From the results of this experiment, it showed that the implanted tendon was gradually extruded from the tibia hole and attached to the periosteum. The dominant breeding of tissue cells, cytodynamics, the perimeter ratio of tendon/bone and the effect of revascularization were discussed in detail.
In order to observe the role of genetically modified Schwann cell (SC) with pSVP0Mcat in the regeneration of injured spinal cord, the cells were implanted into the spinal cord. Ninety SD rats were used to establish a model of hemi-transection of spinal cord at the level of T8, and were divided into three groups, randomly, that is, pSVP0Mcat modified SC implantation (Group A), SC implantation (Group B) and without cell implantation as control (Group C). After three months the presence of axonal regeneration of the injured spinal cord was examined by means of horseradish peroxidase (HRP) retrograde labelling technique and stereography. The results indicated that HRP labelled cells in Group A and B could be found in the superior region of injured spinal cord and the brain stem such as the red nuclei and oculomotor nuclei. The density of ventral hom neurons of the spinal cord and the number of myelinated axons in 100 microns of the white matter was A gt; B gt; C group. In brief, the pSVP0Mcat modified SC intraspinal implantation could promote regeneration of the injured spinal cord.
OBJECTIVE: To evaluate the clinical effect of drilling procedure following the hydroxyapatite orbital implantation. METHODS: From February 1996 to April 2000, 146 consecutive patients who received hydroxyapatite orbital implant were drilled and inserted a motility peg 6 to 16 months after hydroxyapatite implantation. Among them, there were 97 males and 49 females, aged from 18 to 60 years old, of the 146 motility pegs, 36 were sleeved pegs and 110 were nonsleeved. Goldman visual field analyzer was applied to measure the degree of artificial eye’s movement before and after drilling. RESULTS: Followed up for 1 to 40 months, no secondary infection occurred. The mobility of the prosthesis increased from (18.7 +/- 3.8) degrees preoperatively to (42.3 +/- 3.7) degrees postoperatively. CONCLUSION: The delayed drilling procedure and motility peg insertion improve the range of movement and the sensitivity of the artificial eye with a low rate of complications.
Objective To design a novel stentless porcine aortic bioprosthesis and test the feasibility and its function in vitro after the valve was implanted by a modified method. Methods Six stentless porcine aortic bioprosthesis were divided into two groups according to different implantation, single layer suture group: new improvement stentless porcine aortic bioprosthesis sutured with single layer was implanted; double layer suture group: stentless porcine aortic bioprosthesis developmented by our laboratory used double layer suture was implanted. Each group contained three scales: 23 mm ,25 mm and 27 mm. Analogue ex vivo aortic valve replacement was performed , the feasibility of the new implantation was detected. Effective orifice area, transvalvular pressure gradient and regurgitation ratio were recorded at the cardiac output of 2.0 L/min, 3.5 L/min, 5.0 L/min and 7.0 L/min under the guideline of International Organization for tandardization （ISO）5840. Results The average aortic valve implantation time used for single layer suture and tradition double layer suture were 50 min and 70 min respectively. The transvalvular pressure gradient in the single layer suture group were significantly lower than those in double layer suture group under the flow of 5.0 L/min in 23 mm valve and 27 mm valve （13.51±0.51 mm Hg vs. 14.44±0.99 mm Hg, 7.36±0.19 mm Hg vs. 7.53±0.28 mm Hg；P＜0.01）;and the effective orifice area in the single layer suture group were larger than those in double layer suture group in the same case(1.87±0.06 cm2 vs. 1.76±0.08 cm2, 2.26±0.07 cm2 vs. 2.16±0.05 cm2；P＜0.01). There was no statistically difference in other parameters between both groups. Conclusion The novel design of new improvement stentless porcine aortic bioprosthesis used single layer suture has good hemodynamic characteristics as the nature structure . The modified suture method decrease the implantation time.Nemerical data of the evaluation in vitro show that the difference between single layer suture group and double layer suture group in effective orifice area,transvalvular pressure gradient and regurgitation ratio haveno statistical significance. This experiment is the foundation of the animal and clinical experiment in the future.
