Anionic chlorido(triphenyl)tin(IV) bearing N-phthaloylglycinato or 1,2,4-benzenetricarboxylato 1,2-anhydride ligands: potential cytotoxic and apoptosis-inducing agents against several types of cancer.

Two ionic triphenyltin(IV) chloride carboxylate compounds of the formula [NHEt3 ][Ph3 SnCl(L)] [LH = N-phthaloylglycine (P-GlyH), 1; 1,2,4-benzenetricarboxylic 1,2-anhydride (BTCH), 2] were tested for the in vitro activity against 518A2 (melanoma), FaDu (head and neck carcinoma), HT-29 (colon cancer), MCF-7 (breast carcinoma), and SW1736 (thyroid cancer) cell lines. The ammonium salts of the carboxylic acids are found to be not active, while anionic [Ph3 SnCl(L)]- exhibited high cytotoxicity in nM range, both higher activity and selectivity than cisplatin. Compounds 1 and 2 are inducing apoptosis, which was proved with the morphological and biochemical features such as membrane blebbing, translocation of phosphatidylserine, and DNA fragmentation. Thus, accumulation of cells in sub-G1 phase is observed. Both anionic organotin(IV) compounds showed potent cytotoxic and apoptotic properties against five cancer cell lines of various histogenetic origin.

Titanium dental implant surfaces obtained by anodic spark deposition - From the past to the future.

Commercial titanium-based dental implants are obtained applying various methods such as machining, acid etching, anodization, plasma spraying, grit blasting or combination techniques yielding materials with smooth or micro-roughened surfaces. Those techniques are used to optimize the surface properties and to maximize biocompatibility and bioactivity with bone tissue. Present review is focused on the material surfaces obtained by anodic spark deposition (ASD). From the early 1980s till present, the results of numerous studies have shown that anodically oxidized surfaces with different dopants express a positive effect on osteoblasts behavior in vitro and osseointegration in vivo. Those surfaces demonstrated a high biocompatibility and rapid osseointegration in clinical application. This paper provides an overview of the preparation of implant surfaces by employing ASD process. Moreover, reviewed are clinically used ASD implant surfaces (Ticer, TiUnite, Osstem, etc.). The electrolyte variations in ASD process and their influence on surface properties are given herein. Using different electrolytes, anode voltages and temperatures, the above fabrication process can yield various surface morphologies from smooth to rough, porous surfaces. Furthermore, ASD enables thickening of oxide layers and enrichment with different dopands from used electrolyte, which hinder release of potentially toxic titanium ions in surrounding tissue. Particularly exciting results were achieved by calcium and phosphorus doping of the oxide layer (Ticer, ZL Microdent; TiUnite, Nobel Biocare Holding AB) which significantly increased the osteocompatibility. Ticer, a dental implant with anodically oxidized surface and the first among similar materials employed in clinical practice, was found to promote fast osteoblast cell differentiation and mineralization processes. Moreover, Ticer accelerate the integration with the bone, increase the bone/implant contact and improve primary and secondary stability of the implants. Additionally, potential innovations in this field such as fabrication of nanotubes on the implant surfaces as well as novel approaches (e.g. coating with proteins, nanostructured topography; combining implant body and surface derived from titanium and zirconia) are elaborated in this review. Besides, biochemical aspects on implant surface cell/tissue interaction are summarized. From the clinical point of view implant surfaces fabricated by ASD technology possess fast and improved osseointegration, high healing rates and long term prognosis.

Anticancer Activity of Organogallium(III) Complexes in Colon Cancer Cells.

In vitro antitumor activity of various organogallium(III) complexes (1-8) has been tested against CT26CL25, HCT116, SW480 colon cancer cell lines. CV and MTT assays were used to assess on the antiproliferative effect of investigated organogallium(III) complexes. From the investigated complexes, the most active was found to be tetranuclear compound 8 against CT26CL25 cells. Flow cytometric analysis of the CT26CL25 cells upon the treatment with 8 was performed in order to determine the role of apoptosis, caspase activation, autophagy and proliferation rate on the cell death caused with this compound. Results indicate cytotoxic potential of the tetranuclear complex 8 by inducing caspase independent apoptosis and blocking most of the cells before first division.

