Department of Endocrinology, Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou 510120, PR China.
Department of Polymer and Material Science, School of Chemistry, Key Laboratory for Polymeric Composite and Functional Materials of Ministry of Education, Guangdong Provincial Key Laboratory for High Performance Polymer-based Composites, Sun Yat-sen University, Guangzhou 510275, PR China.
Acta Biomater. 2020 Jan 15;102:298-314. doi: 10.1016/j.actbio.2019.11.005. Epub 2019 Nov 18.
The anomalous high expression of matrix metalloproteinase 9 (MMP-9) is one important factor that impedes diabetic wound healing. Therefore, inhibition of MMP-9 expression in a diabetic wound could be a feasible method to promote wound healing. In this study, we studied the possibility of self-therapy using wound dressings that contain bacterial cellulose-hyperbranched cationic polysaccharide (BC-HCP) derivatives that encapsulate siRNA (BC-HCP/siMMP-9) and have controlled release properties. Herein, we used four HCPs (Gly-DMAPA, Gly-D4, Amyp-DMAPA, Amyp-D4) as gene carriers. Our results showed that all HCP derivatives were minimally toxic to cells in vitro, while the cationic properties of HCP could be used as a complexation agent for MMP-9 siRNA (siMMP-9). Upon exposure to bacterial cellulose (BC), the BC slowly released HCP/siMMP-9. The released siMMP-9 effectively reduced the gene expression and protein levels of MMP-9 in a human immortalized epithelial cell line (HaCAT) and in diabetic rat wounds. Inhibition of MMP-9 in the wounds of diabetic rats resulted in a significant enhancement of wound healing, suggesting that the BC-HCP/siMMP-9 composite dressing could be used as a safe and effective dressing to promote wound healing in diabetic rats. STATEMENT OF SIGNIFICANCE: In this work, we evaluated the possibility of using bacterial cellulose-hyperbranched cationic polysaccharide derivatives (BC-HCP) as a self-therapeutic wound dressing with siRNA encapsulated and controlled release properties. Our results showed that the BC-HCP/siMMP-9 composite dressing slowly released HCP/siMMP-9. The released siMMP-9 effectively reduced the gene expression and protein level of MMP-9 in human immortalized epithelial cell line and in the wound of diabetic rats. The BC-HCP/siMMP-9 composite dressing promoted diabetic wound healing by the unique nanostructure of BC and by releasing siMMP-9 for specific MMP-9 inhibition. Therefore, it could be used as a safe and effective dressing to promote wound healing in diabetic rats. This is the first evidence on the study of using BC as a dressing composite by encapsulating HCP/siRNA complexes for efficient RNAi gene silencing for better wound healing in diabetic rats.
基质金属蛋白酶 9（MMP-9）的异常高表达是阻碍糖尿病伤口愈合的一个重要因素。因此，抑制糖尿病伤口中 MMP-9 的表达可能是促进伤口愈合的一种可行方法。在这项研究中，我们研究了使用含有细菌纤维素-超支阳离子多糖（BC-HCP）衍生物的伤口敷料进行自我治疗的可能性，这些衍生物可以包裹 siRNA（BC-HCP/siMMP-9）并具有控制释放特性。在此，我们使用了四种 HCP（Gly-DMAPA、Gly-D4、Amyp-DMAPA、Amyp-D4）作为基因载体。我们的结果表明，所有 HCP 衍生物在体外对细胞的毒性都很小，而 HCP 的阳离子性质可用作 MMP-9 siRNA（siMMP-9）的复合物形成剂。当暴露于细菌纤维素（BC）时，BC 会缓慢释放 HCP/siMMP-9。释放的 siMMP-9 可有效降低永生化人上皮细胞系（HaCAT）和糖尿病大鼠伤口中 MMP-9 的基因表达和蛋白水平。抑制糖尿病大鼠伤口中的 MMP-9 可显著促进伤口愈合，这表明 BC-HCP/siMMP-9 复合敷料可用作安全有效的敷料，以促进糖尿病大鼠伤口愈合。
在这项工作中，我们评估了使用细菌纤维素-超支阳离子多糖衍生物（BC-HCP）作为具有包裹 siRNA 和控制释放特性的自治疗性伤口敷料的可能性。我们的结果表明，BC-HCP/siMMP-9 复合敷料可缓慢释放 HCP/siMMP-9。释放的 siMMP-9 可有效降低永生化人上皮细胞系和糖尿病大鼠伤口中 MMP-9 的基因表达和蛋白水平。BC-HCP/siMMP-9 复合敷料通过 BC 的独特纳米结构和释放用于特定 MMP-9 抑制的 siMMP-9 来促进糖尿病伤口愈合。因此，它可用作促进糖尿病大鼠伤口愈合的安全有效的敷料。这是第一项关于使用 BC 作为敷料复合材料的研究证据，该复合材料通过包裹 HCP/siRNA 复合物用于高效 RNAi 基因沉默，以改善糖尿病大鼠的伤口愈合。
