含富血小板血浆的纳米纤维支架对软骨缺损的修复作用

发布时间：2018-03-17 11:11

本文选题：富血小板血浆　切入点：乳液电纺　出处：《上海交通大学》2014年博士论文　论文类型：学位论文

【摘要】：目的富血小板血浆（platelet-rich plasma，PRP）含有丰富的生长因子，对多种组织具有再生和修复作用。通过构建含PRP的纳米纤维支架，实现生长因子的缓释效果，并研究其对软骨再生的作用。方法通过乳液电纺的方法构建含PRP的纳米纤维支架和PCL/GE支架，并对支架进行力学、亲水性、降解性等表征。通过ELISA法测定生长因子的缓释曲线。将两种支架体外分别与BMSC共培养，通过SEM、CCK-8、免疫荧光、qPCR和Westernblot等方法研究支架对细胞的增殖、分化的影响。体内将支架植入兔膝关节软骨缺损区，并通过组织学、Micro-CT和IL-1β浓度检测的方法对其修复软骨缺损的效果进行评价。结果PRP加入之后，，使PCL/GE纳米纤维支架的力学参数与软骨组织更为接近，同时具有良好的亲水性和缓释效果，三种生长因子的缓释时间达到一个月左右。体外实验中，细胞在含PRP支架上能良好生长及增殖，且含PRP的支架在体外能对BMSC产生显著的趋化效应。通过免疫荧光法、qPCR、Western blot等方法，验证了含PRP支架可以有效促进BMSC的体外成软骨诱导分化和软骨标志物的表达，II型胶原、aggrecan、Sox9的合成均被上调；I型胶原的表达下降。缺损区植入含PRP支架的关节，软骨细胞有明显再生，且软骨细胞周围可见软骨基质合成增加。结论1.通过乳液电纺的方法能够得到结构稳定均一的含PRP纳米纤维支架，其特点为亲水性好，力学参数与软骨组织接近；支架中生长因子的缓释期近一个月。2.与PCL/GE支架相比，含PRP的纳米纤维支架可以促进细胞的粘附、增殖、趋化，并可以促进干细胞的成软骨分化和软骨标志物的表达。3.含PRP纳米纤维支架能够在体内有效促进软骨缺损的修复。其效果接近于正常软骨组织。这可能与纳米纤维支架自身的三维结构，以及PRP中的多种生长因子有关。
[Abstract]:Objective the platelet-rich plasma-rich plasma-rich platelet-rich plasma-rich plasma-rich platelet-rich plasma-rich plasma-rich plasma-rich platelet-rich plasma-rich plasma-rich plasma-rich platelet-rich plasma-rich plasma-rich plasma-rich plasma has rich growth factors, which can regenerate and repair many kinds of tissues. Methods Nano-fiber scaffolds and PCL/GE scaffolds containing PRP were constructed by emulsion electrospinning, and the scaffolds were mechanical and hydrophilic. Biodegradability and other characterization. The sustained release curves of growth factor were measured by ELISA method. The two scaffolds were co-cultured with BMSC in vitro. The proliferation of the scaffolds was studied by means of SEMCCK-8, immunofluorescence qPCR and Westernblot. In vivo, the scaffold was implanted into the cartilage defect area of rabbit knee joint, and the effect of repairing cartilage defect was evaluated by microCT and IL-1 尾. Results after the addition of PRP, the mechanical parameters of PCL/GE nanofiber scaffolds were closer to those of cartilage tissue, and had good hydrophilicity and sustained release effect. Cells could grow and proliferate well on PRP scaffolds, and PRP scaffolds could produce significant chemotactic effect on BMSC in vitro. It was proved that PRP scaffolds could effectively promote the chondrogenic differentiation of BMSC in vitro and the expression of cartilage markers. The synthesis of type II collagen aggrecann Sox9 was up-regulated, and the expression of type I collagen was down-regulated. The joints with PRP stents were implanted in the defect area. Chondrocytes were regenerated obviously and cartilage matrix synthesis was increased around chondrocytes. Conclusion 1. Stable and homogeneous scaffolds containing PRP nanofibers can be obtained by emulsion electrospinning, which are characterized by good hydrophilicity and close mechanical parameters to cartilage tissue, and the sustained release period of growth factor in the scaffold is nearly one month .2.Compared with that of PCL/GE scaffold. Nanofiber scaffolds containing PRP can promote cell adhesion, proliferation and chemotaxis. It can promote the cartilage differentiation of stem cells and the expression of cartilage markers. 3. The scaffold containing PRP nanofiber can effectively promote the repair of cartilage defect in vivo. The effect is close to that of normal cartilage tissue. This may be similar to that of nano-fiber. The three-dimensional structure of the scaffold itself, And various growth factors in PRP.
【学位授予单位】：上海交通大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R687;R318.08

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