Plasma-electrochemical deposition of porous zirconia on titanium-based dental material and in vitro interactions with primary osteoblasts cells.

Three new porous zirconia-coated titanium materials using anodic plasma-electrochemical oxidation have been fabricated and characterized by scanning electron microscopy, electron probe microanalysis and X-ray diffraction. These ZrO2/TiO2 surfaces contained up to 43 wt% of ZrO2, 49 wt% TiO2 ( M1: - M3: ) and 8 wt% P2O5 ( M2: , M3: ). Zirconium titanate was detected as dominant microcrystalline phase. Primary human osteoblast cells were used for in vitro investigations. Cell proliferation and immunohistochemical analyses of morphology and expression of bone sialoprotein and osteocalcin were performed. Novel coatings M2: and M3: were shown to induce proliferation and expression of osteocalcin and bone sialoprotein to the extent comparable to that of Ticer, a material already employed in clinical practice.

Physical vapour deposition of zirconia on titanium: fabrication, characterization and interaction with human osteoblast cells.

The physical vapor deposition of zirconia was used to prepare two new titanium-based surfaces M1 and M2 with a different layer thickness. These novel surfaces were characterized for chemistry, topography and morphology by surface and solid state techniques. Primary osteoblast cells were used for in vitro studies. DAPI assay was applied for cell proliferation, while for bone sialoprotein (BSP), osteonectin and transforming growth factor-ß (TGF-ß) expression immunohistochemical analyses were employed. Materials M1 and M2 affected cell proliferation accordingly to their surface roughness with their impact on cell number being between the impact of two rough (Ticer, SS) and two smooth surfaces (Ti cp and Cercon). Different influence of the investigated materials on the osteoblastic production of BSP (all materials similar impact), ON (Cercon-higher; SS-lower for others) and TGF-ß (Cercon different) was found.

Improved in vitro antitumor potential of (O,O'-Diisobutyl-ethylenediamine-N,N'-di-3-propionate)tetrachloridoplatinum(IV) complex under normoxic and hypoxic conditions.

(O,O'-Diisobutyl-ethylenediamine-N,N'-di-3-propionate)tetrachloridoplatinum(IV), [PtCl4(iBu2eddp)], shows an improved pharmacological profile in comparison to cisplatin. This is manifested through accelerated dying process led by necrotic cell death, reflected through mitochondrial collapse, strong ATP depletion and reactive oxygen species production. Loss of mitochondrial potential was further followed with intensive apoptosis that finalized with DNA fragmentation. Different dynamic of tumoricidal action could be partly ascribed to less affected repair mechanisms in comparison to cisplatin. Importantly, [PtCl4(iBu2eddp)] did not induce necrosis in primary fibroblasts suggesting different intracellular response of normal vs. tumor cells. This selectivity toward malignant phenotype is further confirmed by retained tumoricidal potential in hypoxic conditions, while cisplatin became completely inefficient.

Zirconia coated titanium for implants and their interactions with osteoblast cells.

The anodic plasma-electrochemical oxidation in aqueous electrolytes of Zr(SO4)2 was used to prepare new zirconia/titania-based surfaces M1 (Ti, Zr and O: 7-10, 22-27 and 65-69 at.%) and M2 (Ti, Zr and O: 11-13, 20-23 and 64-69 at.%). The chemical composition and the microstructure of these coatings were characterized by surface and solid state techniques such as scanning electron microscopy, electron probe microanalysis, Raman spectroscopy and X-ray diffraction. These mixed oxides of ZrO2/TiO2 surfaces consist up to 84% (m/m) of ZrO2 and 16% (m/m) of TiO2. Monoclinic zirconia was detected as the dominant microcrystalline phase. In vitro studies were conducted on primary human osteoblast cells. MTT and DAPI assays were used for assessment on cell proliferation. Immunohistochemical analyses of morphology, cell cluster formation and expression of bone sialoprotein (BSP) and osteocalcin (OC) were performed. Novel surfaces M1 and M2 induced proliferation and expression of OC and BSP similarly to Ticer, used in clinical practice. Furthermore, the presence of zirconia on titanium surface has a higher beneficial effect on the osteoblast morphological changes and cell cluster formation.

First titanium dental implants with white surfaces: preparation and in vitro tests.

