Isolation and cultivation as well as in situ identification of MSCs from equine dental pulp and periodontal ligament.
Abstract: The lifelong eruption places a great demand on the dental pulp and periodontal ligament (PDL) of horse teeth. Cells within the pulp and PDL seem to play a key role during this remodeling. Unassigned: In this study, we isolated and cultivated MSCs (medicinal signaling cells) from dental pulp, PDL and retrobulbar fat of four horses. Subsequently, we analyzed them by flow cytometry and immunohistochemistry to determine and compare their characteristics. In addition, we localized these cells within the tissue structure via immunohistochemistry of histological sections. For these analyses, several surface markers were applied. Unassigned: The described method illustrates a feasible approach to isolate and cultivate MSCs from equine dental pulp and PDL. In the flow cytometry a vast majority of cultivated cells were positive for CD90 and CD40 and negative for CD11a/18, CD45, CD105 and MHCII suggesting that these cells feature characteristics of MSCs. Immunohistochemistry of histological pulp and PDL sections showed the localization of CD90 positive cells especially in the perivascular region and the subodontoblastic layer. Unassigned: Our findings indicate that the isolation and cultivation of MSCs from equine dental pulp and PDL is feasible although an elaborate and complicated harvesting protocol is required. MSCs isolated from dental pulp and PDL are regarded as candidates for new therapeutical approaches in equine dental medicine like regeneration of periodontal lesions, enhancement of periodontal re-attachment after dental replantation and stimulation of pulp-obliteration and apexification in combination with endodontic therapies.
Copyright © 2023 Heilen, Roßgardt, Dern-Wieloch, Vogelsberg and Staszyk.
Publication Date: 2023-03-10 PubMed ID: 36968463PubMed Central: PMC10036573DOI: 10.3389/fvets.2023.1116671Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
Researchers have successfully isolated and cultivated medicinal signaling cells (MSCs) from horse teeth and retrobulbar fat, paving the way for potential therapeutic applications in equine dental medicine.
Methodology
- The researchers collected MSCs from the dental pulp, periodontal ligament (PDL) and retrobulbar fat of four horses for their study.
- Post extraction, the cells were cultivated and studied using techniques such as flow cytometry and immunohistochemistry to establish their characteristics and compare them across different sources.
- Further, immunohistochemistry was used on histological sections to understand the position and distribution of these cells within the tissue structure. During these investigations, several surface markers were utilized.
Findings
- This study indicated the successful isolation and cultivation of MSCs from equine dental pulp and PDL. However, the researchers stress on the fact that the extraction process needs careful execution and is complicated.
- The majority of the cultivated cells showed a positive response for CD90 and CD44 (markers for MSCs), and a negative response for CD11a/18, CD45, CD105 and MHCII. These results suggest that these cells exhibit MSC characteristics.
- The CD90 positive cells were predominant in the perivascular region and the subodontoblastic layer. This location information was determined using immunohistology of pulp and PDL sections.
Potential Applications
- The findings show significant potential for innovative treatments in equine dental medicine.
- Therapeutic possibilities include regeneration of periodontal lesions, improved periodontal re-attachment after dental replantation and stimulation of pulp-obliteration and apexification allied with endodontic therapies.
Cite This Article
APA
Heilen LB, Roßgardt J, Dern-Wieloch J, Vogelsberg J, Staszyk C.
(2023).
Isolation and cultivation as well as in situ identification of MSCs from equine dental pulp and periodontal ligament.
Front Vet Sci, 10, 1116671.
https://doi.org/10.3389/fvets.2023.1116671 Publication
Researcher Affiliations
- Faculty of Veterinary Medicine, Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University, Giessen, Germany.
- Faculty of Veterinary Medicine, Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University, Giessen, Germany.
- Faculty of Veterinary Medicine, Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University, Giessen, Germany.
- Faculty of Veterinary Medicine, Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University, Giessen, Germany.
