Tucci M A, Benghuzzi H A
Department of Orthopedics, University of Mississippi Medical Center, Jackson 39216.
Biomed Sci Instrum. 1994;30:205-12.
The specific objective of this study was to investigate the effect of various biomedical polymers on the adhesion rate of buccal epithelial cells (BEC) as a model for biocompatibility. The BEC in this study were isolated, biotin labeled and seeded by following standard laboratory procedures. A total of 2.5 x 10(5) cells was plated in each microtiter-well pretreated with various concentrations of (0.01, 0.10 and 1% wt/vol) poly-L-valanine (P-Val), poly-L-alanine (P-Ala), poly-glycine (P-Gly), poly-L-tryptophan (P-Trp), poly-L-asparagine (P-Asn) and buffered control. At the end of 1, 4, and 24 hours the assay was developed by utilizing avidin-horseradish peroxidase and o-phenylenediamine dihydrochloride as substrate to measure biotin-labeled-BEC. Cell number was then determined using a standard curve prepared by using known BEC number vs. absorbance units. The data from this study suggest that: (I) the ease of adhesion of BEC was in the following order: P-Val > P-Gly = P-Trp = P-Ala > P-Asn > P-Asp = Control, (II) the rate of BEC spreading was strongly influenced by both incubation time and the polymer concentration, (III) the surface attachment of BEC to the polymer were demonstrated to vary depending on their chemical structure and level of microporosity. Thus, overall observation led us to conclude that the surface reactivity of polymer materials must always be taken into account in discussing their biocompatibility in vivo.
本研究的具体目的是研究各种生物医学聚合物对颊上皮细胞(BEC)黏附率的影响,以此作为生物相容性的模型。本研究中的BEC按照标准实验室程序进行分离、生物素标记和接种。将总共2.5×10⁵个细胞接种到预先用各种浓度(0.01%、0.10%和1%重量/体积)的聚-L-缬氨酸(P-Val)、聚-L-丙氨酸(P-Ala)、聚甘氨酸(P-Gly)、聚-L-色氨酸(P-Trp)、聚-L-天冬酰胺(P-Asn)处理过的每个微量滴定孔中,并设置缓冲对照。在1小时、4小时和24小时结束时,利用抗生物素蛋白-辣根过氧化物酶和盐酸邻苯二胺作为底物来检测生物素标记的BEC,从而开展实验。然后使用通过已知BEC数量与吸光度单位制备的标准曲线来确定细胞数量。本研究的数据表明:(I)BEC的黏附难易程度顺序如下:P-Val > P-Gly = P-Trp = P-Ala > P-Asn > P-Asp = 对照;(II)BEC的铺展速率受到孵育时间和聚合物浓度的强烈影响;(III)已证明BEC在聚合物表面的附着情况因其化学结构和微孔率水平而异。因此,总体观察结果使我们得出结论,在讨论聚合物材料在体内的生物相容性时,必须始终考虑其表面反应性。