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A poly(lactide-co-glycolide)/hydroxyapatite composite scaffold with enhanced osteoconductivity.

Biodegradable polymer/ceramic scaffolds can overcome the limitations of conventional ceramic bone substitutes. However, the conventional methods of polymer/ceramic scaffold fabrication often use organic solvents, which might be harmful to cells or tissues. Moreover, scaffolds fabricated with the conventional methods have limited ceramic exposure on the scaffold surface since the polymer solution envelopes the ceramic particles during the fabrication process. In this study, we developed a novel fabrication method for the efficient exposure of ceramic onto the scaffold surface, which would enhance the osteoconductivity and wettability of the scaffold. Poly(D,L-lactide-co-glycolide)/nanohydroxyapatite (PLGA/HA) scaffolds were fabricated by the gas foaming and particulate leaching (GF/PL) method without the use of organic solvents. Selective staining of ceramic particles indicated that HA nanoparticles exposed to the scaffold surface were observed more abundantly in the GF/PL scaffold than in the conventional solvent casting and particulate leaching (SC/PL) scaffold. Both types of scaffolds were implanted to critical size defects in rat skulls for 8 weeks. The GF/PL scaffolds exhibited significantly enhanced bone regeneration when compared with the SC/PL scaffolds. Histological analyses and microcomputed tomography of the regenerated tissues showed that bone formation was more extensive on the GF/PL scaffolds than on the SC/PL scaffolds. Compared with the SC/PL scaffolds, the enhanced bone formation on the GF/PL scaffolds may result from the higher exposure of HA nanoparticles to the scaffold surface. These results show that the biodegradable polymer/ceramic composite scaffolds fabricated with the novel GF/PL method can enhance bone regeneration compared with those fabricated with the conventional SC/PL method.

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Orbital floor reconstruction with flexible Ethisorb patches: a retrospective long-term follow-up study.

OBJECTIVE: The purpose of the study was to investigate whether a flexible, biodegradable material (Ethisorb) shows better long-term results with regard to diplopia, bulbus motility, and exophthalmos/enophthalmos compared to the use of lyophilized dura-patches and polydioxanone (PDS) foils. METHODS: During a period of 6 years 435 patients with an orbital fracture were investigated retrospectively. Inclusion criteria were patients with fractures of the orbital floor with a maximum size of 2 x 2 cm. Bulbus motility, exophthalmos, enophthalmos, and diplopia were investigated during a period of 2 years. RESULTS: One hundred twenty orbital floors were reconstructed by lyophilized dura-patches, 81 by PDS, and 136 by Ethisorb. An exploration without an implantation was performed in 91 patients. The long-term investigation 12 to 15 months after surgery showed an exophthalmos and enophthalmos incidence of 1%, whereas a reduced bulbus motility and diplopia were found in 5% and 4%, respectively. Fifteen to 24 months after surgery 2% of the patients had an exophthalmos and 1% had an enophthalmos. A reduction of bulbus motility was found in 4% of the patients, and diplopia was found in 3%. The use of Ethisorb resulted in a significantly lower incidence of exophthalmos 3 months after surgery compared to PDS. CONCLUSION: The low rate of acquired bulbus motility demonstrates acceptable results in using Ethisorb in the floor of the orbit.

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Biodegradable polymer grafts for surgical repair of the injured spinal cord.

PURPOSE: Biodegradable polymers have been used in the surgical repair of peripheral nerves, but their potential for use in the central nervous system has not been exploited adequately. This article discusses concepts related to the engineering of a biodegradable polymer graft for surgical repair of the injured spinal cord and explores the potential means by which such a device might promote axon regeneration and functional recovery after spinal cord injury. CONCEPT: A biodegradable polymer implant with controlled microarchitecture can be engineered, and its composition can be optimized for implantation in the spinal cord. RATIONALE: The use of a biodegradable polymer implant has the dual advantages of providing a structural scaffold for axon growth and a conduit for sustained-release delivery of therapeutic agents. As a scaffold, the microarchitecture of the implant can be engineered for optimal axon growth and transplantation of permissive cell types. As a conduit for the delivery of therapeutic agents that may promote axon regeneration, the biodegradable polymer offers an elegant solution to the problems of local delivery and controlled release over time. Thus, a biodegradable polymer graft would theoretically provide an optimal structural, cellular, and molecular framework for the regrowth of axons across a spinal cord lesion and, ultimately, neurological recovery. CONCLUSION: Biodegradable polymer grafts may have significant therapeutic potential in the surgical repair of the injured spinal cord. Further research should be focused on the bioengineering, characterization, and experimental application of these devices.

