Tablin Fern, Walker Naomi J, Hogle Sara E, Pratt Suzanne M, Norris Jeffrey W
Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
Am J Vet Res. 2008 Nov;69(11):1512-9. doi: 10.2460/ajvr.69.11.1512.
To determine whether platelet growth factors are preserved in supernatants obtained from rehydrated trehalose-stabilized, freeze-dried (lyophilized) equine platelets and whether those growth factors stimulate fibroblast proliferation and migration and enhance fibroblast-associated contraction in a collagen gel assay.
6 clinically normal adult horses.
Blood samples were obtained from 6 horses, and washed platelets were prepared via differential centrifugation. Washed platelets were freeze-dried in a physiologic buffer with a mixture of trehalose and polyethylene glycol 4000. Rehydrated platelet supernatants and releasates prepared from fresh washed platelets stimulated with thrombin or platelet-activating factor were evaluated for transforming growth factor beta1 and platelet-derived growth factor-BB by use of ELISAs. Effects of rehydrated freeze-dried platelet supernatants on fibroblast proliferation, migration, and collagen gel contraction were compared with effects of 1%, 2.5%, or 10% fetal bovine serum (FBS).
Supernatants from freeze-dried platelets contained similar amounts of growth factors as thrombin- and platelet-activating factor-stimulated platelet releasates. The supernatants significantly enhanced fibroblast proliferation and migration in a scratch assay, compared with FBS-free control or low (1%) FBS conditions. Additionally, supernatants from freeze-dried platelets enhanced contraction of fibroblast-seeded collagen gels, compared with the effect of 1% FBS.
The preparation technique preserved platelet growth factors, enhanced fibroblast proliferation and migration, and improved fibroblastseeded collagen gel contraction under conditions of low FBS concentration; these platelet supernatant preparations may prove useful as an aid to conventional wound management.
确定血小板生长因子是否保留在从用海藻糖稳定、冻干（冻融）的马血小板复水得到的上清液中，以及这些生长因子在胶原凝胶试验中是否刺激成纤维细胞增殖和迁移并增强成纤维细胞相关的收缩。
6匹临床正常的成年马。
从6匹马采集血样，通过差速离心制备洗涤后的血小板。将洗涤后的血小板在含有海藻糖和聚乙二醇4000混合物的生理缓冲液中冻干。使用酶联免疫吸附测定法评估复水的血小板上清液和由用凝血酶或血小板激活因子刺激的新鲜洗涤血小板制备的释放物中的转化生长因子β1和血小板衍生生长因子-BB。将复水的冻干血小板上清液对成纤维细胞增殖、迁移和胶原凝胶收缩的影响与1%、2.5%或10%胎牛血清（FBS）的影响进行比较。
冻干血小板的上清液中生长因子的含量与凝血酶和血小板激活因子刺激的血小板释放物中的相似。与无FBS对照或低（1%）FBS条件相比，上清液在划痕试验中显著增强了成纤维细胞的增殖和迁移。此外，与1%FBS的作用相比，冻干血小板的上清液增强了接种有成纤维细胞的胶原凝胶的收缩。
该制备技术保留了血小板生长因子，在低FBS浓度条件下增强了成纤维细胞增殖和迁移，并改善了接种有成纤维细胞的胶原凝胶收缩；这些血小板上清液制剂可能被证明有助于传统伤口处理。
