GelMA-Encapsulated hDPSCs and HUVECs for Dental Pulp Regeneration
GelMA-Encapsulated hDPSCs and HUVECs for Dental Pulp Regeneration
复制标题
DOI：
10.1177/0022034516682005
复制
发表时间：
2017-02-01
影响因子：
7.6
通讯作者：
Yelick, P. C.
中科院分区：
文献类型：
--
作者：
Khayat, A.;Monteiro, N.;Yelick, P. C.
牙髓血管重建术是牙科临床上常用的方法，用于根尖封闭牙本质壁较薄的未成熟恒牙。虽然有时会成功，但牙根尖周围的刺激性出血可能是具有挑战性的，反过来可能会有害地影响牙根成熟。我们的目标是确定可靠的方法来再生牙根段(RSS)中的牙髓样组织。将人牙髓干细胞(HDPSCs)和包裹在5%甲基丙烯酸明胶(GelMA)水凝胶中的人脐静脉内皮细胞(HUVECs)分别注入G1代RSS。以单纯注射脱细胞GelMA的G2 RSS和G3空白RSS为对照。用白色三氧化二铝骨料封闭牙根段的一端，而另一端则保持开放。标本在成骨介质(OM)中体外培养13d，然后植入裸鼠皮下4wk和8wk。每个试验组至少使用5个样本重复。对收集的标本进行分析发现，G1组形成了健壮的牙髓样组织，GelMA包裹了hDPSC/HUVEC填充的RSS，而G2去细胞GelMA组和G3空RS组的宿主细胞来源的牙髓样组织较少。重要的是，只有G1hDPSC/HUVEC包裹的GelMA构建形成的牙髓细胞附着在RS的内牙本质表面并渗透到牙本质小管中。免疫荧光(IF)组织化学分析显示，GelMA支持hDPSC/HUVEC细胞贴壁和增殖，并为浸润性宿主细胞提供贴壁。人细胞接种的GelMA水凝胶促进了组织良好的新生血管的形成。相反，去细胞的GelMA和空的RS结构支持较少组织的宿主来源的血管形成。综上所述，这些结果表明，GelMA水凝胶与hDPSC/HUVECs相结合是一种有希望的临床相关的新的牙髓血管重建治疗方法，用于再生人类牙髓组织。
Pulpal revascularization is commonly used in the dental clinic to obtain apical closure of immature permanent teeth with thin dentinal walls. Although sometimes successful, stimulating bleeding from the periapical area of the tooth can be challenging and in turn may deleteriously affect tooth root maturation. Our objective here was to define reliable methods to regenerate pulp-like tissues in tooth root segments (RSs). G1 RSs were injected with human dental pulp stem cells (hDPSCs) and human umbilical vein endothelial cells (HUVECs) encapsulated in 5% gelatin methacrylate (GelMA) hydrogel. G2 RSs injected with acellular GelMA alone, and G3 empty RSs were used as controls. White mineral trioxide aggregate was used to seal one end of the tooth root segment, while the other was left open. Samples were cultured in vitro in osteogenic media (OM) for 13 d and then implanted subcutaneously in nude rats for 4 and 8 wk. At least 5 sample replicates were used for each experimental group. Analyses of harvested samples found that robust pulp-like tissues formed in G1, GelMA encapsulated hDPSC/HUVEC-filled RSs, and less cellularized host cell-derived pulp-like tissue was observed in the G2 acellular GelMA and G3 empty RS groups. Of importance, only the G1, hDPSC/HUVEC-encapsulated GelMA constructs formed pulp cells that attached to the inner dentin surface of the RS and infiltrated into the dentin tubules. Immunofluorescent (IF) histochemical analysis showed that GelMA supported hDPSC/HUVEC cell attachment and proliferation and also provided attachment for infiltrating host cells. Human cell-seeded GelMA hydrogels promoted the establishment of well-organized neovasculature formation. In contrast, acellular GelMA and empty RS constructs supported the formation of less organized host-derived vasculature formation. Together, these results identify GelMA hydrogel combined with hDPSC/HUVECs as a promising new clinically relevant pulpal revascularization treatment to regenerate human dental pulp tissues.