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Antitumor efficacy and local distribution of doxorubicin via intratumoral delivery from polymer millirods.

The purpose of this study was to evaluate the antitumor efficacy and local drug distribution from doxorubicin-containing poly(D,L-lactide-co-glycolide) (PLGA) implants for intratumoral treatment of liver cancer in a rabbit model. Cylindrical polymer millirods (length 8 mm, diameter 1.5 mm) were produced using 65% PLGA, 21.5% NaCl, and 13.5% doxorubicin. These implants were placed in the center of VX2 liver tumors (n = 16, 8 mm in diameter) in rabbits. Tumors were removed 4 and 8 days after millirod implantation, and antitumor efficacy was assessed using tumor size measurements, tumor histology, and fluorescent measurement of drug distribution. The treated tumors were smaller than the untreated controls on both day 4 (0.17 +/- 0.06 vs. 0.31 +/- 0.08 cm(2), p = 0.048) and day 8 (0.14 +/- 0.04 vs. 1.8 +/- 0.8 cm(2), p = 0.025). Drug distribution profiles demonstrated high doxorubicin concentrations (>1000 microg/g) at the tumor core at both time points and drug penetration distances of 2.8 and 1.3 mm on day 4 and 8, respectively. Histological examination confirmed necrosis throughout the tumor tissue. Biodegradable polymer millirods successfully treated the primary tumor mass by providing high doxorubicin concentrations to the tumor tissue over an eight day period.

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Experimental study of bone morphogenetic proteins-2 slow release from an artificial trachea made of biodegradable materials: evaluation of stenting time.

We manufactured an artificial trachea that slowly releases bone morphogenetic protein 2 (BMP-2) and used it to replace a section of the canine trachea. We made a three-layered prosthesis composed of an outer layer of gelatin sponge, a middle layer of collagen sponge, and an inner silicone tube. BMP-2 solution was soaked into the gelatin sponge layer. An approximately 3 cm length of the canine trachea was resected, and the artificial trachea was inserted into the resulting gap and anastomosed. The implanted portion was covered by periosteum. At 2, 4, and 8 weeks after surgery, the inner silicone tube was removed. Soon after removal of the silicone tube at 2 and 4 weeks, the dogs died of choking because of collapse of the trachea. One dog whose silicone tube was removed at 8 weeks was able to survive without choking. At 6 months after removal of the silicone tube, the bronchoscopic findings revealed that the gap in the trachea had been closed by regenerated tissue and covered by mucosa. We have demonstrated that our artificial trachea slowly releasing BMP-2 requires at least 8 weeks to achieve regeneration of solid tissue to support the tracheal gap.

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Intrauterine autogenous foetal bone transplantation for the repair of cleft-like defects in the mid-gestational sheep model.

AIM: The success of intrauterine surgery in treating non-life-threatening malformations such as myelomeningocoele, has also renewed strong interest in using this technique for treating craniofacial malformations. Nevertheless, the only experimental cleft-like defect models known, are those concerning wound healing of soft tissues. MATERIAL AND METHODS: Attempts were made to repair artificial cleft-like defects including transplantation of 11 autogenous foetal bone grafts from the iliac crest or ulna, and were randomly assigned to three study groups, using the mid-gestational sheep model. In a 4th study group, lyophilized collagen, a bone-regenerating bioresorbable implant material, was used to fill the alveolar defect. RESULTS: In all groups, there was a slight degree of asymmetry and thinning of the lip. Radiological studies demonstrated a variable degree of abnormality of the maxilla, ranging from none to a mild deviation. Three-dimensional computer tomography, two-dimensional maximal intensity projection findings, and histological analysis confirmed bony healing of the alveolar cleft-like defect. DISCUSSION/CONCLUSION: Intrauterine autogenous foetal bone transplantation for the repair of cleft-like defects in the sheep is feasible. This is a reliable and valuable model toward a possible clinical application for intrauterine treatment of clefts.

