Castronuovo J J, Ghobrial I, Giusti A M, Rudolph S, Smiell J M
Department of Surgery, Morristown Memorial Hospital, New Jersey 07962-1956, USA.
Am J Surg. 1998 Aug;176(2A Suppl):61S-67S. doi: 10.1016/s0002-9610(98)00175-5.
In this study, the effects of chronic wound fluid on the structure and biological activity of becaplermin (recombinant human platelet-derived growth factor-BB [rhPDGF-BB]) were evaluated. Wound fluid was collected from 12 subjects with diabetic ulcers or pressure ulcers. Wound fluid +/- becaplermin was added to cell cultures before- and after incubation for 12 hours at 37 degrees C or after 12 hours' topical treatment. Biological activity, concentration, and immunogenicity were determined by [3H]thymidine incorporation into quiescent human foreskin fibroblasts, enzyme-linked immunosorbent assay (ELISA), and Western blot analysis, respectively. No PDGF-BB or mitogenic activity was detected in chronic wound fluid alone. Mitogenic activity was present in post-treatment samples from becaplermin-treated subjects but not placebo-treated subjects. Exposure to chronic wound fluid for 12 hours did not alter the amount, banding pattern, or mitogenic activity of becaplermin. Biologically active becaplermin remains in wound fluid 12 hours after topical application of becaplermin gel.
在本研究中，评估了慢性伤口渗出液对贝卡普勒明（重组人血小板源性生长因子-BB [rhPDGF-BB]）结构和生物活性的影响。从12名患有糖尿病溃疡或压疮的受试者收集伤口渗出液。在37℃孵育12小时之前和之后或在12小时局部治疗之后，将伤口渗出液±贝卡普勒明添加到细胞培养物中。分别通过将[3H]胸苷掺入静止的人包皮成纤维细胞、酶联免疫吸附测定（ELISA）和蛋白质印迹分析来测定生物活性、浓度和免疫原性。单独的慢性伤口渗出液中未检测到PDGF-BB或促有丝分裂活性。贝卡普勒明治疗受试者的治疗后样本中存在促有丝分裂活性，而安慰剂治疗受试者的样本中则没有。暴露于慢性伤口渗出液12小时不会改变贝卡普勒明的量、条带模式或促有丝分裂活性。在局部应用贝卡普勒明凝胶12小时后，具有生物活性的贝卡普勒明仍存在于伤口渗出液中。
