Zhao Zeqing, Liu Jin, Weir Michael D, Zhang Ning, Zhang Li, Xie Xianju, Zhang Charles, Zhang Ke, Bai Yuxing, Xu Hockin H K
Department of Orthodontics, School of Stomatology, Capital Medical University Beijing China
Department of Advanced Oral Sciences and Therapeutics, University of Maryland Dental School Baltimore MD 21201 USA.
RSC Adv. 2019 Dec 13;9(70):41161-41172. doi: 10.1039/c9ra08336g. eCollection 2019 Dec 9.
Human periodontal ligament stem cells (hPDLSCs) are promising for tissue engineering applications but have received relatively little attention. Human platelet lysate (HPL) contains a cocktail of growth factors. To date, there has been no report on hPDLSC seeding on scaffolds loaded with HPL. The objectives of this study were to develop a calcium phosphate cement (CPC)-chitosan scaffold loaded with HPL and investigate their effects on hPDLSC viability, osteogenic differentiation and bone mineral synthesis for the first time. hPDLSCs were harvested from extracted human teeth. Scaffolds were formed by mixing CPC powder with a chitosan solution containing HPL. Four groups were tested: CPC-chitosan + 0% HPL (control); CPC-chitosan + 2.66% HPL; CPC-chitosan + 5.31% HPL; CPC-chitosan + 10.63% HPL. Scanning electron microscopy, live/dead staining, CCK-8, qRT-PCR, alkaline phosphatase and bone minerals assay were applied for hPDLSCs on scaffolds. hPDLSCs attached well on CPC-chitosan scaffold. Adding 10.63% HPL into CPC increased cell proliferation and viability ( < 0.05). ALP gene expression of CPC-chitosan + 10.63% HPL was 7-fold that of 0% HPL at 14 days. Runx2, OSX and Coll1 of CPC-chitosan + 10.63% HPL was 2-3 folds those at 0% HPL ( < 0.05). ALP activity of CPC-chitosan + 10.63% HPL was 2-fold that at 0% HPL ( < 0.05). Bone minerals synthesized by hPDLSCs for CPC-chitosan + 10.63% HPL was 3-fold that at 0% HPL ( < 0.05). This study showed that CPC-chitosan scaffold was a promising carrier for HPL delivery, and HPL in CPC exerted excellent promoting effects on hPDLSCs for bone tissue engineering for the first time. The novel hPDLSC-CPC-chitosan-HPL construct has great potential for orthopedic, dental and maxillofacial regenerative applications.
人牙周膜干细胞（hPDLSCs）在组织工程应用方面前景广阔，但相对而言受到的关注较少。人血小板裂解物（HPL）含有多种生长因子的混合物。迄今为止，尚无关于将hPDLSCs接种在负载HPL的支架上的报道。本研究的目的是首次开发一种负载HPL的磷酸钙骨水泥（CPC）-壳聚糖支架，并研究其对hPDLSCs活力、成骨分化和骨矿物质合成的影响。hPDLSCs从拔除的人牙齿中获取。通过将CPC粉末与含有HPL的壳聚糖溶液混合形成支架。测试了四组：CPC-壳聚糖+0%HPL（对照组）；CPC-壳聚糖+2.66%HPL；CPC-壳聚糖+5.31%HPL；CPC-壳聚糖+10.63%HPL。对支架上的hPDLSCs进行扫描电子显微镜、活/死染色、CCK-8、qRT-PCR、碱性磷酸酶和骨矿物质测定。hPDLSCs在CPC-壳聚糖支架上附着良好。向CPC中添加10.63%HPL可增加细胞增殖和活力（P<0.05）。在第14天时，CPC-壳聚糖+10.63%HPL组的碱性磷酸酶（ALP）基因表达是0%HPL组的7倍。CPC-壳聚糖+10.63%HPL组的Runx2、OSX和Coll1是0%HPL组的2至3倍（P<0.05）。CPC-壳聚糖+10.63%HPL组的ALP活性是0%HPL组的2倍（P<0.05）。hPDLSCs为CPC-壳聚糖+10.63%HPL组合成的骨矿物质是0%HPL组的3倍（P<0.05）。本研究表明，CPC-壳聚糖支架是一种有前景的HPL递送载体，并且CPC中的HPL首次对hPDLSCs在骨组织工程中发挥了优异的促进作用。新型hPDLSC-CPC-壳聚糖-HPL构建体在骨科、牙科和颌面再生应用方面具有巨大潜力。
