Clinical translational research and application prospects of mesenchymal stem cells in periodontal tissue regeneration / 中华口腔医学杂志

Periodontitis is an oral infectious disease that occurs in the tooth supporting tissues, which damages the soft and hard tissues of the periodontium and eventually results in tooth mobility and loss. Traditional clinical treatment can effectively control the periodontal infection and inflammation. However, as the therapeutic effect depending on the local condition of periodontal defect and the systemic factors of patients, it's hard to achieve satisfactory and stable periodontal tissue regeneration for damaged periodontal tissues. Recently, as a promising therapeutic strategy in periodontal regeneration, mesenchymal stem cells (MSC) play an important role in modern regenerative medicine. Combined with the clinical translational researches of MSC in periodontal tissue engineering and the research results of our group in the past decade, it is summarized and explained the mechanism of MSC promoting periodontal regeneration, the preclinical and the clinical transformation researches as well as the future application prospects of MSC in periodontal regeneration therapy in this paper.

Advances in Research on Stem Cell-Based Pulp Regeneration

Although root canal therapy is the most common and widely used treatment at clinical presentation, there are still some postoperative complications. As cell biology and tissue engineering techniques advance rapidly, the use of biological therapy to regenerate dental pulp has become a new trend; Relevant literatures in recent five years were searched using key words such as "root canal therapy", "Dental pulp stem cells", "Dental pulp regeneration", and "Cell homing" in PubMed, Web of Science, etc; Dental pulp stem cells (DPSCs) have multi-differentiation potential, self-renewal capability, and high proliferative ability. Stem cell-based dental pulp regeneration has emerged as a new research hot spot in clinical therapy. Recently, dental pulp-like structures have been generated by the transplantation of exogenous DPSCs or the induction of homing of endogenous DPSCs. Studies on DPSCs are important and significant for dental pulp regeneration and dental restoration; In this review, the existing clinical treatment methods, dental pulp regeneration, and DPSC research status are revealed, and their application prospects are discussed. The stem cell-based pulp regeneration exerts promising potential in clinical therapy for pulp regeneration.

Clinical and genetic features of Chinese kindreds with Van der Woude syndrome caused by interferon regulatory factor 6 mutation / 华西口腔医学杂志

OBJECTIVE@#This study aimed to investigate the clinical phenotype and genetic characteristics of Chinese families with Van der Woude syndrome (VWS).@*METHODS@#Clinical manifestations between 14 families and within each family were recorded. Possible inheritance modes and pathogenic genes were analyzed. Phenotypic distribution and gene frequencies were calculated.@*RESULTS@#Of the pedigrees investigated, an autosomal dominant inheritance pattern was suggested. All patients had typical symptoms. The pathogenic gene was interferon regulatory factor 6 (IRF6). Phenotypic distribution frequencies were as follows: lip pits (91.9%), cleft lip and/or palate (73.0%), and hyperdontia (8.1%). There were significant differences in clinical phenotypes among individuals of different families and individuals of the same family.@*CONCLUSIONS@#VWS in a Chinese population was dominantly inherited with high penetrance and variable expressivity. The pathogenic gene was IRF6. VWS in a Chinese population was genotyped as VWS1.

Application of computer-assisted surgery techniques in the management of zygomatic complex fractures / 中华创伤杂志(英文版)

Patients suffering from zygomatic complex fractures always present facial deformity and dysfunctions, and thereafter develop psychological and physiological problems. It is really hard to get an ideal prognosis for the zygomatic complex fractures because of the complicated anatomical structures. Computer-assisted surgery techniques, as the new emerging auxiliary methods, can optimize the surgical protocol, predict operation outcomes, and improve the accuracy and quality of the operation. Meanwhile the postoperative complications can be reduced effectively. This review aims to provide a comprehensive overview of the application of computer-assisted surgery techniques in the management of zygomatic complex fractures.

