review article applications of mesenchymal stem cells and...

Review ArticleApplications of Mesenchymal Stem Cells and Neural Crest Cellsin Craniofacial Skeletal Research

Satoru Morikawa12 Takehito Ouchi12 Shinsuke Shibata2 Takumi Fujimura2

Hiromasa Kawana1 Hideyuki Okano2 and Taneaki Nakagawa1

1Department of Dentistry and Oral Surgery Keio University School of Medicine 35 Shinanomachi Shinjuku-kuTokyo 160-8582 Japan2Department of Physiology Keio University School of Medicine 35 Shinanomachi Shinjuku-ku Tokyo 160-8582 Japan

Correspondence should be addressed to Satoru Morikawa morikawakeiojp

Received 11 December 2015 Accepted 2 February 2016

Academic Editor Jiabing Fan

Copyright copy 2016 Satoru Morikawa et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Craniofacial skeletal tissues are composed of tooth and bone together with nerves and blood vessels This composite material ismainly derived from neural crest cells (NCCs)The neural crest is transient embryonic tissue present during neural tube formationwhose cells have high potential for migration and differentiation Thus NCCs are promising candidates for craniofacial tissueregeneration however the clinical application ofNCCs is hindered by their limited accessibility In contrastmesenchymal stemcells(MSCs) are easily accessible in adults have similar potential for self-renewal and can differentiate into skeletal tissues includingbones and cartilage Therefore MSCs may represent good sources of stem cells for clinical use MSCs are classically identifiedunder adherent culture conditions leading to contamination with other cell lineages Previous studies have identified mouse- andhuman-specific MSC subsets using cell surface markers Additionally some studies have shown that a subset of MSCs is closelyrelated to neural crest derivatives and endothelial cells These MSCs may be promising candidates for regeneration of craniofacialtissues from the perspective of developmental fate Here we review the fundamental biology of MSCs in craniofacial research

1 Introduction

Developmental origins are beginning to be elucidatedthrough rigorous studies in stem cell biology Recent studieshave demonstrated that the basis of regenerative medicinecan be found in developmental biology Indeed many appli-cations in regenerative medicine mimic the development andhealing of specific tissues

Mesenchymal stem cells (MSCs) are commonly usedin both basic and clinical studies because they can beeasily identified in adult tissues MSCs were first identifiedas fibroblast-like cells in the bone marrow [1] resemblecolony forming unit-fibroblasts (CFU-Fs) at clonal densityand have the capacity for differentiation into mesenchymallineages such as bone cartilage and fat [2] Notably MSCsand neural crest cells (NCCs) are both used in variousapproaches in craniofacial biology because of their devel-opmental similarities Indeed the craniofacial mesenchyme

developmentally originates from NCCs [3ndash5] and NCCs aredevelopmentally identified at the embryonic stages [6] It isdifficult to isolate NCCs because of their limited accessibilityand ethical concerns therefore it is difficult to directly useNCCs in patients In contrast MSCs are present in easilyaccessible adult tissues such as bone marrow fat tissues andsynovium enabling facile isolation Importantly MSCs andNCCs have similar self-renewal and differentiation potentialandMSCs are capable of differentiating into neuronal cells [78] Furthermore MSCs can also differentiate into endothelialcells [9ndash11] and are indispensable for tissue formation [12 13]Similar findings have also been reported in skeletal tissues[14 15] suggesting that adult MSCs may be useful in clinicalapplications associated with the regeneration of skeletaltissues particularly because of the necessity for synchronizedneural tissue formation and vascularization

Skeletal stem cells (SSCs) which were recently identified[16 17] have been shown to contribute to the construction

Hindawi Publishing CorporationStem Cells InternationalVolume 2016 Article ID 2849879 8 pageshttpdxdoiorg10115520162849879

2 Stem Cells International

of skeletal tissues during development and wound healingHowever the formation and regeneration of skeletal tissuesinvolve not only construction of bone but also vasculariza-tion and neural synchronization [18ndash20] Despite this factfew studies have evaluated these processes with regard toSSCs

NCCs MSCs and SSCs are all isolated using culture andexhibit overlapping self-renewal and multipotent differentia-tion potential Thus clarifying the specific characteristics ofeach cell type will improve the clinical application of thesetypes of stem cells In this review we discuss the fundamentalbiology of stem cells in craniofacial research

