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Lei Cui

Publications and source records attributed to Lei Cui.

69 records · Page 4Linked to original sources

[Experimental study on constructing small-caliber artery by tissue engineering approach].

OBJECTIVE: To investigate the possibility of constructing small-caliber artery by means of tissue engineering. METHODS: Cell-PGA mixtures were made by separately seeding 1 x 10(7) smooth muscle cells and 5 x 10(6) endothelial cells isolated from neonate umbilicus onto PGA scaffold, the cell-PGA constructs were wrapped around a silicone tube before its implantation subcutaneously to nude mice and the mice were sacrificed in 2 and 6 weeks. The tissue engineered artery (TEA) were examined both grossly and immunohistochemically. RESULTS: The gross appearance of TEA was similar to that of the natural counterparts; histologic and immunohistochemical analyses of the neoformed tissues revealed a typical artery structure, including the presence of EC at the luminal surface and the presence of SMC and collagen in the wall. CONCLUSION: TEA with histology similar to natural vessel can be constructed by tissue engineering.

Animals↗

[Inhibition effect of CIITA anti-sense cDNA on the expression of MHCII molecules of Hela cells].

OBJECTIVE: To investigate the inhibiting effect of classII major histocompatibility complex transactivator (CIITA) anti-sense cDNA on MHCII molecules expression. METHODS: CIITA antisense cDNAs (arII1, arII2 and arII3) were obtained from Raji cell by RT-PCR, and then inserted into the pcDNA3.1B plasmid. The stable transfectants of Hela cell were tested for CIITA protein by immunohistochemistry, and classic MHCII molecules (HLA-DR, -DP, -DQ) induction through recombinant human IFN-gamma by flow cytometry. mRNAs of CIITA, invariant chain and classic MHCII molecules were detected by RT-PCR. RESULTS: The effect of arII2 was the best. When induced with IFN-gamma, the expression of CIITA protein was inhibited by 87.23%, and that of HLA-DR, -DP and -DQ was almost totally inhibited on arII2 positive Hela cells. The mRNA contents of CIITA, HLA-DR, -DP, -DQ and invariant chain were decreased significantly (P < 0.05). CONCLUSIONS: arII2 inhibits CIITA and thus the family of MHCIImolecules regulated by CIITA, therefore, it provides a novel approach to graft versus host disease study in hematopoietic stem cell transplantation.

DNA, Antisense↗

[The cloning of mouse FasL gene and expression in chondrocytes].

AIM: To clone mFasL cDNA, then insert it into retrovirus expression system pLNCX(2), and express mFasL cDNA in chondrocytes to study its function. METHODS: RT-PCR was applied to amplify mFasL cDNA from the total RNA of activated mouse spleen lymphocytes. The cDNA was inserted into a T-cloning vector, then subcloned into pLNCX(2). After transfection of chondrocytes, expression of FasL cDNA was detected by FACS, and activity of expressed product was analyzed by single mixed lymphocyte reaction(SMLC). RESULTS: mFasL cDNA fragment of 0.855 kb was successfully cloned and verified by sequencing. The transfection efficiency of mFasL cDNA in chondrocytes determined by FACS was 60.64%. Expressed product could evidently inhibit SMLC of allogeneic lymphocytes. It's stimulating index (SI) reduced to 11.71% as compare with the untransfected chondrocytes. CONCLUSION: Cloned recombinant pLNCX(2)-FasL can effectively express mFasL in chondrocytes, and can evidently inhibit SMLC of allogeneic lymphocytes.

Animals↗

Cardiac-specific overexpression of GLUT1 prevents the development of heart failure attributable to pressure overload in mice.

