Sheela R Damle1, Bernard A Rawlins2, Oheneba Boachie-Adjei2, Ronald G Crystal3, Chisa Hidaka4, Matthew E Cunningham2. 1. Hospital for Special Surgery, 535 East 70th Street, New York, NY 10021 USA. 2. Hospital for Special Surgery, 535 East 70th Street, New York, NY 10021 USA ; Weill Cornell Medical College, New York, NY 10065 USA. 3. Weill Cornell Medical College, New York, NY 10065 USA ; Department of Genetic Medicine and Belfer Gene Therapy Core Facility, Weill Cornell Medical College, New York, NY 10065 USA. 4. Hospital for Special Surgery, 535 East 70th Street, New York, NY 10021 USA ; Department of Genetic Medicine and Belfer Gene Therapy Core Facility, Weill Cornell Medical College, New York, NY 10065 USA.
Abstract
BACKGROUND: Basic research toward understanding and treating disc pathology in the spine has utilized numerous animal models, with delivery of small molecules, purified factors, and genes of interest. To date, gene delivery to the rat lumbar spine has only been described utilizing genetically programmed cells in a matrix which has required partial disc excision, and expected limitation of treatment diffusion into the disc. PURPOSE: This study was designed to develop and describe a surgical technique for lumbar spine exposure and disc space preparation, and use of a matrix-free method for gene delivery. METHODS: Naïve or genetically programmed isogeneic bone marrow stromal cells were surgically delivered to adolescent male Lewis rat lumbar discs, and utilizing quantitative biochemical and qualitative immunohistological assessments, the implanted cells were detected 3 days post-procedure. RESULTS: Statistically significant differences were noted for recovery of the β-galactosidase marker gene comparing delivery of naïve or labeled cells (10(5) cells per disc) from the site of implantation, and between delivery of 10(5) or 10(6) labeled cells per disc at the site of implantation and the adjacent vertebral body. Immunohistology confirmed that the β-galactosidase marker was detected in the adjacent vertebra bone in the zone of surgical implantation. CONCLUSIONS: The model requires further testing in larger cohorts and with biologically active genes of interest, but the observations from the pilot experiments are very encouraging that this will be a useful comparative model for basic spine research involving gene or cell delivery, or other locally delivered therapies to the intervertebral disc or adjacent vertebral bodies in rats.
BACKGROUND: Basic research toward understanding and treating disc pathology in the spine has utilized numerous animal models, with delivery of small molecules, purified factors, and genes of interest. To date, gene delivery to the rat lumbar spine has only been described utilizing genetically programmed cells in a matrix which has required partial disc excision, and expected limitation of treatment diffusion into the disc. PURPOSE: This study was designed to develop and describe a surgical technique for lumbar spine exposure and disc space preparation, and use of a matrix-free method for gene delivery. METHODS: Naïve or genetically programmed isogeneic bone marrow stromal cells were surgically delivered to adolescent male Lewis rat lumbar discs, and utilizing quantitative biochemical and qualitative immunohistological assessments, the implanted cells were detected 3 days post-procedure. RESULTS: Statistically significant differences were noted for recovery of the β-galactosidase marker gene comparing delivery of naïve or labeled cells (10(5) cells per disc) from the site of implantation, and between delivery of 10(5) or 10(6) labeled cells per disc at the site of implantation and the adjacent vertebral body. Immunohistology confirmed that the β-galactosidase marker was detected in the adjacent vertebra bone in the zone of surgical implantation. CONCLUSIONS: The model requires further testing in larger cohorts and with biologically active genes of interest, but the observations from the pilot experiments are very encouraging that this will be a useful comparative model for basic spine research involving gene or cell delivery, or other locally delivered therapies to the intervertebral disc or adjacent vertebral bodies in rats.
Authors: Satoshi Sobajima; John F Kompel; Joseph S Kim; Corey J Wallach; Douglas D Robertson; Molly T Vogt; James D Kang; Lars G Gilbertson Journal: Spine (Phila Pa 1976) Date: 2005-01-01 Impact factor: 3.468
Authors: Jari Salo; Zygmunt Mackiewicz; Aage Indahl; Yrjö T Konttinen; Allison Kaigle Holm; Antti Sukura; Sten Holm Journal: Spine (Phila Pa 1976) Date: 2008-04-15 Impact factor: 3.468
Authors: Matthew E Cunningham; Natalie H Kelly; Bernard A Rawlins; Oheneba Boachie-Adjei; Marjolein C H van der Meulen; Chisa Hidaka Journal: Sci Rep Date: 2022-10-07 Impact factor: 4.996