羟基磷灰石/聚氨酯复合骨诱导再生膜的细胞相容性及自身降解性能

Cell biocompatibility and degradation of hydroxyapatite/polyurethane composite membrane in inducing osseous tissue regeneration

目的：骨诱导再生膜材料对骨组织损伤后的重建和再生起着屏蔽和诱导的作用。拟进一步验证羟基磷灰石（Hydroxyapatite,HA）/聚氨酯（Polyurethane,PU）复合骨诱导再生膜体内外的细胞相容性和降解性能。方法：实验于2007-03/07在四川大学华西口腔医学国家重点实验室及四川大学纳米生物材料研究中心完成。①HA/PU复合膜制备成10mm×10mm大小,选择成熟的成骨细胞株MG63,体外细胞培养,在1,4,7d内观察细胞在膜上的生长和增殖情况,并采用MTT法测定细胞的增殖率,判断其细胞的毒性。②把HA/PU复合膜制备成直径1.5mm大小,选择健康的SD雌性大鼠3只,在脊柱一侧肌肉内埋植HA/PU复合膜,在1,4,12周后取出。以苏木精-伊红染色和Masson染色,组织切片观察材料的细胞相容性和自身降解性。结果：①HA/PU复合膜上细胞增殖良好,无坏死和悬浮细胞出现,细胞在膜上的增殖能力均高于MG63的增殖能力（P〈0.05）,细胞毒性为0级。②组织学切片观察,1～4周时材料在体内依旧是有形固体,材料被周围组织包裹,有巨噬细胞聚集,未见多核细胞,胶原纤维逐渐变得致密。12周时,材料发生降解,有纤维组织长入,周围与组织结合良好,未见有炎症发生。结论：HA/PU复合膜具有良好的细胞相容性和自身降解性能,可作为骨组织诱导再生膜材料。

AIM： The membrane materials for inducing bone regeneration play an important role in screening and inducing the rebuilding and regeneration of osseous tissue after wound. This study was designed to further verify cell compatibility and degradation of hydroxyapatite （HA）/polyurethane （PU） composite membrane to induce osseous tissue regeneration in vitro and in vivo. METHODS： The experiment was conducted in the National Key Laboratory of West China College of Stomatology in Sichuan University and the Research Center for Nano-biomaterials in Sichuan University from March to July in 2007.①HA/PU composite membrane with a size of 10 mm× 10mm was prepared, and mature osteoblast strain MG63 were cultured in vitro, then the cell growth and proliferation on the surface of materials were observed at days l, 4, 7. Cell proliferation rate by MTT method was evaluated so as to justify cytotoxicity.②HA/PU composite membrane with a diameter of 1.5 mm was prepared, and three healthy SD female rats were selected. HA/PU composite membrane was bedded at one side of vertebral column＇s muscle and fetched out at weeks 1, 4, and 12. The cell compatibility and degradation were detected with hematoxylin-eosin stain and Masson stain. RESULTS： ①Cells proliferated well on the HA/PU composite membrane, no necrotic or suspended cells appeared, and the proliferation ability of cells on the composite membrane was higher than MG63 cells （P 〈 0.05）, with the cytotoxicity of zero grade.②Histological sections revealed that, materials were at solid state and wrapped up by the surrounding around tissues in vivo in 1-4 weeks. Some macrophages aggregated around the materials and no apocytes appeared. Collagen fibers turned to dense gradually. In weeks 12, HA/PU composite membrane generated to degradation, some fibrous tissues ingrew. The surrounding tissues combine well with the materials and no inflammation was found. CONCLUSION： HA/PU composite membrane has a good cell compatibility and degradation, and may be used as a membrane material to induce osseous tissue regeneration.