Human osteoblast cultures from osteoporotic and healthy bone: biochemical markers and cytokine expression in basal conditions and in response to 1,25(OH)2D3.
The development of methods to culture bone cells has enhanced in vitro studies and allowed researchers to investigate bone cell metabolism in healthy tissue and in various different bone diseases. Greater knowledge of cultures of pathologic bone tissue-derived osteoblasts may be helpful in performing in vitro experiments that test biomaterials and therapies to be used in the orthopedic field, since this kind of approach better reflects the conditions of clinical relevance to many patients. In the present study primary cultures of human osteoblastic cells were isolated from donors with osteoporosis (HOB, Human Osteopenic Bone) and their respective controls (HNB, Human Normal Bone). They were then characterized in baseline conditions and after stimulation with 10(-9) M 1,25(OH2)D3. Specific biochemical markers of bone cells and cytokines involved in bone turnover were evaluated to assess cell metabolism and any possible differences between osteoblasts derived from healthy and osteopenic bone tissue. Under baseline conditions, HNB and HOB in vitro cultures showed some differences in proliferation (MTT test), PICP, OC and IL-6. The HNB response to 1,25-(OH2)D3 stimulation differed significantly from that of the HOB cultures but only with regard to the MTT test, and ALP and PICP levels; the other selected parameters showed a similar behavior for both cultures. The current findings should be taken into account when cultures derived from human bone are used for in vitro experiments.
Torricelli
Paola
P
Experimental Surgery Department, Research Institute Codivilla-Putti, Bologna, Italy. paola.torricelli@ior.it
Fini
Milena
M
Giavaresi
Gianluca
G
Giardino
Roberto
R
D000469:Alkaline Phosphatase / Q000378:metabolism; D015415:Biomarkers; D002117:Calcitriol / Q000494:pharmacology; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis*; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis; D010006:Osteoblasts / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism* / Q000473:pathology; D015675:Osteocalcin / Q000378:metabolism; D010024:Osteoporosis / Q000276:immunology* / Q000378:metabolism* / Q000473:pathology; D010446:Peptide Fragments / Q000378:metabolism; D011347:Procollagen / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000096:biosynthesis; D053773:Transforming Growth Factor beta1
D015415:Biomarkers; D016207:Cytokines; D015850:Interleukin-6; D010446:Peptide Fragments; D011347:Procollagen; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; C042813:procollagen type I carboxy terminal peptide; D015675:Osteocalcin; D000469:Alkaline Phosphatase; D002117:Calcitriol