BACKGROUND:Local administration of anti-tuberculosis drugs is a commonly used therapy. Due to the rapid absorption, the drugs cannot have the durable therapy effect; therefore, it is necessary to seek an optimal carrier material for the agents. OBJECTIVE:To review the new development for the carrier materials of anti-tuberculosis drugs. METHODS:A computer-based search of PubMed and VIP databases was performed by the first author to search articles related to sustained-released anti-tuberculosis drugs published from January 1990 to December 2014. The key words were osteoarticular tuberculosis; anti-tuberculosis; sustained-released drugs in English and Chinese, respectively. RESULTS AND CONCLUSION:Inorganic materials (calcium phosphate, calcium sulfate), polymer materials (polylactic acid, polyglycolic acid, polylactic-co-glycolic acid) and biomaterials (protein, glutin, alginates, chitin, demineralized bone matrix) are the main three kinds of carrier materials for anti-tuberculosis drugs. These carrier materials have their own advantages and disadvantages, which cannot be the optimal carrier materials. However, the complex of these materials is a promising technology for the optimal carrier materials in the future.

Objective To investigate the effect of two kinds of injected biomaterial combined with bone morphological protein (BMP) on tendon-to-bone healing after anterior cruciate ligament reconstruction (ACL) in rabbits. Methods ACL reconstruction was performed bilaterally in 50 skeletally mature rabbits using long digital extensor tendon grafts. Injected fibrin glue (FG) or injected calcium phosphate cements (CPC) combined with BMP were implanted into the treated knee of each rabbit,with the contralateral knees as controls. Every three rabbits were killed at 2,6 and 12 weeks postoperatively for routine histological studies. The samples were processed through Micro CT and subsequent HE and toludine blue staining. The remaining 16 rabbits were sacrificed at 6 and 12 weeks. Their femur-tendon graft-tibia complexes were harvested for subsequent mechanical testing. Results Imageological results showed that 6 weeks after operation,bone mineral density (BMD) values in FG-bBMP group were higher than those in CPC-bBMP group. However,12 weeks after operation,BMD values in CPC-bBMP group were the highest in the three groups. Histological findings of the interface between the tendon and bone differed in the three groups. Two weeks after operation,more chondrocyte-like cells that were fairly disorganized were noted between the tendon and bone in FG-bBMP group. Six weeks after operation,mature bone formation around the tendon was observed in FG-bBMP group while more immature bone ingrew toward implanted tendon in CPC-bBMP group; at 12 weeks,more chondrocyte cells and new bone formation were seen in CPC-bBMP group while a small quantity of mature bone was around tendon in FG-bBMP group. In biomechanical evaluation,the maximum pull-out load of the tendon from the bone tunnel was significantly higher in the two treatment groups than in the control group 6 and 12 weeks after operation (P

Objective A new type of bone graft material was constructed by combining calcium phosphate cement(CPC) with bone morphogenetic protein(BMP). In order to evaluate the feasibility to use this material to repair the segmental bone defect in clinic, the ability of CPC was compared with CPC/BMP in bone defects reconstruction by animal experiments. Methods The model of 15 mm bone defect was established in the middle shaft of the radius in 60 rabbits, of which 30 defects were implanted with CPC/BMP composites, 22 implanted with CPC, and the other 8 rabbits served as control group. The specimens were harvested separately at the end of 2, 4, 8, 16 and 24 weeks after operation. In order to observe the formation of new bone and the degradation process of the material, a series of examinations were carried out including of radiography, histomorphology, serum detection, energy dispersion analysis X-ray(EDAX), scanning electron microscope(SEM),bone density detection, mechanical measurement and inorganic substance detection. The results of CPC group and CPC/BMP group were compared on the same condition. Results All the animals survived after operation, and no reactions of toxicity were found. New bone formation was observed to be increasing significantly in CPC/BMP group with the time of implantation. Only little new bone formed in CPC group and no healing was found in the control group. By the end of 24 weeks, new bone had bridged the gap between the proximal and distal fragments in CPC/BMP group. In histomorphological detection, chondrocytes were found at the 2nd week, and woven bone at 4th week in CPC/BMP group. Remodeling of new lamellar bone and absorption of the composite material were observed at the 16th week, and the mechanical strength of the composite material reached almost to normal level at the 24th week. Calcification was significantly higher in CPC/BMP group than that in CPC group examined by EDAX, new bone density detection and measurement of inorganic substance in specimens. During the repairing process of bone defect, the material degraded while new bone formed, the speed of degradation of CPC/BMP was evidently higher than that of CPC group. Moreover, in the process of CPC degradation, the concentration of calcium in serum increased, and the concentration of phosphate in serum kept unchanged. Conclusion The CPC/BMP composite has great potential in bone defects repairing and could be used as a material for bone graft substitute in clinical patients.

