Bhuptani Ronak S, Patravale Vandana B
Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, N.P. Marg, Matunga (E), Mumbai 400019, Maharashtra, India.
Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, N.P. Marg, Matunga (E), Mumbai 400019, Maharashtra, India.
Int J Pharm. 2016 Dec 30;515(1-2):555-564. doi: 10.1016/j.ijpharm.2016.10.040. Epub 2016 Oct 27.
The collective power of stem cells due to their evident advantages is incessantly investigated in regenerative medicine to be the next generation exceptional remedy for tissue regeneration and treatment of diseases. Stem cells are highly sensitive and a 3D culture environment is a requisite for its successful transplantation and integration with tissues. Porous microscaffolds can create a 3D microenvironment for growing stems cells, controlling their fate both in vitro and in vivo. In the present study, interconnected porous PLGA microscaffolds were fabricated, characterized and employed to propagate human dental pulp mesenchymal stem cells (DPMSCs) in vitro. The porous topography was investigated by scanning electron microscopy and the pore size was controlled by fabrication conditions such as the concentration of porogen. DPMSCs were cultured on microscaffolds and were evaluated for their morphology, attachment, proliferation, cell viability via MTT and molecular expression (RT-PCR). DPMSCs were adequately proliferated and adhered over the microscaffolds forming a 3D cell-microscaffold construct. The average number of DPMSCs grown on PLGA microscaffolds was significantly higher than monolayer 2D culture during 5th and 7th day. Moreover, cell viability and gene expression results together corroborated that microscaffolds maintained the viability, stemness and plasticity of the cultured dental pulp mesenchymal stem cells. The novel porous microscaffold developed acts as promising scaffold for 3D culture and survival and transplantation of stem cells for tissue engineering.
由于干细胞具有明显优势,其集体力量在再生医学中不断得到研究,有望成为组织再生和疾病治疗的下一代卓越疗法。干细胞高度敏感,三维培养环境是其成功移植并与组织整合的必要条件。多孔微支架可为干细胞生长创造三维微环境,在体外和体内控制其命运。在本研究中,制备了相互连接的多孔聚乳酸-羟基乙酸共聚物(PLGA)微支架,对其进行了表征,并用于体外扩增人牙髓间充质干细胞(DPMSCs)。通过扫描电子显微镜研究多孔形貌,并通过致孔剂浓度等制备条件控制孔径。将DPMSCs接种在微支架上,通过MTT法评估其形态、黏附、增殖、细胞活力,并通过逆转录聚合酶链反应(RT-PCR)评估其分子表达。DPMSCs在微支架上充分增殖并黏附,形成三维细胞-微支架构建体。在第5天和第7天,PLGA微支架上生长的DPMSCs平均数量显著高于单层二维培养。此外,细胞活力和基因表达结果共同证实,微支架维持了培养的牙髓间充质干细胞的活力、干性和可塑性。所开发的新型多孔微支架有望成为用于组织工程的干细胞三维培养、存活及移植的支架。
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