不同抗凝剂和激活剂联合应用对富血小板血浆凝胶释放生长因子影响的比较

背景:富血小板血浆凝胶生物效应的发挥受多种因素的影响,如富血小板血浆制备的方法、血小板的完整性、抗凝剂及激活剂的选择等.目的:比较不同抗凝剂与激活剂联合应用对富血小板血浆凝胶释放生长因子影响的差异.方法:抽取新西兰兔全血制备富血小板血浆,再用牛凝血酶和Ⅰ型胶原激活,实验共分4组:依地酸钠钙-凝血酶组,依地酸钠钙-Ⅰ型胶原组,肝素-凝血酶组,肝素-Ⅰ型胶原组.分别计数各组富血小板血浆血小板数目.在激活富小板血浆后2 h,1 d,3 d,5 d使用酶联免疫吸附法测定空白对照组(全血)和各组富血小板血浆凝胶中转化生长因子β1及血小板源性生长因子AB的浓度,比较各组间2种生长因子释放方式和浓度的差异.结果与结论:依地酸钠钙-Ⅰ型胶原组合制备的富血小板血浆凝胶中转化生长因子β1和血小板源性生长因子AB累积释放量最大(P < 0.05);使用Ⅰ型胶原作为激活剂的富血小板血浆凝胶中上述2种生长因子的释放方式均为持续缓慢,并且转化生长因子β1的释放与激活时间呈正相关关系(r=0.873);而凝血酶激活的富血小板血浆凝胶释放生长因子的速度则较为快速(P > 0.05).结果证实,依地酸钠钙与Ⅰ型胶原制备的富血小板血浆凝胶所释放生长因子的浓度较大.

Comparative study of the release of growth factors from platelet-rich plasma gel with different combinations of anticoagulants and activators

BACKGROUND: The biological effects of platelet-rich plasma gel are affected by many factors, such as the preparation method of platelet-rich plasma, blood platelet integrity, the choice of anticoagulants and activators and so on. OBJECTIVE: To compare the effects on the release of growth factors from platelet-rich plasma gel with different combinations of anticoagulants and activators. METHODS: The whole blood was extracted from New Zealand rabbits to prepare the platelet-rich plasma. The platelet-rich plasma was activated with bovine thrombin and type I collagen in different groups. There were five groups in this experiment: ethylene diamine tetraacetic acid (EDTA)-thrombin group, EDTA-typeⅠcollagen group, heparin-thrombin group, heparin-typeⅠ collagen group, as well as a blank control group (the whole blood). The platelets were counted before and after centrifugation; Transforming growth factor β1 (TGF-β1) and platelet-derived growth factor AB (PDGF-AB) were measured in the platelet-rich plasma gel of all the five groups using the enzyme linked immunosorbent assay at 2 hours, 1, 3, 5 days after the platelet-rich plasma was activated, and release modes of these two growth factors were compared at the same time. RESULTS AND CONCLUSION: The cumulative release of TGF-β1 and PDGF-AB was maximal in the EDTA-type Ⅰ collagen group, and significant differences were noted between the EDTA-type Ⅰ collagen group and the other groups (P<0.05). The use of type I collagen as an activator resulted in sustained release of TGF-β1 and PDGF-AB, and the release of TGF-β1 was in a time-dependent manner (r=0.873); while the aliquots of platelet-rich plasma clotted with thrombin had an immediate release of TGF-β1 and PDGF-AB (P>0.05). The concentration and methods of the release of growth factors in the EDTA-type Ⅰ collagen group is more conducive to the seed cells of tissue engineering nucleus pulposus differentiated into nucleus pulposus-like cells.