- Faculty of Veterinary Medicine, Institute of Veterinary Anatomy, Histology and Embryology, Justus-Liebig-University, Giessen, Germany.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
References
This article includes 44 references
- Ten Cate AR. The development of the periodontium--a largely ectomesenchymally derived unit.. Periodontol 2000 1997 Feb;13:9-19.
- Berkovitz BK. Periodontal ligament: structural and clinical correlates.. Dent Update 2004 Jan-Feb;31(1):46-50, 52, 54.
- Warhonowicz M, Staszyk C, Rohn K, Gasse H. The equine periodontium as a continuously remodeling system: morphometrical analysis of cell proliferation.. Arch Oral Biol 2006 Dec;51(12):1141-9.
- Rou00dfgardt J, Heilen LB, Bu00fcttner K, Dern-Wieloch J, Vogelsberg J, Staszyk C. The Equine Dental Pulp: Analysis of the Stratigraphic Arrangement of the Equine Dental Pulp in Incisors and Cheek Teeth.. Vet Sci 2022 Oct 30;9(11).
- Gronthos S, Brahim J, Li W, Fisher LW, Cherman N, Boyde A, DenBesten P, Robey PG, Shi S. Stem cell properties of human dental pulp stem cells.. J Dent Res 2002 Aug;81(8):531-5.
- Hosoya A, Hiraga T, Ninomiya T, Yukita A, Yoshiba K, Yoshiba N, Takahashi M, Ito S, Nakamura H. Thy-1-positive cells in the subodontoblastic layer possess high potential to differentiate into hard tissue-forming cells.. Histochem Cell Biol 2012 Jun;137(6):733-42.
- Chai Y, Jiang X, Ito Y, Bringas P Jr, Han J, Rowitch DH, Soriano P, McMahon AP, Sucov HM. Fate of the mammalian cranial neural crest during tooth and mandibular morphogenesis.. Development 2000 Apr;127(8):1671-9.
- Rothovu00e1 M, Peterkovu00e1 R, Tucker AS. Fate map of the dental mesenchyme: dynamic development of the dental papilla and follicle.. Dev Biol 2012 Jun 15;366(2):244-54.
- Rothovu00e1 M, Feng J, Sharpe PT, Peterkovu00e1 R, Tucker AS. Contribution of mesoderm to the developing dental papilla.. Int J Dev Biol 2011;55(1):59-64.
- Caplan AI. Mesenchymal Stem Cells: Time to Change the Name!. Stem Cells Transl Med 2017 Jun;6(6):1445-1451.
- Caplan AI. New MSC: MSCs as pericytes are Sentinels and gatekeepers.. J Orthop Res 2017 Jun;35(6):1151-1159.
- Ishikawa S, Horinouchi C, Murata D, Matsuzaki S, Misumi K, Iwamoto Y, Korosue K, Hobo S. Isolation and characterization of equine dental pulp stem cells derived from Thoroughbred wolf teeth.. J Vet Med Sci 2017 Jan 20;79(1):47-51.
- Staszyk C, Gasse H. Primary culture of fibroblasts and cementoblasts of the equine periodontium.. Res Vet Sci 2007 Apr;82(2):150-7.
- Mensing N, Gasse H, Hambruch N, Haeger JD, Pfarrer C, Staszyk C. Isolation and characterization of multipotent mesenchymal stromal cells from the gingiva and the periodontal ligament of the horse.. BMC Vet Res 2011 Aug 2;7:42.
- Bertone AL, Reisbig NA, Kilborne AH, Kaido M, Salmanzadeh N, Lovasz R, Sizemore JL, Scheuermann L, Kopp RJ, Zekas LJ, Brokken MT. Equine Dental Pulp Connective Tissue Particles Reduced Lameness in Horses in a Controlled Clinical Trial.. Front Vet Sci 2017;4:31.
- Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop Dj, Horwitz E. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement.. Cytotherapy 2006;8(4):315-7.
