Fibroblastic cells from human periapical granulation tissue preferentially form calcified matrices in decalcified boiled rat bone.
We have been studying the potential of human fibroblastic cells (HFC) from periapical granulation tissue to form a calcified matrix. Recently, we reported that inflamed periapical granulation tissue contains osteogenic cells. In the present study, we tested the hypothesis that HFC, cultured with decalcified bone (DB) of rat, might form much greater calcified matrices than with rat decalcified boiled bone (DBB), which was originally prepared as a negative control. HFC were cultured with DB or DBB in the presence or absence of 2 mM beta-glycerophosphate (beta-GP) and 50 microg/ml ascorbic acid. After six weeks of culture, a number of von Kossa-positive globular structures were unexpectedly observed inside DBB, but not DB. Without HFC, such structures were never seen in DBB incubated with 2 mM beta-GP and 50 microg/ml ascorbic acid. DB cultured with HFC under the same conditions did not show these structures. Electron-microscopic observation revealed that matrix vesicles aggregated on collagen fibrils around globular structures in DBB. Energy dispersive X-ray microanalysis confirmed that these structures were calcified matrices composed of calcium and phosphate. These results suggest that human periapical granulation tissue contains cells responsible for the formation of calcified matrices in DBB, and that DBB could serve as an excellent scaffold for the calcification of HFC, rather than DB.
Maeda
Hidefumi
H
Department of Endodontology and Operative Dentistry, Division of Oral Rehabilitation, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi, Fukuoka, 812-8582, Japan. hide@dent.kyushu-u.ac.jp
Wada
Naohisa
N
Fujii
Shinsuke
S
Akamine
Akifumi
A
D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D001842:Bone and Bones / Q000166:cytology* / Q000648:ultrastructure; D002113:Calcification, Physiologic / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D004577:Electron Probe Microanalysis; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D005994:Glycerophosphates / Q000494:pharmacology; D006097:Granulation Tissue / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D010486:Periapical Tissue / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D005994:Glycerophosphates; D001205:Ascorbic Acid; C031463:beta-glycerophosphoric acid