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

Abstract

Cervical atresia is a rare congenital Mullerian duct anomaly that manifests as the absence or deformed nonfunctional presence of the cervix. Herein, a multi-layered biodegradable stent is fabricated using a homogeneous blend of silk fibroin with polycaprolactone using hexafluoroisopropanol as a common solution. Briefly, a concentric cylinder of 3D honeycomb layer is sandwiched within electrospun sheets for fixing at the cervico-uterine junction to pave the way of cervical reconstruction. An average length of 40 mm with 3 mm diameter is fabricated for the hybrid stent design. SEM evidences an evenly distributed pore architecture of the electrospun layer, and mechanical characterization of stent reveals a tensile strength of 1.7 ± 0.2 MPa, with a Young's modulus of 5.9 ± 0.1 MPa. Physico-chemical characterization confirms the presence of silk fibroin and poly caprolactone within the engineered stent. Following 14 days of pepsin enzymatic degradation, 18% degradation and a contact angle measurement of 97°are observed. In vitro cytocompatibility studies are performed using site-specific primary human cervical squamous, columnar epithelial cells, and human endometrial stromal cells. The study demonstrates non-cytotoxic cells' viability (no significant toxicity), improved cell anchoring, adherence among the stent layers, and proliferation in the 3D microenvironment. Furthermore, in vivo subcutaneous studies in the rodent model indicate that the implanted stent undergoes constructive remodeling, neo-tissue creation, neo-vasculature formation, and re-epithelialization while maintaining patency for 2 months.

Temporal control in shell–core structured nanofilm for tracheal cartilage regeneration: synergistic optimization of anti-inflammation and chondrogenesis

“…Considering the single layer of BMSCs-loaded nanofilm were ultrathin, failing to match the cartilage thickness and mechanical strength requirements of normal tracheal cartilage. Previous studies have demonstrated that stacking BMSCs and nanofilms using a ‘sandwich model’ helps achieve closer contact between BMSCs and nanofilms, enabling drugs and factors to fully exert their functions [ 63 ]. Hence, this study proposes to form a BMSCs-nanofilm construct via the four-layer superposition of BMSCs-loaded nanofilm based on ‘sandwich model’, aiming to increase the thickness of the resultant BMSCs-nanofilm construct.…”

Electrophoretic Deposition of 58S Bioactive Glass- Polymer Composite Coatings on 316L Stainless Steel: An Optimization for Corrosion, Bioactivity, and Cytocompatibility

“…The chorioallantoic membrane (CAM) assay was performed to determine the angiogenesis capability of the optimized BG-polymer composite coating and control (bare SS). Briefly, the fertilized white Leghorn chicken eggs were procured from an authorized government poultry farm, and procedures were performed as described elsewhere . The eggs were cleaned with sterile water and incubated upright in a humidified atmosphere maintained at 37 °C.…”

“…Briefly, the fertilized white Leghorn chicken eggs were procured from an authorized government poultry farm, and procedures were performed as described elsewhere. 42 The eggs were cleaned with sterile water and incubated upright in a humidified atmosphere maintained at 37 °C. On day 5, the upper side of the eggshell was cracked, and the eggshell membrane was removed at approximately 1.5 cm in diameter to expose the CAM.…”

Biochemical and immunomodulatory insights of extracellular matrix from decellularized human whole cervix: recellularization and in vivo ECM remodeling interplay

“…Primary site specific hCCECs were isolated from endocervical tissues of healthy subjects reporting for ova donation under ethical agreement [37] and cell proliferation assessment was performed over the scaffold. Briefly, MTT assay was executed to evaluate the anti-proliferative potential of the crosslinked DHC.…”