The Experimental Study On Therapy Of Venous Malformation By Tissue Engineering Technology

The therapy of venous malformations (VMs) is always seen as considerable difficulty confronting medical circle. At present the usual therapeutic methods are surgical operation, laser therapy, medical therapy, physical therapy, etc. But they all have certain defects. In recent years, with the rapid development of the technic of tissue engineering and much further study of VM, it is possible to solve the problem to our satisfaction.VMs which often were seen on oramaxillofacial part of the patients are congenital lesions in clinic. Lesions tend to enlarge slowly, in parallel with normal growth, but can dilate and become symptomatic. The indications are unreceptive appearance, severe bleeding, and embarrassment of swallow and breath, which so badly affect the patients'life quality that having come to the public's attention. According to the present research, the mechanism of VM is still a question. But most of scholars consider it is caused by the abnormal conductive ways which adjust the normal vascular tree. And there hasn't been an ideal therapeutic method that is universally accepted up to now. Furthermore, there are still some difficulties for complex VMs that can involve large permeative soft tissue lesions and frequently cross multiple tissue planes in clinical application.Fibrin glue (FG) that was distilled from the extinguished virus blood plasma is a kind of natural biologic protein, which can form three-dimensional poly material in solid. It has a good histocompatibility and degradation with not immunorepulsive response. In early days it is only used in hemostasia. In the last 20 years, FG is increasingly attracted by people because it is useful in healing wound, closing tissue, filling default, preventing tissue conglutination, especially in some medicines'slowgoing carrier. Nowadays FG is widely used in the foreign surgical field and has got perfect effect.The experiment conducts some relevant studies on FG material made by the biotechnological company of Guang Zhou PeiXiu and uses it as the carrier material of VM therapy to do a series of experiments to explore whether FG material is appropriate for scaffold material in vascular vessel. The combination of FG material and autologous fibroblasts (FB) is used to embolize rabbit's jugular vein to provide basis for the choice of scaffold material and the therapy of VM by applying tissue engineering technology. What's more, it also provides experimental foundation for further clinical application.1. The study on the culture and identification of fibroblasts in adult rabbit skin in vitroObjective: To learn about the efficient primary culture method of FB in adult rabbit skin in vitro. Methods: The rabbit FB were obtained by digesting collagen hypoderm and purified by differential speed attachment. Draw the growth carve of FB of the third generation. Identify FB by vimentin immunostaing and chromosome analysis. Results: FB has good expanding form and proliferating trend. The cells appear the typical FB appearance. The growth carve appears the typical"S". The results of vimentin immunostaing and chromosome analysis approve the culture cells are rabbit FB. Conclusion: The method obtained high pure adult rabbit FB is simple and effective. It's a preparation for the next experiment.2. The study on compounding of rabbit FB and FG material in vitroObjective: To learn about the compound situation when rabbit FB and FG material co-culture in vitro, and to further verify the feasibility of FG material as scaffold material in vein. Methods: Cultivated the compound of rabbit FB and FG material in vitro; observed the compound situation of cells and material through inverted microscope, SEM; tested the influence that the material has upon cells multiplication differentiation by vimentin immunostaing analysis. Results: FB had a good quality of adherency, transference, multiplication and differentiation on the surface and in the vivo of FG material, and FG material accelerated multiplication and differentiation in some aspect. Conclusion: The FG material is a good scaffold for FB.3. The study on the embolization of rabbit jugular vein with FG material compound autoallergic FB of rabbitsObjective: To examine the ability of the embolization of rabbit jugular vein by tissue engineering embolism compounded in vitro by FG and autoallergic FB in short terms. Methods: Implanted the compound of FG and BrdU labeled autoallergic FB to the rabbit jugular vein; divided the FG group as the control group; observed and compared the embolization situation of jugular vein and the end-result of seed cells through gross observation, HE staining, immunohistochemistry dyeing after 10 days of operation respectively. Results: FG+ autoallergic FB group had a strong ability in the embolism of rabbit jugular vein. FG group's embolism ability was weaker. Conclusion: The FG is a good scaffold material of tissue engineering and has a good ability in embolizing vein when applied in the construction of tissue engineering embolism in vitro. So it is hopeful to be applied in the clinical therapy on large permeative soft tissue lesions of VM.