Correlation between Condylar Fracture Pattern after Parasymphyseal Impact and Condyle Morphological Features: A Retrospective Analysis of 107 Chinese Patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The treatment of the condylar fractures is difficult. Factors that result in the fractures are complex. The objective of this morphometric study was to investigate the relationship between condylar fracture patterns and condylar morphological characteristics.</p><p><b>METHODS</b>We conducted a retrospective analysis of 107 patients admitted to the West China Hospital of Stomatology for bilateral condylar fractures caused by parasymphyseal impact. The patients were divided into five groups according to the type of condylar fracture. Ten parameters were evaluated on three-dimensional (3D) reconstruction mandible models through the Mimics 16.0 (Materialize Leuven, Belgium) anthropometry toolkit. Each parameter of the 3D models was analyzed using multivariate analysis. Multinomial logistic regression analyses were used to examine the relationships between the five groups.</p><p><b>RESULTS</b>The results showed that the differences of condylar head width (M1), condylar neck width (M3), the ratio of condylar head width to condylar anteroposterior diameter (M1/M2), the ratio of condylar head width to condylar neck width (M1/M3), the ratio of condylar height to ramus height (M8/M7), and mandibular angle (M10) were statistically significant (p < 0.05). Type A condylar head fractures were positively associated with M1 (compared to Type B: OR =1.627, 95% CI: 1.123, 2.359; compared to Type C: OR = 1.705, 95% CI: 1.170, 2.484) and M1/M2 (compared to Type B: OR =1.034, 95% CI: 0.879, 2.484). Type B condylar head fractures were negatively associated with M10 (compared to Type C: OR = 0.909, 95% CI: 0.821, 1.007). Condylar neck fractures were negatively associated with M3 (compared to condylar head: OR = 0.382, CI: 0.203, 0.720 ; compared to condylar base: OR = 0.436, 95% CI: 0.218, 0.874), and positively associated with M1/M3 (compared to condylar head: OR = 1.229, 95% CI: 1.063, 1.420 compared to condylar base: OR = 1.223, 95% CI: 1.034, 1.447). Condylar base fractures were positively associated with M10 (OR = 1.095, 95% CI: 1.008, 1.189) and negatively associated with M8/M7 (OR = 0.855, 95% CI: 0.763, 0.959) as compared with condylar head fractures.</p><p><b>CONCLUSIONS</b>Condylar fracture pattern is associated with the anatomical features of the condyles when a fracture occurs from parasymphyseal impact.</p>

Construction of recombinant gene adenovirus encoding enhanced green fluorecence protein-peroxisome proliferator-activated receptor gamma2 fusion protein and its expression in bone marrow mesenchymal stem cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To construct mouse enhanced green fluorecence protein (EGFP) -peroxisome proliferator-activated receptor (PPAR)gamma2, and to detect EGFP-PPARgamma2 expression in infected mouse bone marrow mesenchymal stem cells (BMSC).</p><p><b>METHODS</b>Cut the fragment of PPARgamma2 from the expression plasmid pcDNA flag PPARgamma2, then cloned the gene fragment into pEGFP-C1 and pEGFP-N1 vector. Subsequently, subclone the fragment EGFP-PPARgamma2 from pEGFP-C1-PPARgamma2 into the shuttle plasmid DC315. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid DC315-EGFP-PPARgamma2 and large adenovirus helper plasmid pBHGlox deltaE1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus Ad-EGFP-PPARgamma2 was confirmed. Then it was propagated in HEK293 cells. After the BMSC were transfected for 72 h, adipogenic differentiation was demonstrated.</p><p><b>RESULTS</b>HEK293 cells were transfected with the pEGFP-C1-PPARgamma2 or pEGFP-N1-PPARgamma2 in mediation of liposome. The former green fluorescence protein was better than the latter by fluorescence microscope. The recombinant plasmids were digested and identified. Western blot analysis showed the expression of EGFP-PPARgamma2 in vitro. EGFP-PPARgamma2 protein was detectable in the nucleus of BMSC.</p><p><b>CONCLUSION</b>The recombinant adenovirus encoding EGFP-PPARgamma2 fusion protein was successfully constructed, which provided a basis for application of EGFP-PPARgamma2 gene to adenovirus-mediated gene therapy.</p>

Beta -transduction repeat containing protein expressed in tooth germs and ameloblast and odontoblast of different stage of tooth development / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The Sonic hedgehog signalling peptide has been demonstrated to play important roles in the growth and patterning of the tooth development. This study aims on whether the antagonist beta-transduction repeat containing protein of Sonic hedgehog signal transduction expressed in tooth germs ameloblast and odontoblast or not.</p><p><b>METHODS</b>The mouse embryo heads of different developmental stages of E10.5, E13.5, E14.5, E16.5, E18.5 and P0, P3, P6 after birth were acquired fixed with 4% paraformaldehyde for 48 hours, embeded with Paraffin and examined using LsAB (labelled streptavidin-biotin) method to observe the beta-TrCP expression pattern in tooth germs, ameloblast and odontoblast.</p><p><b>RESULTS</b>It was demonstrated that beta-TrCP expressed in oral epithelium, tooth bud, mesenchymal cell cytoplasm of ameloblast and odontoblast of different stage of tooth development.</p><p><b>CONCLUSION</b>beta-TrCP expressed from early stage to later stage of murine tooth development. And these findings provide the evidence of antagonist regulatory pathways for shh in teeth development.</p>

