Prakash Ravi, Mishra Rakesh Kumar, Ahmad Anas, Khan Mohsin Ali, Khan Rehan, Raza Syed Shadab
Laboratory for Stem Cell & Restorative Neurology, Department of Biotechnology, Era's Lucknow Medical College and Hospital, Sarfarazganj, Lucknow 226003, Uttar Pradesh, India.
Department of Nano-Therapeutics, Institute of Nano Science and Technology, Habitat Centre, Phase 10, Sector 64, Mohali, Punjab 160062, India.
Mater Sci Eng C Mater Biol Appl. 2021 Jan;120:111700. doi: 10.1016/j.msec.2020.111700. Epub 2020 Nov 6.
Stroke remains the leading cause of morbidity and mortality. Stem cell-based therapy offers promising hope for survivors and their families. Despite the clinical translation of stem cell-based therapies in stroke patients for almost two decades, results of these randomized controlled trials are not very optimistic. In these lines, an amalgamation of nanocarriers based drug delivery with stem cells holds great promise in enhancing stroke recovery. In the present study, we treated oxygen-glucose deprivation (OGD) exposed dental pulp stem cells (DPSCs) and mesenchymal stem cells (MSCs) with sivelestat-loaded nanostructured lipid carriers (NLCs). Various physicochemical limitations associated with sivelestat drug applications and its recent inefficacy in the clinical trials necessitates the development of novel delivery approaches for sivelestat. Therefore, to improve its efficacy on the survival of DPSCs and MSCs cell types under OGD insult, the current NLCs were formulated and characterized. Resulting NLCs exhibited a hydrodynamic diameter of 160-180 nm by DLS technique and possessed good PDI values of 0.2-0.3. Their shape, size and surface morphology were corroborated with microscopic techniques like TEM, SEM, and AFM. FTIR and UV-Vis techniques confirmed nanocarrier's loading capacity, encapsulation efficiency of sivelestat, and drug release profile. Oxidative stress in DPSCs and MSCs was assessed by DHE and DCFDA staining, and cell viability was assessed by Trypan blue exclusion test and MTT assay. Results indicated that sivelestat-loaded NLCs protected the loss of cell membrane integrity and restored cell morphology. Furthermore, NLCs successfully defended human DPSCs and MSCs against OGD-induced oxidative and inflammatory stress. In conclusion, modulation of oxidative and inflammatory stress by treatment with sivelestat-loaded NLCs in DPSCs and MSCs provides a novel strategy to rescue stem cells during ischemic stroke.
中风仍然是发病和死亡的主要原因。基于干细胞的疗法为幸存者及其家人带来了充满希望的前景。尽管基于干细胞的疗法在中风患者中进行临床转化已近二十年，但这些随机对照试验的结果并不十分乐观。在这种情况下，将基于纳米载体的药物递送与干细胞相结合在促进中风恢复方面具有巨大潜力。在本研究中，我们用载有西维来司他的纳米结构脂质载体（NLCs）处理了暴露于氧葡萄糖剥夺（OGD）的牙髓干细胞（DPSCs）和间充质干细胞（MSCs）。与西维来司他药物应用相关的各种物理化学限制及其近期在临床试验中的无效性，使得有必要开发西维来司他的新型递送方法。因此，为了提高其对OGD损伤下DPSCs和MSCs细胞类型存活的功效，制备并表征了当前的NLCs。通过动态光散射（DLS）技术得到的NLCs流体动力学直径为160 - 180nm，且具有0.2 - 0.3的良好多分散指数（PDI）值。它们的形状、大小和表面形态通过透射电子显微镜（TEM）、扫描电子显微镜（SEM）和原子力显微镜（AFM）等显微技术得到证实。傅里叶变换红外光谱（FTIR）和紫外可见光谱（UV - Vis）技术证实了纳米载体的负载能力、西维来司他的包封效率以及药物释放曲线。通过二氢乙啶（DHE）和2',7'-二氯二氢荧光素二乙酸酯（DCFDA）染色评估DPSCs和MSCs中的氧化应激，并通过台盼蓝排斥试验和MTT法评估细胞活力。结果表明，载有西维来司他的NLCs保护了细胞膜完整性的丧失并恢复了细胞形态。此外，NLCs成功地保护了人DPSCs和MSCs免受OGD诱导的氧化和炎症应激。总之，用载有西维来司他的NLCs处理DPSCs和MSCs来调节氧化和炎症应激，为在缺血性中风期间挽救干细胞提供了一种新策略。
