OBJECTIVE: To observe the effects of silks on attachment, shape and function of chondrocytes cultured in vitro. METHODS: The silks from silk worm cocoons were digested by trypsin and coated with polylactic acid to from three dimensional scaffolds for rabbit rib chondrocyte culture. The growth and shape of chondrocytes were observed with phase contrast microscopy, scanning electron microscopy. RESULTS: The chondrocytes were adhered to silks slowly after chondrocytes were seeded into silk scaffolds and cells fixed on silks well 1 or 2 days later. Cells began to proliferate after 3 days and multiplicative growth was observed on the 6th day. Microholes of silk scaffolds were filled with chondrocytes 2 weeks later. Scanning electron microscopy showed that there was a lot of extracellular matrix surrounding cells. CONCLUSION: Silks are ideal for attachment, growth and function maintenance of chondrocytes, and silks can be used as scaffolds for chondrocytes in three dimensional culture.
OBJECTIVE This paper aims to investigate the suitable cell density and the best formation time of tissue engineered autologous cartilage and to provide theoretical basis and parameters for clinical application. METHODS The chondrocytes isolated from mini swines’ ears were mixed with injectable biocompatible matrix (Pluronic), and the density of cell suspensions were 10, 20, 30, 40, 50, 60, 70 x 10(4)/ml. The chondrocyte-polymer constructs were subcutaneously injected into the abdomen of autologous swine. The specimens were observed grossly and histologically after 6 weeks, and investigated the suitable cell density. Then the chondrocyte-polymer constructs with suitable cell density were transplanted into the abdomen of autologous swine and evaluated grossly and histologically in 1, 3, 6, 9, 15 weeks after transplantation to investigate the best formation time of tissue engineered cartilage. RESULTS The experiments demonstrated that the tissue engineered autologous cartilage was similar to the natural cartilage on animals with normal immune system in histological characteristics. The optimal chondrocyte density is 50 x 10(6)/ml, and the proper harvest time is the sixth week. CONCLUSION With tissue engineering skills, we have identified the optimal chondrocyte density and the proper harvest time.
OBJECTIVE: To investigate the influence of tissue engineered tendon on subgroup of T lymphocytes and its receptor in Roman chickens. METHODS: The flexor digitorum profundus of the third toes of right feet in 75 Roman chickens were resected and made 2.5 cm defects as experimental model. They were randomly divided into five groups according to five repair methods: no operation (group A), autograft (group B), fresh allograft (group C), polymer combined with allogenous tendon cells (group D), derived tendon materials combined with allogenous tendon cells (group E). The proliferation and transformation of lymphocytes and contribution of CD4+, CD8+, CD28 and T cell receptor (TCR) were detected to study the immune response. RESULTS: The CD4+, CD8+ and TCR of group D and E were increased slightly than that of group B after 7 days, while after 14 days, those data decreased gradually and no significant difference between tissue engineered tendon and autografts (P gt; 0.05), and there was significant difference between fresh allograft and tissue engineered tendon (P lt; 0.05). Lymphocytes transformation induced by conA also showed no significant difference between tissue engineered tendon and autografts (P gt; 0.05). CONCLUSION: Tendon cells are hypoantigen cells, there are less secretion of soluble antigen or antigen chips dropped out from cells. Tissue engineered tendon has excellent biocompatibility.
Objective To compare the difference of preparing the acellular larynx scaffold between perfusion method and immersion method, and find better way to make acellular larynx scaffold for tissue engineering. Methods Twenty 6-month-old male New Zealand rabbits, weighing 2.0-2.5 kg, were divided into perfusion group (n=10) and immersion group (n=10) at random. All the larynxes were excised in a sterile fashion. The acellular larynx scaffold was obtained by perfusionmethod and immersion method respectively, and then comparative examinations were performed by the macroscopicview, histological view, scanning electron microscope (SEM), cartilage vital ity assay and toluidine blue staining. ResultsMacroscopic view showed that the larynxes perfused by sodium dodecyl sulphate (SDS) became transparent after 2 hoursof perfusion, but the larynxes immersed by SDS over 16 hours still appeared pink-white. Histology and SEM indicated thatcompared with immersion group, perfusion group showed better acellular effect, more ventages and collagen fibers wereretained, no intact cell or nuclei remained in acellular matrix and chondrocytes were still survival. The porosity was 85.39% ± 3.16% in perfusion group and 34.72% ± 4.51% in immersion group, showing significant difference (P lt; 0.01). The chondrocyte vital ity rate of perfusion group (86.93% ± 1.52%) was higher than that of immersion group (77.73% ± 1.66%), showing significant difference (P lt; 0.01). Toluidine blue staining showed that the chondrocyte heterochromaty was ber in perfusion group than that in immersion group. Conclusion Compared with immersion method, perfusion method is a better way to construct acellular larynx scaffold because it can achieve better acellular effect and retain chondrocyte vital ity at the greatest extent in the acellular larynx scaffold.