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The use of resorbable collagen membrane in conjunction with autogenous bone graft and inorganic bovine mineral for buccal/labial alveolar ridge augmentation: a pilot study.

STATEMENT OF PROBLEM: No study provides human histologic evidence regarding the use of resorbable collagen membrane for a 2-stage localized alveolar augmentation procedure. PURPOSE: The purpose of this pilot study was to evaluate the potential of use of a resorbable collagen membrane in conjunction with an autogenous bone graft and inorganic bovine mineral (IBM) for labial/buccal alveolar ridge augmentation prior to placing dental implants. Material and methods Seven consecutively treated human patients participated in the study. All patients received labial/buccal alveolar ridge augmentation. An autogenous block graft was secured at the recipient site with fixation screws and a mixture of autogenous particulate with IBM was placed at the periphery. Resorbable collagen membrane was used as a barrier. Radiographic and laboratory measurements were made to quantify ridge augmentation and resorption rate. Preoperative and postoperative stone casts were used to quantify alveolar ridge augmentation. Volumetric evaluation was measured in mL whereas linear laboratory evaluation was measured in millimeters. Measurements were made 1 and 6 months after bone grafting. Histologic and histomorphometric analysis from the grafted area evaluated new bone formation, and osteoconductivity of IBM. RESULTS: For all patients Type II to III bone quality was achieved at the augmented sites. The implant survival rate was 100% at second-stage surgery. No complication was observed at the recipient sites. Radiographic evaluation revealed 4.65 mm labial/lingual augmentation, whereas laboratory analysis revealed 4.57 mm. Volumetric laboratory analysis demonstrated 1.00 (+/- 0.29) mL alveolar ridge augmentation 6 months after bone grafting and 13.79% resorption between months 1 and 6. Histomorphometric analysis revealed that on average, the area occupied by bone was 34.28% (range 24 to 50; +/-9.05),] soft tissue 46.00% (+/-9.20%; range 30% to 55%), and IBM particles 19.71% (+/-11.74%, range 3% to 42%). The proportion of the surface of the IBM particles in contact with bone was 47.14% (range 15% to 64%; SD 17.21%). CONCLUSIONS: Resorbable collagen membranes may be used as barriers for labial/buccal alveolar ridge augmentation procedures.

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A new biological matrix for septal occlusion.

The ideal septal occluder scaffold should promote the healthiest and most complete healing response possible while eventually facilitating the full resorption of the material, leaving "native" tissue behind. An excellent biocompatibility of the scaffold tissue is a prerequisite for quick, complete, and firm ingrowth of the device, optimizing outcomes and minimizing the potential for complications. Intestinal collagen layer (ICL) is a highly purified (acellular) bioengineered type-1 collagen derived from porcine submucosa. It is gradually resorbed by the host organism and subsequently replaced by the host tissue. CardioSEAL occluders were modified by substituting the conventional polyester fabric for an intestinal collagen layer (ICL). Percutaneous transcatheter closure of interventionally created atrial septal defects was performed in lambs using these modified occluders. A complete pathomorphological investigation including histology was carried out after 2, 4, and 12 weeks follow-up. Standard CardioSEAL implants served as a control group. After 2 weeks in vivo the devices were already covered completely by neo-endothelium. Compared with the conventional synthetic scaffold, ICL devices showed a quicker endothelialization, decreased thrombogenicity, and superior biocompatibility with no significant cellular infiltration observed in the histology of explants with ICL fabrics. After 3 months in vivo the collagen layer remained mechanically intact, but began to show the first histological signs of mild disintegration, gradual resorption, and remodeling. In conclusion, short-term results from preliminary in vivo experiments using a bioengineered collagen matrix as the occluder tissue scaffold showed excellent biocompatibility. This resulted in superior overall results: quicker endothelialization, a decreased thrombogenicity, and decreased immunological host response.

