Titanium alloys (AoN) and their involvement in osseointegration
Identifieur interne : 002B77 ( Pmc/Corpus ); précédent : 002B76; suivant : 002B78Titanium alloys (AoN) and their involvement in osseointegration
Auteurs : Matteo Danza ; Ilaria Zollino ; Valentina Candotto ; Francesca Cura ; Francesco CarinciSource :
- Dental Research Journal [ 1735-3327 ] ; 2012.
Abstract
Osseointegration is essential for a long-term successful and inflammation-free dental implant. Such a result depends on osteoblastic cells growth and differentiation at the tissue-implant interface. The aim of this study was to compare two different AoN titanium layers (GR4 and GR5) to investigate which one had a greater osteoconductive power using human osteoblasts (HOb) culture at two different time-points.
The expression levels of some bone-related (ALPL, COL1A1, COL3A1, SPP1, RUNX2, and SPARC) were analyzed using real time reverse transcription-polymerase chain reaction (real time RT-PCR).
Real-time RT-PCR data showed that after 3 days of treatment with TiA4GR, the genes up-regulated were COL3A1, ALPL, SPP1, and RUNX2. Moreover, no difference in gene expression was noticed 4 days later. On the other hand, the genes that overexpressed after 3 days of treatment with AoN5GR were ALPL, SPP1, and RUNX2. In both cases, the expression of COL1A1 and SPARC was negatively regulated.
Our data showed that both titanium surfaces led to osteoblasts recruitment, maturation, and differentiation, thus promoting osseointegration at the tissue-implant interface.
Url:
DOI: 10.4103/1735-3327.109756
PubMed: 23814585
PubMed Central: 3692175
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PMC:3692175Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Titanium alloys (AoN) and their involvement in osseointegration</title><author><name sortKey="Danza, Matteo" sort="Danza, Matteo" uniqKey="Danza M" first="Matteo" last="Danza">Matteo Danza</name><affiliation><nlm:aff id="aff1">Department of Oral Science, Nano and Biotechnologies, University of Chieti, Chieti, Italy</nlm:aff></affiliation></author><author><name sortKey="Zollino, Ilaria" sort="Zollino, Ilaria" uniqKey="Zollino I" first="Ilaria" last="Zollino">Ilaria Zollino</name><affiliation><nlm:aff id="aff2">Department of Medical-Surgical Sciences of Communication and Behavior, Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara, Italy</nlm:aff></affiliation></author><author><name sortKey="Candotto, Valentina" sort="Candotto, Valentina" uniqKey="Candotto V" first="Valentina" last="Candotto">Valentina Candotto</name><affiliation><nlm:aff id="aff2">Department of Medical-Surgical Sciences of Communication and Behavior, Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara, Italy</nlm:aff></affiliation></author><author><name sortKey="Cura, Francesca" sort="Cura, Francesca" uniqKey="Cura F" first="Francesca" last="Cura">Francesca Cura</name><affiliation><nlm:aff id="aff3">Department of Histology, Embryology and Applied Biology, University of Bologna, Bologna, Italy</nlm:aff></affiliation></author><author><name sortKey="Carinci, Francesco" sort="Carinci, Francesco" uniqKey="Carinci F" first="Francesco" last="Carinci">Francesco Carinci</name><affiliation><nlm:aff id="aff2">Department of Medical-Surgical Sciences of Communication and Behavior, Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara, Italy</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">23814585</idno><idno type="pmc">3692175</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3692175</idno><idno type="RBID">PMC:3692175</idno><idno type="doi">10.4103/1735-3327.109756</idno><date when="2012">2012</date><idno type="wicri:Area/Pmc/Corpus">002B77</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">002B77</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Titanium alloys (AoN) and their involvement in osseointegration</title><author><name sortKey="Danza, Matteo" sort="Danza, Matteo" uniqKey="Danza M" first="Matteo" last="Danza">Matteo Danza</name><affiliation><nlm:aff id="aff1">Department of Oral Science, Nano and Biotechnologies, University of Chieti, Chieti, Italy</nlm:aff></affiliation></author><author><name sortKey="Zollino, Ilaria" sort="Zollino, Ilaria" uniqKey="Zollino I" first="Ilaria" last="Zollino">Ilaria Zollino</name><affiliation><nlm:aff id="aff2">Department of Medical-Surgical Sciences of Communication and Behavior, Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara, Italy</nlm:aff></affiliation></author><author><name