摘要
目的 采用富血小板血浆裂解液(platelet-rich plasma lysate, PL)复合透明质酸制备微针,通过优化制备工艺及评价血小板生长因子活性,探讨PL微针在治疗伤口愈合领域的应用前景。方法 首先使用透明质酸(hyaluronic acid, HA)作为针体基质材料,通过考察HA溶液流动性、针体形貌、压变性能及穿刺性能等,初步确定HA的浓度。再结合HA在PL中的溶解情况、PL微针的机械性能、皮肤穿刺等结果,优化富血小板血浆裂解液微针(PLMN)的制备工艺。考察PL微针的生长因子活性,并评价PL微针应用于大鼠伤口愈合的效果。结果 当HA质量浓度为30 mg/mL时,溶液具有良好的流动性,制备所得HA微针可刺穿3层封口膜,穿刺率达70%,具有较好的机械性能。HA在4℃溶解于PL中浓度为30 mg/mL时,溶液无结晶,流动性好,所制备的PL微针在高清显微镜和扫描电镜下显示针型良好,无弯折。体外透皮模拟实验结果显示,PL微针能够穿透4层封口膜,前3层穿刺率达85%以上。体内溶解实验显示,PL微针插入体内在10 min之内能够完全溶解;在40 min后微针插入体内后留下的微孔几乎闭合,无红肿红斑等不良反应。体内外透皮实验显示,PL微针穿透皮肤染色后可见清晰蓝色孔洞,说明微针能刺破皮肤屏障。生长因子检测结果显示,血小板裂解液的血小板衍生生长因子(PDGF-BB)含量为(17.67±1.23)ng/mL,转化生长因子(TGFβ1)的含量为(105.95±2.16)ng/mL,而PL微针制备并室温保存1周后,微针的PDGF-BB含量为(15.48±2.12)ng/mL,TGFβ1的含量为(56.26±1.53)ng/mL,与PL相比虽然有所下降,但仍较好地保持了生长因子活性。大鼠伤口愈合实验结果显示,d3 PL微针组的伤口愈合率与对照组相比有差异,分别是52.51±6.15(%)、33.31±3.60(%)(P<0.05)。结论 本实验所制备的富血小板血浆裂解液微针具有良好的机械性能,能够成功地刺穿皮肤角质层,并能较好的保持生长因子活性,对大鼠伤口愈合有较好的效果,为组织修复领域在微创治疗方面提供一种新的思路。
Objective To investigate the application prospect of platelet-rich plasma lysate(PL) microneedles in the treatment of wound healing by optimizing the preparation process and evaluating the activity of platelet growth factor.Methods Firstly, hyaluronic acid(HA) was used as needle matrix material, and the concentration of HA was preliminarily determined by investigating the fluidity of HA solution, shape of needle, pressure change performance and puncture performance.Combined with the results of HA dissolution in PL,mechanical properties of PL microneedles and skin puncture, the preparation process of platelet-rich plasma lysis microneedles(PLMN) was optimized to investigate the growth factor activity of PLMN and evaluate the effect of PLMN on wound healing in rats.Results When the concentration of HA was 30 mg/mL,the solution had good fluidity.The HA microneedles could pierce three layers of sealing film, with the puncture rate at 70%,and the solution had good mechanical properties.When HA was dissolved in PL at a concentration of 30 mg/mL at 4℃,the solution had no crystallization and was with good fluidity.The prepared PLMN showed good needle shape and no bending under HD microscope and scanning electron microscope.In vitro transdermal simulation results showed that PLMN could penetrate 4 layers of sealing membrane, and the puncture rate of the first three layers was more than 85%.In vivo lysis experiment showed that PLMNs could be completely dissolved within 10 min after being inserted into the body.Forty minutes later, the micropores left after the insertion of the microneedle were almost closed, without redness, swelling, erythema and other adverse reactions.In vitro and in vivo transdermal experiments showed that clear blue holes were visible after PLMNs penetrated the skin stain, indicating that the microneedles could pierce the skin barrier.Growth factor detection results showed that the content of platelet-derived growth factor(PDGF-BB) and transforming growth factor(TGFβ1) in platelet lysate were(17.67±1.23) ng/mL and(105.95±2.16) ng/mL,respectively.After PLMNs were prepared and stored at room temperature for 1 week, the content of PDGF-BB(ng/mL) and TGFβ1(ng/mL) were 15.48±2.12 and 56.26±1.53,respectively.Compared with PL(vs 17.67±1.23 and 105.95±2.16),the content of PDGF-BB and TGFβ1 decreased, but still maintained the activity of growth factors.The results of wound healing experiment in rats showed that the wound healing rate(%) of PLMN group on the third day was significantly different from that of the control group(52.51±6.15 vs 33.31±3.60,P<0.05).Conclusion The PLMN prepared in this experiment has good mechanical properties and can successfully pierce the skin cuticle.It can also maintain the activity of growth factors, and has a good effect on wound healing in rats, which provides a new idea for minimally invasive treatment in the field of tissue repair.
作者
索正新
曹晔
郑诗凡
高燚秋
徐莹璨
王红
刘嘉馨
钟锐
SUO Zhengxin;CAO Ye;ZHENG Shifan;GAO Yiqiu;XU Yingcan;WANG Hong;LIU Jiaxin;ZHONG Rui(Institute of Blood Transfusion,Chinese Academy of Medical Sciences&Peking Union Medical College,Chengdu 610052,China)
出处
《中国输血杂志》
CAS
2022年第5期508-514,共7页
Chinese Journal of Blood Transfusion
关键词
富血小板血浆
透明质酸
微针
组织修复
platelet-rich plasma
hyaluronic acid
microneedle
tissue repair
作者简介
通信作者:钟锐(1980.12-),女,副主任技师,硕士研究生导师,主要从事生物医用材料与器具、血液保存等方面的研究,电话:028-61648511,Email:ruijiaol999@163.com。
