title: Construction of a tissue engineered intervertebral disc with high biological activity using an allogeneic intervertebral disc supplemented with transfected nucleus pulposus cells expressing exogenous dopamine beta-hydroxylase.
abstract: This study addressed the in vitro construction and biological activity of tissue engineered intervertebral discs with exogenous human dopamine beta-hydroxylase (DBH) nucleus pulposus cells. pSNAV2.0-DBH expression plasmids were utilized to enhance the survival rates of intervertebral disc tissue cells. Various concentrations of transfected nucleus pulposus cells were injected into the discs, and DBH mRNA expression was determined using polymerase chain reaction amplification. Polysaccharide content and total collagen protein content in the engineered disc nucleus pulposus tissue were determined. The visible fluorescence intensities of the 1 x 10(5) and 1 x 10(6) groups vs the 1 x 10(4) group were significantly increased (P < 0.05); no significant difference was observed between the 1 x 10(5) and 1 x 10(6) groups (P > 0.05) at 7 days after injection. DBH mRNA expression could be detected in the all but the EGFP control group at 14 days culture. No significant difference was observed in the protein content between the 1 x 10(4) and the control groups at various times, while the protein content was significantly higher in the 1 x 10(5) vs the control and the 1 x 10(4) groups at 7-, 14-, and 21-day cultures. These results demonstrate that a tissue engineered intervertebral disc with high biological activity can be constructed by utilizing allogeneic intervertebral discs stored in liquid nitrogen and a 1 x 10(5) transfected nucleus pulposus cell complex with in vitro culture for 14 days. This model can be used in animal experiments to study the biological activity of the engineered discs.
authors.affiliation: Department of Spinal Surgery, The Second Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia, China.
authors.forename: M
authors.initials: M
authors.lastname: Bai
authors.affiliation: Department of Gynecology and Obstetrics, The First Hospital of Hohhot City, Hohhot, Inner Mongolia, China.
authors.forename: Y H
authors.initials: YH
authors.lastname: Wang
authors.affiliation: Department of Spinal Surgery, The Second Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia, China yinhpdmis@163.com.
authors.forename: H P
authors.initials: HP
authors.lastname: Yin
authors.affiliation: Department of Spinal Surgery, The Second Hospital of Inner Mongolia Medical University, Hohhot, Inner Mongolia, China.
authors.forename: S W
authors.initials: SW
authors.lastname: Li
mesh_terms: D000818:Animals; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D004285:Dogs; D004299:Dopamine beta-Hydroxylase / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology* / Q000378:metabolism; D055959:Intervertebral Disc Degeneration / Q000628:therapy; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D046509:Tissue Culture Techniques; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection; D019076:Transgenes; D014184:Transplantation, Homologous
chemical_list: D011509:Proteoglycans; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D003094:Collagen; D004299:Dopamine beta-Hydroxylase