Protein adsorption/desorption dynamics on Ca-enriched titanium surfaces: biological implications

[EN] Calcium ions are used in the development of biomaterials for the promotion of coagulation, bone regeneration, and implant osseointegration. Upon implantation, the time-dependent release of calcium ions from titanium implant surfaces modifies the physicochemical characteristics at the implant-tissue interface and thus, the biological responses. The aim of this study is to examine how the dynamics of protein adsorption on these surfaces change over time. Titanium discs with and without Ca were incubated with human serum for 2 min, 180 min, and 960 min. The layer of proteins attached to the surface was characterised using nLC-MS/MS. The adsorption kinetics was different between materials, revealing an increased adsorption of proteins associated with coagulation and immune responses prior to Ca release. Implant-blood contact experiments confirmed the strong coagulatory effect for Ca surfaces. We employed primary human alveolar osteoblasts and THP-1 monocytes to study the osteogenic and inflammatory responses. In agreement with the proteomic results, Ca-enriched surfaces showed a significant initial inflammation that disappeared once the calcium was released. The distinct protein adsorption/desorption dynamics found in this work demonstrated to be useful to explain the differential biological responses between the titanium and Ca-ion modified implant surfaces.This work was supported by MINECO [MAT2017-86043-R; RTC-2017-6147-1], Generalitat Valenciana [GRISOLIAP/2018/091; APOSTD/2020/036, PROMETEO/2020/069], Universitat Jaume I under [ UJI-B2017-37], the University of the Basque Country under [GIU18/189] and Basque Government under [PRE_2017_2_0044]. The authors would like to thank Raquel Oliver, Jose Ortega and Iraide Escobes for their valuable technical assistance.Romero-Gavilán, F.; Cerqueira, A.; Anitua, E.; Tejero, R.; García-Arnáez, I.; Martínez-Ramos, C.; Ozturan, S.... (2021). Protein adsorption/desorption dynamics on Ca-enriched titanium surfaces: biological implications. JBIC Journal of Biological Inorganic Chemistry. 26(6):1-12. https://doi.org/10.1007/s00775-021-01886-4S11226

Evaluation of the inflammatory responses to sol-gel coatings with distinct biocompatibility levels

[EN] The immune system plays a crucial role in determining the implantation outcome, and macrophages are in the frontline of the inflammatory processes. Further, cellular oxidative stress resulting from the material recognition can influence how cell responses develop. Considering this, the aim of this study was to study oxidative stress and macrophages phenotypes in response to sol-gel materials with distinct in vivo outcomes. Four materials were selected (70M30T and 35M35G30T, with high biocompatibility, and 50M50G and 50V50G, with low biocompatibility). Gene expression, immunocytochemistry and cytokine secretion profiles for M1 and M2 markers were determined. Moreover, oxidative stress markers were studied. Immunocytochemistry and ELISA showed that 50M50G and 50V50G lead to a higher differentiation to M1 phenotype, while 70M30T and 35M35G30T promoted M2 differentiation. In oxidative stress, no differences were found. These results show that the balance between M1 and M2, more than individual quantification of each phenotype, determines a biomaterial outcome.Generalitat Valenciana, Grant/Award Number: GRISOLIAP/2018/091; Ministerio de Economia y Competitividad, Grant/Award Numbers: MAT2017-86043-R, RTC2017-6147-1; Universitat Jaume I, Grant/Award Number: POSDOC/2019/28Cerqueira, A.; Araújo-Gomes, N.; Zhang, Y.; Van Den Beucken, JJJP.; Martínez-Ramos, C.; Ozturan, S.; Izquierdo, R.... (2021). Evaluation of the inflammatory responses to sol-gel coatings with distinct biocompatibility levels. Journal of Biomedical Materials Research Part A. 109(9):1539-1548. https://doi.org/10.1002/jbm.a.37149S15391548109

Laser-assisted laterally positioned flap operation: A randomized controlled clinical trial

The purpose of this randomized, controlled clinical trial was to compare the effectiveness of a new treatment approach, which consisted of laterally positioned flap (LPF) procedure combined with an external vestibular releasing incision made by a diode laser (LALPF) to LPF alone for the treatment of isolated gingival recession defects.status: publishe

