共培养促神经化策略在组织工程骨构建中的研究进展

doi:10.3877/cma.j.issn.2095-1221.2022.05.006

组织工程骨移植是治疗大段骨缺损的重要手段，将选定的组织或细胞采用特定的模式进行共培养，使组织工程骨神经化，可有效增强组织工程骨对损伤骨组织的修复作用。本文回顾国内外组织工程骨构建中共培养促神经化策略相关文献，从组织和种子细胞选择、相互作用分子机制和共培养模式应用3个方面进行综述，以期为该技术进一步发展及其临床应用提供参考。

关键词: 共培养, 神经化, 组织工程骨

Tissue engineering bone transplantation is an important method for the treatment of large-segment bone defects. Co-cultivation of selected tissues or cells in a specific culture mode can make tissue engineering bone more neural, which can effectively enhance the repair effect of tissue engineering bone on the damaged bone tissue. The literature related to co-culture promoting neuralization strategy in tissue engineered bone construction was reviewed from three aspects inthisarticle: tissue and seed cell selection, the molecular mechanism of tissue and seed cell interaction, and application of co-culture mode, in order to provide a reference for the further development of the technology and its clinical applications.

Key words: Co-culture, Neuralization, Tissue engineering bone

表1 不同共培养模式神经化组织工程骨特点比较

共培养组织/细胞	体外检测	体内检测	优势	参考文献
大隐神经/组织工程骨	n.a	NPY	大隐神经支配的组织工程骨具有更佳的促进骨修复能力	覃俊君等^[3]
SCs/自体颅骨组织	骨钙素、骨源性碱性磷酸酶、骨桥蛋白	n.a	SCs与骨组织共培养加强成骨细胞的骨修复能力	李钟鹏等^[4]
BMSCs/壳聚糖多孔支架	n.a	溴脱氧核苷尿嘧啶、胶质纤维酸性蛋白	壳聚糖多孔支架与BMSCs共培养能明显改善大鼠受损的神经功能	谭可可等^[5]
多能干细胞/神经前体细胞	巢蛋白	n.a	多能干细胞与神经前体细胞共培养可诱导未分化人类多能干细胞的神经转化	Rhee等^[6]
SCs/MSCs	磷酸化蛋白激酶B、B淋巴细胞瘤-2蛋白	n.a	SCs与MSCs共培养可抑制SCs的凋亡，对神经病变起修复作用	Zhang等^[7]
BMSCs /新生儿海马细胞	n.a	巢蛋白、微管蛋白	BMSCs与海马细胞共培养促进BMSCs向神经元分化	Majeed等^[8]
隐神经/组织工程骨	n.a	组织工程骨形成	植入隐神经的组织工程骨具有更佳的促进骨缺损修复能力	江汕等^[12]
骨皮质/骨髓	BMP2、TGF-β1	骨折愈合情况	骨皮质与骨髓共培养的MSCs具有更好的促进骨折愈合能力	Zhu等^[18]
SCs/ADSCs	神经元特异性烯醇化酶、微管关联蛋白2、胶质纤维酸性蛋白、神经元特异性核蛋白	n.a	SCs与ADSCs共培养可显著促进ADSCs向神经元样细胞分化	张振辉等^[19]
SCs/同种异体骨	S-100蛋白	n.a	SCs与支架具有良好的组织相融性	周翔等^[20]
骨膜细胞/髓核细胞	骨钙素、I型胶原、RUNX2 、ALP	n.a	骨膜细胞与髓核细胞共培养在低氧环境里成骨分化能力更强	陆海涛等^[50]

