J Korean Assoc Oral Maxillofac Surg.  1997 Jan;23(1):43-62.

Experimental study on osteogenesis capacity of allogeneic bone grafts according to various bone preparation (Defatting and demineralization)

• Lee DK

Abstract

Allogeneic demineralized bone powder has been shown to support the osteogenesis in a variety of bone defect and extra-osseous sites. Previous several experiments have been carried out in animal models which have time-consuming and various problems. Therefore author designed some cytotoxicity tests by using of 5-diphenyl dimethyltetrazolium bromide(MTT) assay, neutral red(NR) assay, lactate dehydrogenase(LDH) activity test, sulforbodamine B (SRB) assay, and alkaline phosphatase(ALP) assay. And these method seemed to be a rapid, repeatable and believable reaction. To establish the osteogenesis capacity of allogeneic bone according to various bone preparation(defatting and demineralization), author studied the effect of allogeneic bone on well-established rat 3T3 fibroblasts and MC3TEl osteoblasts in culture to evaluate a cytotoxicity and examined the surface of allogeneic bone with scanning electron microscopy(SEM) for detecting the porositv. Defatting Demineralized Bone Powder(DDBP0 and Non-Defatting Demineralized Bone Powder(NDDBP) of particle size 212mum were prepared from rat calvarial bone and long bones. 1. Cell counts of rat 3T3 fibroblasts and MC3T3E1 osteoblasts were greater in DDBP enriched medium when compared with medium with NDDBP. Cell counts with DDBP were to different from controls. 2. Author found a little differences in cytotoxicity on 3T3 fibroblast and MC3T3E1 osteoblasts between DDBP and NDDBP with MTT assay, NR assay, LDH activity test, SRB assay, Generally, DDBP was represented less cytotoxicity than NDDBP. 3. ALP assay with MC3T3E1 osteoblasts in culture presented the less cytotoxicity and more crowding around the DDBP than NDDBP. With these findings, defatting procedure on making the demineralized bone powder have a merit for supporting osteogenesis. I should be suggested that cell culture with DDBP and NDDBP is a good method for studying induced osteogenesis.

MeSH Terms

Animals
Cell Count
Cell Culture Techniques
Crowding
Fibroblasts
Lactic Acid
Models, Animal
Osteoblasts
Osteogenesis*
Particle Size
Rats
Transplants*
Lactic Acid