OBJECTIVES: To evaluate first white titanium surfaces developed for improvement of existing clinically used titanium-based implants Ticer. METHODS: The anodic plasma-electrochemical oxidation in aqueous solutions of sodium hydroxide and calcium dihydrogen phospate was used to prepare three novel anodic conversion layers with white titanium oxide surfaces. The surfaces have been characterized by the means of scanning electron microscopy, surface microanalysis and X-ray diffraction. In vitro studies were conducted on primary human osteoblast cells using novel surfaces (M1-M3) as well as commercially pure titanium (Ti cp), Ticer and SS (subtracted surface). An indirect toxicity test using MTT and SRB assays has been carried out. Furthermore, immunohistochemical analysis of cell proliferation, morphology, and expression of non-collagenous bone matrix proteins (sialoprotein, BSP, and osteocalcin, OC) were performed. RESULTS: The basic morphology of the surfaces shows clusters in a size of 100 µm of knob-like structures. The coatings are composed of rutile and monoclinic sodium titanates. Novel white surfaces (M1-M3) induced proliferation rates, morphological changes and influenced the expression of OC and BSP similarly to Ticer. On the other hand, Ti cp and SS exhibited different in vitro behavior. SIGNIFICANCE: The novel surfaces expressed similar in vitro behavior as Ticer, successfully used in clinical practice. Furthermore, due to their white color they are also promising from the esthetic point of view. The results described herein open the door toward a new generation of white titanium dental implants.

A Key Role of Autophagy in Osteoblast Differentiation on Titanium-Based Dental Implants.

Autophagy plays an important role in embryogenesis, for the maintenance of tissue homeostasis and the elimination of damaged subcellular structures. Furthermore, autophagy could be a mode of physiological cell death and also be implicated in cell differentiation. Thus, we hypothesized that autophagy may have an impact on the differentiation of osteoblast cells influenced by various titanium-based surfaces. Interactions between smooth, commercially available pure titanium (Ti cp), rough Ticer, acid-etched Ti cp (SS) and M1-M3 (comprised of the monoclinic phase of sodium-titanium oxides and rutile; M2 contains amorphous calcium phosphates) and human osteoblast cells were investigated. Immunofluorescent staining was used for detecting autophagy, cell cluster formation and collagen type I (Col-1) expression. Flow cytometry was employed to identify autophagy, the production of endogenous nitric oxide (NO) and the size and granularity of the cells. Rough surfaces caused osteoblast differentiation via the autophagic-dependent PI3/Akt signalling pathway. These surfaces induced the formation of discrete populations of large, granular cells, i.e. mature osteoblasts. In addition, M1-M3 provoked the development of a third population of small, granular cells, responsible for cell cluster formation, which are important for the formation of bone noduli and mineralisation. The same surfaces induced faster osteoblast maturation and enhanced NO production, a hallmark of the already mentioned processes. Neither the mature osteoblasts nor the small cells appeared after the inhibition of autophagy. Inhibition of autophagy also prevented cell cluster formation. We demonstrate that autophagy plays an essential role in the osteoblast differentiation on titanium-based surfaces with rough topography.

Organogallium(III) complexes as apoptosis promoting anticancer agents for head and neck squamous cell carcinoma (HNSCC) cell lines.

Organogallium(III) dinuclear (1-9) and tetranuclear (10) complexes present potential therapeutic agents for the treatment of various types of cancer. The antiproliferative activity of 1-10 was evaluated with cell lines of head and neck squamous cell carcinomas, e.g. HN (soft palate), Cal27, Cal33 (tongue) and FaDu (hypopharynx) cell lines. The activity of compound 8 is comparable with that of cisplatin on cell line Cal27 (IC(50) 4.6 µM for both compounds). The mode of cell death induced, caspase activity and cell cycle analysis were evaluated for potential hit compounds 3, 5 and 8 Potential hit compounds 3, 5 and 8 were further evaluated for the mode of cell death, caspase activity and cell cycle analysis. Apoptosis induced by compounds 3, 5 and 8 on Cal27 and FaDu cells was confirmed by DNA laddering , as well as acridine orange (AO) and ethidium bromide (EB) double staining. These compounds (3, 5 and 8) induced caspase-independent apoptosis (within 4 h of action) in cell line Cal27. Extrinsic-mediated apoptosis associated with caspase 8 and 3 activation is the main mode of cytotoxicity induced on FaDu cells by compounds 3, 5 and 8. Cell cycle perturbations caused by these compounds are also observed. Our data suggest that compounds 3, 5 and 8 should be studied further for the treatment of head and neck cancer.

