Hambleton J, Schwartz Z, Khare A, Windeler S W, Luna M, Brooks B P, Dean D D, Boyan B D
University of Texas Health Science Center at San Antonio 78284.
J Orthop Res. 1994 Jul;12(4):542-52. doi: 10.1002/jor.1100120411.
The effect on chondrocyte metabolism of culture surfaces sputter-coated with various materials used for orthopaedic implants was studied and correlated with the stage of cartilage cell maturation. Confluent, fourth-passage chondrocytes from the costochondral resting zone and growth zone of rats were cultured for 6 or 9 days on 24-well plates sputter-coated with ultrathin films of titanium, titanium dioxide, aluminum oxide, zirconium oxide, and calcium phosphate (1.67:1). Corona-discharged tissue culture plastic served as the control. The effect of surface material was examined with regard to cell morphology; cell proliferation (cell number) and DNA synthesis ([3H]thymidine incorporation); RNA synthesis ([3H]uridine incorporation); collagenase-digestible protein, noncollagenase-digestible protein, and percentage of collagen production; and alkaline phosphatase-specific activity, both in the cell layer and in trypsinized chondrocytes. Cell morphology was dependent on surface material; only cells cultured on titanium had an appearance similar to that of cells cultured on plastic. While titanium or titanium dioxide surfaces had no effect on cell number or [3H]thymidine incorporation, aluminum oxide, calcium phosphate, and zirconium oxide surfaces inhibited both parameters. Cells cultured on aluminum oxide, calcium phosphate, zirconium oxide, and titanium dioxide exhibited decreased collagenase-digestible protein, noncollagenase-digestible protein, and percentage of collagen production, but [3H]uridine incorporation was decreased only in those chondrocytes cultured on aluminum oxide, calcium phosphate, or zirconium oxide. Chondrocytes cultured on titanium had greater alkaline phosphatase-specific activity than did cells cultured on plastic, but the incorporation of [3H]uridine and production of collagenase-digestible protein, noncollagenase-digestible protein, and percentage of collagen was comparable. The response of chondrocytes from the growth zone and resting zone to culture surface was comparable, differing primarily in magnitude. Cell maturation-dependent effects were evident when enzyme activity in trypsinized and scraped cells was compared. These results indicate that different surface materials affect chondrocyte metabolism and phenotypic expression in vitro and suggest that implant materials may modulate the phenotypic expression of cells in vivo.
研究了用各种用于骨科植入物的材料进行溅射涂层的培养表面对软骨细胞代谢的影响,并将其与软骨细胞成熟阶段相关联。将来自大鼠肋软骨静止区和生长区的汇合的第四代软骨细胞在溅射涂覆有钛、二氧化钛、氧化铝、氧化锆和磷酸钙(1.67:1)超薄膜的24孔板上培养6天或9天。经电晕放电处理的组织培养塑料用作对照。从细胞形态、细胞增殖(细胞数量)和DNA合成([3H]胸苷掺入)、RNA合成([3H]尿苷掺入)、胶原酶可消化蛋白、非胶原酶可消化蛋白和胶原蛋白产生百分比以及细胞层和胰蛋白酶消化的软骨细胞中的碱性磷酸酶比活性等方面研究了表面材料的影响。细胞形态取决于表面材料;只有在钛上培养的细胞外观与在塑料上培养的细胞相似。虽然钛或二氧化钛表面对细胞数量或[3H]胸苷掺入没有影响,但氧化铝、磷酸钙和氧化锆表面均抑制这两个参数。在氧化铝、磷酸钙、氧化锆和二氧化钛上培养的细胞表现出胶原酶可消化蛋白、非胶原酶可消化蛋白和胶原蛋白产生百分比降低,但只有在氧化铝、磷酸钙或氧化锆上培养的那些软骨细胞中[3H]尿苷掺入减少。在钛上培养的软骨细胞比在塑料上培养的细胞具有更高的碱性磷酸酶比活性,但[3H]尿苷掺入以及胶原酶可消化蛋白、非胶原酶可消化蛋白和胶原蛋白百分比的产生相当。生长区和静止区的软骨细胞对培养表面的反应相当,主要区别在于程度。当比较胰蛋白酶消化和刮擦细胞中的酶活性时,细胞成熟依赖性效应很明显。这些结果表明,不同的表面材料在体外影响软骨细胞代谢和表型表达,并表明植入材料可能在体内调节细胞的表型表达。