sortKey="Candotto, Valentina" sort="Candotto, Valentina" uniqKey="Candotto V" first="Valentina" last="Candotto">Valentina Candotto</name><affiliation><nlm:aff id="aff2">Department of Medical-Surgical Sciences of Communication and Behavior, Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara, Italy</nlm:aff></affiliation></author><author><name sortKey="Cura, Francesca" sort="Cura, Francesca" uniqKey="Cura F" first="Francesca" last="Cura">Francesca Cura</name><affiliation><nlm:aff id="aff3">Department of Histology, Embryology and Applied Biology, University of Bologna, Bologna, Italy</nlm:aff></affiliation></author><author><name sortKey="Carinci, Francesco" sort="Carinci, Francesco" uniqKey="Carinci F" first="Francesco" last="Carinci">Francesco Carinci</name><affiliation><nlm:aff id="aff2">Department of Medical-Surgical Sciences of Communication and Behavior, Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara, Italy</nlm:aff></affiliation></author></analytic><series><title level="j">Dental Research Journal</title><idno type="ISSN">1735-3327</idno><idno type="eISSN">2008-0255</idno><imprint><date when="2012">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec id="st1"><title>Background:</title><p>Osseointegration is essential for a long-term successful and inflammation-free dental implant. Such a result depends on osteoblastic cells growth and differentiation at the tissue-implant interface. The aim of this study was to compare two different AoN titanium layers (GR4 and GR5) to investigate which one had a greater osteoconductive power using human osteoblasts (HOb) culture at two different time-points.</p></sec><sec id="st2"><title>Materials and Methods:</title><p>The expression levels of some bone-related (ALPL, COL1A1, COL3A1, SPP1, RUNX2, and SPARC) were analyzed using real time reverse transcription-polymerase chain reaction (real time RT-PCR).</p></sec><sec id="st3"><title>Results:</title><p>Real-time RT-PCR data showed that after 3 days of treatment with TiA4GR, the genes up-regulated were COL3A1, ALPL, SPP1, and RUNX2. Moreover, no difference in gene expression was noticed 4 days later. On the other hand, the genes that overexpressed after 3 days of treatment with AoN5GR were ALPL, SPP1, and RUNX2. In both cases, the expression of COL1A1 and SPARC was negatively regulated.</p></sec><sec id="st4"><title>Conclusion:</title><p>Our data showed that both titanium surfaces led to osteoblasts recruitment, maturation, and differentiation, thus promoting osseointegration at the tissue-implant interface.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Tsukimura, N" uniqKey="Tsukimura N">N Tsukimura</name></author><author><name sortKey="Kojima, N" uniqKey="Kojima N">N Kojima</name></author><author><name sortKey="Kubo, K" uniqKey="Kubo K">K Kubo</name></author><author><name sortKey="Att, W" uniqKey="Att W">W Att</name></author><author><name sortKey="Takeuchi, K" uniqKey="Takeuchi K">K Takeuchi</name></author><author><name sortKey="Kameyama, Y" uniqKey="Kameyama Y">Y Kameyama</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Scarano, A" uniqKey="Scarano A">A Scarano</name></author><author><name 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Dent Res J (Isfahan)</journal-id><journal-id journal-id-type="iso-abbrev">Dent Res J (Isfahan)</journal-id><journal-id journal-id-type="publisher-id">DRJ</journal-id><journal-title-group><journal-title>Dental Research Journal</journal-title></journal-title-group><issn pub-type="ppub">1735-3327</issn><issn pub-type="epub">2008-0255</issn><publisher><publisher-name>Medknow Publications & Media Pvt Ltd</publisher-name><publisher-loc>India</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">23814585</article-id><article-id pub-id-type="pmc">3692175</article-id><article-id pub-id-type="publisher-id">DRJ-9-207</article-id><article-id pub-id-type="doi">10.4103/1735-3327.109756</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject></subj-group></article-categories><title-group><article-title>Titanium alloys (AoN) and their involvement in osseointegration</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Danza</surname><given-names>Matteo</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Zollino</surname><given-names>Ilaria</given-names></name><xref ref-type="aff" rid="aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Candotto</surname><given-names>Valentina</given-names></name><xref