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Synergistic CO2 Cryotherapy and EGF Delivery for Accelerated Wound Healing through Anti-Inflammatory and Regenerative Pathways

이산화탄소 냉동치료와 표피성장인자 전달의 시너지 효과를 통한 항염증 및 재생 경로를 이용한 상처 치유 가속화

초록/요약

상처 치유는 전 세계적으로 중요한 임상 과제로 남아 있으며, 효과적인 관리 전략은 치료 결과 개선에 필수적이다. 본 연구는 고속 CO₂ 냉동 치료와 표피 성장 인자(EGF)의 진피 내 전달을 결합한 새로운 다기능 장치인 AcuCool™ 시스템의 상처 치유 촉진 치료 잠재력을 조사한다. Sprague Dawley 랫드의 전층 피부 상처 모델을 사용하여, 장치+EGF 치료의 효과를 기존 미세침 기반 EGF 전달 및 무처치 대조군과 비교했다. 육안 평가 결과, 장치+EGF 군에서 상처 폐쇄가 유의하게 가속화되었다. 조직학적 분석에서는 재상피화 증진, 염증 세포 침윤 감소, 콜라겐 침착 증가가 관찰되었다. 분자 수준 평가에서는 전염증성 표지자(TNF-α, IL1β, MCP-1)의 하향 조절과 TGF-β1, Collagen I, Vimentin을 포함한 재형성 관련 유전자의 상향 조절이 추가로 입증되었다. 또한, 아질산염 분석을 통해 국소 산화질소 수준 감소가 확인되어 산화 스트레스 억제를 나타냈다. AcuCool™ 플랫폼은 물리적 및 생화학적 이중 치료 메커니즘 을 갖춘 정밀하고 비침습적인 약물 전달을 제공하여, 염증과 조직 재생의 우수한 제어를 가능하게 한다. 이러한 결과는 AcuCool™이 급성 모델에서 상처 치유를 가속화하는 유망한 치료 전략임을 시사한다. 만성 상처 환경에서의 추가 연구가 필요하지만, 이 접근법은 향후 임상 적용을 위한 중개 잠재력을 가질 수 있다.

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초록/요약

Synergistic CO2 Cryotherapy and EGF Delivery for Accelerated Wound Healing through Anti-Inflammatory and Regenerative Pathways Wound healing continues to represent a substantial clinical challenge on a global scale, and the development of effective therapeutic strategies remains essential for optimizing patient outcomes. The present study examines the therapeutic potential of the AcuCool™ system, an innovative multifunctional platform that integrates high-velocity CO₂ cryotherapy with intradermal administration of epidermal growth factor (EGF), for promoting cutaneous wound repair. Utilizing a full-thickness excisional wound model in Sprague Dawley rats, we systematically compared the therapeutic effects of Device+EGF treatment with those of conventional microneedling-based EGF application and untreated control groups. Macroscopic examination demonstrated significantly expedited wound closure in the Device+EGF cohort. Histopathological analysis revealed enhanced re- epithelialization, attenuated inflammatory cell infiltration, and augmented collagen deposition. Molecular assessments further confirmed downregulation of proinflammatory mediators (TNF-α, IL-1β, MCP-1) alongside upregulation of tissue remodeling-associated genes including TGF-β1, Collagen I, and Vimentin. Additionally, nitrite assays validated diminished local nitric oxide concentrations, substantiating suppression of oxidative stress. The AcuCool™ platform provides precise, minimally invasive pharmaceutical delivery through synergistic physical and biochemical therapeutic pathways, thereby enabling superior modulation of inflammation and tissue regeneration processes. These experimental findings indicate that AcuCool™ constitutes a compelling therapeutic approach for expediting wound healing in acute injury models. Although further investigations are warranted in chronic wound environments, this technology demonstrates considerable translational potential for future clinical implementation. Keywords: wound healing; CO₂ cryotherapy; epidermal growth factor; anti- inflammatory therapy; transdermal drug delivery

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목차

I. Introduction 1
II. Materials and Methods 7
1. System Architecture and Operational Workflow of the AcuCool™ Device 7
2. Animal Experiment 10
2.1 Animal Ethics and Experimental Design 10
2.2 Grouping and Treatment Allocation 12
2.3 Wound Induction and Treatment Protocol 13
2.4 Tissue Collection and Endpoints 14
3. Measurement of the Wound Healing Area 15
4. Histological Analysis 16
4.1 Hematoxylin and Eosin (H&E) Staining 17
4.2 Masson's Trichrome (MT) Staining 18
4.3 Immunofluorescence (IF) Staining 20
5. Protein Preparation 22
6. Enzyme-Linked Immunosorbent Assay (ELISA) 23
7. Nitric Oxide Quantification 24
8. RNA Extraction, cDNA Synthesis, and Quantitative PCR Analysis 26
9. Statistics Analysis 28
III. Results 29
1. Macroscopic and microscopic observation of the wound healing process 29
2. Effect of the anti-inflammation and antioxidation during the wound healing process 34
3. Acceleration of the proliferation and remodeling during the wound healing process 40
IV. Discussion 43
V. Conclusion 53
VI. References 54
국문요약 62

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