Trouillas Marina, Prat Marie, Doucet Christelle, Ernou Isabelle, Laplace-Builhé Corinne, Blancard Patrick Saint, Holy Xavier, Lataillade Jean-Jacques
Stem Cell Res Ther. 2013 Jan 4;4(1):1. doi: 10.1186/scrt149.
This study investigated the promising effect of a new Platelet Glue obtained from Cryoprecipitation of Apheresis Platelet products (PGCAP) used in combination with Mesenchymal Stromal Cells (MSC) loaded on ceramic biomaterials to provide novel strategies enhancing bone repair.
PGCAP growth factor content was analyzed by ELISA and compared to other platelet and plasma-derived products. MSC loaded on biomaterials (65% hydroxyapatite/35% beta-TCP or 100% beta-TCP) were embedded in PGCAP and grown in presence or not of osteogenic induction medium for 21 days. Biomaterials were then implanted subcutaneously in immunodeficient mice for 28 days. Effect of PGCAP on MSC was evaluated in vitro by proliferation and osteoblastic gene expression analysis and in vivo by histology and immunohistochemistry.
We showed that PGCAP, compared to other platelet-derived products, allowed concentrating large amount of growth factors and cytokines which promoted MSC and osteoprogenitor proliferation. Next, we found that PGCAP improves the proliferation of MSC and osteogenic-induced MSC. Furthermore, we demonstrated that PGCAP up-regulates the mRNA expression of osteogenic markers (Collagen type I, Osteonectin, Osteopontin and Runx2). In vivo, type I collagen expressed in ectopic bone-like tissue was highly enhanced in biomaterials embedded in PGCAP in the absence of osteogenic pre-induction. Better results were obtained with 65% hydroxyapatite/35% beta-TCP biomaterials as compared to 100% beta-TCP.
We have demonstrated that PGCAP is able to enhance in vitro MSC proliferation, osteoblastic differentiation and in vivo bone formation in the absence of osteogenic pre-induction. This clinically adaptable platelet glue could be of interest for improving bone repair.
本研究调查了一种新型血小板胶(从单采血小板制品冷沉淀中获得的血小板胶,PGCAP)与负载于陶瓷生物材料上的间充质基质细胞(MSC)联合使用时,在增强骨修复方面的潜在效果,以提供新的策略。
通过酶联免疫吸附测定(ELISA)分析PGCAP生长因子含量,并与其他血小板和血浆衍生产品进行比较。将负载于生物材料(65%羟基磷灰石/35%β-磷酸三钙或100%β-磷酸三钙)上的MSC嵌入PGCAP中,并在有或无成骨诱导培养基的情况下培养21天。然后将生物材料皮下植入免疫缺陷小鼠体内28天。通过增殖和成骨细胞基因表达分析在体外评估PGCAP对MSC的作用,并通过组织学和免疫组织化学在体内进行评估。
我们发现,与其他血小板衍生产品相比,PGCAP能够浓缩大量促进MSC和骨祖细胞增殖的生长因子和细胞因子。接下来,我们发现PGCAP可改善MSC和成骨诱导的MSC的增殖。此外,我们证明PGCAP上调成骨标志物(I型胶原、骨连接蛋白、骨桥蛋白和Runx2)的mRNA表达。在体内,在没有成骨预诱导的情况下,在嵌入PGCAP的生物材料中,异位骨样组织中表达的I型胶原显著增强。与100%β-磷酸三钙相比,65%羟基磷灰石/35%β-磷酸三钙生物材料获得了更好的结果。
我们已经证明,PGCAP能够在没有成骨预诱导的情况下增强体外MSC增殖、成骨细胞分化和体内骨形成。这种临床上适用的血小板胶可能对改善骨修复有意义。