ref-type="aff" rid="aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Cura</surname><given-names>Francesca</given-names></name><xref ref-type="aff" rid="aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Carinci</surname><given-names>Francesco</given-names></name><xref ref-type="aff" rid="aff2">2</xref><xref ref-type="corresp" rid="cor1"></xref></contrib></contrib-group><aff id="aff1"><label>1</label>Department of Oral Science, Nano and Biotechnologies, University of Chieti, Chieti, Italy</aff><aff id="aff2"><label>2</label>Department of Medical-Surgical Sciences of Communication and Behavior, Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara, Italy</aff><aff id="aff3"><label>3</label>Department of Histology, Embryology and Applied Biology, University of Bologna, Bologna, Italy</aff><author-notes><corresp id="cor1"><bold>Address for correspondence:</bold> Prof. Francesco Carinci, Department of D.M.C.C.C., Section of Maxillofacial and Plastic Surgery, University of Ferrara, Corso Giovecca 203,44100 Ferrara, Italy. <bold>E-mail:</bold><email xlink:href="crc@unife.it">crc@unife.it</email></corresp></author-notes><pub-date pub-type="ppub"><month>12</month><year>2012</year></pub-date><volume>9</volume><issue>Suppl 2</issue><fpage>S207</fpage><lpage>S210</lpage><history><date date-type="received"><month>8</month><year>2012</year></date><date date-type="accepted"><month>10</month><year>2012</year></date></history><permissions><copyright-statement>Copyright: © Dental Research Journal</copyright-statement><copyright-year>2012</copyright-year><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc-sa/3.0"><license-p>This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><abstract><sec id="st1"><title>Background:</title><p>Osseointegration is essential for a long-term successful and inflammation-free dental implant. Such a result depends on osteoblastic cells growth and differentiation at the tissue-implant interface. The aim of this study was to compare two different AoN titanium layers (GR4 and GR5) to investigate which one had a greater osteoconductive power using human osteoblasts (HOb) culture at two different time-points.</p></sec><sec id="st2"><title>Materials and Methods:</title><p>The expression levels of some bone-related (ALPL, COL1A1, COL3A1, SPP1, RUNX2, and SPARC) were analyzed using real time reverse transcription-polymerase chain reaction (real time RT-PCR).</p></sec><sec id="st3"><title>Results:</title><p>Real-time RT-PCR data showed that after 3 days of treatment with TiA4GR, the genes up-regulated were COL3A1, ALPL, SPP1, and RUNX2. Moreover, no difference in gene expression was noticed 4 days later. On the other hand, the genes that overexpressed after 3 days of treatment with AoN5GR were ALPL, SPP1, and RUNX2. In both cases, the expression of COL1A1 and SPARC was negatively regulated.</p></sec><sec id="st4"><title>Conclusion:</title><p>Our data showed that both titanium surfaces led to osteoblasts recruitment, maturation, and differentiation, thus promoting osseointegration at the tissue-implant interface.</p></sec></abstract><kwd-group><kwd>Gene expression</kwd><kwd>osteoinduction</kwd><kwd>titanium alloys</kwd></kwd-group></article-meta></front><body><sec id="sec1-1"><title>INTRODUCTION</title><p>Osseointegration is a widely used term in dentistry and medicine fields and refers to the narrow junction between bone and artificial implant.</p><p>A rapid and good outcome in osseointegration is a fundamental pre-requisite for a successful dental implantation and depends on the shape, structure, and composition of the used surface.[<xref ref-type="bibr" rid="ref1">1</xref>] The main requirements for a good material are its ability to promote attraction and adhesion of bone precursor cells and their proliferation and differentiation.[<xref ref-type="bibr" rid="ref2">2</xref>] The ability of the materials to lead osteoinduction can be evaluated by some biological parameters such as alkaline phosphatase activity and up-regulation of several bone-related genes. Several data suggest that prosthesis anchorage to bone tissue can be modulated by surface characteristics.[<xref ref-type="bibr" rid="ref3">3</xref><xref ref-type="bibr" rid="ref4">4</xref><xref ref-type="bibr" rid="ref5">5</xref>] Having observed that smooth surfaces are less suitable to induce a similar behavior,[<xref ref-type="bibr" rid="ref6">6</xref>] different treatments can be used to obtain surface roughness and promote osteoblast adhesion and colonization.