title: Periodontal regeneration with stem cells-seeded collagen-hydroxyapatite scaffold.
abstract: Re-establishing compromised periodontium to its original structure, properties and function is demanding, but also challenging, for successful orthodontic treatment. In this study, the periodontal regeneration capability of collagen-hydroxyapatite scaffolds, seeded with bone marrow stem cells, was investigated in a canine labial alveolar bone defect model. Bone marrow stem cells were isolated, expanded and characterized. Porous collagen-hydroxyapatite scaffold and cross-linked collagen-hydroxyapatite scaffold were prepared. Attachment, migration, proliferation and morphology of bone marrow stem cells, co-cultured with porous collagen-hydroxyapatite or cross-linked collagen-hydroxyapatite, were evaluated in vitro. The periodontal regeneration capability of collagen-hydroxyapatite scaffold with or without bone marrow stem cells was tested in six beagle dogs, with each dog carrying one sham-operated site as healthy control, and three labial alveolar bone defects untreated to allow natural healing, treated with bone marrow stem cells - collagen-hydroxyapatite scaffold implant or collagen-hydroxyapatite scaffold implant, respectively. Animals were euthanized at 3 and 6 months (3 animals per group) after implantation and the resected maxillary and mandibular segments were examined using micro-computed tomography scan, H&E staining, Masson's staining and histometric evaluation. Bone marrow stem cells were successfully isolated and demonstrated self-renewal and multi-potency in vitro. The porous collagen-hydroxyapatite and cross-linked collagen-hydroxyapatite had average pore sizes of 415 ± 20 µm and 203 ± 18 µm and porosity of 69 ± 0.5% and 50 ± 0.2%, respectively. The attachment, proliferation and migration of bone marrow stem cells were satisfactory on both porous collagen-hydroxyapatite and cross-linked collagen-hydroxyapatite scaffolds. Implantation of bone marrow stem cells - collagen-hydroxyapatite or collagen-hydroxyapatite scaffold in beagle dogs with experimental periodontal defects resulted in significantly enhanced periodontal regeneration characterized by formation of new bone, periodontal ligament and cementum, compared with the untreated defects, as evidenced by histological and micro-computed tomography examinations. The prepared collagen-hydroxyapatite scaffolds possess favorable bio-compatibility. The bone marrow stem cells - collagen-hydroxyapatite and collagen-hydroxyapatite scaffold - induced periodontal regeneration, with no aberrant events complicating the regenerative process. Further research is necessary to improve the bone marrow stem cells behavior in collagen-hydroxyapatite scaffolds after implantation.
authors.affiliation: State Key Laboratory of Oral Diseases and Department of Orthodontics, Sichuan University West China College of Stomatology, Chengdu, China Chengdu Women and Chidren Central Hospital, 1617 Riyue Avenue, Qingyang District, Chengdu, China.
authors.forename: Zeping
authors.initials: Z
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Oral Diseases and Department of Orthodontics, Sichuan University West China College of Stomatology, Chengdu, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Yin
authors.affiliation: State Key Laboratory of Oral Diseases and Department of Orthodontics, Sichuan University West China College of Stomatology, Chengdu, China School of Dentistry, University of Queensland, Brisbane, Australia.
authors.forename: Qingsong
authors.initials: Q
authors.lastname: Ye
authors.affiliation: State Key Laboratory of Oral Diseases and Department of Orthodontics, Sichuan University West China College of Stomatology, Chengdu, China.
authors.forename: Wulin
authors.initials: W
authors.lastname: He
authors.affiliation: State Key Laboratory of Oral Diseases and Department of Orthodontics, Sichuan University West China College of Stomatology, Chengdu, China.
authors.forename: Mengke
authors.initials: M
authors.lastname: Ge
authors.affiliation: The Number Seventeen Middle School of Chengdu, China.
authors.forename: Xiaofu
authors.initials: X
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Oral Diseases and Department of Oral and Maxillofacial Surgery, Sichuan University West China College of Stomatology, Chengdu, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Oral Diseases and Department of Orthodontics, Sichuan University West China College of Stomatology, Chengdu, China drzsj@scu.edu.cn.
authors.forename: Shujuan
authors.initials: S
authors.lastname: Zou
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry*; D004285:Dogs; D017886:Durapatite / Q000737:chemistry*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D016557:Guided Tissue Regeneration, Periodontal / Q000295:instrumentation* / Q000379:methods; D008297:Male; D010510:Periodontal Diseases / Q000473:pathology* / Q000628:therapy*; D010519:Periodontium / Q000166:cytology / Q000254:growth & development; D016062:Porosity; D033581:Stem Cell Transplantation / Q000295:instrumentation* / Q000379:methods; D013234:Stem Cells / Q000166:cytology / Q000502:physiology; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods; D054457:Tissue Scaffolds; D016896:Treatment Outcome
chemical_list: D003094:Collagen; D017886:Durapatite
keywords: Tissue engineering; bone marrow stem cells; collagen-hydroxyapatite; composite scaffold; micro-computed tomography; periodontal regeneration