Central Leather Research Institute, Department of Biotechnology, Chennai, India.
IET Nanobiotechnol. 2010 Sep;4(3):51-60. doi: 10.1049/iet-nbt.2009.0014.
Delivery of growth factor for the differentiation of stem cells into lineage specific cells holds great potential in regenerative medicine. Stem cell differentiation is governed by cytokines and growth factors secreted upon the organelle injury and, however, their short half-life necessitates exogenous supply. Development of suitable nanodevices using biodegradable polymers to deliver therapeutic proteins to the targeted site in a sustainable manner attracts scientists and clinicians. Here, for the first time, hepatocyte growth factor (HGF) was incorporated into chitosan nanoparticles (CNP) by ionotrophic gelation method. An average size of nanoparticles prepared was 100 nm, showing sustainable release of HGF. Cytotoxicity study did not reveal any adverse effect on bone marrow mesenchymal stem cells (MSC) up to 4 mg CNP/ml culture medium. To evaluate the effect of HGF incorporated CNP (HGF-CNP) on hepatic differentiation in in vitro, MSC were incubated with HGF-CNP and other supplements. After 21 days, fibroblast-like morphology of MSC became round-shape, a typical characteristic of hepatocyte cell. Immunofluorescence study for albumin expression confirmed the hepatic differentiation. In conclusion, HGF released from the HGF-CNP can differentiate MSC into hepatocytes, and this novel technique could also be extended to deliver therapeutic proteins for a variety of tissue regeneration.
生长因子的传递对于将干细胞分化为特定谱系的细胞在再生医学中具有巨大的潜力。干细胞分化受细胞因子和生长因子的调控，这些因子在细胞器损伤时分泌，但它们的半衰期短，需要外源性供应。使用可生物降解聚合物开发合适的纳米器件，以可持续的方式将治疗性蛋白质递送到靶向部位，吸引了科学家和临床医生的关注。在这里，首次通过离子凝胶化方法将肝细胞生长因子（HGF）掺入壳聚糖纳米颗粒（CNP）中。所制备的纳米颗粒的平均粒径为 100nm，表现出 HGF 的可持续释放。细胞毒性研究表明，在 4mg CNP/ml 培养基中，CNP 对骨髓间充质干细胞（MSC）没有任何不良影响。为了评估 HGF 掺入 CNP（HGF-CNP）对体外肝分化的影响，将 MSC 与 HGF-CNP 和其他补充剂一起孵育。21 天后，MSC 的成纤维细胞样形态变为圆形，这是肝细胞的典型特征。白蛋白表达的免疫荧光研究证实了肝分化。总之，从 HGF-CNP 释放的 HGF 可以将 MSC 分化为肝细胞，这项新技术也可以扩展到用于多种组织再生的治疗性蛋白质的传递。
