Microneedle Patches Integrated with Immunomoudulators for anti-Cancer Immunotherapy = 항암 면역치료를 위한 면역조절제 탑재 마이크로니들 패치

다국어 초록 (Multilingual Abstract)

Advancements in micro-resolution 3D printers have significantly facilitated the development of complex mass-producible drug delivery platforms. Conventionally, due to the limitations of micro-milling machineries, dissolvable microneedles (MNs) were mainly fabricated in cone-shaped geometry with limited drug delivery accuracy. Herein, to overcome the limitations of conventional microneedle, a novel projection micro-stereolithography 3D printer-based self-locking microneedle (SLMN) for precise skin insertion, adhesion, and transcutaneous microdose drug delivery is presented. The geometry of self-locking microneedle consists of a sharp skin-penetrating tip, a wide skin interlocking body, and a narrow base with mechanical supports fabricated over a flexible hydrocolloid patch to improve the accuracy of skin penetration into irregular surfaces. Melanoma, a type of skin cancer, was selected as the model for the investigation of self-locking microneedle due to its irregular and uneven surface. In vivo immunotherapy efficacy was evaluated by integrating SD-208, a novel transforming growth factor-β (TGF-β) inhibitor that suppresses the proliferation and metastasis of tumors, and PD-L1 antibody (Anti-PD-L1), an immune checkpoint inhibitor that induces T cell-mediated tumor cell death, into self-locking microneedle and compared with intratumoral injection. Evaluation of Anti-PD-L1/SD-208 delivery effectiveness in B16F10 melanoma-bearing mice model confirmed significantly improved dose efficacy of self-locking microneedle compared with intratumoral injection

목차 (Table of Contents)

• Abstract 6
• 1. Introduction 8
• 2. Material and Methods 13
• 2.1 Reagents 13
• 2.2 Cell culture 14
• 2.3 Confocal microscopy imaging of B16F10 14
• 2.4 Western blotting 15
• 2.5 Wound healing assay 15
• 2.6 3D Spheroid growth assay 16
• 2.7 PD-L1 expression analysis 17
• 2.8 Fabrication of dissolvable Microneedles 17
• 2.9 Characterization of microneedles 18
• 2.10 Skin penetration analysis based on OCT 19
• 2.11 Ex-vivo cutaneous permeation kinetics of microneedles 19
• 2.12 Transepidermal water loss (TEWL) analysis 20
• 2.13 Fluorescence distribution analysis 20
• 2.14 In vivo immunotherapy of B16F10 tumor-bearing C57BL/6 mice 21
• 2.15 Immunofluorescence staining for imaging of CD8+ and granzyme B in the tumor microenvironment 22
• 2.16 TIL assay 23
• 2.17 Statistical analysis 24
• 3. Results and Discussion 25
• 3.1 TGF-β receptor inhibiting effects of SD-208 in B16F10 melanoma cells 25
• 3.2 Anti-proliferative and anti-EMT effects of SD-208 in B16F10 cancer cells 27
• 3.3 PD-L1 expression inhibiting effects of SD-208 in B16F10 cells 30
• 3.4 Microneedle fabrication methods and geometrical properties 31
• 3.5 Geometrical advancements of Self-locking Microneedles 35
• 3.6 Cutaneous penetrating and dissolving properties of Self-locking Microneedles in C57BL/6 mice 38
• 3.7 Anti-tumoral efficacies of Anti-PD-L1/SD-208 Self-locking Microneedle (SLMN) in orthotopic melanoma mouse model 41
• 3.8 Immunomodulation efficacies of Anti-PD-L1/SD-208 Self-locking Microneedle (SLMN) in orthotopic melanoma mouse model 43
• 4. Conclusion 48
• 5. References 52
• 국문 요지 59