Rochkind Shimon, El-Ani Dalia, Nevo Zvi, Shahar Abraham
Division of Peripheral Nerve Reconstruction, Tel Aviv Sourasky Medical Center, Tel Aviv University, Tel Aviv 64239, Israel.
Lasers Surg Med. 2009 Apr;41(4):277-81. doi: 10.1002/lsm.20757.
The present study focuses on the effect of 780 nm laser irradiation on the growth of embryonic rat brain cultures embedded in NVR-Gel (cross-linked hyaluronic acid with adhesive molecule laminin and several growth factors). Dissociated neuronal cells were first grown in suspension attached to cylindrical microcarriers (MCs). The formed floating cell-MC aggregates were subsequently transferred into stationary cultures in gel and then laser treated. The response of neuronal growth following laser irradiation was investigated.
Whole brains were dissected from 16 days Sprague-Dawley rat embryos. Cells were mechanically dissociated, using narrow pipettes, and seeded on positively charged cylindrical MCs. After 4-14 days in suspension, the formed floating cell-MC aggregates were seeded as stationary cultures in NVR-Gel. Single cell-MC aggregates were either irradiated with near-infrared 780 nm laser beam for 1, 4, or 7 minutes, or cultured without irradiation. Laser powers were 10, 30, 50, 110, 160, 200, and 250 mW.
780 nm laser irradiation accelerated fiber sprouting and neuronal cell migration from the aggregates. Furthermore, unlike control cultures, the irradiated cultures (mainly after 1 minute irradiation of 50 mW) were already established after a short time of cultivation. They contained a much higher number of large size neurons (P<0.01), which formed dense branched interconnected networks of thick neuronal fibers.
780 nm laser phototherapy of embryonic rat brain cultures embedded in hyaluronic acid-laminin gel and attached to positively charged cylindrical MCs, stimulated migration and fiber sprouting of neuronal cells aggregates, developed large size neurons with dense branched interconnected network of neuronal fibers and, therefore, can be considered as potential procedure for cell therapy of neuronal injury or disease.
本研究聚焦于780 nm激光照射对包埋于NVR -凝胶（交联透明质酸与黏附分子层粘连蛋白及多种生长因子）中的胚胎大鼠脑培养物生长的影响。首先将解离的神经元细胞悬浮培养于附着于圆柱形微载体（MCs）上。随后将形成的漂浮细胞 - MC聚集体转移至凝胶中的固定培养物中，然后进行激光处理。研究了激光照射后神经元生长的反应。
从16日龄的斯普拉格 - 道利大鼠胚胎中取出全脑。使用细吸管机械解离细胞，并接种于带正电荷的圆柱形MCs上。悬浮培养4 - 14天后，将形成的漂浮细胞 - MC聚集体接种于NVR -凝胶中作为固定培养物。单细胞 - MC聚集体要么用近红外780 nm激光束照射1、4或7分钟，要么不进行照射培养。激光功率分别为10、30、50、110、160、200和250 mW。
780 nm激光照射加速了纤维萌发以及神经元细胞从聚集体中的迁移。此外，与对照培养物不同，照射后的培养物（主要是50 mW照射1分钟后）在短时间培养后就已形成。它们含有数量多得多的大尺寸神经元（P<0.01），这些神经元形成了由粗大神经纤维组成的密集分支相互连接的网络。
对包埋于透明质酸 - 层粘连蛋白凝胶中并附着于带正电荷圆柱形MCs上的胚胎大鼠脑培养物进行780 nm激光光疗，可刺激神经元细胞聚集体的迁移和纤维萌发，发育出具有密集分支相互连接神经纤维网络的大尺寸神经元，因此可被视为神经元损伤或疾病细胞治疗的潜在方法。