Objective To explore the effective autologous bone marrow stem cell dosage for treatment of severe lower limb ischemia. Methods From December 2003 to December 2004, 22 cases of bilateral lower limb ischemia were treated with autologous bone morrow cell transplantation. All the patients were randomly divided into two groups according to ischemia degree. In group A(severe ischemia side), the amount of transplanted autologous bone marrow cells was more than 1×108, and ingroup B(mild ischemia side), the amount was less than 1×105. A series of subjective indexes, such as improvement of pain, cold sensation and numbness, and objective indexes, such as increase of ankle/brachial index (ABI) and transcutaneous oxygen pressure (TcPO2), angiography, amputation rate, and improvement of foot wound healing were used to evaluate the effect of autologous bone marrow stem cells implantation. Results The rates of pain relief were 90.0% in group A and 16.7% in group B (Plt;0.01); the rates of cold sensation relief were 90.5% in group A and 5.3% in group B（Plt;0.01）;the improvement of numbness was 62.5% in group A and 9.1% in group B（Plt;0.01）. Increase of ABI was 31.8% and 0 in groups A and B respectively（Plt;0.01） at 4 weeks after implantation. Increase of TcPO2was 94.4% and 11.1% in groups A and B respectively（Plt;0.01） at 4 weeks after implantation. Twelve cases of angiography showed rich new collateral vessels in 100% of the limbs in group A while no remarkable new collateral vessel in group B. The amputation rates were 4.5% in group A and 27.3% in group B（Plt;0.05） at 4 weeks after implantation. The rate of improvement of foot wound healing was 75% in group A and there was no changein wound healing in group B after 4 weeks of implantation. Conclusion The effectiveness of autologous bone marrow stem cell implantation depends on the number of implanted stem cells. Effectiveness is expected in most patients if the implanted stem cell is more than 1×108, whereas there would be little effect if the cell number is less than 1×105.
Objective To investigate the biocompatibility of acellular urinary bladder submucosa (AUBS). Methods The acellular collagen matrix of human urinary bladder submucosa was developed using freeze-thawed enzymatic treatment and freeze-drying technique. Human oral keratinocytes were cultured and seeded on AUBS at a density of 2×106/ml in vitro.The proliferation of the cells were observed. Pockets were created in the abdominal muscle wall of 18 SD rats. AUBS in size 1 cm×1 cm was implanted into the pocket. The grafts were observed by light microscope 3, 6, 10, 14, 21 and 28 days after operation. Results AUBSmainly consisted of collagen fibers with a three-dimensional network structure. After the oral keratinocytes were seeded, continous oral epithelium layer was formed on the surface of AUBS after 10 days in vitro. Histological observation of the grafted AUBS showed progressive cell infiltration at 6 days. New capillaries formed at 14 days. The collagen fibers arranged regularly at 28 days after implantation. Conclusion Freeze-dried AUBS may be used as a suitable scaffold for tissue regeneration, which can induce cell proliferation both in vivo and in vitro and has good biocompatibilty.
Objective To investigate the feasibility and characteristic of tissue engineered testicular prosthesis with highdensity polyethylene(HDPE，trade name: Medpor) and polyglycolic acid(PGA). Methods The chondrocytes were isolated from the swine articular.The PGA scaffold was incorporated with medpor which semidiameters were 6mmand 4mm respectively.Then, the chondrocytes (5×10 7/ml) were seeded onto Medpor-PGA scaffold and cultured for 2 weeks. The ten BALB／C mice were divided into two groups randomly(n=5). In the experimental group, the cell-scaffold construct was implanted into subcutaneous pockets on the back of nude mice. In the control group, the Medpor-PGA scaffold was implanted. The mice of two groups were sacrificed to harvest the newly formed cartilage prosthesis after 8 weeks. Macroscopy, histology and immunohistochemistry observations were made. Results The gross observation showed that on changes were in shape and at size, the color and elasticity were similar to that of normal cartilage and that the cartilage integrated with Medpor in the experimental group; no cartilage formed and fiberlike tissue was found in the control group. HE staining showed that many mature cartilage lacuna formed without blood vessel and some PGA did not degradated completely. Toluidine blue staining showed extracellular matrix had metachromia. Safranin O-fast green staining showed that many proteoglycan deposited and collagen type Ⅱ expression was bly positive. In the control group, Medpor was encapsulated by fiber tissue with rich blood vessel. Conclusion The newly formed complex of Medpor-PGA and cells was very similar to testicle in gross view and to normal cartilage in histology. This pilot technique of creating testicular prosthesis by incorporating tissue-engineered cartilage with Medpor demonstrated success.