[<xref ref-type="bibr" rid="ref7">7</xref><xref ref-type="bibr" rid="ref8">8</xref>]</p><p>Different biocompatible materials are currently employed as scaffold. Among these, titanium is considered a “gold standard” because of its biocompatibility and good corrosion resistance.[<xref ref-type="bibr" rid="ref9">9</xref><xref ref-type="bibr" rid="ref10">10</xref>]</p><p>Titanium can be characterized by several degree of purity, depending on the relative percentage of different elements as iron, aluminium, vanadium, and molybdenum. Moreover, its excellent corrosion resistance and biocompatibility, have been largely shown.[<xref ref-type="bibr" rid="ref11">11</xref>]</p><p>Recently, a new type of implant with a spiral form has been produced (Ultimate, AoN, Grisignano di Zocco, VI). The aim of this work was to compare two different AoN titanium layers (GR4 and GR5) and to investigate which one had a greater osteoconductive power using human osteoblasts (HOb) culture at two different time-points. The expression levels of some bone-related (ALPL, COL1A1, COL3A1, SPP1, RUNX2, and SPARC) were analyzed using real time reverse transcription-polymerase chain reaction (real time RT-PCR).</p></sec><sec sec-type="materials|methods" id="sec1-2"><title>MATERIALS AND METHODS</title><sec id="sec2-1"><title>AoN titanium implants</title><p>In this study, we used two types of AoN alloy disk (named AoN4GR and AoN5GR), with a diameter of 5 mm that differed for chemical processing. AoN4GR was treated with ortophosphoric acid and coated with calcium phosphate. AoN5GR, on the other hand, having a different purity degree underwent a double sand-blasting process (corundum and aluminium).</p></sec><sec id="sec2-2"><title>Primary human osteoblast cells culture</title><p>Fragments of bone derived from skull of healthy volunteers were collected during operation. The pieces were transferred in 75 cm<sup>2</sup> culture flasks containing DMEM (Dulbecco's Modified Eagle Medium) (Sigma Aldrich, Inc., St Louis, MO, USA) supplemented with 20% fetal calf serum, antibiotics (penicillin 100 U/ml and streptomycin 100 μg/ml; Sigma Aldrich, Inc.).</p><p>Cells were incubated in a humified atmosphere of 5% CO<sub>2</sub> at 37°C. The medium was changed the next day and twice a week. After 15 days, the pieces of bone tissue were removed from the culture flask. Cells were harvested after 30 days of incubation.</p></sec><sec id="sec2-3"><title>Cells culture</title><p>For the investigation, HOb at the second passage were seeded on two different types of AoN titanium dishes (4GR and 5GR). A set of untreated cells were used as controls. The medium was changed three times a week and the cells were maintained in a humified atmosphere of 5% CO<sub>2</sub> at 37°C. Cells were trypsinized and lysed for RNA extraction, after 3 and 7 days of treatment.</p></sec><sec id="sec2-4"><title>RNA processing and real time PCR</title><p>Reverse transcription to cDNA and gene expression quantification were performed as previously discussed.[<xref ref-type="bibr" rid="ref4">4</xref>] In <xref ref-type="table" rid="T1">Table 1</xref> were listed primer and probe sequences for the amplification of the bone related genes.</p><table-wrap id="T1" position="float"><label>Table 1</label><caption><p>Primer and probes used in real time polymerase chain reaction</p></caption><graphic xlink:href="DRJ-9-207-g001"></graphic></table-wrap></sec></sec><sec sec-type="results" id="sec1-3"><title>RESULTS</title><p>Osteoinductive properties of two different titanium disks were evaluated by measuring the gene expression levels of bone-related genes in treated HOb, at two time-points (3 and 7 days).</p><p>Real-time RT-PCR data showed that after 3 days of treatment with TiA4GR, the genes up-regulated were COL3A1, ALPL, SPP1, and RUNX2. Moreover, no difference in gene expression was noticed 4 days later. On the other hand, the genes that overexpressed after 3 days of treatment with AoN5GR were ALPL, SPP1, and RUNX2. In both cases, the expression of COL1A1 and SPARC was negatively regulated [Figure <xref ref-type="fig" rid="F1">1a</xref> and <xref ref-type="fig" rid="F1">1b</xref>].</p><fig id="F1" position="float"><label>Figure 1</label><caption><p>Human osteoblasts gene expression profile after 3 days (a) and 7 days (b) of treatment with AoN 4GR</p></caption><graphic xlink:href="DRJ-9-207-g002"></graphic></fig><p>HOb after 7 days of treatment with AoN5GR showed overexpression of COL3A1, RUNX2, SPP1, and SPARC. Instead ALPL and COL1A1 were the under-regulated genes [Figure <xref ref-type="fig" rid="F2">2a</xref> and <xref ref-type="fig" rid="F2">b</xref>].