BACKGROUND: Increased rates of glucose uptake and glycolysis have been repeatedly observed in cardiac hypertrophy and failure. Although these changes have been considered part of the fetal gene reactivation program, the functional significance of increased glucose utilization in hypertrophied and failing myocardium is poorly understood. METHODS AND RESULTS: We generated transgenic (TG) mice with cardiac-specific overexpression of insulin-independent glucose transporter GLUT1 to recapitulate the increases in basal glucose uptake rate observed in hypertrophied hearts. Isolated perfused TG hearts showed a greater rate of basal glucose uptake and glycolysis than hearts isolated from wild-type littermates, which persisted after pressure overload by ascending aortic constriction (AAC). The in vivo cardiac function in TG mice, assessed by echocardiography, was unaltered. When subjected to AAC, wild-type mice exhibited a progressive decline in left ventricular (LV) fractional shortening accompanied by ventricular dilation and decreased phosphocreatine to ATP ratio and reached a mortality rate of 40% at 8 weeks. In contrast, TG-AAC mice maintained LV function and phosphocreatine to ATP ratio and had <10% mortality. CONCLUSIONS: We found that increasing insulin-independent glucose uptake and glycolysis in adult hearts does not compromise cardiac function. Furthermore, we demonstrate that increasing glucose utilization in hypertrophied hearts protects against contractile dysfunction and LV dilation after chronic pressure overload.

Adenosine Triphosphate↗

[Tissue engineered tendon with skin fibroblasts as seed cells, a preliminary study].

OBJECTIVE: To investigate the effect of tissue engineered tendon using autologous dermal fibroblasts in repairing tendon damage. METHODS: Skin tissues were resected from abdomen of 5 pigs. Fibroblasts were isolated from the skin pieces and cultured in vitro. Bundles of polyglycolic acid (PGA) were arranged in parallel and mixed with the suspension of fibroblasts to form a cell-scaffolded construct, which was further cultured in vitro for 1 week. The tendon of musculus flexor digitorum superficialis of pig's right leg was cut with a defect 3 cm long and then bridged with the cultured construct. Six weeks later, specimens of the regenerated tendon of right musculus flexor digitorum superficialis and specimens of corresponding left leg were taken for gross examination, microscopy and biomechanical analysis. RESULTS: After implantation of the fibroblast-biomaterial complex the wounds healed up and the pigs moved well. The histology of the implanted tendon was similar to that of natural tendon. The breaking strength and maximum tensile force were 173.0 +/- 18.2 N and 18.9 +/- 1.9 MPa, reaching 57.4% and 51.9% of those of normal tendon respectively. CONCLUSION: Skin fibroblasts can be used as seed cells to regenerate tendon with normal structure and function.

Animals↗

[Repair of meniscal defects with autologous tissue-engineered fibrocartilage].

OBJECTIVE: To repair critical-sized meniscal defects in an immunocompetent mammal by tissue engineering approach. METHODS: 15 45-day-old Changfeng crossbred pigs were selected as experimental animals. Autologous fibrochondrocytes were obtained from left knee menisci by modified Klagsbrun's method and were proliferated in vitro to a proper amount. A 1-cm-long full-layer defect of right medial meniscus was created anterior to the medial collateral ligament. PGA-fibrochondrocyte-Pluronic complex, fibrochondrocyte-Pluronic complex or PGA only was respectively implanted into the defects. We used intact menisci and untreated meniscal defects as controls. Samples respectively obtained in 9 w, 16 w and 25 w were appraised by general observation, histology, biochemistry and biomechanics. RESULTS: From the sights of general morphology, histological structure and Young's Modulus (59.7% of that in normal meniscus at 25 w), the PGA-cell-Pluronic complex can form the best quality tissue, which can stabilize the GAG ratio (74.5% of that in normal cartilage at 25 w) of femoral entocondyle cartilage. CONCLUSION: Autologous tissue-engineered fibrocartilage is a promising feasible method to regenerate or reconstruct menisci so as to hold back the degenerative changes of the knee.

Animals↗

Repairing large porcine full-thickness defects of articular cartilage using autologous chondrocyte-engineered cartilage.

