Abstract：
Vascularization plays an important role in dental and craniofacial regenerations. Human periodontal ligament stem cells (hPDLSCs) are a promising cell source and, when co-cultured with human umbilical vein endothelial cells (hUVECs), could promote vascularization. The objectives of this study were to develop a novel prevascularized hPDLSC-hUVEC-calcium phosphate construct, and investigate the osteogenic and angiogenic efficacy of this construct with human platelet lysate (hPL) in cranial defects in rats for the first time.
hPDLSCs and hUVECs were co-cultured on calcium phosphate cement (CPC) scaffolds with hPL. Cell proliferation, angiogenic gene expression, angiogenesis, alkaline phosphatase activity, and cell-synthesized minerals were determined. Bone and vascular regenerations were investigated in rat critical-sized cranial defects in vivo.
hPDLSC-hUVEC-CPC-hPL group had 2-fold greater angiogenic expressions and cell-synthesized mineral synthesis than hPDLSC-hUVEC-CPC group (p < 0.05). Microcapillary-like structures were formed on scaffolds in vitro. hPDLSC-hUVEC-CPC-hPL group had more vessels than hPDLSC-hUVEC-CPC group (p < 0.05). In cranial defects in rats, hPDLSC-hUVEC-CPC-hPL group regenerated new bone amount that was 2.1 folds and 4.0 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05). New blood vessel density of hPDLSC-hUVEC-CPC-hPL group was 2 folds and 7.9 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05).
The hPL pre-culture method is promising to enhance bone regeneration via prevascularized CPC. Novel hPDLSC-hUVEC-CPC-hPL prevascularized construct increased new bone formation and blood vessel density by 4-8 folds over CPC control.
Novel hPDLSC-hUVEC-hPL-CPC prevascularized construct greatly increased bone and vascular regeneration in vivo and hence is promising for a wide range of craniofacial applications.
hPDLSCs and hUVECs were co-cultured on calcium phosphate cement (CPC) scaffolds with hPL. Cell proliferation, angiogenic gene expression, angiogenesis, alkaline phosphatase activity, and cell-synthesized minerals were determined. Bone and vascular regenerations were investigated in rat critical-sized cranial defects in vivo.
hPDLSC-hUVEC-CPC-hPL group had 2-fold greater angiogenic expressions and cell-synthesized mineral synthesis than hPDLSC-hUVEC-CPC group (p < 0.05). Microcapillary-like structures were formed on scaffolds in vitro. hPDLSC-hUVEC-CPC-hPL group had more vessels than hPDLSC-hUVEC-CPC group (p < 0.05). In cranial defects in rats, hPDLSC-hUVEC-CPC-hPL group regenerated new bone amount that was 2.1 folds and 4.0 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05). New blood vessel density of hPDLSC-hUVEC-CPC-hPL group was 2 folds and 7.9 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05).
The hPL pre-culture method is promising to enhance bone regeneration via prevascularized CPC. Novel hPDLSC-hUVEC-CPC-hPL prevascularized construct increased new bone formation and blood vessel density by 4-8 folds over CPC control.
Novel hPDLSC-hUVEC-hPL-CPC prevascularized construct greatly increased bone and vascular regeneration in vivo and hence is promising for a wide range of craniofacial applications.
摘要：
背景：血管化在牙齿和颅面再生中起着重要作用。人牙周膜干细胞(hPDLSCs)是一种很有前途的细胞来源,当与人脐静脉内皮细胞（hUVECs）共培养时，可以促进血管化。本研究的目的是开发新的预血管化hPDLSC-hUVEC-磷酸钙构建体，并首次研究了该构建体与人血小板裂解物（hPL）在大鼠颅骨缺损中的成骨和血管生成功效。
方法:将hPDLSCs和hUVECs在磷酸钙骨水泥(CPC)支架上与hPL共培养。细胞增殖，血管生成基因表达，血管生成,碱性磷酸酶活性，并测定了细胞合成的矿物质。在体内大鼠临界大小的颅骨缺损中研究了骨和血管再生。
结果：hPDLSC-hUVEC-CPC-hPL组的血管生成表达和细胞合成矿物质合成比hPDLSC-hUVEC-CPC组高2倍（p<0.05）。在体外支架上形成微毛细管样结构。hPDLSC-hUVEC-CPC-hPL组比hPDLSC-hUVEC-CPC组有更多的血管(p<0.05)。在大鼠的颅骨缺陷中，hPDLSC-hUVEC-CPC-hPL组再生新骨量分别为2.1倍和4.0倍，分别,hPDLSC-hUVEC-CPC组和CPC对照组（p<0.05）。hPDLSC-hUVEC-CPC-hPL组新血管密度分别为2倍和7.9倍,分别,hPDLSC-hUVEC-CPC组和CPC对照组（p<0.05）。
结论：hPL预培养方法有望通过预血管化CPC增强骨再生。新型hPDLSC-hUVEC-CPC-hPL预血管化构建体比CPC对照增加4-8倍的新骨形成和血管密度。
结论：新型hPDLSC-hUVEC-hPL-CPC预血管化构建体极大地增加了体内骨和血管再生，因此有望用于广泛的颅面应用。
方法:将hPDLSCs和hUVECs在磷酸钙骨水泥(CPC)支架上与hPL共培养。细胞增殖，血管生成基因表达，血管生成,碱性磷酸酶活性，并测定了细胞合成的矿物质。在体内大鼠临界大小的颅骨缺损中研究了骨和血管再生。
结果：hPDLSC-hUVEC-CPC-hPL组的血管生成表达和细胞合成矿物质合成比hPDLSC-hUVEC-CPC组高2倍（p<0.05）。在体外支架上形成微毛细管样结构。hPDLSC-hUVEC-CPC-hPL组比hPDLSC-hUVEC-CPC组有更多的血管(p<0.05)。在大鼠的颅骨缺陷中，hPDLSC-hUVEC-CPC-hPL组再生新骨量分别为2.1倍和4.0倍，分别,hPDLSC-hUVEC-CPC组和CPC对照组（p<0.05）。hPDLSC-hUVEC-CPC-hPL组新血管密度分别为2倍和7.9倍,分别,hPDLSC-hUVEC-CPC组和CPC对照组（p<0.05）。
结论：hPL预培养方法有望通过预血管化CPC增强骨再生。新型hPDLSC-hUVEC-CPC-hPL预血管化构建体比CPC对照增加4-8倍的新骨形成和血管密度。
结论：新型hPDLSC-hUVEC-hPL-CPC预血管化构建体极大地增加了体内骨和血管再生，因此有望用于广泛的颅面应用。
