Benghuzzi H A
Department of Health Sciences, University of Mississippi Med. Center, Jackson 39216, USA.
Biomed Sci Instrum. 1995;31:121-6.
The relationship between various polymers used in surgical implantation and cellular response at that particular site has not been fully elucidated. The objective of this study was to investigate the effect of various biomedical polymers on the adherence and viability of monocyte and monocyte derived macrophages. The monocytes were isolated from human peripheral blood, biotin labeled and seeded at a density of 5 x 10(5) cells/well according to standard laboratory procedures. Cells were considered macrophages after remaining in culture for 24 hours. Cells were then plated in each microtiter well pretreated with various concentrations of (0.01, 0.1 and 1%) poly-1-valine (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 3 hours, 2 and 7 days the viability and cell number of monocyte or monocyte derived macrophages were determined using an established assay (biotin-labeled-macrophages). Cell number was determined in control wells with known amounts of cell number, a standard curve was generated by plotting absorbance units versus cell number. The data from this experiment suggest that: (I) cell number and viability of monocytes and macrophages can be measured concurrently, for the first time, by the use of Alamar Blue methodology, and (II) monocytes and macrophages are capable of surviving on all concentrations (0.01, 0.1 and 1%) of polymers tested over a 7 day period. Information obtained from this study provided new insights on the interrelationship between commercially available polymers, dose effect, incubation time and the possible cell response during chronic inflammation at the site of implantation.
用于外科植入的各种聚合物与该特定部位的细胞反应之间的关系尚未完全阐明。本研究的目的是调查各种生物医学聚合物对单核细胞和单核细胞衍生巨噬细胞的黏附及活力的影响。根据标准实验室程序，从人外周血中分离出单核细胞，用生物素标记，并以5×10(5)个细胞/孔的密度接种。细胞在培养24小时后被视为巨噬细胞。然后将细胞接种到预先用各种浓度（0.01%、0.1%和1%）的聚-1-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，以及缓冲对照孔中。在3小时、2天和7天结束时，使用既定的检测方法（生物素标记的巨噬细胞）测定单核细胞或单核细胞衍生巨噬细胞的活力和细胞数量。在已知细胞数量的对照孔中测定细胞数量，通过绘制吸光度单位与细胞数量的关系生成标准曲线。该实验的数据表明：（I）首次通过使用阿拉玛蓝方法可以同时测量单核细胞和巨噬细胞的细胞数量和活力，并且（II）单核细胞和巨噬细胞能够在7天内测试的所有浓度（0.01%、0.1%和1%）的聚合物上存活。从本研究中获得的信息为市售聚合物之间的相互关系、剂量效应、孵育时间以及植入部位慢性炎症期间可能的细胞反应提供了新的见解。
