Hanuman Srividya, B Harish Kumar, Pai K Sreedhara Ranganath, Nune Manasa
Manipal Institute of Regenerative Medicine, Manipal Academy of Higher Education, Manipal, Karnataka 576104, India.
Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal, Karnataka 576104, India.
ACS Omega. 2024 Aug 1;9(32):34314-34328. doi: 10.1021/acsomega.3c10445. eCollection 2024 Aug 13.
The uterus, a vital organ in the female reproductive system, nurtures and supports developing embryos until maturity. This study focuses on addressing uterine related problems by creating a nanofibrous scaffold to regenerate uterine myometrial tissue, closely resembling the native extracellular matrix (ECM) for enhanced efficacy. To achieve this, we utilized polycaprolactone (PCL) as a biomaterial and employed an electrospinning technique to generate PCL nanofibers in both random and aligned orientations. Due to the inherent hydrophobic nature of PCL nanofibers, a two-step wet chemistry surface modification technique is used, involving the conjugation of galactose onto them. Galactose, a lectin-binding sugar, was chosen to enhance the scaffold's hydrophilicity, thereby improving cell adhesion and fostering l-selectin-based interactions between the scaffold and uterine cells. These interactions, in turn, activated uterine fibroblasts, leading to ECM remodeling. The optimized electrospinning process successfully generated random and aligned nanofibers. Subsequent surface modification was carried out, and the modified scaffold was subjected to various physicochemical characterization, such as the ninhydrin assay, enzyme-linked lectin assay techniques that revealed successful galactose conjugation, and mechanical characterization to assess any changes in material bulk properties resulting from the modification. The tensile strength of random galactose-modified PCL fibers reached 0.041 ± 0.01 MPa, outperforming random unmodified PCL fibers (0.026 ± 0.01 MPa), aligned unmodified PCL fibers (0.011 ± 0.001 MPa), and aligned modified PCL fibers (0.016 ± 0.002 MPa). Cytocompatibility studies with human uterine fibroblast cells showed enhanced viability and proliferation on the modified scaffolds. Initial pilot studies were attempted in the current study involving subcutaneous implantation in the dorsal area of Wistar rats to assess biocompatibility and tissue response before proceeding to intrauterine implantation indicated that the modification did not induce adverse inflammation in vivo. In conclusion, our study introduces a surface-modified PCL nanofibrous material for myometrial tissue engineering, offering promise in addressing myometrial damage and advancing uterine health and reproductive well-being.
子宫是女性生殖系统中的重要器官，它孕育并支持发育中的胚胎直至成熟。本研究致力于通过创建一种纳米纤维支架来解决与子宫相关的问题，该支架可使子宫肌层组织再生，与天然细胞外基质（ECM）极为相似，从而提高功效。为此，我们使用聚己内酯（PCL）作为生物材料，并采用静电纺丝技术制备了随机排列和定向排列的PCL纳米纤维。由于PCL纳米纤维具有固有的疏水性，因此采用了两步湿化学表面改性技术，即将半乳糖缀合到纳米纤维上。半乳糖是一种凝集素结合糖，被选用来增强支架的亲水性，从而改善细胞黏附，并促进支架与子宫细胞之间基于L-选择素的相互作用。这些相互作用进而激活子宫成纤维细胞，导致ECM重塑。优化后的静电纺丝工艺成功制备出了随机排列和定向排列的纳米纤维。随后进行了表面改性，并对改性后的支架进行了各种物理化学表征，如茚三酮测定、酶联凝集素测定技术（结果显示半乳糖成功缀合）以及力学表征，以评估改性对材料整体性能的任何影响。随机排列的半乳糖改性PCL纤维的拉伸强度达到0.041±0.01 MPa，优于随机排列的未改性PCL纤维（0.026±0.01 MPa）、定向排列的未改性PCL纤维（0.011±0.001 MPa）和定向排列的改性PCL纤维（0.016±0.002 MPa）。对人子宫成纤维细胞的细胞相容性研究表明，在改性支架上细胞的活力和增殖能力增强。在本研究中，最初尝试进行了初步试验，即在Wistar大鼠背部皮下植入，以评估生物相容性和组织反应，然后再进行子宫内植入，结果表明该改性在体内未引发不良炎症。总之，我们的研究引入了一种用于子宫肌层组织工程的表面改性PCL纳米纤维材料，有望解决子宫肌层损伤问题，促进子宫健康和生殖福祉。
