富血小板血浆复合软骨细胞构建可注射性组织工程化软骨

Platelet-rich plasma combined with chondrocytes for construction of injectable tissue-engineered cartilage

背景:富血小板血浆中含有大量的生长因子,因此其在骨再生、创伤愈合等方面得到了较多的应用,然而其在组织工程软骨的研究报道较少。目的:观察富血小板血浆对软骨细胞增殖和分化的影响,以及富血小板血浆复合软骨细胞构建组织工程化软骨的可行性。方法:检测兔全血、富血小板血浆及激活富血小板血浆中转化生长因子β、胰岛素样生长因子1、血小板源性生长因子BB及表皮生长因子的浓度。将兔软骨细胞在分别含10%,15%,20%,30%富血小板血浆的DMEM培养液中培养7 d,CCK-8法检测细胞增殖,QT-PCR检测细胞内Ⅱ型胶原、蛋白聚糖、Sox-9的表达。在兔皮下注射自体富血小板血浆与软骨细胞复合物,6周后取材进行组织学检测。结果与结论:富血小板血浆中各生长因子浓度高于全血(P<0.05),激活富血小板血浆中各生长因子浓度高于富血小板血浆(P<0.05)。不同浓度富血小板血浆均能促进软骨细胞的增殖,且20%浓度内呈浓度依赖性。20%浓度组促Ⅱ型胶原表达的能力强于其他浓度组(P<0.05),15%浓度组促Sox-9和蛋白聚糖表达的能力强于其他浓度组(P<0.05)。富血小板血浆-软骨细胞复合物移植后,新生组织呈软骨样并有明显的软骨陷窝,细胞外富含软骨样基质,表明其作为可注射性支架用于软骨组织工程。

BACKGROUND： Owing to containing large amounts of growth factors, platelet-rich plasma has been widely used in bone regeneration, wound healing, and so on, while few studies have been reported on cartilage tissue engineering. OBJECTIVE： To explore the effects of platelet-rich plasma on the proliferation and differentiation of chondrocytes, and the feasibility of constructing tissue-engineered cartilage by the combination of chondrocytes and platelet-rich plasma. METHODS： We detected the concentrations of transforming growth factor-I], insulin-like growth factor-I, platelet-derived growth factor and epidermal growth factor BB in the whole blood, platelet-rich plasma, and activated platelet-rich plasma. The rabbit articular chondrocytes were cultured in 10%, 15%, 20%, 30% platelet-rich plasma for 7 days, then the cell proliferation was tested by cell counting kit-8, and cartilage-related genes （collagen type II, Aggrecan, Sox-9） were determined using QT-PCR. The chondrocytes/platelet-rich plasma composite was implanted subcutaneously into the rabbits, and the samples were harvested after 6 weeks of transplantation for histological examination. RESULTS AND CONCLUSION： The levels of different growth factors in the platelet-rich plasma were higher than those in the whole blood （P 〈 0.05）, but lower than those in the activated platelet-rich plasma （P 〈 0.05）. Platelet-rich plasma at different concentrations promoted the proliferation of chondrocytes. When theconcentration of platelet-rich plasma was no more than 20%, the proliferation of chondrocytes showed a concentration-dependent manner. The 20% platelet-rich plasma showed the best effects to promote the expression of collagen type II （P 〈 0.05）, while the 15% platelet-rich plasma could maximize the expression of Sox-9 and Aggrecan （P 〈 0.05）. After transplantation of chondrocytes/platelet-rich plasma composite, cartilage-like tissue formed with cartilage lacuna-like structures, and rich extracellular matrix was found, which indicates platelet-rich plasma can be used as an injective scaffold in cartilage tissue engineering.