Clinical analysis of midfacial fractures with orbital floor fractures / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the characters of the diagnosis and treatment for midfacial fractures with orbital floor fractures.</p><p><b>METHODS</b>136 cases of midfacial fractures with orbital floor fractures were diagnosed and treated. All patients underwent CT scans and accepted surgical intervention. Orbital floor fracture was restituted in 49 cases. Orbital floor defects were reconstructed with autogenous bone, titanium mesh or porous polyethylene implant (Medpor) in 21 cases.</p><p><b>RESULTS</b>136 cases recovered well, their facial appearance and function were improved significantly. There were no severe complications happened postoperatively in all cases. 2 cases had postoperative wounds infection and 1 case had temporary ablepsia, but healed completely.</p><p><b>CONCLUSION</b>CT is the best method for diagnosing orbital floor fractures. The objective of treatment is restoration of normal orbital floor and orbital capacity, the reduction of orbital contents prolapsed and reconstruction of orbital floor fractures with repair materials.</p>

Retrospective analysis of 3,958 patients with facial injuries / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To determine the causes and incidence of facial injuries by an epidemiologic retrospective study.</p><p><b>METHODS</b>A total of 3 958 patients with facial injuries treated at Department of Oral and Maxillofacial Surgery, West China School of Stomatology, Sichuan University from 1955 to 2001 were investigated. Data regarding age, gender, cause of injury, pattern of fracture and associated systemic injuries were reviewed.</p><p><b>RESULTS</b>The male to female ratio of the patients with facial injury was 4.27:1 and 33.4% of patients were aged between 21 and 30 years. The most common cause of injury was traffic accident (30.6%), followed by falls (21.4%) and collision (15.8%). A total of 794 patients (20.1%) showed only soft tissue injuries. 1 100 patients (27.8%) had multiple fractures in facial bones and 2,064 patients (52.1%) had single fracture. The mandibular fracture was most frequently seen, followed by the maxilla and the zygoma. The most common site of mandible fracture was the body (31.2%), followed by the symphysis (22.7%), the condylar (20.5%) and the angle (13.7%). Accompanied injuries to brain and skull happened in 916 patients (23.1%).</p><p><b>CONCLUSIONS</b>Bone fractures were more common in hospitalized patients with facial injuries. The numbers and sites of fracture were related to the causes of injuries and anatomic structure of the bone. The brain and skull injuries, the most often and seriously accompanied injuries, would not be neglected.</p>

An experimental study on rat bone marrow mesenchymal stem cells transfected with dentin sialophosphoprotein gene in vitro / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To evaluate the expression of dentin sialophosphoprotein (DSPP) in transfected rat bone marrow mesenchymal stem cells (BM-MSC) and the influence of the transfection.</p><p><b>METHODS</b>Plasmid containing mice dentin DSPP was constructed by using the cytomegalovirus (CMV) promoter and then transfected the cultured BM-MSC by lipofectamine; The expression of Pax-9 and dentin matrix protein 1 (DMP1) gene of transfected BM-MSC were detected by RT-PCR. The expression of DSPP was examined by immunocytochemical staining, and the formation ratio of mineralized nodules of transfected BM-MSC was compared with untransfected ones after mineralized induction.</p><p><b>RESULTS</b>The constructed pcDNA3.1(+)/mDSPP could produced 3.0 kb and 5.4 kb fragments, DSPP gene and Pax9 gene were expressed 24 h and 48 h respectively, after BM-MSC were transfected Pax-9 gene was exprssed, but DMP1 gene was not; Immunohistochemical staining showed that DSPP was positive in transfected BM-MSC; The formation ratio of mineralized nodules of transfected BM-MSC was higher than that of untransfected ones after mineralized induction.</p><p><b>CONCLUSIONS</b>The expression of mice DSPP in BM-MSC by gene transfection can induce the expression of tooth development-associated gene Pax9 and enhance the formation of mineralized nodules, which suggests that DSPP gene might induce odontogenic differentiation of BM-MSC.</p>

