Enhanced angiogenesis in grafted skins by laser-induced stress wave-assisted gene transfer of hepatocyte growth factor.

Treatment to increase secretion of growth factors related to angiogenesis by gene transfection is a promising therapeutic solution for improving the outcome of tissue transplantation. We attempted to deliver a therapeutic vector construct carrying the human hepatocyte growth factor (hHGF) gene to skin grafts of rats using laser-induced stress waves (LISWs), with the objective of enhancing their adhesion. First we delivered the hHGF gene to rat native skin in vivo to determine the optimum gene transfer conditions. We then transferred the hHGF gene to excised rat skins, with which autografting was performed. We found that the density and uniformity of neovascularities were significantly enhanced in the grafted skins that were transfected using LISWs. These results suggest the efficacy of this method to improve the outcome of skin grafting. To our knowledge, this is the first experimental demonstration of a therapeutic efficacy based on LISW-mediated gene transfection. Since the present method can be applied not only to various types of tissues but also to bioengineered tissues, this technique has the potential to contribute to progress in transplantation medicine and future regenerative medicine.

[1]  H Green,et al.  Growth of cultured human epidermal cells into multiple epithelia suitable for grafting. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[2]  P. Ballard,et al.  Physical study of laser-produced plasma in confined geometry , 1990 .

[3]  K. Rieger-Christ,et al.  Efficacy of combination gene therapy with multiple growth factor cDNAs to enhance skin flap survival in a rat model. , 2005, DNA and cell biology.

[4]  F. Larcher,et al.  Nonviral transfer of genes to pig primary keratinocytes. Induction of angiogenesis by composite grafts of modified keratinocytes overexpressing VEGF driven by a keratin promoter , 1999, Gene Therapy.

[5]  K. Ishii,et al.  Sterically Stabilized Cationic Liposomes Improve the Uptake and Immunostimulatory Activity of CpG Oligonucleotides1 , 2001, The Journal of Immunology.

[6]  M. Berns,et al.  Direct gene transfer into human cultured cells facilitated by laser micropuncture of the cell membrane. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[7]  J. Morgan,et al.  Transient Hyperproliferation of a Transgenic Human Epidermis Expressing Hepatocyte Growth Factor , 2002, Cell transplantation.

[8]  Nobuyuki Itoh,et al.  Fibroblast growth factors , 2001, Genome Biology.

[9]  G. Huemer,et al.  Adenovirus-mediated transforming growth factor-β ameliorates ischemic necrosis of epigastric skin flaps in a rat model1, 2 , 2004 .

[10]  W. Jiang,et al.  The molecular and clinical impact of hepatocyte growth factor, its receptor, activators, and inhibitors in wound healing. , 2006, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[11]  T Nakamura,et al.  Hepatocyte growth factor (HGF) as a tissue organizer for organogenesis and regeneration. , 1997, Biochemical and biophysical research communications.

[12]  T. Flotte,et al.  Nuclear Transport by Laser-Induced Pressure Transients , 2003, Pharmaceutical Research.

[13]  E. Crescenzi,et al.  Targeted gene transfer in eucaryotic cells by dye-assisted laser optoporation. , 1996, Journal of photochemistry and photobiology. B, Biology.

[14]  A. Doukas,et al.  Cell Loading with Laser-Generated Stress Waves: The Role of the Stress Gradient , 1999, Pharmaceutical Research.

[15]  Minoru Obara,et al.  Gene transfer into mammalian cells by use of a nanosecond pulsed laser-induced stress wave. , 2004, Optics letters.

[16]  Xin-Hua Hu,et al.  Efficient delivery of small interfering RNA to plant cells by a nanosecond pulsed laser-induced stress wave for posttranscriptional gene silencing. , 2006, Plant science : an international journal of experimental plant biology.

[17]  R. Gamelli,et al.  Vascular endothelial growth factor mediates angiogenic activity during the proliferative phase of wound healing. , 1998, The American journal of pathology.

[18]  G. Huemer,et al.  Gene therapy with adenovirus‐mediated VEGF enhances skin flap prefabrication , 2005, Microsurgery.

[19]  H. Kleinman,et al.  Scatter factor induces blood vessel formation in vivo. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Y. Taniyama,et al.  Angiogenesis induced by hepatocyte growth factor in non-infarcted myocardium and infarcted myocardium: up-regulation of essential transcription factor for angiogenesis, ets , 2000, Gene Therapy.

[21]  M. Siemionow,et al.  Enhancement of epigastric skin flap survival by adenovirus-mediated VEGF gene therapy. , 2002, Plastic and reconstructive surgery.

[22]  P. Vogt,et al.  Genetically modified keratinocytes transplanted to wounds reconstitute the epidermis. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Bernhard O Palsson,et al.  Optoinjection for efficient targeted delivery of a broad range of compounds and macromolecules into diverse cell types. , 2006, Journal of biomedical optics.

[24]  Minoru Obara,et al.  In vitro gene transfer to mammalian cells by the use of laser-induced stress waves: effects of stress wave parameters, ambient temperature, and cell type. , 2006, Journal of biomedical optics.

[25]  Minoru Obara,et al.  Targeted DNA transfection into the mouse central nervous system using laser-induced stress waves. , 2005, Journal of biomedical optics.

[26]  P. Walicke,et al.  Fibroblast growth factors. , 1989 .

[27]  M. Urken,et al.  Locally Administered Vascular Endothelial Growth Factor cDNA Increases Survival of Ischemic Experimental Skin Flaps , 1998, Plastic and reconstructive surgery.

[28]  Artium Khatchatouriants,et al.  Femtosecond infrared laser-an efficient and safe in vivo gene delivery system for prolonged expression. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[29]  H. Kleinman,et al.  Scatter factor (hepatocyte growth factor) is a potent angiogenesis factor in vivo. , 1993, Symposia of the Society for Experimental Biology.

[30]  M. Aoki,et al.  A vascular modulator, hepatocyte growth factor, is associated with systolic pressure. , 1996, Hypertension.

[31]  T J Flotte,et al.  Stress‐wave‐assisted transport through the plasma membrane in vitro , 1997, Lasers in surgery and medicine.

[32]  J. Laws State-of-the-art burn treatment. , 1996, Occupational health & safety.

[33]  Jeffrey R Morgan,et al.  FGF-7 expression enhances the performance of bioengineered skin. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[34]  Kunio Matsumoto,et al.  Gene transfer of human hepatocyte growth factor into rat skin wounds mediated by liposomes coated with the sendai virus (hemagglutinating virus of Japan). , 2002, The American journal of pathology.

[35]  Herbert Schneckenburger,et al.  Laser-assisted optoporation of single cells. , 2002, Journal of biomedical optics.

[36]  C. Garlanda,et al.  Involvement of endothelial PECAM-1/CD31 in angiogenesis. , 1997, The American journal of pathology.

[37]  Minoru Obara,et al.  In vivo targeted gene transfer in skin by the use of laser‐induced stress waves , 2004, Lasers in surgery and medicine.

[38]  Y. Taniyama,et al.  Acceleration of wound healing by combined gene transfer of hepatocyte growth factor and prostacyclin synthase with Shima Jet , 2006, Gene Therapy.

[39]  Y. Ikawa,et al.  A novel method of DNA transfection by laser microbeam cell surgery , 1984 .

[40]  C. Dubuquoy,et al.  TLR9 pathway is involved in adjuvant effects of plasmid DNA-based vaccines. , 2005, Vaccine.

[41]  Ulrich Kneser,et al.  Tissue engineering of cultured skin substitutes , 2005, Journal of cellular and molecular medicine.