- de Mattos Carvalho A, Alves AL, Golim MA, Moroz A, Hussni CA, de Oliveira PG, Deffune E. Isolation and immunophenotypic characterization of mesenchymal stem cells derived from equine species adipose tissue.. Vet Immunol Immunopathol 2009 Dec 15;132(2-4):303-6.
- Penny J, Harris P, Shakesheff KM, Mobasheri A. The biology of equine mesenchymal stem cells: phenotypic characterization, cell surface markers and multilineage differentiation.. Front Biosci (Landmark Ed) 2012 Jan 1;17(3):892-908.
- Barberini DJ, Freitas NP, Magnoni MS, Maia L, Listoni AJ, Heckler MC, Sudano MJ, Golim MA, da Cruz Landim-Alvarenga F, Amorim RM. Equine mesenchymal stem cells from bone marrow, adipose tissue and umbilical cord: immunophenotypic characterization and differentiation potential.. Stem Cell Res Ther 2014 Feb 21;5(1):25.
- Radtke CL, Nino-Fong R, Esparza Gonzalez BP, Stryhn H, McDuffee LA. Characterization and osteogenic potential of equine muscle tissue- and periosteal tissue-derived mesenchymal stem cells in comparison with bone marrow- and adipose tissue-derived mesenchymal stem cells.. Am J Vet Res 2013 May;74(5):790-800.
- Paebst F, Piehler D, Brehm W, Heller S, Schroeck C, Tu00e1rnok A, Burk J. Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: an approach toward a standardized definition.. Cytometry A 2014 Aug;85(8):678-87.
- Triadan H. [Veterinary dentistry: paradentology in monkeys and beasts of prey].. Schweiz Arch Tierheilkd 1973 Sep;115(9):401-19.
- Floyd MR. The modified Triadan system: nomenclature for veterinary dentistry.. J Vet Dent 1991 Dec;8(4):18-9.
- Raabe O, Reich C, Wenisch S, Hild A, Burg-Roderfeld M, Siebert HC, Arnhold S. Hydrolyzed fish collagen induced chondrogenic differentiation of equine adipose tissue-derived stromal cells.. Histochem Cell Biol 2010 Dec;134(6):545-54.
- Gale AL, Linardi RL, McClung G, Mammone RM, Ortved KF. Comparison of the Chondrogenic Differentiation Potential of Equine Synovial Membrane-Derived and Bone Marrow-Derived Mesenchymal Stem Cells.. Front Vet Sci 2019;6:178.
- Leisengang S, Heilen LB, Klymiuk MC, Nu00fcrnberger F, Ott D, Wolf-Hofmann K, Gerstberger R, Rummel C, Schmidt MJ, Arnhold S, Roth J. Neuroinflammation in Primary Cultures of the Rat Spinal Dorsal Horn Is Attenuated in the Presence of Adipose Tissue-Derived Medicinal Signalling Cells (AdMSCs) in a Co-cultivation Model.. Mol Neurobiol 2022 Jan;59(1):475-494.
- Rou00dfgardt J, Heilen LB, Bu00fcttner K, Dern-Wieloch J, Vogelsberg J, Staszyk C. The Equine Dental Pulp: Histomorphometric Analysis of the Equine Dental Pulp in Incisors and Cheek Teeth.. Vet Sci 2022 May 30;9(6).
- Nakajima K, Kunimatsu R, Ando K, Hiraki T, Rikitake K, Tsuka Y, Abe T, Tanimoto K. Success rates in isolating mesenchymal stem cells from permanent and deciduous teeth.. Sci Rep 2019 Nov 14;9(1):16764.
- Genova T, Cavagnetto D, Tasinato F, Petrillo S, Ruffinatti FA, Mela L, Carossa M, Munaron L, Roato I, Mussano F. Isolation and Characterization of Buccal Fat Pad and Dental Pulp MSCs from the Same Donor.. Biomedicines 2021 Mar 7;9(3).