2 Stem Cells in Craniofacial Research

Skeletal tissues are composed of a network of hard tissuesincluding bone and cartilage The jawbone and teeth com-prise the craniofacial region and many individuals sufferfrom skeletal diseases such as metastasis of oral malignanttumors into the bone congenital craniofacial malformationsevere periodontitis andmedication-related osteonecrosis ofthe jaw [21] These diseases cause eating difficulty aestheticdisorders respiratory distress and speech disorders leadingto decreased quality of life Current fundamental approachesto these diseases include surgery and subsequent reconstruc-tion using artificial materials or xenobiomaterials Howevernatural bone formation and healing using autologous cellsare preferable Therefore development-based medicine andapproaches are desired

During the most recent decade stem cell researchhas made great advances in clarifying the mechanisms oftissue development Indeed many stem cell researchershave focused on developmental biology and regenerativemedicine and applications of stem cells in craniofacialresearch have been proposed In particular MSCs and NCCshave been studied extensively in craniofacial research MSCspartially originate from NCCs [7 22ndash25] therefore someMSCs may also differentiate into neuronal cells [26ndash28]suggesting that specific subsets of MSCs may have the samepotential as NCCs Moreover because MSCs are present inseveral adult tissues [2] they are easy to isolate and expand invitro

Notably dental-specific MSCs have been identified incraniofacial tissues Several research groups have reported thepresence of dental MSCs in dental pulp stem cells (DPSCs)[29] stem cells from exfoliated deciduous teeth (SHED) [30]periodontal ligament stem cells (PDLSCs) [31] and stem cellsfrom apical papilla (SCAP) [32] These dental MSCs mayhave applications in degenerative and intractable diseases[33] Furthermore conditioned medium (CM) from dentalMSCs supplies paracrine factors and may be effective forinjured areas [34 35] Osugi et al reported that SHED-CM promoted the growth of bone mass in a calvarial defectmodel Additionally the use of conditioned medium fromdental tissue-derivedMSCs is a unique approach for craniofa-cial regenerative medicine without cell transplantation Thisapproach may reduce costs and timelabor requirements andmay alleviate safety concerns [36]

Bone marrow MSCs are utilized as a typical modelfor clinical studies and basic biology research and theyare classically defined by conventional culture as MSCsshow vigorous expansion and multipotent differentiationHowever the mechanisms of regeneration in conventionalMSCs cannot be traced back to developmental fate andit is difficult to predict which subsets of a crowded cellpopulation contribute to the development of specific targettissues Thus mixed cultures of conventional MSCs maynot provide consistent predictable therapeutic outcomesfrom a developmental biological viewpoint Further studiesare needed to clarify the development and degeneration ofcraniofacial skeletal tissues using stem cells particularly foranalysis of the clonal phenotype of NCCs and MSCs

3 Purified MSCs Are PartiallyDerived from NCCs

Cranial NCCs constitute a major part of facial tissues [3 37]Moreover NCCs are localized in the neural folds of fetaltissues migrate into various tissues and regulate skeletaltissues [38] Cranial NCCs form the first pharyngeal archinnerved by the trigeminal nerve and the second pharyngealarch innerved by the facial nerve [39ndash41] Then the firstand second pharyngeal arches interact with each other andform craniofacial tissues [42 43] Craniofacial tissues aremainly composed of bone muscle and tendon togetherwith the neurovascular bundle [44] Therefore improvingthe understanding of neurogenesis and vascularization inskeletal tissues is essential for discussions of craniofacialtissue formation To identify and track the process of cranialtissue specification visualization of the neural crest and itsderivatives is needed

Within the last few decades transgenic technologieshave been developed that enable visualization of target cellsfor prospective and retrospective analyses [45] Transgenicanimal studies can be used to elucidate what and how specificcells contribute to form target tissues This approach has alsobeen applied to the craniofacial field where NCCs and theirderivative cells are labeled and their fates are analyzed [4 4647]This transgenic technology provides clear information onthe development ofNCCs in the craniofacial field and enablesthe visualization of NCCs and their derivatives within dentaland craniofacial tissues as shown in Figure 1(a)

