Bioprinting of three-dimensional dentin–pulp complex with local differentiation of human dental pulp stem cells

Numerous approaches have been introduced to regenerate artificial dental tissues. However, conventional approaches are limited when producing a construct with three-dimensional patient-specific shapes and compositions of heterogeneous dental tissue. In this research, bioprinting technology was applied to produce a three-dimensional dentin–pulp complex with patient-specific shapes by inducing localized differentiation of human dental pulp stem cells within a single structure. A fibrin-based bio-ink was designed for bioprinting with the human dental pulp stem cells. The effects of fibrinogen concentration within the bio-ink were investigated in terms of printability, human dental pulp stem cell compatibility, and differentiation. The results show that micro-patterns with human dental pulp stem cells could be achieved with more than 88% viability. Its odontogenic differentiation was also regulated according to the fibrinogen concentration. Based on these results, a dentin–pulp complex having patient-specific shape was produced by co-printing the human dental pulp stem cell–laden bio-inks with polycaprolactone, which is a bio-thermoplastic used for producing the overall shape. After culturing with differentiation medium for 15 days, localized differentiation of human dental pulp stem cells in the outer region of the three-dimensional cellular construct was successfully achieved with localized mineralization. This result demonstrates the possibility to produce patient-specific composite tissues for tooth tissue engineering using three-dimensional bioprinting technology.

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The performance of human dental pulp stem cells on different three-dimensional scaffold materials

Biomaterials ◽

10.1016/j.biomaterials.2006.07.013 ◽

2006 ◽

Vol 27 (33) ◽

pp. 5658-5668 ◽

Cited By ~ 151

Author(s):

Weibo Zhang ◽

X. Frank Walboomers ◽

Toin H. van Kuppevelt ◽

Willeke F. Daamen ◽

Zhuan Bian ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Three Dimensional ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp ◽

Scaffold Materials

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Evaluation of cell viability of human dental pulp stem cells in two dimensional and three dimensional fibrin glue scaffold

Journal of Conservative Dentistry ◽

10.4103/jcd.jcd_439_20 ◽

2020 ◽

Vol 23 (5) ◽

pp. 479

Author(s):

NeedaA Ansari ◽

Abhishek Parmar ◽

Girish Parmar ◽

Amee Krishnakumar

Keyword(s):

Stem Cells ◽

Cell Viability ◽

Fibrin Glue ◽

Dental Pulp ◽

Three Dimensional ◽

Dental Pulp Stem Cells ◽

Two Dimensional ◽

Human Dental Pulp

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Neurogenic Differentiation of Human Dental Pulp Stem Cells on Graphene-Polycaprolactone Hybrid Nanofibers

Nanomaterials ◽

10.3390/nano8070554 ◽

2018 ◽

Vol 8 (7) ◽

pp. 554 ◽

Cited By ~ 8

Author(s):

Hoon Seonwoo ◽

Kyung-Je Jang ◽

Dohyeon Lee ◽

Sunho Park ◽

Myungchul Lee ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Morphological Characteristics ◽

Electrospun Nanofibers ◽

Dental Pulp Stem Cells ◽

Neurogenic Differentiation ◽

Dental Tissues ◽

Differentiated Cells ◽

Human Dental Pulp

Stem cells derived from dental tissues—dental stem cells—are favored due to their easy acquisition. Among them, dental pulp stem cells (DPSCs) extracted from the dental pulp have many advantages, such as high proliferation and a highly purified population. Although their ability for neurogenic differentiation has been highlighted and neurogenic differentiation using electrospun nanofibers (NFs) has been performed, graphene-incorporated NFs have never been applied for DPSC neurogenic differentiation. Here, reduced graphene oxide (RGO)-polycaprolactone (PCL) hybrid electrospun NFs were developed and applied for enhanced neurogenesis of DPSCs. First, RGO-PCL NFs were fabricated by electrospinning with incorporation of RGO and alignments, and their chemical and morphological characteristics were evaluated. Furthermore, in vitro NF properties, such as influence on the cellular alignments and cell viability of DPSCs, were also analyzed. The influences of NFs on DPSCs neurogenesis were also analyzed. The results confirmed that an appropriate concentration of RGO promoted better DPSC neurogenesis. Furthermore, the use of random NFs facilitated contiguous junctions of differentiated cells, whereas the use of aligned NFs facilitated an aligned junction of differentiated cells along the direction of NF alignments. Our findings showed that RGO-PCL NFs can be a useful tool for DPSC neurogenesis, which will help regeneration in neurodegenerative and neurodefective diseases.

