開發可注射式水凝膠系統：控制釋放花青素以促進糖尿病患者傷口癒合

Abstract

糖尿病足潰瘍（Diabetes foot ulcers，DFU）是一種嚴重的糖尿病併發症。在糖尿病患者中，高血糖會導致過量的Reactive oxygen species（ROS）產生，加劇傷口的氧化壓力和發炎反應，損害皮膚的屏障功能，進而導致傷口修復能力受損。糖尿病傷口患者是截肢的高風險族群，疼痛等負面影響，嚴重降低患者的生活品質，因此迫切需要一種新穎且有效的治療策略。 本研究旨於設計一種由海藻酸鈉、明膠以及透明質酸配置成的可注射水凝膠以包覆Cyanidin-3-glucoside（C3G）。在糖尿病傷口處深層滲透、持續釋放，以改善傷口環境並促進癒合。研究中透過DCFDA螢光探針方法證明，C3G能降低H2O2處理後成纖維細胞和角質形成細胞內ROS含量。透過劃痕實驗證實，C3G能回復氧化壓力所抑制的細胞遷移能力，提升患部的癒合。在Western blot結果發現，C3G能顯著性提升成纖維細胞和角質形成細胞的抗氧化蛋白HO-1表達，提升細胞抗氧化能力。此外，本研究發現，C3G水凝膠具有促進傷口治療的功能性，包括可注射性、高自適性、高吸水能力、低細胞毒性等特性。並且C3G 水凝膠亦可清除細胞中 ROS，改善傷口高氧化壓力，提昇內皮細胞遷移速率，從而促進上皮形成和傷口縮合的效能。總結來說，C3G水凝膠藥物釋放系統能夠有效恢復皮膚再生能力，促進傷口癒合，對於治療糖尿病足潰瘍是具有前景的替代方法。

Diabetes foot ulcer(DFU) is a severe diabetes complication, and one of the most common type of chronic wound[1]. In diabetic patients, hyperglycemia causes excessive production of ROS, and accelerates oxidative stress and inflammation, compromising the barrier function of skin that leads to impaired wound repair[2, 3]. Considering the high risk of limb amputation and negative impact on patients’ quality of life, a novel yet effective therapeutic strategy for diabetic wounds is in urgent need. We herein designed and fabricated an injectable, biocompatible sodium gelatin/alginate/hyaluronic acid (G/SA/HA)-base hydrogel that could sustainedly release the therapeutic payload, cyanidin-3-glucoside(C3G). Our primary results reveal that C3G significantly reduced ROS levels in H2O2-treated L929 fibroblast and HaCaT keratinocytes, which were measured by the 2’,7’-dichlorofluorescin diacetate(DCFDA) assay. Furthermore, C3G proved up-regulates heme oxygenase-1(HO-1) protein expressions, protecting L929 and HaCaT cells damage against oxidative damage. Additionally, C3G was found to enhance the proliferation and migration of L929 and HaCaT cells, resulting in an improved epithelialization and wound contraction. In summary, our SA/G/HA hydrogel-based drug delivery system demonstrated multi-functionality, including injectability, self-adapting behavior and low cytotoxicity, suggesting that it is a promising alterative to achieve complete dermal regeneration in patients with DFUs.