Characterization of magnesium doped sol-gel biomaterial for bone tissue regeneration: the effect of Mg ion in protein adsorption

[EN] Magnesium is the fourth most abundant element in the human body with a wide battery of functions in the maintenance of normal cell homeostasis. In the bone, this element incorporates in the hydroxyapatite structure and it takes part in mineral metabolism and regulates osteoclast functions. In this study, sol-gel materials with increasing concentrations of MgCl2 (0.5, 1, and 1.5%) were synthesized and applied onto Ti surfaces as coatings. The materials were first physicochemically characterized. In vitro responses were examined using the MC3T3-E1 osteoblastic cells and RAW264.7 macrophages. Human serum protein adsorption was evaluated employing nLC-MS/MS. The incorporation of Mg did not affect the crosslinking of the sol-gel network, and a controlled release of Mg was observed; it was not cytotoxic at any of the tested concentrations. The cytoskeleton arrangement of MC3T3-E1 cells cultured on the Mg-doped materials changed in comparison with controls; the cells became more elongated, with protruded lamellipodia and increased cell surface. The expression of integrins (ITGA5 and ITGB1) was boosted by Mg-coatings. The ALP activity and expression of TGF-beta, OSX and RUNX2 genes were also increased. In RAW264.7 cells, TNF-alpha secretion was reduced, while TGF-beta and IL-4 expression rose. These changes correlated with the altered protein adsorption patterns. The Mg-doped coatings showed increased adsorption of anti-inflammatory (CLUS, IC1, CFAH, and VTNC), cell adhesion (DSG1, FILA2, and DESP) and tissue regeneration (VTNC and CYTA) proteins. This integrated approach to biomaterial characterization revealed the potential of Mg in bone tissue regeneration.This work was supported by MINECO [MAT2017-86043-R; RTC2017-6147-1], Generalitat Valenciana [GRISOLIAP/2018/091, APOSTD/2020/036, PROMETEO/2020/069], Universitat Jaume I under [UJI-B2017-37, Posdoc/2019/28], the University of the Basque Country under [GIU18/189] and Basque Government under [PRE_2017_2_0044]. The authors would like to thank Raquel Oliver, Jose Ortega, Jos ' e Miguel Pedra and Iraide Escob ' es for their valuable technical assistance and Antonio Coso (GMI-Ilerimplant) for producing the titanium discs.Cerqueira, A.; Romero-Gavilán, F.; García-Arnáez, I.; Martínez-Ramos, C.; Ozturan, S.; Izquierdo, R.; Azkargorta, M.... (2021). Characterization of magnesium doped sol-gel biomaterial for bone tissue regeneration: the effect of Mg ion in protein adsorption. Materials Science and Engineering C: Materials for Biological Applications (Online). 125:1-11. https://doi.org/10.1016/j.msec.2021.112114S11112

A possible use of melatonin in the dental field: Protein adsorption and in vitro cell response on coated titanium