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Initial assessment of a tissue engineered stomach derived from syngeneic donors in a rat model.

The objective of this study is to assess the feasibility of creating a tissue engineered stomach using isolated stomach epithelium organoid unit from syngeneic adult donors and a biodegradable polymer scaffold in a rat model. Despite recent advances in reconstruction techniques, total gastrectomy is still accompanied by various complications. As an alternative treatment, a tissue engineered stomach that replaces the mechanical and metabolic functions of a normal stomach is proposed. Stomach epithelium organoid units were isolated from syngeneic adult rats and seeded onto biodegradable polymers. These constructs were implanted into the omenta of recipient adult rats. All constructs were harvested for histologic and immunohistochemical examination at designated time points. Cyst-like structures were formed that showed the development of vascularized tissue with a neomucosa. Immunohistochemical staining for alpha-actin smooth muscle, gastric mucin, and proton pump indicated the presence of a smooth muscle layer and gastric epithelium, as well as the existence of parietal cells of the stomach mucosa, respectively. Epithelium derived stomach organoid units seeded on biodegradable polymers were transplanted in donor rats and have been shown to vascularize, survive, and regenerate into complex tissue resembling a native stomach. These initial results are encouraging, and studies are currently underway to further assess this approach.

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Evaluation of a porous, biodegradable biopolymer scaffold for mandibular reconstruction.

PURPOSE: Bioresorbable bone graft substitutes could eliminate disadvantages associated with the use of autografts, allografts, and other synthetic materials. The authors investigated the osteoinductive capacity of a bioresorbable bone graft substitute made from the unsaturated polyester poly(propylene glycol-co-fumaric acid) (PPF) for mandibular reconstruction in a rat model. The eventual intention is to use this material either as a stand-alone bone graft substitute or as an extender to autograft harvested from mandibular reconstruction sites. MATERIALS AND METHODS: The PPF bone graft was crosslinked in the presence of a hydroxyapatite filler and effervescent foaming agents to develop porosity in situ by generating carbon dioxide during the effervescent reaction of citric acid and sodium bicarbonate. The latter reagents are responsible for foam formation and expansion, resulting in a polymeric scaffold with pore sizes in the range of 100 to 500 microm. Twenty adult Sprague-Dawley rats had 3-mm-diameter cortical defects decorticated on the outer aspect of their left mandibular ramus using a Hall drill. Animals were divided into 2 groups of 10 animals each. Animals in group A were treated with implantation of the PPF-based bone graft substitute. Implants were applied buccally to defects on the left side. In group B animals with similar defects, the drill holes were left to heal unaided. The amount of new bone formation and the presence of an inflammatory infiltrate were evaluated at 7 weeks postoperatively. RESULTS: Histologic analysis of the healing process revealed enhanced in vivo new bone formation with the PPF bone graft substitute. These findings were corroborated by the histomorphometric analysis of new bone formation. DISCUSSION: Results of this study demonstrated biocompatibility of the porous PPF-based scaffold in a mandibular defect. CONCLUSIONS: These findings may have applicability to the further development of bone graft substitutes for oral/maxillofacial applications.

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The combined use of bioresorbable membranes and xenografts or autografts in the treatment of bone defects around implants. A study in beagle dogs.

