Limited adhesion of biodegradable microspheres to E- and P-selectin under flow
Limited adhesion of biodegradable microspheres to E- and P-selectin under flow
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DOI：
10.1002/bit.1085
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发表时间：
2001-06-20
影响因子：
3.8
通讯作者：
Goetz, DJ
中科院分区：
文献类型：
--
作者：
Dickerson, JB;Blackwell, JE;Goetz, DJ
在多种疾病环境中，内皮选择素E-和p -选择素的表达似乎增加。这一特性使这些分子成为设计定向给药方案的有吸引力的目标。实现这种递送的一种可能方法是使用带有E-和p -选择素人源化单克隆抗体（MAb HuEP5C7.g2）的聚合物可生物降解微球。也许是最简单的“耦合”技术HuEP5C7。G2通过非特异性吸附进入微球。然而，先前的研究表明，在稳定剂如聚乙烯醇（PVA）存在的情况下，蛋白质在微球上的吸附是有限的。目前尚不清楚这种有限水平的HuEP5C7吸附到何种程度。在流动条件下，g2能够支持E-和p -选择素的粘附。为了探讨这一问题，我们以可生物降解的聚合物聚（epsilon -己内酯）（PCL）为原料，以PVA为稳定剂，采用单乳液法制备了微球。然后用HuEP5C7孵育PCL微球。并对所得HuEP5C7的附着力进行了研究。在体外流动条件下，对E-和p -选择素的影响。我们发现HuEP5C7。PCL微球对表达p -选择素（CHO-P）和4-h IL-1 β活化的人脐静脉内皮细胞（HUVEC）的中国仓鼠卵巢细胞具有特异性粘附。相反，HuEP5C7。PCL微球对亲代CHO细胞或未活化的HUVEC的粘附性很小。对选择基质的附着效率很低，只有在低剪切（0.3 dyn/cm(2)）时才会有明显的附着。其他支持数据有力地表明，有限的依恋效率是由于低水平的HuEP5C7。g2吸附在PCL微球上。虽然附件是有限的，但HuEP5C7的显着百分比。g2 PCL微球能够在相对高的剪切（8 dyn/cm(2)）下保持粘附。综合来看，我们的数据表明HuEP5C7。PCL微球对表达E-和p -选择素的细胞底物表现出选择性的有限粘附。(C) 2001约翰威利父子公司
In a variety of disease settings the expression of the endothelial selectins E- and P-selectin appears to be increased. This feature makes these molecules attractive targets around which to design directed drug-delivery schemes. One possible approach for achieving such delivery is to use polymeric biodegradable microspheres bearing a humanized monoclonal antibody (MAb) for E- and P-selectin, MAb HuEP5C7.g2. Perhaps the simplest technique for "coupling" HuEP5C7.g2 to the microspheres is via nonspecific adsorption. Previous studies suggest, however, that the adsorption of protei ns onto microspheres fabricated in the presence of a stabilizer such as poly(vinyl alcohol) (PVA) is limited. It is unclear to what extent this limited level of adsorbed HuEP5C7.g2 would be able to support adhesion to E- and P-selectin under flow conditions. To explore this issue, we prepared microspheres from the biodegradable polymer, poly(epsilon -caprolactone) (PCL), using a single emulsion process and PVA as a stabilizer. We then incubated the PCL microspheres with HuEP5C7.g2 and studied the adhesion of the resulting HuEP5C7.g2 microspheres to E- and P-selectin under in vitro flow conditions. We found that the HuEP5C7.g2 PCL microspheres exhibit specific adhesion to Chinese hamster ovary cells stably expressing P-selectin (CHO-P) and 4-h IL-1 beta -activated human umbilical vein endothelial cells (HUVEC). In contrast, HuEP5C7.g2 PCL microspheres exhibit little adhesion to parental CHO cells or unactivated HUVEC. The attachment efficiency to the selectin substrates was quite low, with appreciable attachment occurring only at low shear (0.3 dyn/cm(2)). Other supporting data strongly suggest that the limited attachment efficiency is due to a low level of HuEP5C7.g2 adsorbed to the PCL microspheres. Although the attachment was limited, a significant percentage of the HuEP5C7.g2 PCL microspheres were able to remain adherent at relatively high shear (8 dyn/cm(2)). Combined, our data suggest that HuEP5C7.g2 PCL microspheres exhibit selective limited adhesion to cellular substrate expressing E- and P-selectin. (C) 2001 John Wiley & Sons, Inc.