Objective To observe histomorphology of neo-cartilage obtained with tissue-engineering by combining calcium alginate gel and in vitro cultrue of chondrocytes. Methods Articular chondrocytes from the knee joints of 2-week-old New Zealand white rabbits were harvested, expanded in cell culture, and following the second generation of the culture, the chondrocyte suspension was mixed with calcium alginate gel resulting in a cell density of 5? 10 6/ml. Finally the mixture, which contained 1％ sodium alginate, 40 mmol/L calcium gluconate, 0.135 mol/L NaCl, 0.1 mol/L K 2HPO 4 and 5? 10 6 chondrocytes per milliliter, were injected into the dorsal subcutaneous tissue in rabbits of experimental groups (A and B). Animals of the control groups (C and D) were injected with calcium alginate without chondrocytes or chondrocytes without calcium alginate. Specimens were harvested at the 2nd and 4th week after injection, and stained with HE and toluidine blue. Results In the HE stained specimens in the experimental groups, proliferation of chondrocytes was demonstrated at the 2nd week and the formation of neo-cartilage at the 4th week after injection of calcium alginate chondrocyte-composite. Toluidine blue stained specimens showed positive staining of chondrocytes and cartilage matrix of neo-cartilage. In some animals injected with chondrocytes without calcium alginate, relative small amount of neo-cartilage was also formed and no neo-cartilage was observed in animals injected with calcium alginate without chondrocytes. Conclusion Injectable calcium alginate-chondrocyte-composite can induce tissue engineered neo-cartilage in allogenic animal.

[Objective]To study the differentiation of adipose-derived adult stem cells(ADASCs)into chondrocytes induced by TGF-?1 in the presence of different concentrations of serum in vitro.[Method]Adipose tissue-derived adult stem cells were isolated from rabbits subcutaneous adipose tissue using enzymatic digestion.The adipose tissue was minced and digested with collagenase type I,and the released cells were collected by density centrifugation and then placed in culture.After being passaged three times,ADASCs were induced to differentiate into chondrocytes by chondrogenic culture media containing 1% or 10% of FBS for 2 weeks.MTT,alkaline phosphatase toluidine blue staining and type Ⅱ collagen immunohistochemistry staining were then tested in both groups.Gray value was analyzed by Leica patho-image analysis system to observe if there was any difference in differentiative status between the two groups.[Result]MTT showed that the cells of 10% FBS group had higher proliferation activity than those of 1% FBS group(P

BACKGROUND: Matrix material for cartilage tissue engineering exhibits too fast or too slow chondrocytes degradation in vivo, affecting tissue regeneration and shaping reconstruction, which has troubled scholars.OBJECTIVE: To amply bone marrow stromal cells (BMSCs) and induce them to chondrocytes in vitro, so as to study the feasibility of repairing articular cartilage defects in rabbits using poly(lactic-co-glycolic acid) (PLGA) loaded with BMSC-derived chondrocytes.DESIGN, TIME AND SETTING: Comparative experiment was performed at the Institute of Orthopaedics in the Fourth Military Medical University of Chinese PLA and the Center Laboratory of the Airforce General Hospital of Chinese PLA between June 2002 and June 2008.MATERIALS: A total of 36 two-month-old New Zealand white rabbits were used, and 4-6 mL bone marrow was aspirated from bilateral femoral trochanters in each animal. Primary culture and subcultures were done. In subcultures, the medium contained bone morphogenetic protein-2 (100 μg/L.), and high polymer hyaluronic acid was spread on bottom of the culture flasks in advance.In this way, the BMSCs were induced into chondrocytes and the third passage of cells at the adjusted density of 2.0×10~(10)/L wereco-cultured with PLGA for 24 hours, then PLGA-cell composites were prepared.METHODS: A defect of 4-mm in diameter and reaching medullary cavity were created in femoral condyles of 36 rabbits, and 36 right knees were treated with PLGA-cell composites, serving as experimental group, while 18 left knees with PLGA only as material group, and the other 18 knees remained untreated, as blank control group.MAIN OUTCOME MEASURES: At 4,8,12, 24 weeks after operation, the animals were euthanized and the newly formed tissues were observed macroscopically and microscopically, graded histologically, and analyzed statistically.RESULTS: Material group and blank control group shared identical outcomes of gross and histological observation, thus assigning into a control group. In the experimental group at 24 weeks, the defects were filled with white translucent cartilage tissue which appeared smooth and tenacious. The color and the luster were similar to that of normal cartilage, and was ill-demarcated from the surrounding normal cartilage. The cells on the surface paralleled to joint surface. Though the cells in the deep layer arranged disorderly, they tended to align vertically. The matrix was extensively stained. The subchondral bone formed.The tide mark basically recovered, and the new cartilage integrated with normal cartilage finely. In the control group, chondrocytes proliferated in the border, but in the bottom, there were mainly fibrous tissues. The histological grade of 12 and 24 weeks was different significantly from that of 4 and 8 weeks (P < 0.01). There were also significant differences between experimental group and control group at each time intervals after operation (P < 0.01).CONCLUSION: BMSCs were successfully induced into chondroncytes by use of bone morphogenetic protein-2 and high polymer hyaluronic acid. PLGA can be degraded and absorbed gradually while new cartilage tissues form. It can be used as a suitable scaffold material for repairing articular cartilage defects in tissue engineering.

