Vendramini Victor Okada, Pouraghaei Sevda, Barbosa Rafael Maza, Aloise Antônio Carlos, Muniz José Ricardo Ferreira, Sperandio Marcelo, Moy Peter Karyen, Pelegrine André Antonio, Moshaverinia Alireza
Department of Implant Dentistry, Faculdade São Leopoldo Mandic, Rua Areias, 37, Campinas, São Paulo 13024-530, Brazil.
Department of Advanced Prosthodontics, University of California at Los Angeles, School of Dentistry, Los Angeles, USA.
Stem Cells Int. 2021 Jun 21;2021:9952401. doi: 10.1155/2021/9952401. eCollection 2021.
To compare two pulp harvesting methods for stem cell expansion, namely, conservative pulpotomy and pulpectomy from exodontia.
Ten freshly extracted sound third molars from five patients were selected. Five were used in the control group, where pulp harvesting was performed by exodontia and the remaining teeth were used in the test group, where the pulp was harvested by conservative pulpotomy (preserving the tooth). This was a split-mouth design study, where a third molar from one side was randomly allocated into the test group and the contralateral tooth in the control group. After pulp harvesting, the following evaluations were performed: cell morphology, sterility test, immunophenotyping, differentiation assays, first pass live cell counts, time to cryopreservation, and total number of expanded cells at the end of the fourth pass.
Regarding morphology, the cells from both groups presented a fibroblastic phenotype. All samples were sterile. Immunophenotyping demonstrated a positive expression for CD105, CD90, and CD73 and negative expression for CD45 in both groups. Differentiation assays were positive for osteogenic and chondrogenic differentiation in both groups. Regarding live cell counts in the first passage, the control group had 95.8% live cells in the total count and the test group 91.2% ( < 0.05). The time required for cryopreservation was equivalent in both groups 51.6 days and 52.6 days, respectively ( > 0.05). The total number of cells at the end of the fourth passage was 5,286,782 and 5,736,862, respectively ( > 0.05).
These results suggest that adult stem cell harvesting from conservative pulpotomy is as effective as the traditional exodontia-based method.
比较两种用于干细胞扩增的牙髓采集方法,即保守性牙髓切断术和拔牙后的牙髓摘除术。
选取5名患者新拔除的10颗健康第三磨牙。5颗用于对照组,通过拔牙进行牙髓采集,其余牙齿用于试验组,通过保守性牙髓切断术(保留牙齿)采集牙髓。这是一项双口设计研究,一侧的第三磨牙随机分配到试验组,对侧牙齿分配到对照组。牙髓采集后,进行以下评估:细胞形态、无菌试验、免疫表型分析、分化测定、首次传代活细胞计数、冷冻保存时间以及第四次传代结束时扩增细胞的总数。
在形态方面,两组细胞均呈现成纤维细胞表型。所有样本均无菌。免疫表型分析显示两组中CD105、CD90和CD73均呈阳性表达,CD45呈阴性表达。两组的分化测定在成骨和软骨分化方面均为阳性。在首次传代的活细胞计数方面,对照组活细胞占总数的95.8%,试验组为91.2%(P<0.05)。两组冷冻保存所需时间相当,分别为51.6天和52.6天(P>0.05)。第四次传代结束时细胞总数分别为5,286,782和5,736,862(P>0.05)。
这些结果表明,通过保守性牙髓切断术采集成人干细胞与传统的基于拔牙的方法同样有效。