| Objective 1.To investigate the effects of hyaluronic acid oligosaccharide on the proliferation,migration and angiogenesis of human umbilical vein endothelial cells(HUVECs).2.To study the effects of hyaluronic acid oligosaccharide on wound healing area,wound blood flow and angiogenesis in diabetic rats.3.To explore the mechanism of hyaluronic acid oligosaccharide on wound healing in diabetic rats.Methods The effects of o-HA on the wound healing of diabetic rats were studied in vivo and in vitro.The effects of o-HA on HUVEC proliferation,migration and tubule formation was measured by MTT,scratches and tubule formation assay at the concentration of 10 ?g/m L in glucose(33 mmol/L)separately.The o-HA ointment was prepared by emulsification method and its local application was performed on the wound of diabetic rats.The wound size and inflammation were observed and recorded.The degree of epithelialization,granulation tissue and fibroblast regeneration were detected by HE staining.The angiogenesis was examined by CD31 immunohistochemical staining.The expression of p-Src,p-ERK and TGF-beta1 protein in the wound tissues was further detected by western blot to study the mechanism of o-HA on diabetic rats wound healing.Results 1.The MTT assay showed that the cell proliferation in o-HA group was higher than that in PBS group(P<0.05).The scratch test showed the migration area of o-HA group was significantly higher than PBS group(P<0.01)The reductionrate was about 58%.The tube formation test showed that the number of lumen formation in o-HA group was higher than that in PBS group,which was statistically significant(P <0.05).2.Hyaluronic acid oligosaccharide ointment was successfully prepared and applied to the wound of diabetic rats.3.HE staining and photographic record observation showed that the wound healing in the treatment of hyaluronic acid oligosaccharide ointment group of was significantly faster than that in the matrix group(P<0.05).The degree of fibroblasts,granulation tissue regeneration and epithelization in the treatment of hyaluronic acid oligosaccharide ointment group were significantly higher than that in the matrix group(P<0.01).Laser Doppler flow imaging(CDFI)and CD31 immunohistochemical staining showed that o-HA ointment significantly increased blood supply and angiogenesis in the rats,compared with matrix ointment(P<0.05).4.Western Blot showed that the expression of p-Src,p-ERK and TGF?1 protein in o-HA ointment group was significantly higher than that in matrix group(P<0.01).Conclusions 1.o-HA promoted HUVEC proliferation,migration and tubule formation in hyperglycemic environment.2.Topical application of o-HA ointment promoted wound healing by increasing endothelial cell proliferation,migration and angiogenesis in diabetic rats. |
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