Large full-thickness defects of articular cartilage remain a major challenge to orthopedic surgeons because of unsatisfactory results of current therapy. Many methods, such as chondrectomy, drilling, cartilage scraping, arthroplasty, transplantation of chondrocytes, periosteum, perichondrium, as well as cartilage and bone, have been tried to repair articular cartilage defects. However, the results are far from satisfactory. In this study, we applied a tissue-engineering approach to the repair of articular cartilage defects of knee joints in a porcine model. Using isolated autologous chondrocytes, polyglycolic acid (PGA), and Pluronic, we have successfully in vivo-engineered hyaline cartilage and repaired articular cartilage defects. The surface of the repaired defects appeared smooth at 24 weeks postrepair. Histological examination demonstrated a typical hyaline cartilage structure with ideal interface healing between the engineered cartilage and the adjacent normal cartilage and underlying cancellous bone. In addition, glycosaminoglycan (GAG) levels in the engineered cartilage reached 80% of that found in native cartilage at 24 weeks postrepair. Biomechanical analysis at 24 weeks demonstrated that the biomechanical properties of the tissue-engineered cartilage were improved compared with those at an earlier stage. Thus, the results of this study may provide insight into the clinical repair of articular cartilage defects.

Animals↗

Bridging tendon defects using autologous tenocyte engineered tendon in a hen model.

Tendon defects remain a major concern in plastic surgery because of the limited availability of tendon autografts. Whereas immune rejection prohibits the use of tendon allografts, most prosthetic replacements also fail to achieve a satisfactory long-term result of tendon repair. The tissue engineering technique, however, can generate different tissues using autologous cells and thus may provide an optimal approach to address this concern. The purpose of this study was to test the feasibility of engineering tendon tissues with autologous tenocytes to bridge a tendon defect in either a tendon sheath open model or a partial open model in the hen. In a total of 40 Leghorn hens, flexor tendons were harvested from the left feet and were digested with 0.25% type II collagenase. The isolated tenocytes were expanded in vitro and mixed with unwoven polyglycolic acid fibers to form a cell-scaffold construct in the shape of a tendon. The constructs were wrapped with intestinal submucosa and then cultured in Dulbecco's Modified Eagle Medium plus 10% fetal bovine serum for 1 week before in vivo transplantation. On the feet, a defect of 3 to 4 cm was created at the second flexor digitorum profundus tendon by resecting a tendon fragment. The defects were bridged either with a cell-scaffold construct in the experimental group ( n= 20) or with scaffold material alone in the control group ( n= 20). Specimens were harvested at 8, 12, and 14 weeks postrepair for gross and histologic examination and for biomechanical analysis. In the experimental group, a cordlike tissue bridging the tendon defect was formed at 8 weeks postrepair. At 14 weeks, the engineered tendons resembled the natural tendons grossly in both color and texture. Histologic examination at 8 weeks showed that the neo-tendon contained abundant tenocytes and collagen; most collagen bundles were randomly arranged. The undegraded polyglycolic acid fibers surrounded by inflammatory cells were also observed. At 12 weeks, tenocytes and collagen fibers became longitudinally aligned, with good interface healing to normal tendon. At 14 weeks, the engineered tendons displayed a typical tendon structure hardly distinguishable from that of normal tendons. Biomechanical analysis demonstrated increased breaking strength of the engineered tendons with time, which reached 83 percent of normal tendon strength at 14 weeks. In the control group, polyglycolic acid constructs were mostly degraded at 8 weeks and disappeared at 14 weeks. However, the breaking strength of the scaffold materials accounted for only 9 percent of normal tendon strength. The results of this study indicated that tendon tissue could be engineered in vivo to bridge a tendon defect. The engineered tendons resembled natural tendons not only in gross appearance and histologic structure but also in biomechanical properties.

Animals↗

[Transposition of pedicled sternocleidomastoid muscle for repair of facial paralysis in late stage].

