Martin J Y, Dean D D, Cochran D L, Simpson J, Boyan B D, Schwartz Z
Wilford Hall Medical Center, Lackland AFB, TX, USA.
Clin Oral Implants Res. 1996 Mar;7(1):27-37. doi: 10.1034/j.1600-0501.1996.070104.x.
This study compared osteoblasts proliferation, differentiation, and protein synthesis on new and used titanium (Ti) disks to test the hypothesis that cleaning and resterilization of previously used Ti disks does not alter cell response to a particular surface. Ti disks of varying roughness were prepared by one of five different treatment regimens. Standard tissue culture plastic was used as a control. Human osteoblast-like cells (MG63) were cultured on the Ti disks and cell proliferation, cell differentiation, RNA synthesis and matrix production (collagen and noncollagen protein; proteoglycans) measured. After their first use, the disks were cleaned, resterilized by autoclaving, and MG63 cells cultured on them as before. At confluence, the same parameters were measured and cell behavior on new and used disks compared. When confluent cultures of cells on plastic were compared to those cultured on new Ti surfaces, cell number was reduced on the roughest surfaces and equivalent to plastic on the other surfaces. Cell number was further reduced when disks with the roughest surfaces were re-used; no differences in cell number could be discerned after cleaning and resterilization. Cell proliferation was inversely related to surface roughness and was less than seen on tissue culture plastic. Re-use of the Ti disks resulted in no change in cell proliferation rate. Alkaline phosphatase specific activity in isolated cells was lowest on the rougher surfaces; no differences between new and used disks were observed. Similarly, enzyme activity in the cell layer was decreased in cultures grown on rougher surfaces, with no effect of prior disk use being noted. RNA synthesis was decreased with respect to plastic in cultures on smoother surfaces and increased on rougher surfaces; prior disk use did not alter RNA synthesis. Collagen production by the cells was decreased on smoother surfaces, but was comparable to tissue culture plastic when grown on rougher surfaces. Non-collagen protein production was unaffected by culture surface and whether or not the disk had been previously used. Proteoglycan synthesis by cells was decreased on all surfaces studied and comparable on both new and used disks. The results of this study indicate that Ti implant surfaces are unaffected by cleaning and resterilization, although rougher surfaces may require more extensive cleaning than smoother ones. This suggests the possibility that implants, in the same patient, could be safely reused. In vivo studies in animals, however, need to be performed before clinical application can be considered.
本研究比较了新的和使用过的钛（Ti）盘上成骨细胞的增殖、分化及蛋白质合成，以验证以下假设：对先前使用过的Ti盘进行清洁和再灭菌不会改变细胞对特定表面的反应。通过五种不同处理方案之一制备了不同粗糙度的Ti盘。使用标准组织培养塑料作为对照。将人成骨样细胞（MG63）培养在Ti盘上，并测量细胞增殖、细胞分化、RNA合成及基质产生（胶原蛋白和非胶原蛋白；蛋白聚糖）。首次使用后，将盘进行清洁，通过高压灭菌进行再灭菌，并像之前一样在上面培养MG63细胞。汇合时，测量相同参数并比较新盘和使用过的盘上的细胞行为。当将塑料上的汇合细胞培养物与新Ti表面上的培养物进行比较时，最粗糙表面上的细胞数量减少，而其他表面上的细胞数量与塑料上的相当。当再次使用最粗糙表面的盘时，细胞数量进一步减少；清洁和再灭菌后细胞数量无差异。细胞增殖与表面粗糙度呈负相关，且低于组织培养塑料上的增殖情况。Ti盘的再次使用导致细胞增殖率无变化。分离细胞中碱性磷酸酶的比活性在较粗糙表面上最低；未观察到新盘和使用过的盘之间存在差异。同样，在较粗糙表面上生长的培养物中细胞层中的酶活性降低，未发现先前盘的使用有影响。相对于塑料，在较光滑表面上的培养物中RNA合成减少，而在较粗糙表面上增加；先前盘的使用未改变RNA合成。细胞产生的胶原蛋白在较光滑表面上减少，但在较粗糙表面上生长时与组织培养塑料相当。非胶原蛋白的产生不受培养表面及盘是否先前使用的影响。在所研究的所有表面上，细胞的蛋白聚糖合成均减少，新盘和使用过的盘上相当。本研究结果表明，Ti植入物表面不受清洁和再灭菌的影响，尽管较粗糙的表面可能比较光滑的表面需要更彻底的清洁。这表明在同一患者体内，植入物有可能安全地重复使用。然而，在考虑临床应用之前，需要在动物身上进行体内研究。
