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Biomedical subjects

Zhuo Xiong

Publications and source records attributed to Zhuo Xiong.

11 recordsLinked to original sources

Generation of three-dimensional hepatocyte/gelatin structures with rapid prototyping system.

Using rapid prototyping technology, three-dimensional (3D) structures composed of hepatocytes and gelatin hydrogel have been formed. This technique employs a highly accurate 3D micropositioning system with a pressure-controlled syringe to deposit cell/biomaterial structures with a lateral resolution of 10 microm. The pressure-activated micro-syringe is equipped with a fine-bore exit needle for which a wide variety of 3D patterns with different arrays of channels (through-holes) were created. More than 30 layers of a hepatocyte/gelatin mixture were laminated into a high spacial structure using this method. The laminated hepatocytes remained viable and performed biological functions in the construct for more than 2 months. The rapid prototyping technology offers potential for eventual high-throughout production of artificial human tissues or organs.

Animals↗

Repair of the radial defect of rabbit with polyester/tricalcium phosphate scaffolds prepared by rapid prototyping technology.

OBJECTIVE: To evaluate the effects of repairing rabbit radial defects with polyester/tricalcium phosphate scaffolds prepared by rapid prototyping technology loaded with bovine bone morphogenetic protein (bBMP), and find new carriers for growth factors. METHODS: Polyester/tricalcium phosphate scaffolds prepared by rapid prototyping technology loaded with and without bovine BMP were used to repair the 15 mm radial defect in rabbit. Then the results of radiography, histology, scaffolds degrade rates and bone mineral density (BMD) were appraised to examine the effects at the 12th week. RESULTS: At the 12th week postoperatively, all defects treated with bBMP were radiographically repaired. No radius implanted polyester/tricalcium phosphate scaffolds without bBMP showed radiographic and histological union. At experimental groups, longitudinal alignment of lamellar structure was observed histologically at the 12th week, indicating that remodeling of regenerated bone was complete in different degree. Of the three experimental groups, the bony regeneration and remodeling of callus in poly lactide-co-glycolide/tricalcium phosphate (PLGA/TCP) group was the best. The BMD values were beyond 70% of normal value at the 12th week while the PLGA/TCP scaffolds group was the highest, and no abnormalities were observed in the surrounding soft tissue in all groups. CONCLUSIONS: Polyester/tricalcium phosphate scaffolds prepared by rapid prototyping technology loaded with bovine BMP can repair a 15 mm radial defect of rabbit. As for the results, the PLGA/TCP scaffold is ideal and better than poly L-lactide-co-D, L-lactide (PDLLA/TCP) scaffold, but the ploy L-lactic acid (PLLA/TCP) is not so good for its low degradation rates.

Animals↗

[Repair of the radial defect of rabbit by polyester/tricalcium phosphate scaffolds prepared by rapid prototyping technology].

OBJECTIVE: To evaluate the repairing effect of the rabbits radial defects of by polyester/tricalcium phosphate scaffolds prepared by rapid forming technology loaded with bovine BMP, and find a new carrier for growth factor. METHODS: Polyester/Tricalcium phosphate scaffolds prepared by rapid prototyping (RP) technology loaded with and without bovine BMP were used to repair the 15 mm radial defect of rabbit. Then results of radiography, histology, scaffolds degrade rates and bone density were appraised to examine the repairing effects of the scaffolds at 12 weeks. RESULTS: At 12 weeks, all defects treated with bBMP were radiographically repaired. No radii implanted polyester/tricalcium phosphate scaffolds alone showed radiographic and historical union. At experimental groups, longitudinal alignment of lamellar structure was observed histologically at 12 weeks, indicating that remodeling of regenerated bone almost completed, the scaffolds degradation rates were different by 12 weeks, and no abnormalities were observed in the surrounding soft tissue in all groups. CONCLUSION: Polyester/tricalcium phosphate scaffolds prepared by rapid prototyping technology loaded with bovine BMP can repair the rabbits radical defects. As for the effects, the poly (L-lactic-co-glycolide)/tricalcium phosphate (PLGA/TCP) scaffold are ideal and better than poly (L-lacide-co-D, L-lactide)/tricalcium phosphate (PDLLA/TCP) scaffold, but the poly (L-lactic acid)/tricalcium phosphate (PLLA/TCP) is not so good for its low degradation rates.

Animals↗

Fabrication of viable tissue-engineered constructs with 3D cell-assembly technique.

We have recently developed an organ manufacturing technique that enables us to form cell/biomaterial complex three-dimensional (3D) architectures in designed patterns. This technique employs a highly accurate 3D micropositioning system with a pressue-controlled syringe to deposit cell/biomaterial structures with a lateral resolution of 10 microm. The pressure-activated micro-syringe is equipped with a fine-bore exit needle using which a wide variety of 3D patterns with different arrays of channels (through-holes) were created. The channels can supply living cells with nutrients and allow removing the cell metabolites. The embedded cells remain viable and perform biological functions as long as the 3D structures are retained. The new technology has the potential for eventual high-throughput production of artificial human tissues and organs.

