Objective To review research progress of corneal tissueengineering.Methods The recent articles on corneal tissue engineering focus on source and selection of corneal cells, the effects of growth factors on culture of corneal cells in vitro. The preparation and selection of three-dimensional biomaterial scaffolds and their b and weak points were discussed. Results The corneal tissue engineering cells come from normal human corneal cells. The embryo corneal cell was excellent. Several kinds of growth factors play important roles in culture, growth and proliferation of corneal cell, and incroporated into matrix.Growth factors including basic fibroblast growth factor, keratinocyte growth factor, transforming growth factor β1 and epidermal growth factor was favor to corneal cell. Collagen, chitosan and glycosaninoglycans were chosen as biomaterial scaffolds. Conclusion Human tissue engineering cornea can be reconstructed and transplanted. It has good tissue compatibility and can be used as human corneal equivalents.
ObjectiveTo summarize the research progress of tissue-engineered bile duct in recent years. MethodsThe related literatures about the tissue-engineered bile duct were reviewed. ResultsIn recent years, the research of tissue-engineered bile duct has made a breakthrough in scaffold materials, seed cells, growth factors etc. However, the tissue-engineered bile duct is still in the research stage of animal experiments, which can not be directly applied to clinical practice. ConclusionsThe research of tissue-engineered bile duct becomes popular at present. With the rapid development of materials science and cell biology, the basic research and clinical application of tissue-engineered duct will be more in-depth research and extension, which might bring new ideas and therapeutic measures for patients with biliary defect or stenosis.
OBJECTIVE To prevent early closure of growth plate and developmental deformities of limbs by allografts of cultured cartilages into growth plate defects of rabbits. METHODS Chondrocytes isolated from articular cartilage of 1-month rabbits formed cartilage after cultivation in centrifuge tubes. The cartilages cultured for two weeks were implanted into growth plate defects of proximal tibiae of 6-weeks rabbits. At 4th and 16th weeks, X-ray, histologic and immunohistochemical examination were performed. RESULTS The tibiae had no marked deformities after 4 weeks of operation. Histologic examinations showed that the defects were filled with cartilage. Immunohistochemical results of type II collagen were positive. The tibiae with allografts of cultured cartilages had no evident deformities after 16 weeks of operation. Histologic examination showed nearly closure of growth plates. On the contrary, the tibiae on control side formed severe deformities and growth plate were closed. CONCLUSION Allograft of cultured cartilages into growth plate defects may replace lost growth plate tissues, maintain normal growth of limbs and prevent developmental deformity.
OBJECTIVE: To review research progress of the relation between growth factor and repair of intervertebral disc. METHODS: The recent articles on growth factor and repair of intervertebral disc were extensively reviewed. The expression of growth factor in intervertebral disc and the effect of growth factor on disc cells were investigated. RESULTS: Some growth factors play roles in the development and degeneration of intervertebral disc. Exogenous growth factor can increase proliferation of disc cells and production of proteoglycans and collagens. Gene of growth factor can be transferred to intervertebral disc cell by adenovirus. CONCLUSION: Growth factor plays an important role in the regulation of development and degeneration of interertebral disc. The above results show that the feasibility of usage of growth factor in the treatment of disc degeneration and in repair and reconstruction of disc.
Objective To study the effect of platelet-rich plasma (PRP) on repairing chronic wounds of lower l imbs. Methods From May 2007 to November 2007, 47 patients suffering from chronic wounds of lower l imbs were treated. There were 41 males and 6 females, aged from 15 to 68 years (43.2 years on average). The disease was caused by tibiofibulafracture in 20 cases, calcaneus fracture in 4 cases, metatarsal fracture in 1 case, multiple open fracture of lower l imbs in 3 cases, tibia osteomyel itis in 10 cases, femur osteomyel itis in 1 case, soft tissue injury of ankle in 4 cases, infection after amputation in 2 cases, infection after foot orthomorphia in 1 case, and infection after calcaneus tendon neoplasty in 1 case. Their chronic wounds did not healed after 2 to 4 months of therapy. Among them, chronic wounds compl icated with fracture nonunion in 23 cases and positive bacterial culture result in 38 cases. Debridement and autogenous PRP gel injection were appl ied every 2 months and for twice. Results The patients were followed up for 4 months after the first PRP injection. Two months after the first PRP injection, chronic wounds contracted significantly in 34 patients with purulence and necrosis tissue cleaned up, circulation of soft tissue improved and exposed bone or muscle tissue covered by neogenetic granulation. No patient was completely cured. Two months after the second PRP injection, the average coverage rate was 79.3% ± 18.0%, the total cure rate was 29.8%. The volume of the chronic wounds decreased by (9.3 ± 4.9) mL after PRP therapy (2.5 ± 2.7) mL when compared with (11.8 ± 5.6) mL of before therapy, showing significant difference (P lt; 0.05). X-ray photograph showed that among the 23 cases of fracture nonunion, fracture healed completely in 9 cases; bony callus formation increased obviously in 12 cases; no significant change was observed in 2 cases. No aggravated sign of osteomyel itis was notified. Positive results of bacterial culture reduced to 15 cases. Conclusion PRP efficiently enhances the recovery of soft tissue defect and speeds up the chronic wounds heal ing oflower l imbs.
