Secreted factors from cultured dental pulp stem cells promoted neurite outgrowth of dorsal root ganglion neurons and ameliorated neural functions in streptozotocin-induced diabetic mice
Secreted factors from cultured dental pulp stem cells promoted neurite outgrowth of dorsal root ganglion neurons and ameliorated neural functions in streptozotocin-induced diabetic mice
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DOI：
10.1111/jdi.13085
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发表时间：
2020-01-01
影响因子：
3.2
通讯作者：
Kamiya, Hideki
中科院分区：
文献类型：
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作者：
Miura-Yura, Emiri;Tsunekawa, Shin;Kamiya, Hideki
目的/介绍干细胞移植以旁分泌方式促进轴突再生和血管生成。在本研究中，我们检测了人脱落乳牙干细胞条件培养基中的分泌因子是否对小鼠糖尿病多发性神经病变有有益的影响。材料与方法取人脱落乳牙干细胞在无血清Dulbecco's modified Eagle's培养基（DMEM）中培养48 h后的条件培养基，按分子量分成4组。用SHED-CM或DMEM培养C57BL/6J小鼠背根神经节神经元，观察其对神经突生长的影响。将链脲霉素诱导的糖尿病小鼠单侧后肢肌肉注射SHED-CM或DMEM 100 μ L，每周2次，连续4周。通过足底试验评估周围神经功能，以及运动和感觉神经传导速度。观察表皮内神经纤维密度、毛细血管数与肌纤维比、毛细血管血流量及腓肠神经形态学变化。结果与DMEM相比，人脱落乳牙干细胞条件培养基能显著促进背根神经节神经元突起的生长。在SHED-CM的4个组分中，只有kDa组分促进背根神经节神经元的突起生长。此外，与DMEM相比，SHED-CM显著阻止了糖尿病小鼠感觉神经传导速度的下降。虽然SHED-CM没有改善表皮内神经纤维密度或腓肠神经形态，但SHED-CM改善了毛细血管数与肌纤维比和毛细血管血流量。结论SHED-CM可能通过促进神经突生长来治疗糖尿病多发性神经病变，其毛细血管的增加可能有助于神经功能的改善。
Aims/Introduction Transplantation of stem cells promotes axonal regeneration and angiogenesis in a paracrine manner. In the present study, we examined whether the secreted factors in conditioned medium of stem cells from human exfoliated deciduous teeth (SHED-CM) had beneficial effects on diabetic polyneuropathy in mice. Materials and Methods Conditioned medium of stem cells from human exfoliated deciduous teeth was collected 48 h after culturing in serum-free Dulbecco's modified Eagle's medium (DMEM), and separated into four fractions according to molecular weight. Dorsal root ganglion neurons from C57BL/6J mice were cultured with SHED-CM or DMEM to evaluate the effect on neurite outgrowth. Streptozotocin-induced diabetic mice were injected with 100 mu L of SHED-CM or DMEM into the unilateral hindlimb muscles twice a week over a period of 4 weeks. Peripheral nerve functions were evaluated by the plantar test, and motor and sensory nerve conduction velocities. Intraepidermal nerve fiber densities, capillary number-to-muscle fiber ratio, capillary blood flow and morphometry of sural nerves were also evaluated. Results Conditioned medium of stem cells from human exfoliated deciduous teeth significantly promoted neurite outgrowth of dorsal root ganglion neurons compared with DMEM. Among four fractions of SHED-CM, the only fraction of kDa promoted the neurite outgrowth of dorsal root ganglion neurons. In addition, SHED-CM significantly prevented decline in sensory nerve conduction velocities compared with DMEM in diabetic mice. Although SHED-CM did not improve intraepidermal nerve fiber densities or morphometry of sural nerves, SHED-CM ameliorated the capillary number-to-muscle fiber ratio and capillary blood flow. Conclusions These results suggested that SHED-CM might have a therapeutic effect on diabetic polyneuropathy through promoting neurite outgrowth, and the increase in capillaries might contribute to the improvement of neural function.