OBJECTIVE: To explore a new surgical approach to repair facial paralysis in late stage, using regional transposition of pedicled sternocleidomastoid muscle for the dynamic reanimation of the paralyzed face. METHODS: Seven cases with facial paralysis in late stage from December 1999 were treated and followed up for 10 months before clinical evaluation. In all of the cases, the sternal and clavicular branches of the sternocleidomastoid muscle were both elevated from their bony attachments, with the mastoid insertion left in situ as the pedicle for blood supply and accessory nerve maintained in it. The muscle strips were transposed and sutured to the orbicularis oris around the mouth corner on the paralyzed side. RESULTS: Static asymmetry of nose and oral commissure on the paralyzed side were corrected immediately after operation, and the movement of the oral commissure recovered one week after operation. Symmetric smiling was observed in one month and all of the oral movements recovered in 10 months postoperatively. CONCLUSION: The new approach to repair facial paralysis in late stage by regional transposition of pedicled sternocleidomastoid muscle is effective in restoration of both static and dynamic symmetry of nose and mouth, and in recovery of the facial expression and the oral commissure.

Adult↗

Blocking transforming growth factor-beta receptor signaling down-regulates transforming growth factor-beta1 autoproduction in keloid fibroblasts.

OBJECTIVE: To study transforming growth factor-beta1 (TGF-beta1) autoproduction in keloid fibroblasts and the regulation effect of blocking TGF-beta intracellular signaling on rhTGF-beta1 autoproduction. METHODS: Keloid fibroblasts cultured in vitro were treated with either rhTGF-beta1 (5 ng/ml) or recombinant adenovirus containing a truncated type II TGF-beta receptor gene (50 pfu/cell). Their effects of regulating gene expression of TGF-beta1 and its receptor I and II were observed with Northern blot. RESULTS: rhTGF-beta1 up-regulated the gene expression of TGF-beta1 and receptor I, but not receptor II. Over-expression of the truncated receptor II down-regulated the gene expression of TGF-beta1 and its receptor I, but not receptor II. CONCLUSIONS: TGF-beta1 autoproduction was observed in keloid fibroblasts. Over-expression of the truncated TGFbeta receptor II decreased TGF-beta1 autoproduction via blocking TGF-beta receptor signaling.

Activin Receptors, Type I↗

[Repair of porcine full-thickness skin defects with autologous tissue engineered skin].

OBJECTIVE: To explore a feasible method to repair full-thickness skin defects with tissue engineered techniques. METHODS: The skin specimens were cut from the Changfeng hybrid swines' abdomen, then keratinocytes and fibroblasts were isolated and harvested by trypsin, EDTA and type II collagenase. The cells were seeded in petri dishes for primary culture. When the cells were in logarithmic growth phase, they were treated with dispase II (keratinocytes) or trypsin (fibroblasts) to separate them from the floor of the tissue culture dishes. A biodegradable material-pluronic F-127 was prefabricated and mixed with these cells, and then the cells-pluronic compounds were seeded evenly into polyglycolic acid (PGA). Tinally the constructs were replanted to autologous animals to repair full-thickness skin defects. Histological changes were observed in 1, 2, 4 and 8 weeks postsurgery. RESULTS: The cells-pluronic F-127-PGA compounds could repair autologous full-thickness skin defects. Histologically, the tissue engineered skin was similar to normal skin with stratified epidermis overlying a moderately thick collageneous dermis. CONCLUSION: Tissue engineered skin can repair autologous full-thickness skin defects with primary-cultured keratinocytes and fibroblasts as seed cells and PGA as a cell carrier.

Animals↗

[In vitro chondrogenic phenotype differentiation of bone marrow stem cells].

OBJECTIVE: To investigate the feasibility of chondrogenic phenotype differentiation of adult swine bone marrow stem cells(MSCs) in a defined medium as seeding cells in cartilage tissue engineering. METHODS: A volume of 5 ml bone marrow was aspirated from swine iliac crest and cultured in the complete medium of DMEM-LG for two weeks. The growth and ultrastructure of the cultured MSCs were observed. Immunohistochemistry and in situ hybridization were applied to detect the expression of collagen type II. RESULTS: The MSCs changed from a spindle-like fibroblastic appearance to a polygonal shape when transferred from the complete medium of DMEM-LG to a defined medium. A large amount of endoplasmic reticulum was observed in large Golgi ccmplex and mitochondria. The differentiation of MSCs toward chondrogenic phenotype was verified by the positive result of collagen type II through immunohistochemistry and in situ hybridization respectively. CONCLUSIONS: Bone marrow stem cells obtained from adult swine can differentiate to be chondrogenic phenotype when cultured in vitro. MSCs can likely be served as optimal autogenous cell source for cartilage tissue engineering.