Objective To investigate the effects of transformin growth factor-beta (TGF-beta;) and interferon-gamma(IFN-gamma;)on collagen synthesis in human retinal pigment epithelial cells(RPE)． Methods TGF-beta;(0.01～10 ng/ml),recombinant IFN-gamma;(100～10000 Ｕ/ml)or a combination of two were added to cultures of RPE and collagen synthesis of the cells were measured by3 H-proline incorporation assay,indirect immunofluorescence staining and dot-blot hybridization. Results TGF-beta; at 10 ng/ml increased cell uptake of 3 H-proline to 130．87% of controls．It intensified Type IV,I and Ⅲ collagen fluorescent staining as well as mRNA expression．IFN-gamma; at 10000 Ｕ/ml caused 54．72% inhibition of 3 H-proline uptake by RPE,and decreased TypeⅣ collagen fluorescent staining as well as mRNA expression of Type Ⅳ,I and Ⅲ collagens. Conclusion TGF-beta; and IFN-gamma; stimulated and inhibited collagen synthesis of human RPE,respectively．The combination of two had antagonistic effects．IFN-gamma; can be used for inhibition of collagen synthesis of RPE. (Chin J Ocul Fundus Dis, 1999, 15: 245-248)
OBJECTIVE :To investigale effect of subretinal fluld(SRF)on proliferalion of the cellular elements of PVR. METHOD:The effect of SRF of 28 patients with rhegmatogenous retinal detachment proliferation of the cultured human retinal pigment epithelial cells(RPE),retinal glial cells (RG),and fibroblast (FB)was observed and detected by the methods of cell-counting and 3H-TdR in DNA synthesis. RESULTS:The range of proliferatinn-stimulating activity was 52.5%~233.3%, 36.4% ~ 177.8%，55.4% ~277.8% above the baseline in 1:10 dilution of these 3 kinds ,of cellular elements,and there was no significant difference among them. CONCLUSION ;The stimulating effect of SRF on the cellular proliferation was thougt to be due to the actions from certain growth factors. （Chin J Ocul Fundus Dis,1996,12: 233-235）
OBJECTIVE: To observe the effects of hyaluronic acid (HA) and basic fibroblast growth factor (bFGF) on the proliferation of the cells from medial collateral ligament (MCL) and anterior cruciate ligament (ACL) cells. METHODS: The MCL cells and ACL cells of mature New Zealand white rabbit were cultured, while HA, bFGF or HA and bFGF were added to the cell culture media, the cellular proliferation was assayed by MTT method. RESULTS: HA only had no effect on the preoliferation of ACL cells, but had a small stimulatory effect on the proliferation of MCL cells. The addition of 1 ng/ml bFGF enhanced the proliferation of both MCL and ACL cells significantly, and this enhancement was maximal in the concentration of 50 ng/ml. However, the enhancement of proliferation of MCL and ACL cells could be achieved when the combination of HA in concentration of 100 micrograms/ml and bFGF in concentration of 100 ng/ml. CONCLUSION: It is evident that bFGF can enhance the proliferation of the ligament cells. HA can maintain the normal growth of ACL cells with no effect on the proliferation of the cells, while HA has a small stimulatory effect on the proliferation of MCL cells. However, when bFGF is coordinated with HA, more improvement of cellular proliferation can be achieved. HA can be used as a potential carrier for bFGF to enhance the healing of ligamentous tissue injuries.
Objective To establish a better method of isolating andculturing ofneural stem cells(NSCs) in neonatal rat brain. Methods Tissue of brain was isolated from neonatal rats. Different medium and culture concentration were used toculture NSCs of neonatal rat. The culture concentration used were 1×10 4, 1×105, 1×106and 1×107/ml respectively. Ingredient of medium was classified into group 1 to 8 respectively according to whether to add 2% B27, epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) as well as the difference in culture concentration. The cells were induced to differentiate asto be confirmed as NSCs, and then were checked by phase contrast microscopy and identified by immunocytochemistry. Results The cells isolated and cultured gathered into neurospheres. The cells were capable of proliferating and maintaining longterm survival in vitro. The cells could be differentiated into neurons and glia.It was to the benefit of the survival of NSCs to add 5% fetal bovine serum(FBS)into the medium at the beginning of the culturing. When 10% FBS was added intothe medium, the neurospheres differentiated quickly. When concentration 1×106/ ml was used, the growth rate of the cells was the highest of all the concentrations. Reasonably higher cell concentration promoted the proliferation of NSCs. It was necessary to add 2% B27, EGF, and bFGF into the medium. The cells had the best growth when 2% B27, 20 ng/ml bFGF and 20 ng/ml EGF were added into the culture medium. EGF and bFGF had cooperative effect. Conclusion A better method of isolating and culturing of NSCs in neonatal rat brain is established and the foundation for future research is laid.
ObjectiveTo investigate effect of small interfering RNA (siRNA) targeting inhibition of growth hormone receptor (GHR) on proliferation and invasion of human liver cancer line SMMC-7721. MethodsSMMC-7721 cells were transfected with siRNA targeting human GHR by GenMuteTM transfection regent.The cells were divided into three groups:blank control group (non-transfected siRNA),negative control group (transfected with non-specific siRNA),and specificity transfected group (transfected with expression specifically interfere by GHR siRNA).the relative expression of GHR mRNA was detected by using real-time PCR.the expression of GHR protein was detected by using Western blot.The cell proliferation was assessed by CCK-8 assay.And the ability of invasion was examined by Transwell assay. ResultsThe expressions of GHR mRNA and GHR protein in the specificity transfected group were significantly lower than those in the blank control group (P<0.05) and the negative control group (P<0.05).Compared with the blank control group and the negative control group,the absorbance value and the number of migrating cells of SMMC-7721 cells were decreased obviously (P<0.05) in the specificity transfected group. ConclusionsiRNA targeting human GHR could reduce capability of proliferation migration and invasion of SMMC-7721 cells.