| Purpose:Venous malformations (VMs) are one type of vascular malformations, formerly referred as "cavernous hemangiomas". The lesions mainly occured in the oral and maxillofacial area and involved multiple anatomical regions including the skin, mucosa, and critical neuromuscular structures. VMs located in the head and neck area and mucosal surfaces may cause significant cosmetic defects, interfere with normal speech and dentition and lead to recurrent bleeding, obstruct the airway. Due to its numerous treatments, modern medicine advocates that the appropriate treatment should be selected according to the patient’s specific condition. Pingyangmycin (bleomycin A5hydrochloride, PYM) sclerotherapy is a preferred method for the treatment of venous malformations in the oral and maxillofacial area, which got an encouraging therapy results. Nevertheless, maxillofacial vascular have an abundant transportion, and the venous reflex quickly, traditional sclerotherapy carries the risk of complications, and cann’t be used to treat some vascular lesions (especially the larger ones). In addition, the side effects of pingyangmycin may be more harmful to the patients with long term application. It is necessary for us to look for other more effective methods with relatively minimal side effects during sclerotherapy. Therefore, we seek out the Hyaluronic acid (HA).HA is a naturally linear glycosaminoglycan (GAG) composed of alternating disaccharide units of D-glucuronic acid and N-acetyl-D-glucosamine. Because of its features:water soluble, biodegradable, and suggested to be biocompatible, non-toxic, non-immunogenic and can be chemically modified, which make it an attractive targeting drug delivery vehicles. Also it is vital to proper cell growth, embryonic development, healing processes, inflammation, and tumor development. Its function that the inhibitory effect on the endothelial cell implies that it may also inhibit the development of venous malformations.Lesions are composed of expansion venous cavity lined with mature endothelial cells. In this study, we applied HVMECs as a research model, and explored the effects of PYM cooperating with HA on the function of HVMECs. Also the mechanism was speculated.Materials and Methods:We obtained primary HVMECs from a surgical resection human venous malformation (VM) specimen and cultured with endothelial cell medium (ECM). Observe the growth and morphological characteristics of the cultured cells with an inverted phase contrast microscope. Identify the cultured cells with endothelial cell-specific antigen:the rabbit anti-human vWF and rabbit anti-human CD34. Then Filter the optimal concentration of HA and PYM on HVMECs. The effects on cells proliferation were studied by MTT assay, angiogenesis were evaluated by tube formation assay and cells apoptosis were assessed by TUNEL method. Results:From the VMs specimen, we acquired the HVMECs. CD34and vwf were both positively expressed within the cultured cells. The optimal concentration of HA and PYM on HVMECs was400ug/ml and lOug/ml. The results of MTT assay showed that significant difference was observed between PYM and PYM combined with HA. There were almost no capillary tube formation in the group of PYM combined with HA, and the difference was significant with control group. The group of PYM combined with HA caused more TUNEL-positive cells than the group of PYM, and the difference was significant.Conclusion:Our results indicate that the combination of PYM and HA was more effective than single PYM on inhibiting HVMECs-growth, angiogenesis and promoting HVMECs-apoptosis. This study suggests that combination of PYM and HA could be a novel attractive agent for treatment of vascular malformation. |
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