Experimental Research On Vascularization Of Tissue-engineered Submandibular Gland With Prefabrication In Vivo

ObjectiveThe ultimate goal of Submandibular gland tissue-engineering study is to have a artificial organs with specific structure and secretory function,that derived from biodegradable materials with a three-dimensional spatial structure implanted with seed cells with biological activity, and cell proliferation and differentiation continue to maintain its function, and eventually formed a artificial organs. As the blood supply of the reconstruction is the basis for tissue regeneration, to get a good new tissue-engineered organizations, must ensure that the seed cells, particularly the inside seed cells of the material able to obtain adequate nutrition timely, which in turn depends on an adequate blood supplies and materials complexed within cells and rebuild the blood supply of the extent and speed. Therefore, the the reconstruction of blood supply has become the basis of tissue-engineered objects from the critical link to the clinical application.This study aims to use tissue engineering techniques to have a capacity of submandibular gland proliferation and differentiation of seeded cells and collagen sponge implantation animal culture, using the method of prefabrication to nurture tissue-engineered complexes and observe the situation of vascularization, to explore the feasibility of constructing tissue-engineered Submandibular gland with vitality in the allogeneic Animals' body.MethodsSubmandibular glands were obtained from three days age SD rat, submandibular gland cells(RSGCs) were cultured with tissue-culture method, and the source of RSGCs was identified; the cells were seeded in gelatin sponge scaffolds, cultured in vitro for 72 hours; In experimental group of prefabricated vascularized epigastric pedicled flap, Parcel complexes of cells and gelatin sponge scaffold, control group of SD rats were implanted directly under the abdominal wall. Different time (4 days,7 days,14 days,21 days) were observed by light microscopy, scanning electron microscopy, histology and vascular perfusion ink marker,BrdU immunohistochemical staining such as testing, and MetaMorph software is used in analysis The proportion of stained regional area to the total area(Pv), SPSS 13.0 software statistical analysis of the data.Observation of submandibular gland cells and gelatin sponge scaffold complex in vivo The growth and blood vessel of the situation.ResultsIn experimental group, the implants has noticeable blood vessels and a large number of India ink staining and dyeing area exists in the sponge materials at the first 7-14 days, submandibular gland cells with the sponge materials adhesively growth, similar to duct-like structure can be observed, vascular has no significant change and maintains stability at 14-21 day.ConclusionsThe method of prefabrication can provide a good blood supply to tissue-engineered submandibular gland, and provide a theoretical and experimental basis to study vascularization of tissue-engineered submandibular gland.