骨髓基質與牙齦造纖維細胞在阻隔膜上的附著與形態表現

Morphologic Characteristics and Cell Attachment on Osteopromotive Barrier Materials

董醒任(S. Z. Dung) ； 藍文謙(W. C. Lan) ； 陳瑞雲(S. W. Chan) ； 楊世芳(S. F. Yang)

《中華民國牙周病醫學會雜誌》 4卷1期 (1999/03) Pp. 25-32

摘要

由於阻隔膜材料的不斷研發及牙周再生技術的成功運用，使引導骨再生技術得以蓬勃發展。最近有學者發現阻隔膜之組織整合能力和牙周再生治療的效果有關。但是阻隔膜之組織整合能力對引導骨再生術的臨床治療效果之影響目前仍不清楚，值得探討。由於細胞附著是組織整合的第一與最重要的一步，本實驗目的在觀察骨髓基質與牙齦造纖維細胞在不同阻隔膜上的形態表現與附著的情形。從雄性大白鼠大腿骨沖出骨髓，以α-MEM培養得到骨髓基質細胞。從病人口內取得牙齦組織，切碎後以DMEM培養得到牙齦造纖維細胞。二種細胞均在37℃、5% CO2的條件下進行培養。培養得到的骨髓基質與牙齦造纖維細胞，在24洞培養皿，與GTAM、TefGen、BioMend、LamBone及Resolut共同培養六或二十四小時，再以PBS沖去沒有附著的細胞。最後，以電子顯微鏡觀察細胞附著於不同阻隔膜上的數目和形態表現。結果骨髓基質細胞附著在TefGen及LamBone的數量最多；牙齦造纖維細胞主要附著在BioMend與LamBone；兩種細胞在Resolut與GTAM上附著的數量很少甚至沒有附著。骨髓基質細胞在TefGen上的形態呈現梭狀或星狀，有一些偽足，比其他阻隔膜上的情形稍好；在LamBone上的形態為圓球狀。牙齦造纖維細胞在BioMend及LamBone上的形態表現最為理想，細胞呈現梭狀、星狀或攤平狀，且有發育良好的觸足。整體而言，不同細胞在不同阻隔膜上附著能力及形態表現不盡相同。細胞附著能力和形態表現對臨床治療效果之影響我們正在進 一步評估中。

關鍵字

牙齦造纖維細胞 ； 骨髓基質細胞 ； 細胞附著 ； 細胞形態 ； 阻隔膜 ； 組織整合

並列摘要

Guided bone regeneration has been successfully used to treat alveolar ridge and pen-implant defects. However, the biologic properties of various baffler materials have seldom been compared. The purpose of the study was to compare the attachment and morphologic characteristics of gingival fibroblasts and bone marrow stromal cells on various baffler membranes. Gingival fibroblasts were derived from human gingiva and grew in DMEM and 10% FBS. Bone marrow was flushed from rat femurs and the adherent bone marrow stromal cells were separated from the non-adherent cells, and were maintained in a -MEM containing 10% FBS and ascorbic acid (50 μg/ml). The confluent cultured cells (1×10^5 cells/well) were passed to 24-well plates with various membranes fixed to the bottom of each well. The baffler membranes tested were GTAM, BioMend, Resolute, TefGen, and LamBone. The wells with no membranes served as controls. After cultured for 6 or 24 h, the non-adherent cells were washed out with phosphate buffer saline and the adhered cells were fixed with 2% paraformaldehyde, processed, and observed under scanning electron microscope. Results showed that different membranes exhibited different ultrastructures. TefGen, Resolut and LamBone had smoother surfaces. Gingival fibroblasts and bone marrow stromal cells appeared to have similar adhesive ability on a particular membrane except that gingival fibroblasts had better adhesive ability on LamBone. The surfaces of Resolute and GTAM attached the fewest number of both cells. The cells on TefGen and BioMend were flat and exhibited more processes. In general, cells after 6 h of incubation demonstrated more processes. Results from the study indicated that the adhesive ability and morphologic characteristics of cells on various barrier membranes were different and its significance on tissue integration and bone regeneration in vivo needs further evaluation.

並列關鍵字

gingival fibroblasts ； bone marrow stromal cells ； cell adhesion ； cell morphology ； barrier membrane ； tissue integration