</p><fig id="F2" position="float"><label>Figure 2</label><caption><p>Human osteoblasts gene expression profile after 3 days (a) and 7 days (b) of treatment with AoN 5GR</p></caption><graphic xlink:href="DRJ-9-207-g003"></graphic></fig></sec><sec sec-type="discussion" id="sec1-4"><title>DISCUSSION</title><p>Osseointegration is essential for a long-term successful and inflammation-free dental implant [<xref ref-type="fig" rid="F3">Figure 3</xref>].[<xref ref-type="bibr" rid="ref12">12</xref><xref ref-type="bibr" rid="ref13">13</xref>] Immediate loading implants are gold standard to improve comfort of edentulous patients.[<xref ref-type="bibr" rid="ref14">14</xref><xref ref-type="bibr" rid="ref15">15</xref>] Such a result depends on osteoblastic cells growth and differentiation at the tissue-implant interface.</p><fig id="F3" position="float"><label>Figure 3</label><caption><p>Dental implant</p></caption><graphic xlink:href="DRJ-9-207-g004"></graphic></fig><p>The aim of this study was to compare osteoblasts behavior, cultured on two different types of AoN titanium disks. AoN4GR was treated with ortophosphoric acid and coated with calcium phosphate. AoN5GR, having a different purity degree (as consequence of having a different chemical composition), underwent a double sand-blasting process (corundum and aluminium).</p><p>After 3 and 7 days of treatment the expression levels of bone-related genes were measured by relative quantification method using real-time RT-PCR in HOb.</p><p>AoN4GR caused up-regulation of ALPL, COL3A1, SPP1, and RUNX2 and down-regulation of COL1A1 and SPARC both after 3 days than after 7 days of treatment.</p><p>Alkaline phosphatase (ALPL) plays a critical role in bone-matrix mineralization. Its activity is required for the normal mineralization of the osteoid during bone modelling and remodelling. An increased expression of this protein can be used as marker of calcification and osteoblasts differentiation.[<xref ref-type="bibr" rid="ref16">16</xref>]</p><p>RUNX2 is a transcription factor required at the early stage of osteoblast differentiation and it can induce ALPL activity.[<xref ref-type="bibr" rid="ref17">17</xref>] Moreover, it has been demonstrated that RUNX2 is involved during cell migration. Its up-regulation may be involved in pre-osteoblasts attraction and in their subsequent maturation and differentiation.</p><p>SPP1 encodes osteopontin, the most important non-collagenic component of extracellular bone matrix. Weber <italic>et al</italic>. demonstrated an involvement of this protein in cell adhesion and migration, suggesting the interest of this protein also, in recruitment of immature cells around the implant surface and in extracellular matrix deposition.[<xref ref-type="bibr" rid="ref18">18</xref>]</p><p>AoN GR4 also modulates the expression of COL3A1. Type 3 collagen is a protein that gives strength and support to several body tissues.</p><p>The other type of titanium disks is AoN which caused an up-regulation of ALP, SPP1, and RUNX2 and a down-regulation of COL1A1 and SPARC on exposition to Hob after 3 days of treatment.</p><p>Expression of ALPL decreased after 7 days of exposure, whereas we registered an increased expression of SPARC and COL3A1.</p><p>Osteonectin (SPARC), a multi-functional protein, plays a double action in modulating cell-cell and cell-matrix interactions. Moreover, it has a recognized role in osteoblastic functional differentiation of bone cells.</p></sec><sec sec-type="conclusion" id="sec1-5"><title>CONCLUSION</title><p>Our data showed that both titanium surfaces led to osteoblasts recruitment, maturation, and differentiation, thus promoting osseointegration at the tissue-implant interface.</p></sec></body><back><ack><title>ACKNOWLEDGEMENTS</title><p>This work was supported by the University of Ferrara (F.C.), Ferrara, Italy and by PRIN 2008 (20089MANHH_004).</p></ack><fn-group><fn fn-type="supported-by"><p><bold>Source of Support:</bold> This work was supported by the University of Ferrara (F.C.), Ferrara, Italy and by PRIN 2008 (20089MANHH_004)</p></fn><fn fn-type="conflict"><p><bold>Conflict of Interest:</bold> None declared</p></fn></fn-group><ref-list><title>REFERENCES</title><ref id="ref1"><label>1</label><element-citation publication-type="journal"><person-group 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