Animals↗

Preparation and evaluation of ammonia-treated collagen/chitosan matrices for liver tissue engineering.

To further enhance the properties of existing collagen/chitosan scaffolds for liver tissue engineering, a very simple method was developed to form noncovalently linked mimic of the liver extracellular matrices. Collagen/chitosan mixtures in various proportions (i.e., 1:0, 3:2, 1:1, 2:3, and 0:1 v/v) were lyophilized or evaporated to form sponges or flat films before they were gelled using an aqueous 25% ammonia solution. The porosities of the obtained sponges were above 90% with various pore sizes. The highest mechanical strength (1.9+/-0.7 MPa) and the lowest degradation time (65+/-1.7 days) were achieved by the collagen/chitosan (1:1) matrices. Hepatocytes cultured on the collagen/chitosan (1:1) matrices exhibited relatively high glutamate-oxaloacetate transaminase and glucose secretion functions 25 days post-seeding. Nuclues of the hepatocytes were more elongated and arranged in certain directions on the 1:1 matrices. The cytocompatibility and enhanced biostability of our new ammonia-treated collagen/chitosan matrices suggest that they could be used as scaffolds for liver tissue engineering.

Amines↗

Preparation and characterization of a collagen/chitosan/heparin matrix for an implantable bioartificial liver.

A new type of collagen/chitosan/heparin matrix, fabricated by gelation of collagen/ chitosan with heparin sodium containing ammonia, was produced to construct livers by tissue engineering and regenerative engineering. The obtained collagen/chitosan/heparin matrix was found to be highly porous, swelled rapidly in PBS solution and was stable in vitro for at least 60 days in collagenase/lysozyme containing buffered aqueous solution (PBS, pH 7.4) at 37 degrees C. The collagen/chitosan/heparin matrix resulted in a superior blood compatibility compared to the ammonia-treated collagen and collagen/chitosan matrices. The morphology and behavior of the cells on the collagen/chitosan/heparin membrane were found to be similar to those on the collagen membrane but different from those on the collagen/chitosan membrane. Hepatocytes cultured on the collagen/chitosan/heparin matrices exhibited highest urea and triglyceride secretion functions 25 days post seeding. These results suggest that this collagen/chitosan/heparin matrix is a potential candidate for liver tissue engineering.

Animals↗

[Tissue engineered bone regeneration of periosteal cells using recombinant human bone morphogenetic protein 2 induce].

OBJECTIVE: To investigate bone regeneration of the cell-biomaterial complex using strategies of tissue engineering based on cells. METHODS: Hydroxyapatite/collagen (HAC) sandwich composite was produced to mimic the natural extracellular matrix of bone, with type I collagen serving as a template for apatite formation. A three-dimensional poly-porous scaffold was developed by mixing HAC with poly(L-lactic acid) (PLA) using a thermally induced phase separation technique (TIPS). The rabbit periosteal cells were treated with 500 ng/ml of recombinant human bone morphogenetic protein 2 (rhBMP-2), followed by seeded into pre-wet HAC-PLA scaffolds. Eighteen 3-month nude mice were implanted subcutaneously cell suspension (group A, n = 6), simple HAC-PLA scaffold (group B, n = 6) and cell-biomaterial complex (group C, n = 6) respectively. RESULTS: Using type I collagen to template mineralization of calcium and phosphate in solution, we get HAC sandwich composite, mimicking the natural bone both in composition and microstructure. The three dimensional HAC-PLA scaffold synthesized by TIPS had high porosity up to 90%, with pore size ranging from 50 microm to 300 microm. SEM examination proved that the scaffold supported the adhesion and proliferation of the periosteal cells. Histology results showed new bone formation 8 weeks after implantation in group C. The surface of group A was smooth without neoplasma. Fibrous tissue invasion occurred in group B and no bone and cartilage formations were observed. CONCLUSION: The constructed tissue engineering bone has emerged as another promising alternative for bone repair.

Animals↗

Effect of type I collagen on the adhesion, proliferation, and osteoblastic gene expression of bone marrow-derived mesenchymal stem cells.

