Manu Krishnan1, Anu Sharma2, Seema Saraswathy3, Brijesh Tiwari4, Gurudatta Ganganahalli5, Sanjay Londhe6, Ajay Kumar Singh7, Velu Nair8. 1. Classified Specialist (Orthodontics), Department of Dental Research & Implantology, Institute of Nuclear Medicine and Allied Sciences (INMAS), Timarpur, Delhi 110054, India. 2. PhD Research Scholar, Department of Dental Research & Implantology, Institute of Nuclear Medicine and Allied Sciences (INMAS), Timarpur, Delhi 110054, India. 3. Department of Biochemistry, Army College of Medical Sciences (ACMS), Delhi Cantt, India. 4. Project Dental Officer, Department of Dental Research & Implantology, Institute of Nuclear Medicine and Allied Sciences (INMAS), Timarpur, Delhi, India. 5. Scientist 'G' & Addl Director, Head, Division of Stem Cell and Gene Therapy Research, Institute of Nuclear Medicine and Allied Sciences (INMAS), Timarpur, Delhi, India. 6. Director General Dental Services (DGDS), IHQ of MoD, L Block, New Delhi, India. 7. Scientist 'H' & Director, Institute of Nuclear Medicine and Allied Sciences (INMAS), Ministry of Defence, Govt of India, Timarpur, Delhi, India. 8. Senior Consultant, Haemato-Oncology & Bone Marrow Transplant, Comprehensive Blood & Cancer Center (CBCC), 632, C-1, Ansals Palam Vihar, Carterpuri, Gurgaon 122017, India.
Abstract
BACKGROUND: Considering the limitations in isolating Bone Marrow Mesenchymal Stem Cells (BMSCs), alternate sources of Mesenchymal Stem Cells (MSCs) are being intensely investigated. This study evaluated dental pulp MSCs (DP-MSCs) isolated from orthodontically extracted premolar teeth from a bone tissue engineering perspective. METHODS: MSCs isolated from premolar teeth pulp were cultured and studied using BMSCs as the control. Flow cytometry analysis was performed for the positive and negative MSC markers. Multilineage differentiation focusing on bone regeneration was evaluated by specific growth induction culturing media and by alkaline phosphatase (ALP) activity. Data were compared by repeated measurement analysis of variance and Student's t-test at a p value <0.05. RESULTS: Proliferation rate, population doubling time, and colony formation of DP-MSCs were significantly higher (p < 0.001) than BMSCs. More than 85% of DP-MSCs expressed CD44, CD73, CD90, CD105, and CD166. Negative reaction was found for CD11b CD33, CD34, and CD45. Positive reaction was displayed by 7.2% of cells for early MSC marker, Stro-1. Both the cell populations differentiated into adipogenic, osteogenic, and chondrogenic lineages, with adequate ALP expression. CONCLUSION: Because DP-MSCs from orthodontic premolars hold a neural crest/ectomesenchymal ancestry, its prudent growth characteristics and multilineage differentiation open up exciting options in craniofacial tissue engineering.
BACKGROUND: Considering the limitations in isolating Bone Marrow Mesenchymal Stem Cells (BMSCs), alternate sources of Mesenchymal Stem Cells (MSCs) are being intensely investigated. This study evaluated dental pulp MSCs (DP-MSCs) isolated from orthodontically extracted premolar teeth from a bone tissue engineering perspective. METHODS: MSCs isolated from premolar teeth pulp were cultured and studied using BMSCs as the control. Flow cytometry analysis was performed for the positive and negative MSC markers. Multilineage differentiation focusing on bone regeneration was evaluated by specific growth induction culturing media and by alkaline phosphatase (ALP) activity. Data were compared by repeated measurement analysis of variance and Student's t-test at a p value <0.05. RESULTS: Proliferation rate, population doubling time, and colony formation of DP-MSCs were significantly higher (p < 0.001) than BMSCs. More than 85% of DP-MSCs expressed CD44, CD73, CD90, CD105, and CD166. Negative reaction was found for CD11b CD33, CD34, and CD45. Positive reaction was displayed by 7.2% of cells for early MSC marker, Stro-1. Both the cell populations differentiated into adipogenic, osteogenic, and chondrogenic lineages, with adequate ALP expression. CONCLUSION: Because DP-MSCs from orthodontic premolars hold a neural crest/ectomesenchymal ancestry, its prudent growth characteristics and multilineage differentiation open up exciting options in craniofacial tissue engineering.
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