Some researchers have focused on cells that are easierto obtain than embryonic NCCs such as bone marrowstem cells or pluripotent stem cells The bone marrow isa typical source of MSCs and contains two distinct typesof stem cells hematopoietic stem cells (HSCs) and MSCs[48] Transgenic animal studies have demonstrated thatMSCs partially originate from NCCs NCCs delaminatefrom neuroepithelial cells which can differentiate into cellsexpressing the MSC marker platelet-derived growth factorreceptor alpha (PDGFR120572) (CD140a) [22] Thus neuroep-ithelial cells are a source of MSC differentiation MoreoverNagoshi et al showed that NCCs migrate though the aorta-gonad-mesonephros region and circulate into the bone mar-row demonstrating how neural crest-derived stem cells cantravel to the bone marrow [23] Additionally Morikawa

Stem Cells International 3

GFP Phase Merge

(a)

Sca-1

PDGFR120572

CD45minusTer119minus gated

(b)

Figure 1 (a) Neural crest lineage labeling mouse (Wnt1-CreGFP) clearly demonstrates green fluorescence-positive NCCs in craniofacialtissues at embryonic day 11 Scale bar 1mm (b) Murine bone marrow cells were analyzed by flow cytometry The chart shows that cellsexpressing hematopoietic lineage markers (CD45 and Ter119) were negatively gated and the highly potent murine MSC marker-expressing(P120572S) subset is indicated by the black-colored box

et al reported a subset of highly potent murine MSCscharacterized by the cell surface marker combination ofPDGFR120572 stem cell antigen-1 (Sca-1lymphocyte activationprotein Ly-6) CD45 and Ter119 (Ly-76)This specific subsetPDGFR120572+Sca-1+CD45minusTer119minus (P120572S) could be found in thebone marrow as shown in Figure 1(b) [49] The P120572S subsetpartially originates from NCCs as shown by developmentalfate analysis using transgenic mice [7] Thus application ofcell tracking systems in mice has clarified the relationshipbetween MSCs and NCCs supporting the application ofMSCs in craniofacial skeletal tissue research

The P120572S subset of MSCs has been identified as theperivascular niche and has differentiation potential to bothmesenchymal and neural crest lineages [7] Indeed theP120572S subset promotes neural crest-derived periodontal tissueregeneration [50] P120572S cells are closely related to SSCs whichare identified by rigorous assays Bone marrow MSCs alsocontain stem cells that can only differentiate into skeletaltissues Thus SSCs represent a reservoir for bone-formingcells and have the potential to shape and regulate the localmicrovascular network in the bone marrow [51]

Worthley et al reported that Gremlin-1 functions as aspecificmarker of skeletal stem cells in the long bonemarrowGremlin-1 is a bone morphogenic peptide (BMP) antagonistand the transforming growth factor (TGF)-120573BMP signalingpathway regulates osteoblastogenesis and bone formation[52] In Gremlin-1-overexpressing transgenic mice Gremlin-1+ cells differentiate into bone cartilage and reticular stromalcells Postnatally Gremlin-1+ cells also contribute to develop-ment Gremlin-1+ cells are not further enriched for P120572S cellsHowever the P120572S subset also contributes to the formation of

skeletal tissues [53] Therefore some MSCs in the P120572S subsetmay have the differentiation potential of SSCs Consistentwith this notion Gremlin regulates developing limbs andGremlin-1+ and P120572S cell subsets have been identified inthe long bone [17 49 54 55] Additionally recent reportshave shown that cranial MSCs are different from long bonemarrow MSCs [56]

Zhao et al reported that glioma-associated oncogenehomolog-1 (Gli-1) is amarker of craniofacial-specificMSCs incranial bones [57] Gli-1+ cells are not coexpressedwith classi-calMSCmarkers in vivo Although the specific characteristicsof these cells have not been defined in vivo Gli-1+ cells showtypical phenotypes of MSCs such as vigorous expansionexpression of classical MSC markers and differentiation tomesenchymal lineages in vitro [44 57] However Gli-1 is notalways expressed in MSCs during development in contrastto PDGFR120572 Notably no direct gene regulation mechanismhas been identified between Gli-1 and PDGFR120572 [58] Gli-1 isinduced by sonic hedgehog (Shh) signaling and is associatedwith transient Sox2 expression during tooth formation [59]Shh signaling has also been detected in Hertwigrsquos epithelialroot sheath (HERS) and found to lead to tooth root formation[60] suggesting that Shh signalingmay promoteGli-1 expres-sion in craniofacial-specific mesenchyme Although Gli-1+cells are not present in the perivascular niche [57] MSCs inlong bones are regulated by the perivascular niche [61ndash65]The leptin receptor (LepR) (CD295) has been reported as anexcellent marker for MSCs LepR+ cells are identified aroundsinusoids [61 66] In contrast MSCs in the cranial bonesuture are found around the midline of the suture structureThis specificity of craniofacial MSCs does not correspond


to MSCs in long bone marrow These differences in murinecraniofacial and long bone MSCs should be examined ingreater detail in future studies

