糖尿病傷口是由糖尿病與持續性發炎所引起，無法順利進入正常的癒合階段。在各類慢性傷口中，糖尿病傷口是最常見的類型，其特徵包括長期的發炎反應與組織再生能力受損。在糖尿病傷口的微環境中，巨噬細胞、成纖維細胞及其他參與癒合的細胞會分泌具發炎性的外泌體（inflammatory exosomes）。巨噬細胞透過由促發炎型（M1）向抗發炎型（M2）的表型轉變來調控傷口癒合；而成纖維細胞則在皮下組織填補與細胞外基質（ECM）重塑中扮演關鍵角色。過量的活性氧（ROS）會引發氧化壓力，進一步損害細胞功能。目前已開發具抗氧化功能的奈米粒子（aoNP，如SMAPoTN），用於治療與ROS相關的疾病。此外，人體脂肪來源的幹細胞（ASCs）也展現出促進傷口癒合的潛力，其來自3D球體培養的粒線體（3D ASC spheroids）在內皮與神經細胞中表現出更強的治療效果。這種增強的效益可能來自其三維結構更貼近自然組織環境，因而提升了治療反應。這些特性使其在組織修復與發炎相關疾病的治療上具有廣泛應用潛力。
在本研究中，我們探討了抗氧化奈米粒子（aoNP）與幹細胞來源粒線體（scMito）聯合使用的治療效果，並假設這種組合有望更有效並加速慢性傷口的癒合。研究結果顯示，aoNP與來自2D培養（ascMito）或3D球體（sphMito）的粒線體共同處理，可降低發炎反應，進而促進發炎型成纖維細胞及高糖發炎誘導成纖維細胞（HGII fibroblasts）的遷移與增殖。此組合也顯著降低ROS濃度，並在HGII條件下有效改善成纖維細胞的存活。此外，aoNP結合ascMito或sphMito皆可上調M2型巨噬細胞標誌物（如CD206）的表現，成功將巨噬細胞向抗發炎M2表型轉化。進一步分析顯示，此療效透過活化STAT/PPAR訊號通路實現。我們也使用來自糖尿病病人傷口液分離出的慢性傷口外泌體（CW-EVs）來驗證發炎性外泌體對巨噬細胞的影響，發現CW-EVs會誘導促發炎的M1型表現。然而，在aoNP與scMito的聯合治療下，則表現出抗發炎效果，並促進巨噬細胞由M1向M2表型轉變。在體內實驗方面，我們以糖尿病小鼠（db/db mice）為模型進行測試，結果顯示聯合治療能顯著促進傷口癒合，顯示其作為糖尿病傷口治療策略的潛力。我也進一步透過組織切片染色來評估傷口床的組織重塑與傷口縮小情形。此外，亦以免疫螢光染色（IF）分析傷口組織中成纖維細胞與巨噬細胞的狀態。結果顯示，聯合治療組相較於單一處理或未治療組別，在傷口閉合、組織再生能力以及M2型巨噬細胞極化表現上均更為顯著，支持aoNP與scMito在糖尿病傷口治療中具有協同效益。

Diabetic wounds, caused by diabetes and persistent inflammation, fail to progress through the normal phases of healing. Among chronic wounds, diabetic wounds are the most common type, characterized by prolonged inflammation and impaired tissue regeneration. In the diabetic wound bed, inflammatory exosomes are secreted by macrophages, fibroblasts, and other cells involved in the healing processes. Macrophages regulate wound healing by shifting from pro-inflammatory (M1) to anti-inflammatory (M2) phenotypes, while fibroblasts play a critical role in subcutaneous tissue filling and extracellular matrix (ECM) remodeling. Excessive reactive oxygen species (ROS) induce oxidative stress to damage cellular functions. Antioxidant nanoparticles (aoNP, SMAPoTN) have been developed for the treatment of ROS-related diseases. Additionally, human adipose-derived stem cells (ASCs) have shown potential in accelerating wound closure, and their mitochondria derived from 3D ASC spheroids exhibit enhanced therapeutic effects in endothelial and neuronal cells. This enhanced efficacy is likely attributed to the 3D architecture, which better recapitulates native tissue environments These properties collectively support their application in tissue repair and inflammation-associated conditions.
In my project, we investigated the therapeutic effects of combining aoNP with stem cell-derived mitochondria (scMito), hypothesizing that this combination would lead to more effective and accelerated healing of chronic wounds. Our results demonstrated that aoNP combined with mitochondria derived from 2D-cultured ASCs (ascMito) or 3D ASC spheres (sphMito) promoted the migration and proliferation of inflammatory fibroblast by reducing inflammation as well as the high-glucose inflammation-induced (HGII) fibroblasts. This combination significantly decreased ROS levels and rescued fibroblasts under HGII conditions. Additionally, the aoNP combined with either ascMito or sphMito enhanced the expression of M2 markers, such as CD206, effectively modulating macrophages toward an anti-inflammatory phenotype. This therapeutic effect was confirmed as the combined treatment promoted M2 macrophage transformation through the activation of the STAT/PPAR pathway. Furthermore, we used chronic wound exosomes (CW-EVs) derived from patient wound fluid to validate the effects of inflammatory exosomes on macrophages and observed that CW-EVs induced the pro-inflammatory M1 expressions. The combined treatment exhibited anti-inflammatory effects and promoted macrophage phenotypic switching toward the M2 phenotype. In the in vivo model, our experiments conducted on diabetic db/db mice demonstrated that the combined therapy significantly promoted wound healing, indicating a promising therapeutic strategy for diabetic wound treatment. I also focused on evaluating tissue remodeling and reduction of wound size in the wound bed using histological examinations. Furthermore, I assessed fibroblast and macrophage status in diabetic wound tissues following aoNP and scMito treatments using immunofluorescence staining. The results showed that the combined treatment group exhibited more effective wound closure, enhanced regenerative properties, and increased M2 macrophage polarization compared to single-treatment or untreated groups, supporting the synergistic effect of aoNP and scMito in diabetic wound healing.

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