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Analysis of growth kinetics and colony-forming unit efficiency of human dental pulp stem cells cultured in different media and supplements

Research Journal of Biotechnology ◽

10.25303/167rjbt20321 ◽

2021 ◽

Vol 16 (7) ◽

pp. 203-210

Author(s):

Kumar Chethan ◽

Shishir Shetty ◽

Basan Gowda Kurkalli ◽

Veena Shetty ◽

Kumar Basavarajappa Mohana

Keyword(s):

Stem Cells ◽

Growth Kinetics ◽

Dental Pulp ◽

Culture Conditions ◽

Dental Pulp Stem Cells ◽

Population Doubling ◽

Animal Origin ◽

Dental Tissues ◽

Colony Forming Unit ◽

Human Dental Pulp

Dental tissues are considered as ideal autologous sources of multipotent stem cells. Presently, human dental pulp stem cells (DPSCs) are largely being isolated and expanded in media containing fetal bovine serum (FBS). However, the use of FBS has limitations due to its animal origin. Therefore, the present study evaluated the morphology, proliferation rate, population doubling time (PDT) and colony-forming unit fibroblast (CFU-F) efficiency of DPSCs cultured in animal serum-containing medium (SCM) and serumfree medium (SFM) in addition to serum-free culture conditions by supplementing human blood-derivatives such as platelet lysate (PL), fresh frozen plasma (FFP) and umbilical cord blood serum (UCS) at 2.5%, 5% and 7.5% concentrations. Established DPSCs had spindle-shape during primary culture but acquired characteristic fibroblast-like features when cultured in PL, FFP and UCS. DPSCs in SCM, SFM and PL had significantly (P<0.05) higher proliferative potential than those in UCS and FFP and these observations were supported by PDT values. The CFU efficiency of DPSCs was confirmed in all culture conditions with a slightly varied clonogenic potential in blood-derived components. Based on the growth kinetics and CFU ability, it is concluded that PL could be considered as a suitable alternative to FBS for the ex vivo expansion of DPSCs.

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PD-1 is required to maintain stem cell properties in human dental pulp stem cells

Cell Death and Differentiation ◽

10.1038/s41418-018-0077-8 ◽

2018 ◽

Vol 25 (7) ◽

pp. 1350-1360 ◽

Cited By ~ 10

Author(s):

Yao Liu ◽

Huan Jing ◽

Xiaoxing Kou ◽

Chider Chen ◽

Dawei Liu ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp

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Viability of Quercetin-Induced Dental Pulp Stem Cells in Forming Living Cellular Constructs for Soft Tissue Augmentation

Journal of Personalized Medicine ◽

10.3390/jpm11050430 ◽

2021 ◽

Vol 11 (5) ◽

pp. 430

Author(s):

Hytham N. Fageeh ◽

Shilpa Bhandi ◽

Mohammed Mashyakhy ◽

Ahmed Al Kahtani ◽

Zahi Badran ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Soft Tissue ◽

Dental Pulp ◽

Donor Site ◽

Dental Pulp Stem Cells ◽

Soft Tissue Augmentation ◽

Background Information ◽

Tissue Augmentation ◽

Dental Pulp Stem Cell

Autogenous gingival grafts used for root coverage or gingival augmentation procedures often result in donor site morbidity. Living cellular constructs as an exogenous alternative have been proven to be associated with lower morbidity. With the available background information, the present study aims to assess if quercetin-induced living cell constructs, derived from dental pulp stem cells, have the potential to be applied as a tool for soft tissue augmentation. The characterized dental pulp stem cells (positive for CD73, CD90, and negative for CD34, HLA-DR) were expanded in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10 mM quercetin. The handling properties of the quercetin-induced dental pulp stem cell constructs were assessed by visual, and tactile sensation. A microscopic characterization using hematoxylin and eosin staining, and qRT-PCR-based analysis for stemness-associated genes (OCT4, NANOG, SOX2, and cMyc) was also performed. Dental pulp stem cells without quercetin administration were used as the control. Dental pulp stem cell constructs induced by quercetin easily detached from the surface of the plate, whereas there was no formation in the control cells. It was also simple to transfer the induced cellular construct on the flattened surface. Microscopic characterization of the constructs showed cells embedded in a tissue matrix. Quercetin also increased the expression of stemness-related genes. The use of quercetin-induced DPSC living constructs for soft tissue augmentation could provide an alternative to autogenous soft tissue grafts to lower patient morbidity and improve esthetic outcomes.

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