Stem cell‐derived conditioned media from human exfoliated deciduous teeth promote bone regeneration

• Published in: Oral diseases Vol. 26; no. 2; pp. 381 - 390
• Main Authors: Hiraki, Tomoka, Kunimatsu, Ryo, Nakajima, Kengo, Abe, Takaharu, Yamada, Sakura, Rikitake, Kodai, Tanimoto, Kotaro
• Format: Journal Article
• Language: English
• Published: Denmark Wiley Subscription Services, Inc 01-03-2020
• Subjects: Alveolar bone; Angiogenesis; Animals; Atelocollagen; Bone grafts; Bone growth; bone regeneration; Bone Regeneration - drug effects; CD34 antigen; Cell culture; Child; Cleft Lip - surgery; Cleft lip/palate; Cleft Palate - surgery; Computed tomography; conditioned media; Culture Media, Conditioned - pharmacology; Defects; Dental pulp; Dental Pulp - physiology; Dentistry; exfoliated deciduous teeth; Growth factors; Humans; Immunodeficiency; Immunohistochemistry; Male; Mesenchyme; Mice; Mice, Inbred BALB C; Osteogenesis; Osteogenesis - drug effects; Paracrine signalling; Regeneration; Secretome; Stem cell transplantation; Stem cells; Stem Cells - physiology; Tooth Exfoliation; Tooth, Deciduous - cytology; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - metabolism; X-Ray Microtomography; exfoliated deciduous teeth; stem cells; bone regeneration; conditioned media
• ISSN: 1354-523X; 1601-0825
• DOI: 10.1111/odi.13244

Objectives
Cleft lip and palate (CL/P) are common congenital orofacial anomalies. Autogenous iliac bone grafting closes alveolar cleft defects but requires surgical intervention. Mesenchymal stem cell culture supernatant can regenerate tissues via paracrine activity. However, little is known about the bone‐regenerative effects of stem cells from human exfoliated deciduous teeth (SHED) and conditioned media (CM). Our aim was to address this.
Materials and methods
Stem cells were isolated from primary tooth pulp and cultured. Defects were made in calvariae of immunodeficient mice and implanted with stem cell‐ or CM‐containing atelocollagen. Regenerated bone was analysed by microcomputed tomography, haematoxylin–eosin and Masson's trichrome staining. Vascular endothelial growth factor, CD31 and CD34 expression were confirmed by immunohistochemistry, and the presence of several proteins and growth factors was verified in SHED‐CM.
Results
Bone regeneration was enhanced in defects treated with stem cells and CM compared to that in controls 8 weeks after transplantation. Mature bone formation and angiogenesis were confirmed with CM but not with stem cells or in controls. Secretome analysis using multiple cytokine assays revealed that SHED‐CM contained tissue‐regenerating factors with roles in angiogenesis and osteogenesis.
Conclusion
CM non‐invasively regenerate bone and might be effective to reconstruct alveolar clefts in CL/P patients.