A study on repairing mandibular defect by means of tissue-engineering and human bone morphogenetic protein-2 gene transfection in osteoporotic rats / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of repairing bone defect with methods of tissue-engineering and human bone morphogenetic protein-2 (hBMP-2) gene transfection in osteoporotic rats.</p><p><b>METHODS</b>Twenty-four 6-month-old female Sprague-Dawley rats underwent ovariectomy, while 8 rats received sham-operations. Three months later, bone mesenchymal stem cells (BMSC) harvested from osteoporotic rats were divided into two groups randomly. Experimental group were transfected by recombinant plasmid carrying hBMP-2 gene, and control group left untreated. All BMSC were seeded into coralhydroxyapatite scaffolds. Then the cell/scaffold constructs were implanted into the defect site created in the ramus of mandible of osteoporotic rats respectively.</p><p><b>RESULTS</b>Positive results were confirmed by immunohistochemistry and in situ hybridization in experimental group. New bone formation was found at the margin of the defect treated with the BMSC modified by hBMP-2 gene transfer at 4 weeks after implantation and appeared mature 8 weeks after the treatment. However, the amount of newly formed bone was much less and there was some adipose tissue at defect margins 8 weeks after implantation in control group.</p><p><b>CONCLUSIONS</b>The results of this experiment indicate that BMSC-mediated rhBMP-2 gene therapy in conjunction with bone tissue engineering may allow for successful treatment of large bone defects in osteoporosis rats.</p>

Expression of human interleukin-1 receptor antagonist in the transfected chondrocytes of temporomandibular joint / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression of the human interleukin-1 receptor antagonist (hIL-1ra) in the transfected chondrocytes of temporomandibular joint (TMJ).</p><p><b>METHODS</b>Chondrocytes of TMJ in vitro were transfected by hIL-1ra gene via cationic liposome as a medium. The stable transfected cells were selected by G418. The proliferations of the transduced cell were examined with the growth curve, cell population doubling time. The protein expressing in different periods was detected by immunocytochemistry and enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The proliferation suppression of gene transfected cells fell significantly with compared to normal cells. The expression of hIL-1ra was detected in the cell plasma and the cell culture supernatant. The highest expression of IL-1ra protein was at the time of 48 hours after gene transfection. The transiently transfected cells were secreted IL-1ra protein continuously 28 days and the stably transduced cells were secreted IL-1ra protein till 72 days.</p><p><b>CONCLUSION</b>This study showed that hIL-1ra protein expressed positively in the cell plasma and the culture supernatant after gene transfection within a certain periods.</p>

Identification of three novel mutations of IRF6 in Chinese families with Van der Woude syndrome / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify mutations of interferon regulatory factor 6 (IRF6) gene in Van der Woude syndrome (VWS) patients in China.</p><p><b>METHODS</b>Three Chinese VWS families were screened to IRF6 gene mutation via PCR and sequence techniques. After amplification of exons 1-8 and their flanking splice junctions and part of exon 9 of the IRF6 gene by polymerase chain reaction, mutations were detected by direct sequencing.</p><p><b>RESULTS</b>Three novel mutations, one in each family, were identified in all the affected members in the three families. There were one missense mutation 1214 (T-->C) in exon 9, two nonsense mutations 981 (T-->A) in exon 7 and 1234 (C-->T) in exon 9. All affected members of the three families were heterozygous for their respective mutation.</p><p><b>CONCLUSION</b>Mutations in IRF6 gene were found in all VWS patients. This observation supports the hypothesis that IRF6 is the gene responsible for VWS across different populations.</p>

Biomechanical study of rigid internal fixation for mandibular angle fracture / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To develop a functional biomechanical mandibular model, and to observe the stress distribution of angle-fractured mandible under different rigid internal fixation (RIF) methods.</p><p><b>METHODS</b>A biomechanical model of mandible was built which include the simulative temporal-mandibular joint (TMJ) and was under mechanical loads of masseter, temporalis, medial pterygoid and lateral pterygoid. The different stress pattern was measured by strain gauges. Under the standard mandibular angle fracture, bilateral molar biting and four pairs of muscles loading, the strains were compared to evaluate the stability of one-or two-miniplate fixation.</p><p><b>RESULTS</b>The fixation of miniplate in the lateral oblique line can recover the main stress on the non-fracture side, but it was broken in the fracture side. The tension increased in the lower border of mandibular angle.</p><p><b>CONCLUSION</b>Through the biomechanical study based on the functional mandibular model, only one miniplate fixation in lateral oblique line for mandible angle fracture was insufficient buttressing of the segments, while the additional miniplate in the lower margin can recover the stress pattern and provide more stability.</p>