Cytotoxicity, apoptosis and study of the DNA-binding properties of bi- and tetranuclear gallium(III) complexes with heterocyclic thiolato ligands.

The reaction of the heterocyclic thiol derivatives, 2-mercapto-1-methylimidazole (SH-imi), 5-mercapto-1-methyltetrazole (SH-tet), 2-mercaptobenzothiazole (SH-ben) and 5-phenyl-1,3,4-oxadiazole-2-thiol (SH-oxa), with trimethylgallium (1:1) afforded the dimeric or tetrameric complexes [Me(2)Ga(S-imi)](2) (1), [Me(2)Ga(S-tet)](2) (2), [Me(2)Ga(S-ben)](2) (3) and [Me(2)Ga(S-oxa)](4) (4), respectively. Molecular structures of 2 and 4 were determined by X-ray diffraction studies. The cytotoxicity of the gallium(III) complexes 1-4 was tested against human cell lines 8505C anaplastic thyroid cancer, A253 head and neck tumor, A549 lung carcinoma, A2780 ovarian cancer, DLD-1 colon carcinoma and compared with those of cisplatin and Ga(NO(3))(3). Compound 4 seems to be slightly more active against 8505C, A253 and A2780 and substantially more active than all the other complexes against DLD-1, with an IC(50) value of 5.49 ± 0.16 µM, very close to that of cisplatin (5.14 ± 0.12 µM). Complexes 1-4 were less toxic on normal human fibroblasts (WWO70327) than on the investigated tumor cell lines and more selective to cancer cells than cisplatin. DNA laddering method showed that treatment of the DLD-1 cell line with IC(90) doses of 1-4 resulted in the induction of apoptosis. Compound 1 caused apoptosis by upregulation of caspases 2, 3 and 8. Since no activity of caspase 9 is observed, complex 1 is causing apoptosis triggered by an extrinsic pathway. DNA-interaction tests have been also carried out. Solutions of all the studied complexes have been treated with different concentrations of fish sperm DNA (FS-DNA). Modifications of the UV spectra which gave intrinsic binding constants of 3.03 × 10(5), 4.44 × 10(5), 3.02 × 10(6) and 5.56 × 10(5) M(-1) for 1-4 were observed, however, no notable interaction with pBR322 plasmid DNA was detected.

Anticancer activity of dinuclear gallium(III) carboxylate complexes.

The reaction of 3-methoxyphenylacetic acid, 4-methoxyphenylacetic acid, mesitylthioacetic acid, 2,5-dimethyl-3-furoic acid and 1,4-benzodioxane-6-carboxylic acid with trimethylgallium (1:1) yielded the dimeric complexes [Me(2)Ga(micro-O(2)CCH(2)C(6)H(4)-3-OMe)](2) (1), [Me(2)Ga(micro-O(2)CCH(2)C(6)H(4)-4-OMe)](2) (2), [Me(2)Ga(micro-O(2)CCH(2)SMes)](2) (3) (Mes=2,4,6-Me(3)C(6)H(2)), [Me(2)Ga{micro-O(2)C(Fur)}](2) (4) (Fur=2,5-dimethylfuran) and [Me(2)Ga{micro-O(2)C(Bdo)}](2) (5) (Bdo=1,4-benzodioxane) respectively. The molecular structure of 5 was determined by X-ray diffraction studies. The cytotoxic activity of the gallium(III) complexes (1-5) was tested against human tumor cell lines 8505C anaplastic thyroid cancer, A253 head and neck tumor, A549 lung carcinoma, A2780 ovarian cancer, DLD-1 colon carcinoma and compared with that of cisplatin. Taking into account the standard deviation, there is no significant difference in the activity for any of the compounds in any cell line. However, complex 5 presents the best IC(50) value against A253 head and neck tumor (6.6+/-0.2 microM), while complex 3 seems to be the most active against A2780 ovarian cancer (12.0+/-0.4 microM) and marginally on DLD-1 colon carcinoma (12.4+/-0.1 microM).