Human Alveolar Bone Cells Research Articles

Proliferation/differentiation of osteoblastic human alveolar bone cell cultures in the presence of stainless steel corrosion products.

Human osteoblastic alveolar bone cells were cultured for 28 days in control conditions and in the presence of three non-lethal concentrations of AISI 316L stainless steel (SS) corrosion products. Cells were exposed to SS corrosion products in two experimental situations: (i) in selected stages of the incubation time (during the first, second, third and fourth week of culture); and (ii) during the 28 days incubation period. Cultures were characterized for cell proliferation, total protein content, alkaline phosphatase activity (ALP) and ability to form mineralized deposits; culture media was analyzed for ionized calcium (Ca) and phosphorus (P) concentrations throughout the incubation period. The presence of SS corrosion products during the different stages of the incubation period did not significantly affect the cell proliferation; however, a significant dose-dependent deleterious effect was observed on the levels and pattern of ALP activity, concentration of ionized Ca and P in the culture medium and, also, ability to form mineralized deposits, especially in cultures exposed during the first and second week of culture (respectively, lag phase and exponential cell growth phase). Similar effects were observed in cultures exposed to the SS corrosion products during the 28 days incubation period. However, the presence of such products during the third week (when the mineralization process occurs) and, also, during the fourth week, resulted in little or no significant effects on the behavior of alveolar bone cells. Results suggested that SS corrosion products above certain non-lethal concentrations may disturb the proliferation/differentiation relationship of osteoblastic human alveolar bone cell cultures.

Activated adult human alveolar bone cells: a new model of matrix mineralization

Activated adult human alveolar bone cells were isolated from 2-wk-old osteogenic tissue recuperated from dental implant surgeries following a two-step procedure. Osteogenic tissues were cultured as explant for 2 months. Cells began to migrate in the first 3 d and were confluent in 3-4 wk. However, adjacent to the explants, multicellular cell layers began to form in 10 d, and matrix mineralization was observed by 4 wk in these areas. These formations enlarged and by the end of the culture period, large diffuse matrix mineralization areas were observed. Light and electron microscopic observations confirmed the presence of a collagen matrix undergoing a mineralization process but showing important differences with the mineralized matrix tissue formed with a rat embryo calvaria bone cell system. This new model, using activated human alveolar bone cells, may provide a tool to investigate alveolar bone development and physiology and to set up new therapeutic approaches.

Cementum, Root Dentin and Bone Extracts Stimulate Chemotactic Behavior in Cells from Periodontal Tissue

In this paper, we used the extracts from cementum, bone, dentin and enamel, and compared the chemotactic behavior of human periodontal ligament (HPDL) cells, human gingival fibroblasts (HGF) and human alveolar bone (HAB) cells using modified Boyden chambers. The extracts, obtained using 4 M guanidine HCl (“G” extract), from cementum, root dentin, and bone had greater chemotactic effects than all the 4 M guanidine HCl/0.5 M EDTA extracts (“E” extract). HPDL cells and HGF exhibited higher chemotactic behavior than osteoblast-like HAB cells in response to cementum and root dentin “G” extracts. On the other hand, HAB cells showed the highest migratory activity in response to the bone “G” extract. The chemotactic activities of these extracts were reduced by heat- and trypsin-treatment. These results suggest that proteinaceous chemotactic factors exist in the cementum, root dentin and bone, and they appear to regulate the migration and orientation of HPDL cells, HGF and HAB cells during periodontal wound healing.

Mineralization in serially passaged human alveolar bone cells.