4 Purified MSCs Can Be Derived fromInduced Pluripotent Stem Cells (iPSCs)

To achieve the transition of stem cell research from thebench to the bedside more studies investigating humancells are required Human MSCs can be found in severaltypes of tissues The classical definition of MSCs is the samein mice and in humans and conventional MSCs can becontaminated by other cell lineages To avoid such problemsmore studies of cell markers and selection of target cellsare needed Previously Mabuchi et al identified the highlyspecific human MSC markers low-affinity nerve growthfactor receptor (LNGFR) and thymocyte antigen 1 (THY-1)[67] Cells expressing LNGFRandTHY-1 have been identifiedin the bone marrow and the combination of LNGFR andTHY-1 cell surface markers characterizes a distinct subset ofMSCs with a hematopoietic lineage LNGFR+THY-1+ cellshave high colony forming potential and differentiate intomesenchymal lineages These specific cells have also beenidentified in decidua fat tissues synovium and dental pulp[67ndash69] The LNGFR+THY-1+ subset shows highly potentself-renewal and differentiation capacity both in vitro andin vivo and is also associated with the expression of otherclassical MSC markers such as CD29 (integrin beta 1)CD44 (homing cell adhesionmolecule HCAM) CD73 (ecto-51015840-nucleotidase) CD105 (endoglin) CD146 (melanoma celladhesionmoleculeMCAM) andCD166 (activated leukocytecell adhesion molecule ALCAM) [67 69] Efficient proce-dures for purification will improve the ability to analyzeMSCs

In craniofacial diseases there is a great need for recon-struction of large areas affected by disease or injury Thusit is necessary to obtain large numbers of MSCs for suchclinical applications This must be achieved without dam-aging original tissues therefore specific cells derived frompluripotent stem cells based on developmental research mayhelp to overcome this problem

Human iPSCs were first generated in 2007 and havebeen used extensively in biomedical studies [70] HumaniPSCs have the capacity for self-renewal and can be expandedrelatively easily The strong potential of iPSCs can also beapplied in craniofacial research MSCs can be derived bothdirectly from iPSCs and indirectly from neural crest like cellsusing specific markers and culture conditions [24 71] MSCsinduced from iPSCs have been used in the regeneration ofperiodontal tissues [72] Thus these findings suggest thatiPSC-derived MSCs have the capacity for use in craniofacialtissue regeneration

5 Application of Human MSCs inCraniofacial Research

Craniofacial connective tissues originate from neural crest-derived ectomesenchyme which is a source of many cranio-facial bone and cartilage structures Umeda et al reported the

generation of ectomesenchymal cells through neural crest-like progeny from human iPSCs [73] Sensory innervation isnecessary for maintaining sound bone In dentistry sensoryinnervation of craniofacial tissues involves the trigeminalnerve Previously several methods were reported for induc-tion of peripheral nerve formation using NCCs [74 75]Application of sensory neuronal cells to craniofacial skele-tal tissues requires neural crest-derived craniofacial-specificsensory neuronal induction Dincer et al reported that thecraniofacial placode can be used to identify the craniofacialtrigeminal nerve [76] However induction procedures forcraniofacial target tissues are a relatively new approach inregenerative medicine and disease-specific iPSC technologyFurther studies are needed to provide clear information onneural crest biology and craniofacial specificity Methods forinduction of target cellsmust take advantage of the generationof a sufficient number of cells and induction based on thebasic knowledge of developmental biology may provide anevidence-based approach for application of stem cells in theclinical field

