The effect of calcium phosphate microstructure on bone-related cells in vitro.
Microstructure is essential for inductive bone formation in calcium phosphate materials after soft tissue implantation. We hereby evaluated activities (cell attachment, proliferation, ALP/DNA and protein/DNA) of three types of cells cultured on three kinds of calcium phosphate ceramic discs to study how microstructure takes its role in inductive bone formation. Three kinds of biphasic calcium phosphate (BCP) ceramic discs with the same chemistry and the same dimension of 10.0 x 1.0 mm3 (BCP1150-P, BCP1150-D and BCP1300), either having similar micropore sizes and surface roughness but different surface area (BCP1150-P vs BCP1150-D) or having similar surface area but different micropore sizes and different roughness (BCP1150-D vs BCP1300), were prepared. Conventionally Culturing C2C12, human bone marrow stromal cells (HBMSC) and MC3T3-E1 cells on BCP discs showed that, surface roughness did not affect cell attachment, cell proliferation and ALP expression of all cell types evaluated, while surface area did affect cell functions. ALP/DNA of C2C12 on BCP1150-P, having larger surface area, was significantly higher than on BCP1300 and BCP1150-D. Furthermore, all cells cultured on all of the three kinds of BCPs pre-soaked in culture medium having additional rhBMP-2 had a higher ALP expression than the conventional cell culture. Comparing with on BCP1300 and BCP1150-D, ALP/DNA of all cells tested increased more on BCP1150-P after the discs were pre-soaked in culture medium with rhBMP-2. The results indicated that increasing surface areas, microstructured calcium phosphate materials might concentrate more proteins (including bone-inducing proteins) that differentiate inducible cells to osteogenic cells that form inductive bone.
Li
Xiaoming
X
Department of Tissue Regeneration, Institute for Biomedical Technology, University of Twente, 7522NB Enschede, The Netherlands. x.m.li@hotmail.com
van Blitterswijk
Clemens A
CA
Feng
Qingling
Q
Cui
Fuzhai
F
Watari
Fumio
F
D016475:3T3 Cells; D000327:Adsorption; D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000008:administration & dosage; D018786:Bone Substitutes / Q000737:chemistry / Q000494:pharmacology*; D002130:Calcium Phosphates / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D020099:Coated Materials, Biocompatible; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D008422:Materials Testing; D051379:Mice; D008855:Microscopy, Electron, Scanning; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects; D010012:Osteogenesis / Q000187:drug effects*; D010316:Particle Size; D011994:Recombinant Proteins / Q000008:administration & dosage; D017550:Spectroscopy, Fourier Transform Infrared; D013499:Surface Properties; D016212:Transforming Growth Factor beta / Q000008:administration & dosage; D014961:X-Ray Diffraction
C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D018786:Bone Substitutes; D002130:Calcium Phosphates; D020099:Coated Materials, Biocompatible; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C020243:calcium phosphate