The aim of the present investigation was to test the effect of a bioresorbable membrane supported by xenografts or autografts in regenerating bone into peri-implant defects. In 3 dogs, the mandibular premolars P2, P3, P4 and M1 were extracted bilaterally. After 4 months of healing, 3 standardized bone defects were prepared on each side of the mandible and 1 implant per defect was placed. The 6 sites in each dog were distributed into 4 different treatment groups: 2 sites received a Bio-Gide membrane alone (BG); 2 sites received a Bio-Gide membrane supported by Bio-Oss (BG + BO); 1 site received the Bio-Gide membrane supported by autogenic bone harvested from the prepared defects (BG + Aut); 1 site received neither membrane nor bone graft and served as control (C). The soft tissue flaps were adapted and sutured for primary healing. No adverse events occurred during the experimental period. After 16 weeks, the dogs were sacrificed and histomorphometric examinations on non-decalcified ground sections were carried out. The vertical bone growth amounted to 45% (SD +/- 13%) of the defect height in the BG group, to 78% (SD +/- 29%) in the BG + BO group, to 69% (SD +/- 9%) in the BG + Aut group, and to 22% (SD +/- 10%) in C group. The horizontal bone growth measured 78% (SD +/- 16%) in the BG group, 81% (SD +/- 21%) in the BG + BO group, 82% (SD +/- 12%) in the BG + Aut group, and 46% (SD +/- 21%) in the C group. The vertical height of bone growth in contact with the implant measured 17% (SD +/- 12%) in the BG group, 20% (SD +/- 12%) in the BG + BO group, 17% (SD +/- 7%) in the BG + Aut group, and 12% (SD +/- 8%) in the C group. The surface fraction of the graft in direct bone contact measured 89% (SD +/- 9%) in the BG + BO group and 93% (SD +/- 3%) in the BG + Aut group. It is concluded that the bioresorbable membrane tested enhances bone regeneration, in particular in conjunction with the use of a supporting graft material. In addition, deproteinized bovine bone mineral and autogenic bone grafts appeared to be equally well integrated into regenerating bone. Finally, no additional effects in the bone growth was observed with the autogenous bone in comparison with the hydroxyapatite.

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Permeable guidance channels containing microfilament scaffolds enhance axon growth and maturation.

Successful peripheral nerve regeneration is still limited in artificial conduits, especially for long lesion gaps. In this study, porous poly(L-lactide-co-DL-lactide, 75:25) (PLA) conduits were manufactured with 16 poly(L-lactide) (PLLA) microfilaments aligned inside the lumen. Fourteen and 18 mm lesion gaps were created in a rat sciatic nerve lesion model. To evaluate the combined effect of permeable PLA conduits and microfilament bundles on axon growth, four types of implants were tested for each lesion gap: PLA conduits with 16 filaments; PLA conduits without filaments; silicone conduits with 16 filaments; and silicone conduits without filaments. Ten weeks following implantation, regeneration within the distal nerve was compared between corresponding groups. Antibodies against the markers S100, calcitonin gene related peptide (CGRP), RMDO95, and P0 were used to identify Schwann cells, unmyelinated axons, myelinated axons, and myelin, respectively. Results demonstrated that the filament scaffold enhanced tissue cable formation and Schwann cell migration in all groups. The filament scaffold enhanced axonal regeneration toward the distal stump, especially across long lesion gaps, but significance was only achieved with PLA conduits. When compared to corresponding silicone conduits, permeable PLA conduits enhanced myelinated axon regeneration across both lesion gaps and achieved significance only in combination with filament scaffolds. Myelin staining indicated PLA conduits supported axon myelination with better myelin quantity and quality when compared to silicone conduits.

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A prostanoid receptor EP4 agonist enhances ectopic bone formation induced by recombinant human bone morphogenetic protein-2.

The anabolic effects of prostaglandin E(2) on bone are effected through the activation of EP4, a G protein-coupled receptor. In the present study, we examined the effects of a prostanoid receptor-selective agonist (ONO-4819) in an experimental system of ectopic bone formation using recombinant human bone morphogenetic protein-2 (rhBMP-2). Collagen pellets containing rhBMP-2 were implanted onto the back muscles of mice and then treated with ONO-4819 administered every 8 h by subcutaneous injection. The ossicles elicited ectopically by rhBMP-2 in mice treated with 30 microg/kg ONO-4819 were significantly larger in size and had a higher bone mineral density and bone mineral content when compared to the controls. We also noted that the anabolic effect of ONO-4819 was seen only in the early phase of the rhBMP-2-induced bone-forming process. These experimental results indicate that the EP4 receptor agonist enhances the rhBMP-2-induced bone formation through a selective effect on early stage mesenchymal cells, which in turn may result in increased responsiveness of the host animals to rhBMP-2.