For application of human iPSC-derived cells in theregeneration of craniofacial tissues prevention of teratomaformation represents amajor challenge iPSC lines show vari-ations in the patterns of teratoma formation in iPSC-derivedneural progenitor cells [77 78] Surprisingly despite theexclusion of pluripotent markers iPSCs may form teratomasin some cases Lee et al reported that iPSC-derived neuralcrest-derived stem cells which exhibit downregulation ofpolysialic acid-neural cell adhesion molecule (PSA-NCAM)tend to form teratomas [79] These findings suggest theimportance of basic and preclinical studies of iPSC-derivedNCCs Recent studies have shown the safety of iPSC-derivedcells in preclinical models [80] Reconstruction of the targetcraniofacial tissues in nonhuman primate models is essentialbefore human clinical studies using iPSCs can be initiatedPrescott et al showed that cis-regulatory divergence is associ-ated with differences in quantitative expression in human andchimpanzee cranial NCCs derived from iPSCs [81] It is quiteimportant that this study demonstrated the novel applicationof iPSC-derived NCCs for analyzing evolutionary cellularanthropology in the context of craniofacial development

Thus iPSC technology may facilitate future applicationsin regenerative medicine if the risk of iPSC-derived tumorformation can be minimized

6 Conclusion

Skeletal tissues are composed of bone cartilage and tendonThese mesenchymal tissues are generated fromNCC-derivedMSCs and exhibit neurogenesis and neovascularizationMSCs are part of the perivascular niche and overlap withneural crest-derived stem cells [25] MSCs have potential fordifferentiating into skeletal cells neuronal cells and endothe-lial cells suggesting that MSCs may be useful for craniofacialtissue regeneration Indeed MSCs have been applied for thetreatment of craniofacial diseases such as periodontitis andosteonecrosis of the jaw in small animal models [82 83] Thepotential of stem cells is typically demonstrated using miceand rats because they are easy to breed and handle and there


is a variety of well-established disease models recognized bythe scientific community However the craniofacial anatomyof these animals is different from that of humans Severalresearch groups reported stem cell approaches using largeranimals such as pigs dogs and chimpanzees [32 81 84] Inanimal studies establishment of live cell imaging systems canbe used to clearly visualize the potential for migration anddifferentiation Such imaging systems have been utilized inthe craniofacial field [69 85] Demonstration of cell trackingin large animal models such as nonhuman primates providesessential evidence for human preclinical studies that expectedto take place in the near future

Human clinical application of stem cells has alreadystarted [86 87] however the current protocols for clinicalapplicationmainly utilize conventionally cultured cells Con-ventionalMSCs contain various types of cells within adherentculture resulting in contamination of the MSCs with othercell lineages Moreover MSCs show great differences incharacteristics between long bones and craniofacial tissueand these differences should be evaluated in detail in futurestudies

For further analyses of the applications of MSCs inhumans the developmental fate of human MSCs must beelucidated Sufficient quantities of purified MSCs from adulttissues for reconstruction of large spaces in the craniofacialregion are difficult to collect Human iPSC technologymay beused to overcome this problem Furthermore more analysesof MSCs NCCs SSCs and iPSCs are required based on adevelopmental biological approach whichwill be expected toprovide evidence-based methods for the treatment of variouscraniofacial diseases

Conflict of Interests

HideyukiOkano is a paid scientific advisor of SanBioCo LtdThe other authors have no conflict of interests to declare

Authorsrsquo Contribution

Satoru Morikawa and Takehito Ouchi contributed equally tothis work

Acknowledgments

Theauthors thank themembers of their laboratory for helpfuldiscussionsThis workwas supported by Japan Society for thePromotion of Science KAKENHI grants numbered 15K11221(Taneaki Nakagawa) and 25463226 (Satoru Morikawa) andby the Program for Intractable Disease Research UtilizingDisease-specific iPS Cells from the Japan Science and Tech-nology Agency (JST) Japan Agency for Medical Researchand Development (A-MED) and the Ministry of EducationScience Sports and Culture (MEXT) to Hideyuki Okano

References

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[56] H Zhao and Y Chai ldquoStem cells in teeth and craniofacialbonesrdquo Journal of Dental Research vol 94 no 11 pp 1495ndash15012015

[57] H Zhao J Feng T Ho W Grimes M Urata and Y ChaildquoThe suture provides a niche for mesenchymal stem cells ofcraniofacial bonesrdquo Nature Cell Biology vol 17 no 4 pp 386ndash396 2015

[58] X-Q Zhang G B Afink X-R Hu K Forsberg-Nilsson andM Nister ldquoGli1 is not required for Pdgfr120572 expression duringmouse embryonic developmentrdquo Differentiation vol 73 no 2-3 pp 109ndash119 2005

