Experimental Study Of Repairing Effect Of HBD₂ Gene Modified Dermal Multipotent Stem Cells Transplantation On Infected Wound

Wound healing is relative difficult under the condition of large area of skin injury and burn, especially when accompanied by radiation injury. This is due to the significant loss of repair cells in wounded skin and the frequently accompanied wound infection as well. In our previous study, my colleagues found that transplantation of rat's dermal multipotent stem cells (dMSCs) and mouse's bone marrow stromal stem cell either by spotted injection or by systemic transfusion could significantly promote wound healing, but wound infection was still a factor hindering wound healing.Human beta defensin 2 (hBD2), one of the antimicrobial peptides family, has wide-spectrum and high efficient antimicrobial activity. hBD2 is positive charged on its surface, which make it can combine with microbes abounding with phospholipid (which is negtive charged), then to form aperture in the microbes'cell membrane and eventually cause the death of microbes due to the loss of important substance in the cells. Its different antimicrobial mechanisms with the traditional antibiotics result in relative low frequency of resistance. The above-mentioned advantages make hBD2 a potential good measure in the treatment of infected wound. But under normal conditions, the expression of hBD2 in normal tissue is very low, so in the present study, in order to exert the repairing effect of dMSCs and the antimictobial effect of hBD2 at the same time, we firstly constructed the adenoviral expression vector of hBD2, and transfected dMSCs with the vector to make dMSCs overexpressed hBD2, then transplanted the gene modified dMSCs to infected wound by spotted injection. The main methods, techniques and results are shown as the following:1. Construction and identification of recombinant adenovirus vector of hBD2. Total RNA was obtained from the injured skin of patients suffered from psoriasis, and full-length cDNA of hBD2 containing NotI and HindIII endonuclease sites was amplified by RT-PCR. Then hBD2 gene was first cloned into shuttle vector pAdTrack-CMV, then and backbone...