Background

We developed a novel, injectable and decellularized human peripheral nerve-based scaffold, named Micronized Human Neural Tissue (hMINT), designed to be used as a supportive matrix for stem cell transplantation in the context of spinal cord injury (SCI).

Materials and methods

Human donated sciatic nerves were micronized at liquid nitrogen temperature prior to decellularization using 3 different procedures of various harshness. hMINT were characterized in terms of particle size, DNA, sulfated glycosaminoglycans (sGAG) and growth factors content. To test the biocompatibility and bioactivity of the various preparations, we used a type of mesenchymal stromal cells (MSCs), termed MIAMI cells, which were placed in contact with hMINT to monitor cell attachment by confocal microscopy and gene expression by RT-qPCR in vitro.

Results

The content of DNA, sGAG and growth factors left in the product after processing was highly dependent on the decellularization procedure used. We demonstrated that hMINT are biocompatible and promoted the attachment and long-term survival of MIAMI cells in vitro. Finally, combination with hMINT increased MIAMI cells mRNA expression of pro-survival and anti-inflammatory factors. Importantly, the strongest bioactivity on MIAMI cells was observed with the hMINT decellularized using the mildest decellularization procedure, therefore emphasizing the importance of achieving an adequate decellularization without losing the hMINT’s bioactivity.

Perspectives and clinical significance

The capacity of hMINT/stem cells to facilitate protection of injured neural tissue, promote axon re-growth and improve functional recovery will be tested in an animal model of SCI and other neurodegenerative disorders in the future.

中文翻译：

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用于生产生物活性人微粉化神经组织 (hMINT) 的三种洗涤/脱细胞程序的表征

背景

我们开发了一种新型、可注射、脱细胞的人外周神经支架，名为微米化人神经组织（hMINT），旨在用作脊髓损伤（SCI）背景下干细胞移植的支持基质。

材料和方法

将人类捐赠的坐骨神经在液氮温度下进行微粉化，然后使用 3 种不同程度的不同程序进行脱细胞。hMINT 的特征包括粒径、DNA、硫酸化糖胺聚糖 (sGAG) 和生长因子含量。为了测试各种制剂的生物相容性和生物活性，我们使用了一种称为 MIAMI 细胞的间充质基质细胞 (MSC)，将其与 hMINT 接触，通过共聚焦显微镜监测细胞附着，通过 RT-qPCR 体外监测基因表达。

结果

加工后产品中残留的 DNA、sGAG 和生长因子的含量很大程度上取决于所使用的脱细胞程序。我们证明 hMINT 具有生物相容性，可促进 MIAMI 细胞的体外贴壁和长期存活。最后，与 hMINT 组合增加了 MIAMI 细胞促生存和抗炎因子的 mRNA 表达。重要的是，使用最温和的脱细胞程序对 MIAMI 细胞进行脱细胞后观察到对 MIAMI 细胞最强的生物活性，因此强调了在不损失 hMINT 生物活性的情况下实现充分脱细胞化的重要性。

观点和临床意义

未来将在 SCI 和其他神经退行性疾病的动物模型中测试 hMINT/干细胞促进保护受损神经组织、促进轴突重新生长和改善功能恢复的能力。