Boyan B D, Lincks J, Lohmann C H, Sylvia V L, Cochran D L, Blanchard C R, Dean D D, Schwartz Z
Department of Periodontics, University of Texas Health Science Center at San Antonio, 78284-7774, USA.
J Orthop Res. 1999 May;17(3):446-57. doi: 10.1002/jor.1100170322.
During endochondral bone formation, as occurs in fracture healing, chondrocytes are one of the first cells to see an implant surface. We tested the hypothesis that chemical composition and surface roughness affect chondrocyte differentiation, matrix synthesis, and local factor production and that the nature of the response is dependent on the state of maturation of the cells. To do this, we harvested rat growth zone and resting zone chondrocytes and examined their response to smooth and rough disk surfaces manufactured from either commercially pure titanium or titanium alloy. Profilometry, scanning electron microscopy, Auger spectroscopy, and Fourier transform infrared spectroscopy were used to characterize the surfaces. Average roughness values were 0.22 microm for smooth titanium surfaces, 0.23 microm for smooth titanium alloy surfaces, 4.24 microm for rough titanium surfaces, and 3.20 microm for rough titanium alloy surfaces. Cells were grown on the different disk surfaces until the cultures had reached confluence on plastic. The effect of the surfaces was determined by assaying cell number and [3H]thymidine incorporation as measures of cell proliferation, cell layer and cell alkaline phosphatase specific activity as markers of differentiation, and collagen production and [35S]sulfate incorporation as indicators of extracellular matrix production. In addition, the synthesis of prostaglandin E2 and transforming growth factor-beta were examined to measure changes in local factor synthesis. In growth zone and resting zone cultures, cell number and [3H]thymidine incorporation were decreased on rough surfaces; however, this effect was greater on commercially pure titanium surfaces. Cell layer and cell alkaline phosphatase specific activity were decreased in resting zone cells grown on rough surfaces. Cell alkaline phosphatase specific activity in growth zone cells was decreased on rough surfaces, whereas cell layer alkaline phosphatase specific activity was increased only in growth zone cells grown on rough commercially pure titanium surfaces. Resting zone cell collagen production was decreased only on rough commercially pure titanium, whereas in growth zone cells, collagen production was increased. Increased prostaglandin E2 release into the media was found for growth zone and resting zone cell cultures on the disks with rough surfaces. The observed effect was greater on rough commercially pure titanium. Production of transforming growth factor-beta by resting zones was similarly affected, whereas an increase in its production by growth zone cells was measured only on rough commercially pure titanium. These results indicate that surface roughness affects chondrocyte proliferation, differentiation, matrix synthesis, and local factor production and that these parameters are also affected by chemical composition. Furthermore, the nature and extent of the cell response is dependent on cell maturation. The overriding variable in response to an implant material, however, appears to be roughness of the surface.
在软骨内骨形成过程中,如骨折愈合时发生的那样,软骨细胞是最早接触植入物表面的细胞之一。我们测试了以下假设:化学成分和表面粗糙度会影响软骨细胞的分化、基质合成和局部因子产生,并且反应的性质取决于细胞的成熟状态。为此,我们采集了大鼠生长区和静止区的软骨细胞,并研究了它们对由商业纯钛或钛合金制成的光滑和粗糙圆盘表面的反应。使用轮廓仪、扫描电子显微镜、俄歇电子能谱和傅里叶变换红外光谱对表面进行表征。光滑钛表面的平均粗糙度值为0.22微米,光滑钛合金表面为0.23微米,粗糙钛表面为4.24微米,粗糙钛合金表面为3.20微米。细胞在不同的圆盘表面上生长,直到培养物在塑料上达到汇合。通过检测细胞数量和[3H]胸腺嘧啶核苷掺入量来测定细胞增殖,以细胞层和细胞碱性磷酸酶比活性作为分化标志物,以胶原蛋白产生和[35S]硫酸盐掺入量作为细胞外基质产生的指标,来确定表面的影响。此外,检测前列腺素E2和转化生长因子-β的合成,以测量局部因子合成的变化。在生长区和静止区培养物中,粗糙表面上的细胞数量和[3H]胸腺嘧啶核苷掺入量减少;然而,这种影响在商业纯钛表面上更大。在粗糙表面上生长的静止区细胞中,细胞层和细胞碱性磷酸酶比活性降低。粗糙表面上生长区细胞的细胞碱性磷酸酶比活性降低,而仅在粗糙商业纯钛表面上生长的生长区细胞中,细胞层碱性磷酸酶比活性增加。静止区细胞的胶原蛋白产生仅在粗糙商业纯钛表面上减少,而在生长区细胞中,胶原蛋白产生增加。在表面粗糙的圆盘上培养的生长区和静止区细胞培养物中,发现前列腺素E2释放到培养基中的量增加。观察到的影响在粗糙商业纯钛上更大。静止区转化生长因子-β的产生受到类似影响,而仅在粗糙商业纯钛表面上测量到生长区细胞中其产生增加。这些结果表明,表面粗糙度会影响软骨细胞的增殖、分化、基质合成和局部因子产生,并且这些参数也受化学成分影响。此外,细胞反应的性质和程度取决于细胞成熟度。然而,对植入材料反应中的首要变量似乎是表面粗糙度。