- van Foreest A. [Veterinary dentistry (9). Classification, nomenclature and identification of animal dentition].. Tijdschr Diergeneeskd 1995 Apr 15;120(8):233-40.
- Schrock P, Lu00fcpke M, Seifert H, Staszyk C. Three-dimensional anatomy of equine incisors: tooth length, enamel cover and age related changes.. BMC Vet Res 2013 Dec 9;9:249.
- Muylle S, Simoens P, Lauwers H. Age-related morphometry of equine incisors.. Zentralbl Veterinarmed A 1999 Dec;46(10):633-43.
- Ru00f3u017cau0144ski P, Slaska B, Ru00f3u017cau0144ska D. Prevalence of yeasts in English full blood mares.. Mycopathologia 2013 Apr;175(3-4):339-44.
- Pascucci L, Curina G, Mercati F, Marini C, Dall'Aglio C, Paternesi B, Ceccarelli P. Flow cytometric characterization of culture expanded multipotent mesenchymal stromal cells (MSCs) from horse adipose tissue: towards the definition of minimal stemness criteria.. Vet Immunol Immunopathol 2011 Dec 15;144(3-4):499-506.
- Kamm JL, Parlane NA, Riley CB, Gee EK, Dittmer KE, McIlwraith CW. Blood type and breed-associated differences in cell marker expression on equine bone marrow-derived mesenchymal stem cells including major histocompatibility complex class II antigen expression.. PLoS One 2019;14(11):e0225161.
- Schnabel LV, Pezzanite LM, Antczak DF, Felippe MJ, Fortier LA. Equine bone marrow-derived mesenchymal stromal cells are heterogeneous in MHC class II expression and capable of inciting an immune response in vitro.. Stem Cell Res Ther 2014 Jan 24;5(1):13.
- Kenmotsu M, Matsuzaka K, Kokubu E, Azuma T, Inoue T. Analysis of side population cells derived from dental pulp tissue.. Int Endod J 2010 Dec;43(12):1132-42.
- Sano Y, Sugiuchi A, Mitomo K, Yanagisawa A, Kambe R, Furusawa M, Muramatsu T. Changes of CD90 expression and immunoreactive cell localisation in rat dental pulp after cavity preparation.. Aust Endod J 2019 Aug;45(2):189-195.
- Esteves CL, Sheldrake TA, Dawson L, Menghini T, Rink BE, Amilon K, Khan N, Pu00e9ault B, Donadeu FX. Equine Mesenchymal Stromal Cells Retain a Pericyte-Like Phenotype.. Stem Cells Dev 2017 Jul 1;26(13):964-972.
- Zhao J, Faure L, Adameyko I, Sharpe PT. Stem cell contributions to cementoblast differentiation in healthy periodontal ligament and periodontitis.. Stem Cells 2021 Jan;39(1):92-102.
- Lin NH, Menicanin D, Mrozik K, Gronthos S, Bartold PM. Putative stem cells in regenerating human periodontium.. J Periodontal Res 2008 Oct;43(5):514-23.
- Caplan AI. All MSCs are pericytes?. Cell Stem Cell 2008 Sep 11;3(3):229-30.
- Crisan M, Yap S, Casteilla L, Chen CW, Corselli M, Park TS, Andriolo G, Sun B, Zheng B, Zhang L, Norotte C, Teng PN, Traas J, Schugar R, Deasy BM, Badylak S, Buhring HJ, Giacobino JP, Lazzari L, Huard J, Pu00e9ault B. A perivascular origin for mesenchymal stem cells in multiple human organs.. Cell Stem Cell 2008 Sep 11;3(3):301-13.
- Imai C, Sano H, Quispe-Salcedo A, Saito K, Nakatomi M, Ida-Yonemochi H, Okano H, Ohshima H. Exploration of the role of the subodontoblastic layer in odontoblast-like cell differentiation after tooth drilling using Nestin-enhanced green fluorescent protein transgenic mice.. J Oral Biosci 2022 Mar;64(1):77-84.