OBJECTIVE: To investigate the effects of porous poly lactide-co-glycolide (PLGA) modified by type I collagen on the adhesion, proliferation, and differentiation of rabbit marrow-derived mesenchymal stem cells (MSCs). METHODS: The third generation MSCs isolated from mature rabbits by density gradient centrifugation were cultured at different initial concentrations on 0.3 cm x 1.2 cm x 2.0 cm 3-D porous PLGA coated by type I collagen in RPMI 1640 containing 10% fetal calf serum, while cultured on PLGA without type I collagen as control. The cells adhesive and proliferative behavior at 7, 14, and 21 days after inoculation was assessed by determining the incorporation rate of [(3)H]-TdR. In order to examine MSCs differentiation, the expression of osteoblasts marker genes, osteocalcin (OCN), alkaline phosphatase (ALP), osteopontin (OPN) mRNA, were evaluated by reverse transcription-polymerase chain reaction (RT-PCR), and further more, the cell morphology at 21 days was also observed by scanning electron microscope (SEM). RESULTS: Type I collagen promoted cell adhesion on PLGA. The valve was significantly higher than controls (6 h, 2144 cpm+/-141 cpm vs. 1797 cpm+/-118 cpm, P=0.017; 8 h, 2311 cpm+/-113 cpm vs. 1891 cpm+/-103 cpm, P=0.01). The cells which cultured on PLGA coated with type I collagen showed significantly higher cell proliferation than controls on the 7 th day (1021 cpm+/-159 cpm vs. 451 cpm+/-67 cpm, P=0.002), the 14th day (1472 cpm+/-82 cpm vs. 583 cpm+/-67 cpm, P<0.001) and 21 th day (1728 cpm+/-78 cpm vs. 632 cpm+/-55 cpm, P<0.001). Osteoblasts markers, OCN, ALP, OPN mRNA, were all detected on PLGA coated by type I collagen on the 21 th day, but OCN, OPN mRNA could not be found in controls. Spindle and polygonal cells well distributed on the polymer coated by type I collagen while cylindric or round cells in controls. CONCLUSIONS: Type I collagen is effective in promoting the adhesion, proliferation and differentiation of MSCs on PLGA.

Biocompatible Materials↗

Development of a porous poly(L-lactic acid)/hydroxyapatite/collagen scaffold as a BMP delivery system and its use in healing canine segmental bone defect.

A hydroxyapatite/collagen (HAC) composite was produced to mimic the natural extracellular matrix of bone, with the collagen serving as a template for apatite formation. A three-dimensional highly porous scaffold was developed by mixing HAC with poly(L-lactic acid) (PLA) using a thermally induced phase separation technique. Naturally derived bovine bone morphogenetic protein (bBMP) was incorporated into the porous HAC-PLA scaffolds, and the composite then was implanted in diaphyseal defects (2 cm in radius) of adult beagle dogs. Controls were implanted with scaffolds without BMP. The dogs were sacrificed at 6 months, at which time biocompatibility, biodegradability, and osteoinduction were evaluated by histologic and radiologic examination and by bone mineral density (BMD) measurements. All defects healed after treatment with BMP combined with HAC-PLA, and BMD at the site of the defect was higher than the BMD of the intact radius. Fibrous union developed in the control group animals. Histologic observation indicated that the presence of BMP not only promoted osteogenesis but that it also accelerated degradation of the biomaterials. Optimized design parameters of a three-dimensional porous biomaterial would give full scope to the role of BMP as an osteoinductive growth factor.

Animals↗

[Effects of collagen I coating on the porous poly-lactide-co-glycolid on adhesion, proliferation, and differentiation of mesenchymal stem cells].

OBJECTIVE: To investigate the effects of collagen I on the adhesion, proliferation, and differentiation of MSCs on PLGA. METHODS: Collagen I was added onto the surface of pores in pieces of 3-D porous poly-lactide-co-glycolid (PLGA). Bone marrow-derived mesenchymal stem cells (MSCs) were obtained from New Zealand rabbits and were cultured for 3 generations, inoculated into the pores of PLGA pieces with the volume of 0.3 cm x 1.2 cm x 2.0 cm, and then cultured in solution with [(3)H]-thymidine deoxyribose (TdR). PLGA pieces not coated by collagen I were used as controls. The incorporation rate of [(3)H]-TdR was detected 2, 4, 6, and 8 hours, and 7, 14, and 21 days after culture, shown in count per minute (CPM) value, to determine the adhesion and proliferation of the MSCs. RT-POCR was used to examine the expressions of mRNA of the osteoblast markers: osteocalcin (OCN), alkaline phosphatase (ALP), and osteopontin (OPN). Scanning electron microscopy (SEM) was used to observe the morphology of MSCs. RESULTS: The CPM value since 6 hours after culture between the experimental group and control group began to be significantly different (both P < 0.05) The CPM values 7, 14, and 21 days after culture between the experimental group and control groups (P < 0.05 or P < 0.01). OCN, ALP, and OPN mRNA were expressed in MSCs of the experimental group and only ALP mRNA was weakly expressed in the control group. SEM showed the distribution of spindle and polygonal cells in the pores of the 3-D PLGA pieces and distribution of cylindrical or round cells in the control group. CONCLUSION: Collagen I is effective in promoting the adhesion, proliferation, and differentiation of MSCs on PLGA.

Animals↗

[Rapid manufacturing of degradable porous polymer bone scaffold].

Making bone scaffold through tissue engineering method presents a new choice for both the patients and the doctors of orthopaedics. The biodegradable polymer PLA is chosen to make porous fundus scaffold jetting through special designed nozzle on multi-functional rapid prototyping machine controlled by computer according to the CT data CAD model. The scaffold is then chemically aggregated to compound with collagen-hydroxyapatite, and the ideal bone repair material is obtained. Animal experiment has indicated the correctness of this conclusion.

Absorbable Implants↗