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Geometrically structured implants for cranial reconstruction made of biodegradable polyesters and calcium phosphate/calcium carbonate.

The aim of this study was the development of a processing pathway for manufacturing of biodegradable skull implants with individual geometry. The implants on the basis of polylactide and calcium phosphate/calcium carbonate were prepared by a combination of hot pressing and gas foaming. On the inside, the implant consists of a macroporous and faster degradable material (poly(D,L-lactide)+CaCO3) to allow the ingrowth of bone cells. The pore size is in the range of 200-400 microm. On the outside, the implant consists of a compact and slower biodegradable material (poly(L-lactide) and calcium phosphate) to ensure mechanical stability and protection. To overcome problems like inflammatory reactions caused by acidic degradation products of polylactide, the polyester was combined with basic filling materials (calcium salts). The filler neutralises the lactic acid produced during polymer degradation and increases the bioactivity of the material. The stabilised pH was demonstrated by long-term in vitro pH studies. Over a time period of 250 d in demineralised water, the pH was in the physiological range. The in vitro biocompatibility was shown by cell cultures with human osteoblasts. A good proliferation of the cells was observed over the whole test period of 4 weeks.

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Tailoring biomaterial compatibility: in vivo tissue response versus in vitro cell behavior.

Biocompatibility relies essentially on surface phenomena, represented by cell-cell, cell-material and material (polymer)-protein interactions. An in vivo and in vitro experimental investigation was carried out on the biomaterials of two different classes with a good potential for in situ utilisation. Non-resorbable (Polypyrrole, Polyaniline, Polyimide) and resorbable (PLLA-PDXO-PLLA) materials for tissue engineering were studied for their overall tissue tolerance and cellular interactions. These non-resorbable polymers conceived for biosensor applications and implantable drug-delivery systems are intrinsically conductive. The PLLA-PDXO-PLLA triblock copolymer showed interesting tensile properties for bone and cartilage tissue engineering due to the presence of 1,5-dioxepan-2-one. In vitro and in vivo parallel studies showed an interesting correspondence: a) the cells in contact with the resorbable material that appeared to be capable of migratory-regenerative aspects in vitro exhibited good compatibility in vivo; whereas b) the non-resorbable materials, which are designed to remain in situ in vivo, were seen to have the potential to represent an adverse factor (inflammation, fibrotic reactions) that correlated with some aspects of cell behaviour in vitro.

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Resorbable plate osteosynthesis of sagittal split osteotomies with major bone movement.

