Acute skeletal injury is necessary for human adipose-derived stromal cell-mediated calvarial regeneration.
Acute skeletal injury is necessary for human adipose-derived stromal cell-mediated calvarial regeneration.
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DOI：
10.1097/prs.0b013e318205f274
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发表时间：
2011-03
影响因子：
3.6
通讯作者：
Longaker MT
中科院分区：
文献类型：
--
作者：
Levi B;James AW;Nelson ER;Peng M;Wan DC;Commons GW;Lee M;Wu B;Longaker MT
研究表明，人类脂肪源性基质细胞（hASCs）能够修复急性颅骨损伤。然而，更为临床相关的修复已建立的骨骼缺陷尚未解决。我们试图确定hASCs是否可以治愈慢性（已建立的）颅骨缺陷。创建了临界尺寸（4mm）的小鼠顶骨缺陷。hASCs要么在术后立即植入（急性缺损），要么在缺损形成后8周植入（已建立缺损）。分析方法包括显微ct扫描、组织学和原位杂交。最后，体外用PRP处理hASCs，模拟急性伤口环境；通过碱性磷酸酶、茜素红和qRT-PCR检测细胞增殖和成骨分化。当颅骨缺损立即植入hASCs时，可以观察到近乎完全的骨愈合。相比之下，当hASCs被移植到已建立的缺陷中时，很少发生骨形成。组织学分析证实了显微ct的发现，显示急性碱性磷酸酶和小天狼星红的染色比已建立的hASC移植缺陷更强。原位杂交和qRT - PCR显示，在颅骨缺损形成后，BMP （Bmp2、Bmp4和Bmp7）的表达急剧增加。最后，体外用PRP处理hASCs可促进成骨分化并增加Bmp2表达。虽然hASCs可用于治疗急性小鼠颅骨缺损，但hASCs不能促进已建立（或慢性）缺损的愈合。内源性BMP信号在损伤后激活可以解释这些愈合差异。富血小板血浆增强体外造血干细胞成骨分化，可能是未来骨组织工程的一种有前途的治疗方法。
Studies have demonstrated that human adipose derived stromal cells (hASCs) are able to repair acute calvarial injuries. However, the more clinically relevant repair of an established skeletal defect has not been addressed. We sought to determine whether hASCs could heal chronic (established) calvarial defects. Critical-sized (4mm) mouse parietal defects were created. hASCs were either engrafted immediately postoperatively (acute defect), or 8 weeks following defect creation (established defect). Methods of analysis included microCT scans, histology, and in situ hybridization. Finally, hASCs were treated in vitro with PRP to simulate an acute wound environment; proliferation and osteogenic differentiation were assessed (Alkaline phosphatase, Alizarin red, and qRT-PCR). Near complete osseous healing was observed when calvarial defects were immediately engrafted with hASCs. In contrast, when hASCs were engrafted into established defects, little bone formation occurred. Histological analysis affirmed findings by microCT, showing more robust staining for alkaline phosphatase and picrosirius red in an acute than in a established hASC engrafted defect. In situ hybridization and qRT PCR showed an increase in BMP expression (Bmp2, Bmp4 and Bmp7) acutely following calvarial defect creation. Finally, in vitro treatment of hASCs with PRP enhanced osteogenic differentiation and increased Bmp2 expression. While hASCs can be utilized to heal an acute mouse calvarial defect, hASCs do not enhance healing of an established (or chronic) defect. Endogenous BMP signaling activated post-injury may explain these differences in healing. Platelet rich plasma enhances osteogenic differentiation of hASCs in vitro and may prove a promising therapy for future skeletal tissue engineering efforts.