表1 不同共培养模式神经化组织工程骨特点比较

共培养组织/细胞	体外检测	体内检测	优势	参考文献
大隐神经/组织工程骨	n.a	NPY	大隐神经支配的组织工程骨具有更佳的促进骨修复能力	覃俊君等^[3]
SCs/自体颅骨组织	骨钙素、骨源性碱性磷酸酶、骨桥蛋白	n.a	SCs与骨组织共培养加强成骨细胞的骨修复能力	李钟鹏等^[4]
BMSCs/壳聚糖多孔支架	n.a	溴脱氧核苷尿嘧啶、胶质纤维酸性蛋白	壳聚糖多孔支架与BMSCs共培养能明显改善大鼠受损的神经功能	谭可可等^[5]
多能干细胞/神经前体细胞	巢蛋白	n.a	多能干细胞与神经前体细胞共培养可诱导未分化人类多能干细胞的神经转化	Rhee等^[6]
SCs/MSCs	磷酸化蛋白激酶B、B淋巴细胞瘤-2蛋白	n.a	SCs与MSCs共培养可抑制SCs的凋亡，对神经病变起修复作用	Zhang等^[7]
BMSCs /新生儿海马细胞	n.a	巢蛋白、微管蛋白	BMSCs与海马细胞共培养促进BMSCs向神经元分化	Majeed等^[8]
隐神经/组织工程骨	n.a	组织工程骨形成	植入隐神经的组织工程骨具有更佳的促进骨缺损修复能力	江汕等^[12]
骨皮质/骨髓	BMP2、TGF-β1	骨折愈合情况	骨皮质与骨髓共培养的MSCs具有更好的促进骨折愈合能力	Zhu等^[18]
SCs/ADSCs	神经元特异性烯醇化酶、微管关联蛋白2、胶质纤维酸性蛋白、神经元特异性核蛋白	n.a	SCs与ADSCs共培养可显著促进ADSCs向神经元样细胞分化	张振辉等^[19]
SCs/同种异体骨	S-100蛋白	n.a	SCs与支架具有良好的组织相融性	周翔等^[20]
骨膜细胞/髓核细胞	骨钙素、I型胶原、RUNX2 、ALP	n.a	骨膜细胞与髓核细胞共培养在低氧环境里成骨分化能力更强	陆海涛等^[50]

图1 CGRP相关信号分子对成骨-破骨作用信号通路注：CGRP为降钙素基因相关肽；TRPV1为辣椒素受体；RANKL为核因子活化因子受体配体；cAMP为环磷酸腺苷；PKA为蛋白激酶A；IGF1为胰岛素样生长因子1；GSK-3β为糖原合成酶激酶；SP7/Osterix为锌指结构转录因子；CREB1为成骨相关蛋白环磷酸腺苷应答元件结合蛋白1；BMP2为骨形态发生蛋白2；TNF-α为肿瘤坏死因子α；β-catenin为β-连环蛋白

图2 Sema3A相关信号分子对成骨-破骨作用信号通路注：Sema3A为信号素家族的一种可扩散蛋白；PlexinA1/TREM2/DAP12为Sema3A-Nrp1轴干扰/髓系细胞触发受体2/自然杀伤激活受体相关蛋白12；GEF为鸟嘌呤核苷酸交换因子；Rac1为C3肉毒素底物1；RANKL为核因子活化因子受体配体；RhoA为Ras同源基因家族成员A重组蛋白阻止破骨前体细胞分化；β-catenin为β-连环蛋白

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[1]	张宁, 刘萍, 郝翠芳, 王昕荣. 白细胞介素－1β，－1Rα对胚胎－内膜共培养体系分泌基质金属蛋白酶－9/细胞间黏附分子－1的影响[J]. 中华妇幼临床医学杂志(电子版), 2012, 08(02): 159-162.
[2]	杨园园, 朱新贤, 靳霞, 童晓文. 人间充质干细胞对乳腺癌细胞生长的影响研究[J]. 中华细胞与干细胞杂志(电子版), 2020, 10(04): 195-203.
[3]	张新媛, 王麒雲, 陈晓思. 糖尿病视网膜病变血管内皮细胞与神经细胞藕联二维体外共培养模型的实验研究[J]. 中华眼科医学杂志(电子版), 2023, 13(01): 6-11.

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Advances in co-culture promoted neuralization strategy in tissue engineering bone construction

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Advances in co-culture promoted neuralization strategy in tissue engineering bone construction