title: Establishment of a 3D-dynamic osteoblasts-osteoclasts co-culture model to simulate the jawbone microenvironment in vitro.
abstract: OBJECTIVE
  We aimed to establish a 3D osteoblasts/osteoclasts co-culture system requiring limited amounts of human primary cells and useful as platform to 1. recapitulate an "oral bone microenvironment" in healthy or pathological condition, and 2. produce potential implantable cell constructs for regeneration of jawbone which can be negatively affected by bisphosphonates (BPs).
  
  
  METHODS
  Osteoblasts from normal bone chips (hOBs) or from jawbone of patients taking BPs (hnOBs) were co-cultured with monocytes (hMCs) either in static (3D-C) or dynamic (3D-DyC) condition using the RCCS-4™ bioreactor for 3weeks. Cell aggregates were characterized for viability, histological features and specific osteoclastic and osteogenic markers.
  
  
  RESULTS
  In all tested conditions hOBs supported the formation of mature osteoclasts (hOCs), without differentiating agents or exogenous scaffolds. 3D-DyC condition associated with a ground based condition (Xg) rather than modeled microgravity (μXg) produced aggregates with high level of osteogenic markers including Osteopontin (OPN), Osterix (OSX), Runx2 and appreciable bone mineral matrix. hnOBs co-cultured with hMCs in 3D-Dyc/Xg condition generated OPN and mineral matrix positive aggregates.
  
  
  CONCLUSIONS
  We optimized a 3D co-culture system with a limited amount of cells preserving viability and functionality of bone cellular components and generating bone-like aggregates also by using cells from jawbone necrotic tissue. The feasibility to obtain from poor-quality bone sites viable osteoblasts able to form aggregates when co-cultured with hMCs, allows to study the development of autologous implantable constructs to overcome jawbone deficiency in patients affected by MRONJ (Medication-Related Osteonecrosis of the Jaws).
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
authors.forename: Letizia
authors.initials: L
authors.lastname: Penolazzi
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Lolli
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Sardelli
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Angelozzi
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Lambertini
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Trombelli
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Ciarpella
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy.
authors.forename: Renata
authors.initials: R
authors.lastname: Vecchiatini
authors.affiliation: Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Italy. Electronic address: piv@unife.it.
authors.forename: Roberta
authors.initials: R
authors.lastname: Piva
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D050071:Bone Density Conservation Agents / Q000494:pharmacology; D001842:Bone and Bones / Q000166:cytology; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques; D004164:Diphosphonates / Q000494:pharmacology; D004777:Environment; D005260:Female; D006801:Humans; D007568:Jaw / Q000166:cytology*; D008297:Male; D008334:Mandible / Q000166:cytology; D008875:Middle Aged; D009000:Monocytes / Q000187:drug effects; D009336:Necrosis; D010006:Osteoblasts / Q000187:drug effects / Q000502:physiology*; D010010:Osteoclasts / Q000187:drug effects / Q000502:physiology*; D014893:Weightlessness
chemical_list: D015415:Biomarkers; D050071:Bone Density Conservation Agents; D004164:Diphosphonates
keywords: 3D cell co-culture; MRONJ; Osteoblasts; Osteoclasts