OBJECTIVE: To explore the relationship between characteristics of transformed cell and tumorigenicity. METHODS: Documents about transformed cell and tumorigenicity were reviewed in detail. RESULTS: Normal biological characteristics and cell function could be maintained in non-tumorigenic transformed cell, but it was changed markedly in malignant transformed cell. CONCLUSION: Non-tumorigenic transformed cell can be served as a standard cell line to study the function and growth characteristics of normal cell.
Objective To investigate bio characteristics of bone stromal cells (MSC) in different concentrations of alginate combined with xenograft. Methods The configuration and secretion of MSC in different concentrations of alginate combined with xenograft were observed by scanning electron microscope and inverted microscope. Results When the concentration of alginate was 0.25% or 1%, alginate was equally combined in xenograft, 4% and 8% only on the surface of xenograft. After cultured for 4 days, alginate of 0.25# came off from xenograft. But alginate of 1% was equally combined in xenograft with cell secreting well in alginate. The growth of cells in alginate of 4% was restricted and no cell was seen in alginate of 8%. Conclusion Alginate of 1% is suitable fro constructing carrier of tissue engineering bone.
OBJECTIVE: To prevent the senescence of ’seed cells’ for tissue engineering, the life span of human fibroblasts is extended by reconstitution of telomerase activity, and the osteogenic potential of these fibroblasts are tested. METHODS: The pGRN145 plasmids encoding human telomerase reverse transcriptase (hTERT) were introduced into the normal human primary fibroblasts by electroporation. Telomerase activity was analyzed by TRAP-PCR assay. The beta-galactosidase stain was used to indicate the signs of cell senescence. The hTERT positive fibroblasts were then induced to form bone nodules. The bone nodules were stained by tetracycline and Alizarin Red S. RESULTS: Stable telomerase activity could be detected in the transfected fibroblasts and no signs of cell senescence were found in the fibroblasts cultured for more than 50 doublings. The hTERT positive fibroblasts could form bone nodules when they were cultured in vitro induced by bone morphogentic protein 2 and tumor necrosis factor-alpha. CONCLUSION: The fibroblasts with reconstituted telomerase activity reserve their osteogenic potential.
Objective To comment on the recent advances of production and application of the bio-derived scaffold in the tissue engineered peripheral nerve. Methods The recent articles were systematically analyzed, and then the production methods of the bio-derived scaffold and its application to the tissue engineered peripheral nerve were evaluated and prospected. Results B iological tissues were processed by some methods to produce the bio-derived materials. These mat erials could maintain the structure and components of the tissues. Moreover, the immunogenicity of these materials was reduced. Conclusion Application of the bio-derived materials is a trend in the fabricating scaffold of the tissue en gineered peripheral nerve.
OBJECTIVE To study the biocompatibility of skin reproductive membrane. METHODS According to ISO’s standards, the extractions of the skin reproductive membrane were prepared, and the acute systematic toxicity test, primary skin irritant test, cytotoxicity test, gene expression of type I collagen and fibronectin were detected to evaluate the biocompatibility of skin reproductive membrane. RESULTS All of those tests showed negative results. CONCLUSION The skin reproductive membrane has excellent biocompatibility in the level of the systematic, cellular and molecular biology.
Objective To study the difference of repairing segmental bone defect with bio-derived bone preserved by various methods.Methods Freeze-dried biomaterials had been stored in two different preservation solutions for three months,while the biomaterials stored for same period were observed as control group. The experimental model of 15 mm radial segmentaldefect was made in 60 New Zealand white rabbits, which were divided into groups A,B and C according to transplant materials preserved by various methods. Groups A and B were deeply divided into A1 and A2 subgroups, B1 and B2 subgroups according to whether materials were cocultured with osteoblasts. Tissue engineered bone was used to repair bone defects of left limbs in A1 and B1 subgroups, while simple material to repair defects of right limbs in A2 and B2 subgroups. Group C was divided into C1 and C2 subgroups. Freeze-dried material was used to repairbone defects of the left limbs, while defects of the right limbs as blank control group. The samples were harvested and observed by the roentgenographical, histomorphological, biomechanical and computerized graphical analysis at 4,8 and 16 weeks. Results All of the defects treated with implants exhibited new bone formation 4, 8 and 16 weeks postoperatively, increasing with time. The radiological, histomorphological and biomechanical evaluation showed that the ability of new bone formation was arranged in 6 subgroups as follows:A1gt;A2gt;C1gt;B1gt;B2gt;C2, the difference was significant between them (P＜0.001, P＜0.05).The ability of new bone formation was best and at 16 weeks the defect was bridged with the appearance of marrow cavities in A1 subgroup, the biomechanicalproperties in implants approached to those of normal bone. Conclusion The choice of proper preservation solution can improve the ability of repairing bone defect.