation group showed the largest degree of effect followed by the periosteum excision group. The indices of the mucosal flap excision group, which retained the structures of the periosteum layer, had the most approximate values to the control group; （2） Different histological changes among the experimental groups were detected. The fibers penetrated into the palatal bone as Sharpey＇s fibers in the mucoperiosteal denudation group. The pattern of bone deposition was the bundle type. Sharpey＇s fibers were not found in the mucosal flap and periosteum excision groups and the depositions of palatal bone were the lamellar type as those in the control group; （3） The rates of bone deposition in the experimental groups decreased compared with the     

Crystallogeny fundamentals of the cholesterol gallstone

The nucleation mechanism and crystal growth process of the cholesterol gallstone are studied and a systematic theory expounded by crystallogeny is proposed. Normal feed and stone-forming feed were used to raise guinea pigs in the control and stone-causing groups respectively. The state and transformation of liquid crystal vesicles, the appearance of crystal nuclei, and the formation of microcrystal grains were observed under a polarizing microscope during the experimental period. It was found that the liquid crystal vesicles in the bile of the control group were small, scattered, and always existed as single forms, and no shaped gallstone crystals were formed. While in the stone-causing group, liquid crystal vesicles grew to larger ones, and then aggregated to form large liquid crystal cells. Solid crystal growth along the edge of these liquid crystal cells formed microcrystal grains. These demonstrated that bile liquid crystal vesicles form the basic nuclei of cholesterol gallstone. Heterogeneous nucleation is the common process in the formation of crystal nuclei and crystal growth.     

Biporous nanocarbon foams and the effect of the structure on the capillary performance

Biporous nanocarbon foams (NC foams) which have both nano-pores and micro-pores,were fabricated and used to explore the liquid transportation in the porous structures formed by nanomaterials.Scanning electron microscopy was utilized to observe the structure of the NC foams.Capillary rise tests using acetone as the working fluid were conducted by the IR thermal imaging method.The results show that the capillary rise height of the NC foam with 65μm micro-pore size was 47%higher than that with 9μm micro-pore size in 50 s.The capillary performance of the NC foams was reduced when its density was higher than 60 mg/cm~3.A model is built to illustrate the effect of both nano-structure and micro-structure on the transportation of working fluid in the biporous foam.The data indicates that the nano-structure generated high capillary pressure while the microstructure provided low resistant pathways for fast liquid transportation.The size of micro-pores and the ratio of micro-pores to nano-pores were two important factors.It is demonstrated that the NC foams had large sorption capacity up to 878 mg/cm~3and their capillary performance was further enhanced when using a substrate.     

Crystallogeny fundamentals of the cholesterol gallstone

The nucleation mechanism and crystal growth process of the cholesterol gallstone are studied and a systematic theory expounded by crystallogeny is proposed. Normal feed and stone-forming feed were used to raise guinea pigs in the control and stone-causing groups respectively. The state and transformation of liquid crystal vesicles, the appearance of crystal nuclei, and the formation of microcrystal grains were observed under a polarizing microscope during the experimental period. It was found that the liquid crystal vesicles in the bile of the control group were small, scattered, and always existed as single forms, and no shaped gallstone crystals were formed. While in the stone-causing group, liquid crystal vesicles grew to larger ones, and then aggregated to form large liquid crystal cells. Solid crystal growth along the edge of these liquid crystal cells formed microcrystal grains. These demonstrated that bile liquid crystal vesicles form the basic nuclei of cholesterol gallstone. Heterogeneous nucleation is the common process in the formation of crystal nuclei and crystal growth.     

Effect of tunnel structure on the specific capacitance of etched aluminum foil

The morphology of etched aluminum foil was observed using scanning electron microscopy, which led to the establishment of a cylindrical model and two merged models, considering the fixed weight loss of etching. The maximum of specific capacitance and the corresponding optimum values for tunnel sizes at various anodization voltages were predicted. The increased size distribution and taper of tunnels were demonstrated to decrease the specific capacitance, whereas the addition of polymeric additive into the tunnel widening solution was demonstrated to increase the capacitance. The formation of merged tunnels on the etched aluminum surface, irrespective of the presence of row-merged tunnels or cluster-merged tunnels, resulted in a dramatic decrease in the specific capacitance. It is concluded that, enhancing the uniformity of tunnel size and distribution and avoiding the formation of merged tunnels are the effective approach to achieving the higher capacitance for the tunnel etched and formed aluminum foil.     

Immobilization of biomacromolecules on poly-L-lactide surface via a layer-by-layer method for the improving of its cytocompatibility to bone marrow stromal cells

Hyaluronic acid （HA） and chitosan （CS） were immobilized on the surface of poly-L-lactide （PLLA） by the following procedure： Firstly, PLLA was aminolyzed with 1, 6-hexanediamine, and part of the PLLA surface ester groups were converted to free amino groups. Then negatively charged hyaluronic acid and positively charged chitosan were deposited onto the surface of aminolyzed PLLA film in a layer-by-layer assembly manner. The effect of the layer-by- layer deposition was evaluated by ATRoFTIR spectroscopy, Raman spectroscopy and static contact angle measurements. The cytocompatibility of PLLA sample to bone marrow stromal cells （BMSCs） was improved after modification with chitosan and HA. The cell attachment, activity, and proliferation on CS/HA modified PLLA films were enhanced comparing with the control. The cells cultured on the modified PLLA samples excreted abundant cytoplasm and can differentiate to vascular smooth muscle （SM）-like （SM-like） cells. A macroporous three-dimensional PLLA scaffold was prepared by integrating both the technique of freeze-drying and particle leaching. Layer-by-layer modification by HA/CS and cell culture was also applied on this scaffold. The scaffold cultured with BMSCs for 2 weeks has been tested successfully in vivo as a patch for repairing the artificial incision on canine pulmonary artery.