[59] J Li J Feng Y Liu et al ldquoBMP-SHH signaling networkcontrols epithelial stem cell fate via regulation of its niche in thedeveloping toothrdquoDevelopmental Cell vol 33 no 2 pp 125ndash1352015

[60] Y Liu J Feng J Li H Zhao T Ho and Y Chai ldquoAn Nfic-hedgehog signaling cascade regulates tooth root developmentrdquoDevelopment vol 142 no 19 pp 3374ndash3382 2015

[61] B O Zhou R Yue M M Murphy J G Peyer and S JMorrison ldquoLeptin-receptor-expressing mesenchymal stromalcells represent the main source of bone formed by adult bonemarrowrdquo Cell Stem Cell vol 15 no 2 pp 154ndash168 2014

[62] M Crisan S Yap L Casteilla et al ldquoA perivascular origin formesenchymal stem cells in multiple human organsrdquo Cell StemCell vol 3 no 3 pp 301ndash313 2008

[63] B Sacchetti A Funari S Michienzi et al ldquoSelf-renewingosteoprogenitors in bone marrow sinusoids can organize ahematopoietic microenvironmentrdquo Cell vol 131 no 2 pp 324ndash336 2007

[64] J Street M Bao L DeGuzman et al ldquoVascular endothelialgrowth factor stimulates bone repair by promoting angiogenesisand bone turnoverrdquo Proceedings of the National Academy ofSciences of the United States of America vol 99 no 15 pp 9656ndash9661 2002

[65] N Ono W Ono T Mizoguchi T Nagasawa P S Frenetteand H M Kronenberg ldquoVasculature-associated cells express-ing nestin in developing bones encompass early cells in theosteoblast and endothelial lineagerdquo Developmental Cell vol 29no 3 pp 330ndash339 2014

[66] Y Matsuzaki Y Mabuchi and H Okano ldquoLeptin receptormakes its mark on MSCsrdquo Cell Stem Cell vol 15 no 2 pp 112ndash114 2014

[67] Y Mabuchi S Morikawa S Harada et al ldquoLNGFR+THY-1+VCAM-1hi+ cells reveal functionally distinct subpopulationsin mesenchymal stem cellsrdquo Stem Cell Reports vol 1 no 2 pp152ndash165 2013

[68] Y Ogata Y Mabuchi M Yoshida et al ldquoPurified humansynovium mesenchymal stem cells as a good resource forcartilage regenerationrdquo PLoS ONE vol 10 no 6 Article IDe0129096 2015

[69] T Yasui Y Mabuchi H Toriumi et al ldquoPurified human dentalpulp stem cells promote osteogenic regenerationrdquo Journal ofDental Research vol 95 no 2 pp 206ndash214 2016

[70] K Takahashi K Tanabe M Ohnuki et al ldquoInduction ofpluripotent stem cells from adult human fibroblasts by definedfactorsrdquo Cell vol 131 no 5 pp 861ndash872 2007

[71] M Giuliani N Oudrhiri Z M Noman et al ldquoHuman mes-enchymal stem cells derived from induced pluripotent stemcells down-regulate NK-cell cytolytic machineryrdquo Blood vol118 no 12 pp 3254ndash3262 2011

[72] K Hynes D Menicanin J Han et al ldquoMesenchymal stem cellsfrom iPS cells facilitate periodontal regenerationrdquo Journal ofDental Research vol 92 no 9 pp 833ndash839 2013

[73] KUmedaHOdaQ Yan et al ldquoLong-term expandable SOX9+chondrogenic ectomesenchymal cells from human pluripotentstem cellsrdquo Stem Cell Reports vol 4 no 4 pp 712ndash726 2015

[74] G Lee H Kim Y Elkabetz et al ldquoIsolation and directeddifferentiation of neural crest stem cells derived from humanembryonic stem cellsrdquo Nature Biotechnology vol 25 no 12 pp1468ndash1475 2007

[75] LMenendez T A Yatskievych P B Antin and S Dalton ldquoWntsignaling and a Smad pathway blockade direct the differentia-tion of human pluripotent stem cells tomultipotent neural crestcellsrdquo Proceedings of the National Academy of Sciences of theUnited States of America vol 108 no 48 pp 19240ndash19245 2011

[76] Z Dincer J Piao L Niu et al ldquoSpecification of functionalcranial placode derivatives from human pluripotent stem cellsrdquoCell Reports vol 5 no 5 pp 1387ndash1402 2013

