Experimental Design for the Innervation of Tooth Forming from Implanted Cell Re-associations

Kökten, Tunay; Lesot, Hervé; Kuchler‐Bopp, Sabine

doi:10.5772/59340

Cited by 5 publications

(20 citation statements)

References 110 publications

(159 reference statements)

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“…Several populations of glial cells positive for anti‐S100β and anti‐GFAP were present in the dental pulp of Sema3A −/− bioengineered teeth after 15 days of coimplantation with a TG. The same results were observed for reassociations implanted in “Nude” mice, where glial cells were identified in the dental pulp (Kökten, Lesot, & Kuchler‐Bopp, ).…”

Section: Discussionsupporting

confidence: 70%

“…We previously developed an in vivo biomimetic model in which E14 tooth germ were implanted under immunosuppressive conditions (“Nude” mice, cyclosporin A model) to allow their innervation (Kökten, Bécavin, et al, ; Kökten, Lesot, & Kuchler‐Bopp, ). To test the possibility to obtain tooth innervation when inhibiting Sema3A, we adapted tooth germ of mutant mice to our biomimetic model.…”

Section: Resultsmentioning

confidence: 99%

“…They were then associated with the cultured teeth or with the cell reassociations for one night, before subcutaneous implantation for 2 weeks in adult ICR mice (Kökten, Bécavin, et al, ; Figure S4). To study the role of NRP1, 100 μl of the TM peptide (3.10 −7 M diluted in sterile PBS) were injected subcutaneously every 2 days for 2 weeks at the site of implantation of cultured ICR reassociations between dental dissociated mesenchymal and epithelial cells (Kökten, Bécavin, et al, ; Kökten, Lesot, & Kuchler‐Bopp, ; Nait Lechguer et al, ) and TG on one side. LDS (3.10 −7 M diluted in sterile PBS) was injected on the other side, for control experiments (Figure S4).…”

Section: Methodsmentioning

confidence: 99%

“…Tissue engineering is a vast field aiming to fully regenerate functional tissues and organs (Langer & Vacanti, ). Different models for tooth engineering were proposed using reassociations of dental mesenchymal and epithelial cells (Honda, Tsuchiya, Sumita, Sagara, & Ueda, ; Kökten et al, ; Kökten, Lesot, & Kuchler‐Bopp, ; Kuchler‐Bopp et al, ; Nait Lechguer et al, ; Nait Lechguer, Kuchler‐Bopp, Hu, Haïkel, & Lesot, ; Nakao et al, ; Oshima et al, ) or using stem cells (Ohazama, Modino, Miletich, & Sharpe, ; Volponi, Pang, & Sharpe, ). In mouse, the first trigeminal axons reach the mandibular first molar tooth germ region at the bud stage at embryonic day‐13 (E13; Kettunen et al, ).…”

Section: Introductionmentioning

confidence: 99%

“…Contrary to vascularization, the innervation never occurs spontaneously (Keller, Kuchler‐Bopp, & Lesot, ; Kökten, Bécavin, et al, ). Because clinical observations showed that immunodepression stimulate innervation (Siemionow, Gharb, & Rampazzo, ), cultured reassociations were implanted with TG in “Nude” mice or in cyclosporin A (CsA)‐treated ICR mice (Kökten, Bécavin, et al, ; Kökten, Lesot, & Kuchler‐Bopp, ). Under these conditions, the innervation of the dental mesenchyme did occur and the axons coming from the TG reached the odontoblasts 2 weeks after implantation (Kökten, Bécavin, et al, ; Kökten, Lesot, & Kuchler‐Bopp, ; Kuchler‐Bopp et al, ).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Semaphorin 3A receptor inhibitor as a novel therapeutic to promote innervation of bioengineered teeth

Kuchler‐Bopp

Bagnard

Van‐Der‐Heyden

et al. 2018

J Tissue Eng Regen Med

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The sensory innervation of the dental pulp is essential for tooth function and protection. It is mediated by axons originating from the trigeminal ganglia and is spatio-temporally regulated. We have previously shown that the innervation of bioengineered teeth can be achieved only under immunosuppressive conditions. The aim of this study was to develop a model to determine the role of Semaphorin 3A (Sema3A) in the innervation of bioengineered teeth. We first analysed innervation of the dental pulp of mandibular first molars in newborn (postnatal day 0: PN0) mice deficient for Sema3A (Sema3A ), a strong inhibitor of axon growth. While at PN0, axons detected by immunostaining for peripherin and NF200 were restricted to the peridental mesenchyme in Sema3A mice, they entered the dental pulp in Sema3A mice. Then, we have implanted cultured teeth obtained from embryonic day-14 (E14) molar germs of Sema3A mice together with trigeminal ganglia. The dental pulps of E14 cultured and implanted Sema3A teeth were innervated, whereas the axons did not enter the pulp of E14 Sema3A cultured and implanted teeth. A "Membrane Targeting Peptide NRP1," suppressing the inhibitory effect of Sema3A, has been previously identified. The injection of this peptide at the site of implantation allowed the innervation of the dental pulp of bioengineered teeth obtained from E14 dental dissociated mesenchymal and epithelial cells reassociations of ICR mice. In conclusion, these data show that inhibition of only one axon repellent molecule, Sema3A, allows for pulp innervation of bioengineered teeth.