ObjectiveTo construct large block of engineered liver tissue by co-culture of fibroblasts and hepatocytes on collagen hydrogels in vitro and do in vivo implantation research.
MethodsSilastic mould was prepared using three-dimensional printing technology. The collagen hydrogel scaffold was prepared by collagen hydrogel gel in the silicone mould and was removed. Sprague Dawley rat lung fibroblasts were co-cultured with primary hepatocytes at a ratio of 0.4:1 on the collagen hydrogel scaffold to construct large block of engineered liver tissue in vitro (group B), and primary hepatocytes cultured on the collagen hydrogel scaffold served as control group (group A). The cell morphology was observed, and the liver function was tested at 1, 3, 7, 14, and 21 days after culture. The rat model (n=24) of hepatic cirrhosis was made by subcutaneous injection of carbon tetrachloride. And in vivo implantation study was carried in cirrhosis rat model. The phenotypic characteristics and functional expression of hepatocytes were evaluated at 3, 7, 14, 21, and 28 days after implantation.
ResultsIn vitro results indicated that hepatocytes in group B exhibited compact polyhedral cells with round nuclei and high expression of liver function. Moreover, cells aggregated to the most at 7 days. Album production and urea synthesis incresed significantly when compared with group A (P<0.05). In vivo results showed hepatocytes in group B survived for 28 days, and albumin production and urea synthesis were significantly increased. In addition, hepatocytes showed an aggregated distribution and cord-like structures, which was similar to normal liver tissue.
ConclusionThe large block of engineered liver tissue constructed by co-cultured cells can form tissue similar to normal liver tissue in vivo, and survive for a long time, laying foundations for building more complete engineered liver tissue in the future.
Objective To investigate the possibility of repairing defected tendon with a tissue engineering tendon, combined culture of allogenous tenocyte and derived tendon. Methods Macaca tenocytes labelled by BrdU were seeded on the derived tendon. The flexor digitorum profundus of five fingers of left hand in 15 Macaca mulatta were resected and made 2.5cm defects as experimental model. They were divided into three groups according to repair methods (Group A: Combined culture of derived tendon materials and alloggenous tendon cells; Group B; Derived tendon materials; Group C; Autograft). In different stages, the labeled BrdU of tendon cells were observed. Results In Groupo A, after iin vivo implantation, the tenocytes could proliferate and synthesize collagen; the new tissue was white and glossy and the collagen fibers fused to form dense tendon structure as several weeks passed. Twelve weeks after implantation, the tenocytes still survived and synthesized collagen, the results of labelled cells were positive by immunothistochemical methods. By scanning electron microscopic observation, the tenocytes arraged regularly and evely among the derived tendon; the collagen fibers formed a network and its main direction was accord with that of the derived tendon. Normal nucleus, nucleolus, and cell organelles were seen under transmission electron microscope. Conclusion Combined culture of tenocytes with derived tendon is able to make tendon like tissue. The structure of tissue engineering tendon in similar to that of normal tendon.
The aim of this experiment was to study the osteogenesis in vivo of allogenic osteoblast combined culture with calcium phosphate composites. The osteoblasts were obtained by enzymatic digestion of periosteum from fibula subcultured to 13 generations, the cells were combined culture with hydroxyapatite and biphasic calcium phosphate. Subseguently, the composite was implanted into rabbits subcutaneously or intramuscularly. The blank material was implanted in the contralateral side as control. Four weeks later, all animals were sacrificed. All the implants were examined by gross observation, histological examination and EDXA. The results showed: 1. obvious ingrowth of connective tissue with very little inflammatory reaction; 2. new bone formation in the composites with deposit of Ca and P on the surface of osteoblast, but none in the blank materials; 3. no significant difference of new bone formation between the different sites of implantation or different materials, but those implanted intramuscularly had lamellae form of new bone while those implanted subcutaneously had only mineralization of extracellular matrix. The conclusion were: 1. the composites are biocompatible with prior osteogenesis property; 2. periosteal-derived allogenic osteoblasts obatined by enzymatic digestion could survive following implantation with bioactivity; 3. rich blood supply might be advantageous to new bone formation and its maturation.