[77] K Miura Y Okada T Aoi et al ldquoVariation in the safety ofinduced pluripotent stem cell linesrdquo Nature Biotechnology vol27 no 8 pp 743ndash745 2009

[78] H Okano M Nakamura K Yoshida et al ldquoSteps toward safecell therapy using induced pluripotent stem cellsrdquo CirculationResearch vol 112 no 3 pp 523ndash533 2013

[79] D R Lee J Yoo J Lee et al ldquoPSA-NCAM-negative neuralcrest cells emerging during neural induction of pluripotent stemcells cause mesodermal tumors and unwanted graftsrdquo Stem CellReports vol 4 no 5 pp 821ndash834 2015

[80] Y Kobayashi Y Okada G Itakura et al ldquoPre-evaluated safehuman iPSC-derived neural stem cells promote functionalrecovery after spinal cord injury in commonmarmoset withouttumorigenicityrdquo PLoS ONE vol 7 no 12 Article ID e527872012

[81] S L Prescott R Srinivasan M C Marchetto et al ldquoEnhancerdivergence and cis-regulatory evolution in the human andchimp neural crestrdquo Cell vol 163 no 1 pp 68ndash83 2015

[82] H Yang R M Aprecio X Zhou et al ldquoTherapeutic effect ofTSG-6 engineered iPSC-derived MSCs on experimental peri-odontitis in rats a pilot studyrdquo PLoS ONE vol 9 no 6 ArticleID e100285 2014

[83] K Ogata W Katagiri M Osugi et al ldquoEvaluation of thetherapeutic effects of conditioned media from mesenchymalstem cells in a rat bisphosphonate-related osteonecrosis of thejaw-like modelrdquo Bone vol 74 pp 95ndash105 2015

[84] T Iwata M Yamato H Tsuchioka et al ldquoPeriodontal regen-eration with multi-layered periodontal ligament-derived cellsheets in a caninemodelrdquo Biomaterials vol 30 no 14 pp 2716ndash2723 2009


[85] J Dixon N C Jones L L Sandell et al ldquoTcof1Treacle isrequired for neural crest cell formation and proliferation defi-ciencies that cause craniofacial abnormalitiesrdquo Proceedings ofthe National Academy of Sciences of the United States of Americavol 103 no 36 pp 13403ndash13408 2006

[86] P Voss S Sauerbier M Wiedmann-Al-Ahmad et al ldquoBoneregeneration in sinus lifts comparing tissue-engineered boneand iliac bonerdquoBritish Journal of Oral andMaxillofacial Surgeryvol 48 no 2 pp 121ndash126 2010

[87] M Griffin D M Kalaskar P E Butler and A M SeifalianldquoThe use of adipose stem cells in cranial facial surgeryrdquo StemCell Reviews and Reports vol 10 no 5 pp 671ndash685 2014

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

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International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

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Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



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ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


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Stem CellsInternational



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of skeletal tissues during development and wound healingHowever the formation and regeneration of skeletal tissuesinvolve not only construction of bone but also vasculariza-tion and neural synchronization [18ndash20] Despite this factfew studies have evaluated these processes with regard toSSCs

NCCs MSCs and SSCs are all isolated using culture andexhibit overlapping self-renewal and multipotent differentia-tion potential Thus clarifying the specific characteristics ofeach cell type will improve the clinical application of thesetypes of stem cells In this review we discuss the fundamentalbiology of stem cells in craniofacial research

2 Stem Cells in Craniofacial Research

Skeletal tissues are composed of a network of hard tissuesincluding bone and cartilage The jawbone and teeth com-prise the craniofacial region and many individuals sufferfrom skeletal diseases such as metastasis of oral malignanttumors into the bone congenital craniofacial malformationsevere periodontitis andmedication-related osteonecrosis ofthe jaw [21] These diseases cause eating difficulty aestheticdisorders respiratory distress and speech disorders leadingto decreased quality of life Current fundamental approachesto these diseases include surgery and subsequent reconstruc-tion using artificial materials or xenobiomaterials Howevernatural bone formation and healing using autologous cellsare preferable Therefore development-based medicine andapproaches are desired