A study on the chondrogenesis of the compound of alginate gelatin and bone marrow stromal cells in vivo / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the chondrogenisis by alginate gelatin and rats' bone marrow stromal cells (BMSCs) chondrogenicly induced in vitro.</p><p><b>METHODS</b>Thirty-two male adult SD rats were assigned randomly to experimental and control groups. In experimental group, bone marrow was obtained from the right tibias of all the rats. After expanding and culturing 3 passages, induced BMSCs by chondrogenic culture medium for 10 days. Suspended induced cells in alginate gelatin, and injected the complex into the hypodermic tissue of the backs of rats autogenously. In control group only alginate gelatins were injected. The grafts were taken out for examinations 4 and 8 weeks after the operations.</p><p><b>RESULTS</b>Considerable cartilage appeared in experimental group 8 weeks after operations. Regular HE staining and alcian blue staining showed a great deal of cartilage holding chondrocyte masses surrounded by abundant matrix. Alginate gelatin decompounded obviously, and the rest distributed among newly formed cartilage. No cartilage appeared in control group all through.</p><p><b>CONCLUSION</b>BMSCs and alginate gelatin have a beautiful future in cartilage tissue engineering.</p>

The study of cell biocompatibility of new pattern biphasic calcium phosphate nanocomposite in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the cell biocompatibility of porous biphasic calcium phosphate nanocomposite in vitro.</p><p><b>METHODS</b>Bone marrow mesenchymal cell (BMSCs) obtained from SD rat bone marrow were in vitro induced and proliferated. Afler their osteoblast phenotypes were verified, BMSCs were seeded onto prepared porous biphasic calcium phosphate nanocomposite (Experiment group) and common porous hydroxyapatite (Control group). The cell adhesion was evaluated by scanning electron microscope. Synthesis of alkaline phosphatase enzyme (ALP) and osteocalcin were detected and cell cycle was detected by flow cytometry.</p><p><b>RESULTS</b>BMSCs could fully attach to and extend on the material in experiment and control group, Moreover, experiment group were superior to control group in adhesion, proliferative abilities and osteogenic activity.</p><p><b>CONCLUSION</b>BMSCs can differentiate to osteoblast phenotype; the porous biphasic calcium phosphate nanocomposite as bone tissue engineering scaffold has good cell biocompatibility.</p>

A study on transfecting green fluorescent protein gene to rat bone marrow mesenchymal stem cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study an efficient method to transfect green fluorescent protein gene (GFP) to rat bone marrow mesenchymal stem cells (MSCs) and to determine the biological properties and differentiation potency of transfected MSCs.</p><p><b>METHODS</b>SD rats' bone marrow MSCs were separated and purified in vitro. After subculture and expansion, MSCs infected with Adenoviral vector (Ad-GFP) or transfected with liposome were observed, and their transfection efficiency was assessed with flow cytometry. The MSCs expressing GFP gene were induced to differentiate to osteoblast, and non-transfected MSCs were set as control.</p><p><b>RESULTS</b>Ad-GFP delivered GFP gene with high efficiency to rat MSCs. (41.3 +/- 1.4)% of MSCs infected with Ad-GFP expressed GFP gene, which was much higher than the control (12.5%). Expression of GFP gene of infected MSCs maintained stable from 1 to 6 weeks after infection. Infected MSCs possessed the same alkaline phosphatase activation as non-infected MSCs, and formed mineralized mouldes.</p><p><b>CONCLUSIONS</b>The infected MSCs with Ad-GFP expressed GFP with much higher efficiency than liposome transfection, and maintained the same ability of proliferation and differentiation as non-infected MSCs. Transfection with Ad-GFP is a highly effective method for labeling MSCs.</p>