[EN] Melatonin (MLT) is widely known for regulating the circadian cycles and has been studied for its role in bone regeneration and inflammation. Its application as a coating for dental implants can condition the local micro environment, affecting protein deposition on its surface and the cellular and tissue response. Using sol-gel coatings as a release vehicle for MLT, the aim of this work was to assess the potential of this molecule in improving the osseointegration and inflammatory responses of a titanium substrate. The materials obtained were physicochemically characterized (scanning electron microscopy, contact angle, roughness, Fourier-transform infrared spectroscopy, nuclear magnetic resonance, Si release, MLT liberation, and degradation) and studied in vitro with MC3T3-E1 osteoblastic cells and RAW264.7 macrophage cells. Although MLT application led to an increased gene expression of RUNX2 and BMP2 in 10MTL, it did not improve ALP activity. On the other hand, MLT-enriched sol-gel materials presented potential effects in the adsorption of proteins related to inflammation, coagulation and angiogenesis pathways depending on the dosage used. Using LC-MS/MS, protein adsorption patterns were studied after incubation with human serum. Proteins related to the complement systems (CO7, IC1, CO5, CO8A, and CO9) were less adsorbed in materials with MLT; on the other hand, proteins with functions in the coagulation and angiogenesis pathways, such as A2GL and PLMN, showed a significant adsorption pattern.This work was supported by MINECO [MAT MAT2017-86043-R; RTC-2017-6147-1], Universitat Jaume I under [UJI-B2017-37; POSDOC/2019/28], Generalitat Valenciana [GRISOLIAP/2018/091], University of the Basque Country under [UFI11/56] and Basque Government under [PRE_2017_2_0044]. CIC bioGUNE is supported by Basque Department of Industry, Tourism and Trade (Etortek and Elkartek programs), the Innovation Technology Department of the Bizkaia County; The ProteoRed-ISCIII (Grant PRB3 IPT17/0019); CIBERehd Network and Severo Ochoa Grant (SEV-2016-0644). Authors would like to thank Antonio Coso (GMI-Ilerimplant) for their inestimable contribution to this study, and Raquel Oliver, Jose Ortega (UJI) and Iraide Escobes (CIC bioGUNE) for their valuable technical assistance.Cerqueira, A.; Romero-Gavilán, F.; Araujo-Gomes, N.; García-Arnáez, I.; Martínez-Ramos, C.; Ozturan, S.; Azkargorta, M.... (2020). A possible use of melatonin in the dental field: Protein adsorption and in vitro cell response on coated titanium. Materials Science and Engineering C: Materials for Biological Applications (Online). 116:1-10. https://doi.org/10.1016/j.msec.2020.111262S110116Smeets, R., Stadlinger, B., Schwarz, F., Beck-Broichsitter, B., Jung, O., Precht, C., … Ebker, T. (2016). 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Influence of calcium ion-modified implant surfaces in protein adsorption and implant integration

[EN] Background Calcium (Ca) is a well-known element in bone metabolism and blood coagulation. Here, we investigate the link between the protein adsorption pattern and the in vivo responses of surfaces modified with calcium ions (Ca-ion) as compared to standard titanium implant surfaces (control). We used LC-MS/MS to identify the proteins adhered to the surfaces after incubation with human serum and performed bilateral surgeries in the medial section of the femoral condyles of 18 New Zealand white rabbits to test osseointegration at 2 and 8 weeks post-implantation (n=9). Results Ca-ion surfaces adsorbed 181.42 times more FA10 and 3.85 times less FA12 (p<0.001), which are factors of the common and the intrinsic coagulation pathways respectively. We also detected differences in A1AT, PLMN, FA12, KNG1, HEP2, LYSC, PIP, SAMP, VTNC, SAA4, and CFAH (p<0.01). At 2 and 8 weeks post-implantation, the mean bone implant contact (BIC) with Ca-ion surfaces was respectively 1.52 and 1.25 times higher, and the mean bone volume density (BVD) was respectively 1.35 and 1.13 times higher. Differences were statistically significant for BIC at 2 and 8 weeks and for BVD at 2 weeks (p<0.05). Conclusions The strong thrombogenic protein adsorption pattern at Ca-ion surfaces correlated with significantly higher levels of implant osseointegration. More effective implant surfaces combined with smaller implants enable less invasive surgeries, shorter healing times, and overall lower intervention costs, especially in cases of low quantity or quality of bone.This work was supported by Universitat Jaume I under [POSDOC/2019/28], Generalitat Valenciana [GRISOLIAP/2018/091], University of the Basque Country under [UFI11/56], and Basque Government under [PRE_2017_2_0044]. CIC bioGUNE is supported by Basque Department of Industry, Tourism and Trade (Etortek and Elkartek programs), the Innovation Technology Department of the Bizkaia County; The ProteoRed-ISCIII (Grant PRB3 IPT17/0019); and CIBERehd Network and Severo Ochoa Grant (SEV-2016-0644).Anitua, E.; Cerqueira, A.; Romero-Gavilán, F.; García-Arnáez, I.; Martínez-Ramos, C.; Ozturan, S.; Azkargorta, M.... (2021). Influence of calcium ion-modified implant surfaces in protein adsorption and implant integration. International Journal of Implant Dentistry. 7(1):1-11. https://doi.org/10.1186/s40729-021-00314-1S1117