This study evaluates resorbable miniplate osteosyntheses in sagittal split osteotomies with major bone repositioning. Two resorbable 2.0-mm miniplate systems, MacroSorb (Macropore, San Diego, Calif.) and PolyMax (Synthes, Oberdorf, Switzerland), were compared consecutively. Amorphous 70:30 poly-L/DL-lactide copolymer plates sustain continuous hydrolysis through water penetration into the implant body during the first 6 months in situ. This breaks copolymer chains into smaller particles, which later become degraded through phagocytotic cells. Eighteen patients, 10 women and eight men, 16 to 57 years old (average, 27 years) were examined. They had severe dysgnathia caused by congenital craniofacial malformations, systemic disorders, trauma, amelogenesis imperfecta, oligodontia, and other conditions, and they needed five 8- to 10-mm and 13 major 10- to 12-mm repositions. Twelve sagittal split osteotomies were fixed with 12 MacroSorb plates in six patients, and 24 osteotomies were filled with 32 PolyMax plates in 12 patients. Ten mandibular plate, screw, hard-tissue, and soft-tissue specimens were taken at 3, 6, 9, or 12 months postoperatively in secondary operations (e.g., dental implant placement).Follow-up ranged from 4 to 19 months; all osteosyntheses reossified. Four patients showed proximal fragments rotated up to 5 mm sagittally anteriorly and nonaligned burr holes on the postoperative radiogram, suggesting plate fractures or screw pullout. When plate fracture was noted, guided occlusion was maintained 4 weeks after surgery. Occlusal, radiologic, and skeletal results remained stable. After starting fixation with two plates on each side, no more plate fractures were seen. In three other patients, minor skeletal relapses up to 3 mm horizontally resulted. Local histologic inspection of specimens showed thorough osseous union. Screw remnants embedded in bone made screw pullout unlikely; rather, screw-head or plate fractures were found as multiple degraded particles. Microscopy showed a chronic foreign body reaction. Two patients (11 percent) developed a sterile fistula 3 and 4 months after surgery, draining implant debris. Here, the biopsies showed a granulocytic infiltrate that subsided clinically after excisional biopsy. The assignment of MacroSorb plates followed by PolyMax plates was done in an otherwise unchanged treatment protocol. Comparison of the number of patients in each group with stable osteosyntheses and regular healing showed no significant differences by Fisher's exact test (p = 0.1516); therefore, the authors focused on the combined results for both treatments. The current osteosynthesis systems showed sufficient stability for mandibular fixation after sagittal split osteotomy and repositioning more than 10 mm distant when two plates were applied to each side; however, 27 percent of patients had complications, including relapses. Disadvantages were the cost, breakability, diameter, and need to place the screws vertically to the plate, necessitating a bent instrument or transbuccal incisions.

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[Construct tissue-engineered bone by co-seeding marrow stromal cells and endothelial cells].

To identify the role of co-seeding marrow stromal cells (MSCs) and endothelial cells (ECs) onto biocomposite in constructing tissue-engineered bone for promoting osteogenesis and angiogenesis, we seeded rat marrow stromal cells (rMSCs) and human umbilical vein endothelial cells (hUVECs) onto poly (L-lactic acid)/beta-tricalcium phosphate (PLLA/beta-TCP) macroporous composite (the test group), and seeded rMSCs onto the same kind of composite (the control group). We implanted these biocomposites into the thighs of nude mice intramuscularly and sacrificed these mice at 1, 4, 8, 12, 16 weeks after implantation, respectively. Biocomposites were taken out and histomorphometry was performed. Image manipulation technology was used to calculate the percentages of new bone area and material area in the test group and control group. We found that the percentage of new bone area of test group increased faster than that of control group, the percentage of material of test group decreased faster than that of control group, and the growth of capillary network of test group was much better than that of control group. These results suggest that co-seeding MSCs and ECs onto scaffold for constructing tissue-engineered bone is beneficial to osteogenesis, to angiogenesis in the prosthesis, and to the degradation of scaffold, and it is of significance in the research of bone tissue engineering.

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Biodegradable hyaluronic acid/n-carboxyethyl chitosan/protein ternary complexes as implantable carriers for controlled protein release.

An ampholytic N-carboxyethyl chitosan (CEC), with various isoelectric points (IPs), was synthesized by grafting acrylic acid on chitosan utilizing Michael's reaction. Compared to native chitosan, CEC has enhanced water solubility and dramatically accelerated enzymatic degradation; the rate of degradation is proportional to the degree of substitution (DS). The results from turbidimetric titration and fluorescence studies revealed that CEC formed complexes with either hyaluronic acid (HA) or bovine serum albumin (BSA) within a certain pH range. The HA/CEC/BSA ternary complexes could be prepared by colloid titration with quantitative yield and BSA entrapment. The rate of BSA release from the complexes was affected by pH, ionic strength, DS of CEC, and the molecular weight (MW) of HA. The endurance of BSA release from the complexes could be extended up to 20 d by formulating them with high-MW HA and CEC with low DS.BSA release profiles from HA/CEC-2/BSA complexes.

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