Objective: To prepare surface with collagen, so as to increase the histocompatibility of deantigenic bovine cancellous bone. Methods: Bovine cancellous bone was degreased and macerated by H 2O 2 and decalcified with 0.3 mol/L HCl to form scaffold, surface of which was treaed with collagen, then cultured rabbit osteoblasts were seeded into the scaffold and cultured in vitro . 4, 10, and 21 days later, the cells on the composite materials were observed under sanning election microscope. Results: On day 4, the cells grew in the surface of the scaffold; on day 21, the cells grow into the pores and on all of the surace of the scaffold.Conclusion: Deantigenic bovine cancellous bone with surfaces treated by collagen is histocompatable with rabbit osteoblasts.

Normal cells have limited proliferation ability.After certain cycles of proliferation,they will lose the response ability to growth factors and finally cease division and start the course of aging. In current opinion,lacking of the terminal end of a chromosome(telomere)is the cause for cells to loss the proliferation ability and leads cells to aging and death.The human telomerase catalytic subunit 1(hTERT)can activate telomemse which prolong DNAs of the terminal end of chmmosome and help cells gain genomic stabilization.The discoveries of telomere,telomerase and hTERT provide new idea for studying of cell aging and the findings are also applied in the establishment of immortal cell line. Also they may play an important role in the studv of biological feature of seed cell in tissue engineering and the establishment of cell bank.

To develop a biodegradable implant with both the ossific and anti infectious abilities. Antigen extracted bovine cancellous bone combined with bBMP as the center carrier was coated with either gelatin or poly ? caprolaton (PCL). They were then compounded with gentamicin. Their in vivo gentamycin releasing characteristics were assayed. There was an initial phase of rapid release of gentamycin within 24 hours, followed by a second phase of sustained but gradual diminution of drug release. Effective release of gentamicin from the PCL encapsulated RBX G LDDS may last 30 days in vivo . There was no significant difference in releasing rate between 30% gelatin and 40% gelatin in statistic. The results showed that RBX gentamycin possessed a fine in vivo gradual release property. Collaborated with its ossification activity, it could be considered as an ideal repair material for open fracture with bone defect.

[Objective]To compare the purity and extraction rate of four purecollagen type I products prepared from various biomaterials by a limited enzyme digestion method for the use in tissue engineering.[Method]Bovine cortical bone, bovine achilles tendon, porcine achilles tendon and porcine skin were splitted into pieces of 0.2-0.5 mm. After being immersed in glacial acetic acid, they were extracted with pepsin. Then the crude products were dissolved, centrifuged, dialyzed and freeze drying to prepare pure collagen type I. The final products were confirmed by absorbance, amino acid analysis and'SDS-PAGE electrophoresis comparing them with the products of Sigma Company.[Result]The wave length of maximum absorbance of the final products was 230 nm, and the amino acid analysis and SDS-PAGE electrophoresis confirmed that the final products were collagen type I. The purity of product extracted from bovine cortical bone was the highest (96.12%) and higher than that from Sigma Company. The extraction rate of bovine achilles tendon collagen was the highest (75.34%).[Conclusion]Collagen type I of higher purity and higher extraction rate can be prepared using a limited enzyme digestion method.And the product from bovine cortical bone is better than the others,which has a promising prospect.