Biodegradable Multi-layered Silk Fibroin-PCL Stent for the Management of Cervical Atresia: In Vitro Cytocompatibility and Extracellular Matrix Remodeling In Vivo

Cervical atresia is a rare congenital Mu''llerianduct anomalythat manifests as the absence or deformed nonfunctional presence ofthe cervix. Herein, a multi-layered biodegradable stent is fabricatedusing a homogeneous blend of silk fibroin with polycaprolactone usinghexafluoroisopropanol as a common solution. Briefly, a concentriccylinder of 3D honeycomb layer is sandwiched within electrospun sheetsfor fixing at the cervico-uterine junction to pave the way of cervicalreconstruction. An average length of 40 mm with 3 mm diameter is fabricatedfor the hybrid stent design. SEM evidences an evenly distributed porearchitecture of the electrospun layer, and mechanical characterizationof stent reveals a tensile strength of 1.7 & PLUSMN; 0.2 MPa, with aYoung's modulus of 5.9 & PLUSMN; 0.1 MPa. Physico-chemical characterizationconfirms the presence of silk fibroin and poly caprolactone withinthe engineered stent. Following 14 days of pepsin enzymatic degradation,18% degradation and a contact angle measurement of 97 & DEG; are observed.In vitro cytocompatibility studies are performed using site-specificprimary human cervical squamous, columnar epithelial cells, and humanendometrial stromal cells. The study demonstrates non-cytotoxic cells'viability (no significant toxicity), improved cell anchoring, adherenceamong the stent layers, and proliferation in the 3D microenvironment.Furthermore, in vivo subcutaneous studies in the rodent model indicatethat the implanted stent undergoes constructive remodeling, neo-tissuecreation, neo-vasculature formation, and re-epithelialization whilemaintaining patency for 2 months.