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Section: Discussionsupporting

confidence: 70%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Semaphorin 3A receptor inhibitor as a novel therapeutic to promote innervation of bioengineered teeth

Kuchler‐Bopp

Bagnard

Van‐Der‐Heyden

et al. 2018

J Tissue Eng Regen Med

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Three-dimensional Micro-culture System for Tooth Tissue Engineering

et al. 2016

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The arrangement of cells within a tissue plays an essential role in organogenesis, including tooth development. Progress is being made to regenerate teeth by reassociating dissociated embryonic dental cells and implanting them in vivo. In the present study, we tested the hanging drop method to study mixed epithelial-mesenchymal cell reorganization in a liquid instead of semisolid medium to see whether it could lead to tooth histogenesis and organogenesis. This method allowed the control of the proportion and number of cells to be used, and the forming microtissues showed homogeneous size. The liquid environment favored cell migrations as compared with collagen gels. Three protocols were compared. The one that sequentially combined the hanging drop and semisolid medium cultures prior to in vivo implantation gave the best results. Indeed, after implantation, teeth developed, showing a well-formed crown, mineralization of dentin and enamel, and the initiation of root formation. Vascularization and the cellular heterogeneity in the mesenchyme were similar to what was observed in developing molars. Finally, after coimplantation with a trigeminal ganglion, the dental mesenchyme, including the odontoblast layer, became innervated. The real advantage of this technique is the small number of cells required to make a tooth. This experimental model can be employed to study the development, physiology, metabolism, or toxicology in forming teeth and test other cell sources.

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Bone Marrow Stromal Cells Promote Innervation of Bioengineered Teeth

et al. 2018

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Transplantation of bone marrow mesenchymal stem cells (BMDCs) into a denervated side of the spinal cord was reported to be a useful option for axonal regeneration. The innervation of teeth is essential for their function and protection but does not occur spontaneously after injury. Cultured reassociations between dissociated embryonic dental mesenchymal and epithelial cells and implantation lead to a vascularized tooth organ regeneration. However, when reassociations were coimplanted with a trigeminal ganglion (TG), innervation did not occur. On the other hand, reassociations between mixed embryonic dental mesenchymal cells and bone marrow-derived cells isolated from green fluorescent protein (GFP) transgenic mice (BMDCs-GFP) (50/50) with an intact and competent dental epithelium (ED14) were innervated. In the present study, we verified the stemness of isolated BMDCs, confirmed their potential role in the innervation of bioengineered teeth, and analyzed the mechanisms by which this innervation can occur. For that purpose, reassociations between mixed embryonic dental mesenchymal cells and BMDCs-GFP with an intact and competent dental epithelium were cultured and coimplanted subcutaneously with a TG for 2 wk in ICR mice. Axons entered the dental pulp and reached the odontoblast layer. BMDCs-GFP were detected at the base of the tooth, with some being present in the pulp associated with the axons. Thus, while having a very limited contribution in tooth formation, they promoted the innervation of the bioengineered teeth. Using quantitative reverse transcription polymerase chain reaction and immunostainings, BMDCs were shown to promote innervation by 2 mechanisms: 1) via immunomodulation by reducing the number of T lymphocytes (CD3+, CD25+) in the implants and 2) by expressing neurotrophic factors such as NGF, BDNF, and NT3 for axonal growth. This strategy using autologous mesenchymal cells coming from bone marrow could be used to innervate bioengineered teeth without treatment with an immunosuppressor such as cyclosporine A (CsA), thus avoiding multiple side effects.

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Experimental Design for the Innervation of Tooth Forming from Implanted Cell Re-associations

Cited by 5 publications

References 110 publications

Semaphorin 3A receptor inhibitor as a novel therapeutic to promote innervation of bioengineered teeth

Semaphorin 3A receptor inhibitor as a novel therapeutic to promote innervation of bioengineered teeth

Three-dimensional Micro-culture System for Tooth Tissue Engineering

Bone Marrow Stromal Cells Promote Innervation of Bioengineered Teeth

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