MSC/ECM Cellular Complexes Induce Periodontal Tissue Regeneration
MSC/ECM Cellular Complexes Induce Periodontal Tissue Regeneration
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DOI：
10.1177/0022034517708770
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发表时间：
2017-05
影响因子：
7.6
通讯作者：
Manabu Takewaki;M. Kajiya;K. Takeda;Shinji Sasaki;Souta Motoike;Nao Komatsu;S. Matsuda;K. Ouhara;N. Mizuno;T. Fujita;H. Kurihara
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文献类型：
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作者：
Manabu Takewaki;M. Kajiya;K. Takeda;Shinji Sasaki;Souta Motoike;Nao Komatsu;S. Matsuda;K. Ouhara;N. Mizuno;T. Fujita;H. Kurihara
间充质干细胞（MSCs）具有自我更新和多能性，移植到牙周缺损被认为是牙周组织再生的一个有用的选择。然而，仍然需要开发更可靠和可预测的植入技术。最近，我们产生了MSC/细胞外基质（ECM）复合物（C-MSC）的团块，其由细胞和自身产生的ECM组成。C-MSCs可以在体外调节其细胞功能，并且可以移植到缺损部位，而无需任何人工支架，以诱导骨再生。因此，本研究旨在评估C-MSC移植对Beagle犬牙周组织再生的影响。采用7只比格犬建立前磨牙III类根分叉缺损模型。将从狗髂骨分离的MSC以7.0 × 104个细胞/孔的密度接种于24孔板中，并在补充有50 μg/mL抗坏血酸的生长培养基中培养4 d。为了获得C-MSC，使用微量移液管尖端刮擦汇合的细胞，然后作为细胞片撕下。将薄片卷起以形成圆形细胞团。将C-MSCs在生长培养基或骨诱导培养基（OIM）中培养5或10 d。体外评价C-MSCs的生物学特性，并采用犬III类根分叉缺损模型检测其牙周组织再生活性。免疫荧光分析显示I型胶原可诱导C-MSCs的形成。OIM在第10天显著增加C-MSCs中的钙沉积，表明其成骨分化能力。C-MSCs和C-MSCs培养OIM移植到狗分叉缺损没有人工支架诱导牙周组织再生成功相比，没有移植，而成骨分化的C-MSCs导致快速牙槽骨再生。这些结果表明，使用C-MSCs精制的自产生ECM可能代表一种新的可预测的牙周组织再生治疗。
Transplantation of mesenchymal stem cells (MSCs), which possess self-renewing properties and multipotency, into a periodontal defect is thought to be a useful option for periodontal tissue regeneration. However, developing more reliable and predictable implantation techniques is still needed. Recently, we generated clumps of an MSC/extracellular matrix (ECM) complex (C-MSC), which consisted of cells and self-produced ECM. C-MSCs can regulate their cellular functions in vitro and can be grafted into a defect site, without any artificial scaffold, to induce bone regeneration. Accordingly, this study aimed to evaluate the effect of C-MSC transplantation on periodontal tissue regeneration in beagle dogs. Seven beagle dogs were employed to generate a premolar class III furcation defect model. MSCs isolated from dog ilium were seeded at a density of 7.0 × 104 cells/well into 24-well plates and cultured in growth medium supplemented with 50 µg/mL ascorbic acid for 4 d. To obtain C-MSCs, confluent cells were scratched using a micropipette tip and were then torn off as a cellular sheet. The sheet was rolled up to make round clumps of cells. C-MSCs were maintained in growth medium or osteoinductive medium (OIM) for 5 or 10 d. The biological properties of C-MSCs were evaluated in vitro, and their periodontal tissue regenerative activity was tested by using a dog class III furcation defect model. Immunofluorescence analysis revealed that type I collagen fabricated the form of C-MSCs. OIM markedly elevated calcium deposition in C-MSCs at day 10, suggesting its osteogenic differentiation capacity. Both C-MSCs and C-MSCs cultured with OIM transplantation without an artificial scaffold into the dog furcation defect induced periodontal tissue regeneration successfully compared with no graft, whereas osteogenic-differentiated C-MSCs led to rapid alveolar bone regeneration. These findings suggested that the use of C-MSCs refined by self-produced ECM may represent a novel predictable periodontal tissue regenerative therapy.