Well-characterized human bone cell cultures have been regarded as a useful tool to study bone control mechanisms and also to analyse bone/biomaterials interactions. In the present study, human alveolar bone cells were cultured in alpha-minimal essential medium (alpha-MEM) containing 10% foetal bovine serum (FBS), 50 microg/ml ascorbic acid, 10 mM sodium beta-glycerophosphate and either in the presence or in the absence of 10 nM dexamethasone (Dexa). Cultures were characterized concerning cell viability/proliferation, alkaline phosphatase (ALP), acid phosphatase (ACP) and tartraric acid resistant phosphatase (TRAP) activities, and formation of mineralized areas. Cell proliferation increased gradually for approximately 20 days. In the presence of Dexa, cells formed isolated or interconnected multilayered clusters that increased with culture time. Histochemical assays revealed strong positive reactions for ALP and calcium and phosphates deposits, mainly in relation t! o ce lls associated with the clusters. High levels of ALP activity (biochemical determination) were observed. Cells cultured in the absence of Dexa showed significantly lower ALP activity and no calcium and phosphates deposits were present. Serially passaged cells kept the proliferation rate constant but a decrease in ALP activity was observed either in the presence or in the absence of Dexa. The ability to form mineralized areas (cultures fed with Dexa) also decreased on serial subculture.

Characterization of Cementum Derived Growth Factor as an Insulin-Like Growth Factor-I Like Molecule

Cementum is the thin calcified outer layer through which tooth-root surfaces are anchored to soft periodontal connective tissues. A variety of growth factors and adhesion molecules are sequestered in the extracellular matrix of cementum, and we have purified and characterized one of the growth factors. This growth factor, the cementum derived growth factor (CGF), was purified from bovine cementum by acetic acid extraction followed by heparin affinity chromatography and HPLC using cation exchange, molecular sieve, and reverse-phase columns. NaDodSO4-polyacrylamide gel electrophoresis of purified CGF preparation revealed the presence of two major protein bands migrating with Mr 18,000–22,000 and 14,000–16,000. The latter was associated with the major part of the mitogenic activity. The activity of CGF was inhibited by antibodies to insulin-like growth factor-I (IGF-I) and IGF-I receptor. Both CGF and IGF-I were mitogenic to human gingival fibroblasts and alveolar bone cells, but the bone cells responded better to CGF than to IGF-I. The IGF-I did not bind to heparin-sepharose, while CGF bound to it and was eluted with 0.6M NaCl from heparin-sepharose columns. Heparin-sepharose 0.2M NaCl fractions of cementum extracts contained IGF-I migrating with Mr 7,500, but its mobility was not affected by N-glycosidase treatment. Western analysis using anti-IGF-I antibodies showed that CGF preparations contained cross-reacting species migrating with Mr 18,000–22,000, 14,000–16,000 and 11,000–12,000, however after treatment with N-glycosidase the Mr 18,000–22,000 component was absent. Internal amino acid sequences of six tryptic peptides of CGF were determined by microsequencing. The sequence of one 15-amino acid long peptide was the same as the receptor binding domain of IGF-I, and another 9-amino acid peptide had 78% homology to a sequence derived from an untranslated region of sheep IGF-I exon 1. Four other peptides had no apparent homology with IGF-I. From these results we conclude that the CGF is an IGF-I like molecule.

In vitro osteogenesis by human alveolar bone cells in the presence of dexamethasone

In vitro osteogenesis by human alveolar bone cells in the presence of dexamethasone

Alkaline phosphatase of human periodontal ligament fibroblast-like cells.

Cultures of human periodontal ligament fibroblasts (HPLF) and human alveolar bone cells (HABC) were established, maintained, and characterized with respect to alkaline phosphatase (ALPase) activity, a marker enzyme of bone cells. During the proliferative phase, ALPase of HPLF and HABC were not detectable. But at the confluent stage, HPLF synthesized levels of ALPase similar to HABC. Also, ALPase activity of HPLF was markedly stimulated (7.5-fold) by 5 x 10-9 mol/L 1,25 (OH)2-vitamin D3 (1,25 (OH)2-D3). Therefore, since ALPase activity of HPLF resembles that present in osteoblasts, human periodontal ligament fibroblasts may function as "osteoblastic" fibroblasts.