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MV-mimicking micelles loaded with PEG-serine-ACP nanoparticles to achieve biomimetic intra/extra fibrillar mineralization of collagen in vitro
Authors:Minjuan Shen  Mingli Lin  Mengqi Zhu  Wenxin Zhang  Danyang Lu  Huanhuan Liu  Jingjing Deng  Kehua Que  Xu Zhang
Institution:School of Stomatology, Hospital of Stomatology, Tianjin Medical University, 12 Observatory Road, Tianjin 300070, China
Abstract:Since their discovery, matrix vesicles (MVs) containing minerals have received considerable attention for their role in the mineralization of bone, dentin and calcified cartilage. Additionally, MVs' association with collagen fibrils, which serve as the scaffold for calcification in the organic matrix, has been repeatedly highlighted. The primary purpose of the present study was to establish a MVs–mimicking model (PEG-S-ACP/micelle) in vitro for studying the exact mechanism of MVs-mediated extra/intra fibrillar mineralization of collagen in vivo. In this study, high-concentration serine was used to stabilize the amorphous calcium phosphate (S-ACP), which was subsequently mixed with polyethylene glycol (PEG) to form PEG-S-ACP nanoparticles. The nanoparticles were loaded in the polysorbate 80 micelle through a micelle self-assembly process in an aqueous environment. This MVs–mimicking model is referred to as the PEG-S-ACP/micelle model. By adjusting the pH and surface tension of the PEG-S-ACP/micelle, two forms of minerals (crystalline mineral nodules and ACP nanoparticles) were released to achieve the extrafibrillar and intrafibrillar mineralization, respectively. This in vitro mineralization process reproduced the mineral nodules mediating in vivo extrafibrillar mineralization and provided key insights into a possible mechanism of biomineralization by which in vivo intrafibrillar mineralization could be induced by ACP nanoparticles released from MVs. Also, the PEG-S-ACP/micelle model provides a promising methodology to prepare mineralized collagen scaffolds for repairing bone defects in bone tissue engineering.
Keywords:Matrix vesicles  Extra/intra mineralization of collagen fibrils  Biomimetic mineralization  Amorphous calcium phosphate  Micelle  MVs  matrix vesicles  PEG  polyethylene glycol  ACP  amorphous calcium phosphate  HAP  hydroxyapatite  NCPs  non-collagenous proteins  PASP  polyaspartic acid  PAA  polyacrylic acid  PAH  poly(allylamine)hydrochloride  CMC  carboxylmethly chitosan  ECM  extracellular matrix  DPPC  dipalmitoylphosphatidylcholine  DMPC  dimyristoylphosphatidylcholine  DLPA  dilauroyl phosphatidic acid  AAs  amino acids  Glu  Asp  Lys  Ser  PBS  phosphate-buffered saline  EDC  1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide  NHS  IPA  isopropanol  DLS  dynamic light scattering  XRD  x-ray diffraction  CMC  critical micelle concentration  TEM  transmission electron microscope  SAED  selected area electron diffraction  ATR-FTIR  attenuated total reflection–fourier transform infrared spectroscopy  ICP-OES  inductively coupled plasma-atomic emission spectroscopy  SEM  scanning electron microscopy  EE  encapsulation efficiency  SD  standard deviation  EEs  encapsulation efficiencies  ANOVA  analysis of variance  SBE  sigmoid-boltzman equation  
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