Animals↗

[Intra-uterine model of cleft lip and scarless wound healing of fetal rats].

OBJECTIVE: To properly evaluate the benefits of in utero repair of cleft lip, an animal model of cleft lip was developed and the fetal scarless wound healing was observed in SD rat fetus. METHODS: Through a maternal celiotomy and hysterotomy, twenty excisional cleft was created in the upper lip of rat fetus. Another group of 20 rat fetus were given full-thickness incisional wounds in the upper lips. The fetus with cleft lip were returned into utero for further development till birth.Wound specimens from the incisional wounds in the other group were removed successively 12, 24, 48, 72h postwounding for histological examination. RESULTS: The fetus with excisional wound demonstrated a complete cleft lip after birth. In contrast, the wound in the fetus with incisional wounds regenerated with normal architecture histologically, indistinguishable from the surrounding normal skin. CONCLUSION: The results of this preliminary study indicates that the fetal rat cleft lip is technically feasible with an excellent survival rate. Healing occurs without scar formation. This model will be used to document facial growth following in utero repair of a cleft lip.

English Abstract↗

[Experimental study of the inducing BMSCs into osteogenesis].

OBJECTIVE: To study the bone tissue osteogenesis ability of bone marrow stromal cells (BMSCs) in vitro and in vivo. METHODS: The BMSCs were separated by gradient centrifugation on Percoll, culture-expanded and induced with a conditional medium. The cells were examined by transmission electron microscopy and histochemistry or immunochemistry stains technique including AKP,Cbfa1,OCN, OPN, BSP and collagen type I. The mRNA of collagen type I in cultured cell was also examined by in situ hybridization. RESULTS: Histochemical studies showed positive stain of AKP,Cbfa1, OCN, OPN,BSP and collagen type I,and in situ hybridization also showed the positive express of mRNA of collagen type I. The excretion of collagen and deposition of calcium salt were observed under transmission electron microscopy. CONCLUSION: BMSCs cultured in conditional medium shows osteoblast-like functions in vitro, which provides an ideal seed cells for bone tissue engineering.

English Abstract↗

Tissue engineering of nearly transparent corneal stroma.

We determined whether a polyglycolic acid (PGA) scaffold bearing an adherent corneal stromal cell insert could be integrated into the ultrastructure of rabbit corneal stroma without compromising tissue transparency. Stromal cells were isolated from 10 newborn rabbits and expanded by tissue culture. After reaching confluence, the cells were harvested and mixed with nonwoven PGA fibers to form cell-scaffold constructs. After 1 week of culturing, they were implanted into the corneal stroma of female rabbit recipients. Green fluorescent protein (GFP) expression in transplanted corneal stromal cells was monitored at the protein level to determine cellular origin in the reconstructed stroma. Eight weeks after implantation, transmission electron microscopy and histological evaluation were performed on stromal tissue. Insertion of PGA scaffold alone served as a sham control. After stromal implantation, implants gradually became transparent over an 8-week period. During this time stromal histology was gradually restored, as collagen fibril organization approached that of their normal counterpart. GFP-labeled corneal stromal cells were preponderant in the regions bearing inserted scaffolds, suggesting that they were derived from the implants rather than from neighboring corneal stromal cells. No reconstructed stroma developed in regions where naked PGA was implanted instead. We conclude that intrastromal implantation of PGA fiber scaffold implants bearing corneal stromal cells is a useful procedure for corneal stromal tissue reconstruction because, over an 8-week period, the implants become progressively more transparent. Marked losses of translucence during this period combined with restoration of ultrastructure indicate that the implants provide the functions needed for deturgescing initially swollen stroma.

Animals↗