Construction Of Vascularized Inguinal Hernia Patch And Its Pro-vascularization Mechanism By Silencing The Int6 Gene Composite With Bioscaffold

Inguinal hernia is the most common type of hernia.Hernia is a common surgical disease.There are some gaps in the abdominal ca vity of the human body,abdominal wall defects or some weak positions.The abdominal organs or tissues leave the original normal part and slide in other locations which can cause hernia.According to the anatomical site,the inguinal hernia is divided into inguinal hernia,femoral hernia,umbilical hernia,incisional hernia,ctc.;according to the cause of the disease:congenital and acquired;the following picture:inguinal hernia,femoral hernia,umbilical hernia,etc.;According to clinical manifestations:easy rcnaturation,difficult to reversible sputum(sliding sputum),incarcerated sputum(enveloped spulum),Richter's hernia,Litter's hernia,Maydl's hernia,strangulation hernia.Adult inguinal hernia is now thought to be caused by a disorder of the extracellular r matrix.For example,changes in matrix metalloproteinases and their inhibitors have been observed in diseases,and collagen mctabolism in patients has been disturbed inaunique manner.Compared with the control group,the degradation of immature type ?collagen was reduced in patients with inguinal hernia,and the conversion of type ?collagen in the basement membrane was inereased.Clinically,the main treatment for inguinal hernia is torepair hernia by directsuturing or artificial patching.More than 20 million pitients worldwide undergo inguinal hernia repair every year.Most of the inguinal hernias can be repaired withartifieial patches.Inguinal hernia repair is the most common genera surgery in industrialized countrice,and about 200 out of every 100,000 pcople undergoing surgeryis an inguinal hernia repair.Surgery based suture and patch techniques can be used andthe procedure can be open or minimally invasive.There are many different approaches to inguinal hernia repair,and indications for treatment and a range of techniques require standardized guidance to minimize theprobability of complications and improve prognosis.The main treatment goal is to improve the patient's treatment effeet,especially to reduce the infection and recurrence rate,reduce chronic pain,which is the most common problem after inguinal hernia irepair.Although the repair of this patch significantly reduces the recurrence rate alter inguinal hernia repair,the rate of replacement of the patch into autologous tissue does not match the rate of degradation.The patch's own anti-inlective ability leads to postoperative complications such as infection.Acellular vascular patch,through chemical or physical methods to remove the cells,DNA,RNA,and protein components wlth strong antigenieity in the porcine aorta,and then only a series of operations are completed,leaving only the elastic liber and collagen.The basic exlracellular matrix eomponent,such as prolein,provides a suilable thre-dimensilonal structure for the growth of blood vessels,adhesion of tissue cells,and entry of nerve fibers,and preserves the mechanical properties of the original large blood vessels very well.Polymeric polymer patch of polycaprolactone PCL/gelatin electrospinning patch,PCL/gelatin polymer can he used as a scatltbld to help cell adhesion,maintain cell function and support cell growth,provide presence for cells and extracellular matrix place.However,the cellular affinity of the synthesized polymers is generally poor.In a previous study,PCL/gelatin nanofiber scaffolds were used as skin reconstruction.This experiment used PCL/gelatin nanofiber seaffold for theprep aration of inguinal hernia patch.We prepared a two-ayer composite patch of acvellular vaseular-PCL/gelatin.Preparation of decellularized blood vessel-PCL/gelatin electrospinning composite patch.Based on the preparation of the above two materials,the prepared decellularized vascular material is placed on the spinning platform of the electrospinning machine,and the static electricity is directly applied thereto.Spin-spray PCL/gelatin solution.A double-sided decellularized blood vessel-PCL/gelatin electrospinning composite patch was formed.This material was compared with a simple decellularized vascular material and a simple PCL/gelatin nanofiber scaffold to determine the acellular vascular-PCL/gelatin electrospun composite patch as the mainseaffold carrier for the next in vivo experiment.Insuffieient vascularization is an important cause of infection.In prophylaetie applications,optimizing wound perfusion and providing perative supplementation with O2 can reduce the incidence of postoperative infection.Tissues in the near hypoxic area are prone to necrosis,which in turn may cause secondary tissue damage and infection.Therefore,we thought of making a tissue engineering patch that would light infection and increase the accumulation of extracellular matrix.One of the most direct methods of giving it long-term anti-infective function in the body is to promote its own vascularization.The operation of promoting HIF-2? expression is a promising method for promoting vascularization of grafts in vivo,as demonstrated by the previous studies.HIF-2? is mainly expressed in vascular endothelial cells.Under hypoxic conditions,HIF-2a is stabilized and forms heterodimers,which activate other vascular production factors.Chen et al reporting that endogenous int6 silencing is possible even in an environment of normoxia.It also induces an increase in HIF-2a expression,sufficient to induce expression of other angiogenic factors.Interfering with int6 gene expression,up-regulating HIF-2a and promoting functional angiogenesis,promoting accelerated skin wound healing.Since the eukaryotic initiation factor 3 subunit e(eIF3e)/INT6 specifically intcracls with HIF-2? and induces proteasome inhibitor-sensitive HIF-2? degradation,independent of hypoxia and von Hippel-Lindau protein,eIF3e/INT6 shRNA treatment of cells,even under normoxie conditions,cells can stably exert HIF-2? activity and induce the expression of several angiogenic factors at levels sufficient to produce functional artcrics and veins in vivo.If lentivirus is used to promote the stable expression of HIF2?,it can help the tissue engineering patch transplanted into the inguinal region to undergo more vascularization to fight in fection,and promote tissue regeneration and fusion of tissue engineering patches and tisstues.The delivery of lentiviruss in the body requires a slow release system.In this study,carboxymethyl chitosan hydrogel was chosen as the carrier for the sustained releasesystem.Chitosan is one of the most reported mon-viral natural source cationic gene polymers due to its biodegradability,biodegradability,non-sensitization,mucoadhesiveness and strong affinity for DNA.Chitosan has been shown to have low cytotoxicity in a series of experiments on experimental animals and humans.Its excellent soft tissue compatibility has also been proven.Binding of lentivirus to hydroxyapatite HA increased the activity of the complex,and hydroxyapatite HA increased viral activity to 72 hours.The cells inoculated onto the hydrogel loaddd with lentivirus-HA were less inlected with the hydrogel outside the hydrogel with hydroxyapatite HA than the hydrogel without hydroxyapatite HA.In vivo studies with collagen hydrogels loaded with lentivirus and HA lentivirus showed increased expression of sustained and local transgene expression for at least 4 weeks using the lentiviral-HA complex.This strategy of stably binding and retaining the nanoparticles is widely applicable to hydrogels,and can provide a combination of gene therapy and biomterials for regenerative medicine.The strategy of combining and retaining lentiviral vectors in hydrogels can be used to enhance and control gene transler,for regenerative medicine and to study tissue formation models,providing the basis for this research.Both in vivo and in vitro experimenls,it was demonstrated that the hypoxieenvironmental insult Int6 can induce the expression of HIF-2a,which has the effect of promoting angiogenesis,which is sufficient to promote the expression of other angiogenic factors,and its effect depends on endothelial cells.Our results also indicate that the int6 gene is a critical determinant of HIF-2a-mediated angiogenesis.And by stabilizing the long-term silence of int6,it may he an effective straiegy to promote angiogenesis.