Objective

Per-implantitis is one of the implant treatment complications. Dentists have failed to restore damaged periodontium by using conventional therapies. Tissue engineering (stem cells, scaffold and growth factors) aims to reconstruct natural tissues. The paper aimed to isolate both periodontal ligament stem cells (PDLSCs) and bone marrow mesenchymal stem cells (BMMSCs) and use them in a co-culture method to create three-layered cell sheets for reconstructing natural periodontal ligament (PDL) tissue.

Materials and methods

BMMSCs were isolated from rabbit tibia and femur, and PDLSC culture was established from the lower right incisor. The cells were co-cultured to induce BMMSC differentiation into PDL cells. Cell morphology, stem cells and PDL-specific markers (CD90, CD34, and periostin) were also detected using immunofluorescent assay. Co-cultured cell monolayers were detached using temperature-responsive tissue culture dishes and collagen graft to create the three-layer construct. The 3D-engineered tissue was examined histologically and by field emission scanning electron microscopy (FESEM).

Results

BMMSCs co-cultured with PDLSCs successfully induced more PDL cells. The newly induced PDL cells exhibited periostin and CD90 expression. Fluorescence green intensity was measured for the co-cultured cells that were stained with periostin, the mean fluorescence green intensity (periostin expression) was significantly higher for the newly induced PDL cells after 1, 2, and 3 weeks when compared with control (BM-MSCs), at 21 days non-significant difference was measured when compared with control (PDLSCs).

The results showed the successful formation of 3D multilayer PDL tissue. Histological cross-section showed cell sheets and the stable adhesion between them. FESEM examination was conducted for the cross-section, showing three-layered cell sheets with stable adhesion between cells.

Conclusions

The results of this paper report that the three layered-cell sheets were successfully constructed by the novel use of collagen graft as a scaffold to be used in treatment of periodontitis and to envelop the dental implants to create biohybrid implant.

中文翻译：

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通过共培养方法体外扩增牙周膜细胞并形成多层牙周膜衍生细胞片。

客观的

种植体周围炎是种植体治疗并发症之一。牙医未能通过使用传统疗法来恢复受损的牙周组织。组织工程（干细胞、支架和生长因子）旨在重建天然组织。该论文旨在分离牙周膜干细胞 (PDLSCs) 和骨髓间充质干细胞 (BMMSCs)，并将它们用于共培养方法，以创建用于重建天然牙周膜 (PDL) 组织的三层细胞片。

材料和方法

从兔胫骨和股骨中分离BMMSCs，从右下切牙建立PDLSC培养物。将细胞共培养以诱导BMMSC分化为PDL细胞。还使用免疫荧光测定法检测细胞形态、干细胞和 PDL 特异性标志物（CD90、CD34 和骨膜素）。使用温度响应性组织培养皿和胶原移植物分离共培养的细胞单层以产生三层结构。通过组织学和场发射扫描电子显微镜 (FESEM) 检查 3D 工程组织。

结果

与 PDLSCs 共培养的 BMMSCs 成功地诱导了更多的 PDL 细胞。新诱导的PDL细胞表现出periostin和CD90表达。测量用骨膜素染色的共培养细胞的荧光绿色强度，与对照相比，新诱导的 PDL 细胞在 1、2 和 3 周后的平均荧光绿色强度（骨膜素表达）显着高于对照（BM- MSCs)，在 21 天时测量到与对照 (PDLSCs) 相比无显着差异。

结果表明成功形成了 3D 多层 PDL 组织。组织学横截面显示细胞片和它们之间的稳定粘附。对横截面进行FESEM检查，显示细胞之间具有稳定粘附的三层细胞片。

结论

本文的研究结果表明，通过新型胶原移植物作为支架，成功构建了三层细胞片，用于治疗牙周炎并包裹牙种植体以制造生物混合种植体。