Multipotent postnatal stem cells from human periodontal ligament and uses thereof
Inventors
Shi, Songtao • Seo, Byoung-Moo • Miura, Masako
Assignees
GOVERNMENT OF United States, HEALTH AN HUMAN SERVICES, Secretary of, Department of • National Institutes of Health NIH • US Department of Health and Human Services
Publication Number
US-9210925-B2
Publication Date
2015-12-15
Expiration Date
2024-11-22
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Abstract
The invention generally relates to postnatal periodontal ligament stem cells and methods for their use. More specifically, the invention relates in one aspect to postnatal periodontal ligament multipotent stem cells, use of the cells to generate periodontium, differentiation of the cells and methods of tissue cryopreservation.
Core Innovation
The invention provides isolated human postnatal periodontal ligament multipotent stem cells (PDLSCs) that are capable of extensive proliferation and multipotential differentiation into cementoblasts, cementocytes, adipocytes, and fibroblasts. The invention includes methods for isolating these PDLSCs, culturing and expanding them ex vivo, differentiating them, cryopreserving tissue containing them, and implanting them to generate periodontal tissue including cementum and periodontal ligament both in vitro and in vivo.
Periodontal ligament is a specialized connective tissue that connects teeth to alveolar bone, supporting function and maintaining tissue homeostasis. Periodontal diseases like gingivitis and periodontitis cause destruction of the periodontium, including PDL, cementum, alveolar bone, and gingiva. Existing treatment methods for periodontal defects such as guided tissue regeneration and growth factors lack consistently predictable outcomes, creating a need for new regenerative methods.
The invention addresses this need by providing PDLSCs capable of generating functional periodontal tissue in vivo, including cementum and PDL-like connective tissue, demonstrated by transplantation into immunocompromised animals forming structures resembling native tissue. The cells express mesenchymal stem cell markers, including STRO-1 and CD146/MUC18, and a tendon-specific transcription factor, scleraxis. The invention also provides methods for cryopreserving human PDL tissue with retention of functional PDLSC recovery and tissue regeneration capability after thawing, facilitating tissue storage for later therapeutic use.
Claims Coverage
The patent contains multiple independent claims covering compositions and methods involving isolated human periodontal ligament multipotent stem cells and their use in generating periodontal tissue.
Composition of isolated clonal periodontal ligament multipotent stem cells with biocompatible carrier
A composition consisting of an effective amount of an isolated clonal population of human periodontal ligament multipotent stem cells that proliferate, express STRO-1 and MUC18, form colonies in vitro, and differentiate into cells producing collagen fibers and cementum in vivo, combined with a biocompatible three dimensional carrier such as a polymer, gelatin, polyvinyl sponges, collagen matrix, or combination of hydroxyapatite and tricalcium phosphate, wherein cells were expanded ex vivo.
Method to generate periodontal tissue or cells using the composition
A method of generating periodontal tissue or cells in a subject by implanting the above composition into tissue of the subject, wherein the generated tissue includes periodontal ligament, cementocytes, cementoblasts, or cementum expressing markers such as bone sialoprotein, osteocalcin, matrix extracellular protein, human mitochondrial protein, type I collagen, and fibromodulin, to reduce or ameliorate trauma including periodontal disease or physical injury such as dental procedures.
Composition of clonal periodontal ligament multipotent stem cells in hydroxyapatite or tricalcium phosphate carrier
A composition consisting of isolated clonal human periodontal ligament multipotent stem cells expanded ex vivo in combination with a carrier of hydroxyapatite or tricalcium phosphate, wherein the cells proliferate, express STRO-1 and MUC18, form colonies in vitro, and differentiate into cells producing collagen fibers and cementum in vivo.
Method to generate periodontal tissue using hydroxyapatite or tricalcium phosphate carrier composition
A method of generating periodontal tissue or cells by implanting the above composition into tissue of a human subject, wherein the generated tissue comprises periodontal ligament, cementocytes, cementoblasts, or cementum expressing characteristic markers, to reduce or ameliorate trauma including periodontal disease and injuries such as dental procedures.
Composition of periodontal ligament multipotent stem cells expressing scleraxis in hydroxyapatite/tricalcium phosphate carrier
A composition consisting of an effective amount of a carrier combining hydroxyapatite and tricalcium phosphate and an isolated clonal population of human periodontal ligament multipotent stem cells expanded ex vivo expressing STRO-1, MUC18, and scleraxis that proliferate, form colonies in vitro, and differentiate into collagen-producing cells and cementum forming cells in vivo.
Method to generate periodontal tissue using composition with scleraxis-expressing PDLSCs
A method to generate periodontal tissue or cells by implanting the above described composition into tissue of a human subject, generating periodontal ligament, cementocytes, cementoblasts, or cementum expressing defined markers, to reduce or ameliorate periodontal trauma including disease and physical injury.
Composition of cryopreserved isolated periodontal ligament multipotent stem cells
A composition consisting of a cryopreservation solution and an effective amount of isolated clonal human periodontal ligament multipotent stem cells expanded ex vivo that proliferate, express STRO-1 and MUC18, form colonies in vitro, and differentiate into periodontal ligament cells producing collagen fibers and cementum in vivo, wherein the isolated population is cryopreserved in the cryopreservation solution, preferably in liquid nitrogen.
Genetically modified isolated clonal periodontal ligament multipotent stem cells
An isolated clonal population of human periodontal ligament multipotent stem cells expanded ex vivo that proliferate, express STRO-1 and MUC18, form colonies in vitro, differentiate into collagen and cementum producing cells in vivo, and are transfected with a preselected nucleic acid segment encoding hormones, chemokines, growth factors, growth factor receptors or cytokines.
The claims cover isolated human periodontal ligament multipotent stem cells characterized by marker expression and functional differentiation, compositions with various carriers, methods for generating periodontal tissue in subjects by implanting these compositions, cryopreserved forms of these cells, and genetically modified forms expressing therapeutic factors. These inventive features enable periodontal tissue regeneration for treatment of trauma including periodontal disease and dental injuries.
Stated Advantages
Provides a restorative method to generate lost periodontium biologically, including cementum, alveolar bone, and periodontal ligament.
Allows minimally invasive placement of stem cells to produce regenerative periodontal tissue, potentially reducing pain and costs associated with current surgical treatments.
Enables preservation and later recovery of functional periodontal ligament stem cells from cryopreserved human tissue, facilitating clinical use and stem cell therapies.
Documented Applications
Treatment of periodontal disease and other injuries to the periodontium by regenerating periodontal tissue including cementum and periodontal ligament.
Generation of adipose tissue in vivo by implantation of periodontal ligament stem cells for conditions requiring fat formation or replacement.
Cryopreservation of human periodontal ligament tissue as a practical approach for preserving stem cells for future therapeutic applications.
Potential use in repair of trauma from dental procedures or physical injury to periodontal tissues.
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