title: Osteoclastic bioresorption of biomaterials: two- and three-dimensional imaging and quantification.
abstract: OBJECTIVE
  Bioresorbable materials have been developed in the hope that the body will replace them with newly formed tissue. The first step of this remodeling process in bone is the bioresorption of the material by osteoclasts. The aim of this study was to analyze osteoclastic resorption of biomaterials in vitro using the commonly used two-dimensional methods of light-microscopy (LM) and scanning electron microscopy (SEM) in comparison with infinite focus microscopy (IFM), a recently developed imaging method allowing for three-dimensional surface analysis.
  
  
  METHODS
  Human hematopoietic stem cells were cultivated in the presence of the cytokines M-CSF and RANK-L for 4 weeks directly on dentin and a calcium phosphate cement. Osteoclast development was surveyed with standard techniques. After removal of the cells, resorption was characterized and quantified by LM, SEM and IFM.
  
  
  RESULTS
  Osteoclast cultures on the biomaterials presented the typical osteoclast-specific markers. On dentin samples LM, SEM as well as IFM allowed for discrimination of resorption. Quantification of the resorbed area showed a linear correlation between the results (LM vs. SEM: r=0.996, p=0.004; SEM vs. IFM: r=0.989, p=0.011; IFM vs. LM: r=0.995). It was not possible to demarcate resorption pits on GB14 using LM or SEM. With IFM, resorption on GB14 could be visualized and quantified two- and three-dimensionally.
  
  
  CONCLUSIONS
  In this paper we introduce IFM as a technology for three-dimensional visualization and quantification of resorption of biomaterials. Better understanding of the bioresorption of biomaterials may help in the design of better materials and might therefore constitute an important step on the avenue to the development of artificial bone.
authors.affiliation: Biomechanics Section, Hamburg University of Technology, Hamburg - Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Winkler
authors.forename: Elisa
authors.initials: E
authors.lastname: Hoenig
authors.forename: Gerd
authors.initials: G
authors.lastname: Huber
authors.forename: Rolf
authors.initials: R
authors.lastname: Janssen
authors.forename: Daniel
authors.initials: D
authors.lastname: Fritsch
authors.forename: Renate
authors.initials: R
authors.lastname: Gildenhaar
authors.forename: Georg
authors.initials: G
authors.lastname: Berger
authors.forename: Michael M
authors.initials: MM
authors.lastname: Morlock
authors.forename: Arndt F
authors.initials: AF
authors.lastname: Schilling
mesh_terms: D000042:Absorption; D001672:Biocompatible Materials / Q000737:chemistry*; D001843:Bone Cements; D001862:Bone Resorption / Q000503:physiopathology; D002130:Calcium Phosphates / Q000737:chemistry; D002478:Cells, Cultured; D003804:Dentin / Q000737:chemistry; D006412:Hematopoietic Stem Cells; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008422:Materials Testing / Q000379:methods; D010010:Osteoclasts / Q000378:metabolism*; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods
chemical_list: D001672:Biocompatible Materials; D001843:Bone Cements; D002130:Calcium Phosphates; C020243:calcium phosphate