During the most recent decade stem cell researchhas made great advances in clarifying the mechanisms oftissue development Indeed many stem cell researchershave focused on developmental biology and regenerativemedicine and applications of stem cells in craniofacialresearch have been proposed In particular MSCs and NCCshave been studied extensively in craniofacial research MSCspartially originate from NCCs [7 22ndash25] therefore someMSCs may also differentiate into neuronal cells [26ndash28]suggesting that specific subsets of MSCs may have the samepotential as NCCs Moreover because MSCs are present inseveral adult tissues [2] they are easy to isolate and expand invitro

Notably dental-specific MSCs have been identified incraniofacial tissues Several research groups have reported thepresence of dental MSCs in dental pulp stem cells (DPSCs)[29] stem cells from exfoliated deciduous teeth (SHED) [30]periodontal ligament stem cells (PDLSCs) [31] and stem cellsfrom apical papilla (SCAP) [32] These dental MSCs mayhave applications in degenerative and intractable diseases[33] Furthermore conditioned medium (CM) from dentalMSCs supplies paracrine factors and may be effective forinjured areas [34 35] Osugi et al reported that SHED-CM promoted the growth of bone mass in a calvarial defectmodel Additionally the use of conditioned medium fromdental tissue-derivedMSCs is a unique approach for craniofa-cial regenerative medicine without cell transplantation Thisapproach may reduce costs and timelabor requirements andmay alleviate safety concerns [36]

Bone marrow MSCs are utilized as a typical modelfor clinical studies and basic biology research and theyare classically defined by conventional culture as MSCsshow vigorous expansion and multipotent differentiationHowever the mechanisms of regeneration in conventionalMSCs cannot be traced back to developmental fate andit is difficult to predict which subsets of a crowded cellpopulation contribute to the development of specific targettissues Thus mixed cultures of conventional MSCs maynot provide consistent predictable therapeutic outcomesfrom a developmental biological viewpoint Further studiesare needed to clarify the development and degeneration ofcraniofacial skeletal tissues using stem cells particularly foranalysis of the clonal phenotype of NCCs and MSCs

3 Purified MSCs Are PartiallyDerived from NCCs

Cranial NCCs constitute a major part of facial tissues [3 37]Moreover NCCs are localized in the neural folds of fetaltissues migrate into various tissues and regulate skeletaltissues [38] Cranial NCCs form the first pharyngeal archinnerved by the trigeminal nerve and the second pharyngealarch innerved by the facial nerve [39ndash41] Then the firstand second pharyngeal arches interact with each other andform craniofacial tissues [42 43] Craniofacial tissues aremainly composed of bone muscle and tendon togetherwith the neurovascular bundle [44] Therefore improvingthe understanding of neurogenesis and vascularization inskeletal tissues is essential for discussions of craniofacialtissue formation To identify and track the process of cranialtissue specification visualization of the neural crest and itsderivatives is needed

Within the last few decades transgenic technologieshave been developed that enable visualization of target cellsfor prospective and retrospective analyses [45] Transgenicanimal studies can be used to elucidate what and how specificcells contribute to form target tissues This approach has alsobeen applied to the craniofacial field where NCCs and theirderivative cells are labeled and their fates are analyzed [4 4647]This transgenic technology provides clear information onthe development ofNCCs in the craniofacial field and enablesthe visualization of NCCs and their derivatives within dentaland craniofacial tissues as shown in Figure 1(a)

Some researchers have focused on cells that are easierto obtain than embryonic NCCs such as bone marrowstem cells or pluripotent stem cells The bone marrow isa typical source of MSCs and contains two distinct typesof stem cells hematopoietic stem cells (HSCs) and MSCs[48] Transgenic animal studies have demonstrated thatMSCs partially originate from NCCs NCCs delaminatefrom neuroepithelial cells which can differentiate into cellsexpressing the MSC marker platelet-derived growth factorreceptor alpha (PDGFR120572) (CD140a) [22] Thus neuroep-ithelial cells are a source of MSC differentiation MoreoverNagoshi et al showed that NCCs migrate though the aorta-gonad-mesonephros region and circulate into the bone mar-row demonstrating how neural crest-derived stem cells cantravel to the bone marrow [23] Additionally Morikawa


GFP Phase Merge

(a)

Sca-1

PDGFR120572


(b)


















6 Conclusion










Acknowledgments


References



































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


GFP Phase Merge

(a)

Sca-1

PDGFR120572


(b)


















6 Conclusion










Acknowledgments


References



































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology












6 Conclusion










Acknowledgments


References



































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology









Acknowledgments


References



































































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

















































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

review article applications of mesenchymal stem cells and...

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