Cultivation and induced differentiation of bone marrow stromal cells of SD rats with type I osteoporosis in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the biological features and osteoblast/adipocyte phenotypes of bone marrow stromal cells (BMSCs) of Sprague-Dawley (SD) rats with Type I osteoporosis by induced culture.</p><p><b>METHODS</b>Six-month-old SD rats were used in this study. 16 female rats were randomly divided into two groups. Eight rats were ovariectomied as experimental group to establish the modle of Type I osteoporosis, while other rats received sham-operation. Three month later BMSCs of 16 rats were isolated by discontinueous gradient centrifugation and then plated in alpha-MEM medium as primary culture. Secondary harvested cells were cultured for 14 days in alpha-MEM medium supplemented with dexamethasone, ascorbic acid, vitamin D3, beta-glycerophosphate or dexamethasone, 3-isobutyl-1-methylxanthine, insuline, and indomethine. The cells were screened by inverted microscope each day and cell growth was studied with cell counting. The osteoblast and adipocyte phenotypes were verified by cytochemistry staining, counted the percentage of positive stained cells.</p><p><b>RESULTS</b>The weight and bone mineral density of rats were statistically different between experimental group and control group. Gomori and Von Kossa's staining demonstrated positive osteoblast phenotypes of alkaline phosphatase and mineralized nods by osteogenic inducer, while Oil Red O staining identified BMSCs treated with adipogenic medium resulted in adipocyte formation and there was no significant difference in the percentage of positive stained cells between two groups.</p><p><b>CONCLUSION</b>The model of Type I osteoporosis has been established successfully. BMSCs from SD rats with osteoporosis maintain their differentiation potential.</p>

Study on multi-lineage potential of bone marrow mesenchymal stem cells derived from green fluorescent protein transgenic mice / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the multi-lineage potential of bone marrow mesenchymal stem cells (MSCs) derived from transgenic mice with green fluorescent protein (GFP) gene in vitro.</p><p><b>METHODS</b>A 6-week-old GFP transgenic mouse was executed by dislocation of cervical vertebra, and the marrow in tibia and thighbone was washed out with asepsis. The limited cell strains of MSCs derived from GFP transgenic mice (GFP-MSCs) were obtained with density gradient centrifugation. The passage 3 GFP-MSCs were induced to differentiate into osteoblast, adippcyte, neuron with solution of calcium induction medium, adipogenic medium and neural induction medium respectively. After being calcium-induced, the activity of alkaline phosphatase on GFP-MSCs was determined by micro-plate reader, and alizarin red staining was performed to test the formation of calcium concentration. The adipo-induced MSCs were detected with oil red O staining. Immunocytochemical staining was performed to detect the expression of NSE on neuron-induced MSCs.</p><p><b>RESULTS</b>The ALP activity of GFP-MSCs heightened gradually along with being calcium-induced, and alizarin red staining showed positive. Oil red O staining of adipo-induced cells and NSE immunocytochemical staining of neuron-induced cells demonstrated positive.</p><p><b>CONCLUSION</b>The limited cell strain of GFP-MSCs possesses multi-lineage potential, which can be used as an efficient tracking facility for studying the mechanism of multi-lineage potential on the MSCs.</p>

Expression and secretion of human tumor necrosis factor gene transfected on human embryo myoblasts / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe human tumor necrosis factor-alpha (hTNF-alpha) expression and secreting level of human embryo myoblasts transfected by hTNF-alpha gene.</p><p><b>METHODS</b>Human embryo myoblasts were transfected with shuttle plasmid pSV23SHTNF containing hTNF-alpha gene by cationic liposomes DOSPER. The control group was only given equivalent liposomes except plasmid. After culturing for 24, 48, 72 and 96 hours, hTNF-alpha expression level of human embryo myoblasts was observed with immunocytochemistry staining, and hTNF-alpha secreting of human embryo myoblasts was analyzed by ELISA.</p><p><b>RESULTS</b>After transfected by hTNF-alpha gene for 24, 48, 72 and 96 hours, the human embryo myoblasts displayed significant secretion of hTNF-alpha in the cultural supernatant (P < 0.05), and overexpression in cytoplasma and cell membrane.</p><p><b>CONCLUSION</b>Transfection of hTNF-alpha gene to human myoblasts made myoblasts secrete high concentration of hTNF-alpha, implying it is feasible that transfecting muscle cells surrounding tongue carcinoma lesion with hTNF-alpha gene can prevent tongue carcinoma from intruding into deeper muscle tissue.</p>