1 Department of Urology and Andrology, Affiliated Drum Tower Hospital, Nanjing University School of Medicine , Nanjing, China .
2 Department of Urology, Nanjing First Hospital, Nanjing Medical University , Nanjing, China .
Tissue Eng Part A. 2018 Jul;24(13-14):1066-1081. doi: 10.1089/ten.TEA.2017.0376. Epub 2018 Mar 1.
Tissue engineering techniques provide a great potential to de novo construct a histological bladder. Smooth muscle regeneration is extremely important for the functional recovery of engineered neobladder. However, many challenges remain for the use of bladder smooth muscle cells (SMCs) as the cell sources. Recent evidences showed that smooth muscle progenitor cells (SPCs) in the peripheral blood have the capacity of differentiating into SMCs, while their use for bladder regeneration has not yet been reported. The aim of our study was to investigate the effect and mechanism of autologous SPCs on bladder regeneration in a rabbit model. In this study, autologous SPCs were isolated and cultured from the peripheral blood, labeled with CM-DiI, and then seeded into a porcine bladder acellular matrix (BAM) to construct a SPC-BAM complex, which was finally implanted to substitute the partial bladder with an equivalent size. In the results, SPCs demonstrated the phenotype of stem/progenitor cells, expressed SMs markers (alpha-smooth muscle actin [α-SMA], desmin, calponin, SM22α, and smooth muscle myosin heavy chain [SMMHC]), and displayed carbachol-induced contraction. Compared with cell-free BAM, the SPC-BAM was able to improve histological regeneration (smooth muscle regeneration, vascularization, and nerve formation) and functional recovery (urodynamic function and smooth muscle contraction) of the engineered neobladder. Cell tracing indicated that seeded SPCs could survive and directly integrated into the regenerated neobladder. In addition, SPCs could also promote proliferation and migration of rabbit bladder SMCs through the paracrine platelet-derived growth factor-BB (PDGF-BB). In conclusion, our study first demonstrated that SPCs from the peripheral blood could enhance histological regeneration and functional recovery of the tissue-engineered neobladder through both the direct integration and indirect paracrine effect, supporting the use of SPCs as the cell sources for tissue engineering of the bladder.
组织工程技术为从头构建组织学膀胱提供了巨大的潜力。平滑肌再生对于工程化新膀胱的功能恢复极其重要。然而，使用膀胱平滑肌细胞 (SMC) 作为细胞来源仍然存在许多挑战。最近的证据表明，外周血中的平滑肌祖细胞 (SPC) 具有分化为 SMC 的能力，但其用于膀胱再生尚未见报道。我们的研究目的是在兔模型中研究自体 SPC 对膀胱再生的作用和机制。在这项研究中，我们从外周血中分离和培养自体 SPC，用 CM-DiI 标记，然后接种到猪膀胱去细胞基质 (BAM) 中构建 SPC-BAM 复合物，最后将其植入以替代具有同等大小的部分膀胱。结果表明，SPC 表现出干细胞/祖细胞的表型，表达 SMC 标志物（α-平滑肌肌动蛋白 [α-SMA]、结蛋白、钙调蛋白、SM22α 和平滑肌肌球蛋白重链 [SMMHC]），并显示出乙酰胆碱诱导的收缩。与无细胞 BAM 相比，SPC-BAM 能够改善工程化新膀胱的组织学再生（平滑肌再生、血管生成和神经形成）和功能恢复（尿动力学功能和平滑肌收缩）。细胞示踪表明，接种的 SPC 可以存活并直接整合到再生的新膀胱中。此外，SPC 还可以通过旁分泌血小板衍生生长因子-BB (PDGF-BB) 促进兔膀胱 SMC 的增殖和迁移。总之，我们的研究首次表明，外周血中的 SPC 可以通过直接整合和间接旁分泌作用增强组织工程化新膀胱的组织学再生和功能恢复，支持将 SPC 用作膀胱组织工程的细胞来源。
