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Article ; Online: Proteomics as a tool to study the osteoimmunomodulatory role of metallic ions in a sol-gel coating.

García-Arnáez, Iñaki / Romero-Gavilán, Francisco / Cerqueira, Andreia / Azkargorta, Mikel / Elortza, Félix / Suay, Julio / Goñi, Isabel / Gurruchaga, Mariló

Journal of materials chemistry. B

2023 Volume 11, Issue 34, Page(s) 8194–8205

Abstract: The success of bone implants depends on the osteoimmunomodulatory (OIM) activity of the biomaterials in the interactions with the periimplantary tissues. ... ...

Abstract	The success of bone implants depends on the osteoimmunomodulatory (OIM) activity of the biomaterials in the interactions with the periimplantary tissues. Many
MeSH term(s)	Coated Materials, Biocompatible/pharmacology ; Proteomics ; Metals ; Bone Regeneration ; Ions
Chemical Substances	Coated Materials, Biocompatible ; Metals ; Ions
Language	English
Publishing date	2023-08-30
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2702241-9
ISSN	2050-7518 ; 2050-750X
ISSN (online)	2050-7518
ISSN	2050-750X
DOI	10.1039/d3tb01204b
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Article ; Online: Proteomic Analysis of Mesenchymal Stem Cells and Monocyte Co-Cultures Exposed to a Bioactive Silica-Based Sol-Gel Coating.

Cerqueira, Andreia / Romero-Gavilán, Francisco / Helmholz, Heike / Azkargorta, Mikel / Elortza, Félix / Gurruchaga, Mariló / Goñi, Isabel / Willumeit-Römer, Regine / Suay, Julio

ACS biomaterials science & engineering

2023 Volume 9, Issue 6, Page(s) 3306–3319

Abstract: New methodologies capable of extensively analyzing the cell-material interactions are necessary to improve current in vitro characterization methods, and proteomics is a viable alternative. Also, many studies are focused on monocultures, even though co- ... ...

Abstract	New methodologies capable of extensively analyzing the cell-material interactions are necessary to improve current in vitro characterization methods, and proteomics is a viable alternative. Also, many studies are focused on monocultures, even though co-cultures model better the natural tissue. For instance, human mesenchymal stem cells (MSCs) modulate immune responses and promote bone repair through interaction with other cell types. Here, label-free liquid chromatography tandem mass spectroscopy proteomic methods were applied for the first time to characterize HUCPV (MSC) and CD14
MeSH term(s)	Humans ; Monocytes ; Coculture Techniques ; Proteomics ; Mesenchymal Stem Cells/metabolism ; Integrins/metabolism ; Silicon Dioxide/chemistry ; Silicon Dioxide/metabolism ; Glutathione Transferase/metabolism
Chemical Substances	Integrins ; Silicon Dioxide (7631-86-9) ; GSTO1 protein, human (EC 2.5.1.18) ; Glutathione Transferase (EC 2.5.1.18)
Language	English
Publishing date	2023-05-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2373-9878
ISSN (online)	2373-9878
DOI	10.1021/acsbiomaterials.3c00254
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Article ; Online: Enhancing the correlation between

Romero Gavilán, Francisco / Cerqueira, Andreia / Anitua, Eduardo / Muñoz, Fernando / García Arnáez, Iñaki / Azkargorta, Mikel / Elortza, Félix / Gurruchaga, Mariló / Goñi, Isabel / Suay, Julio / Tejero, Ricardo

Journal of materials chemistry. B

2024 Volume 12, Issue 11, Page(s) 2831–2842

Abstract: This study delves into the osteogenic potential of a calcium-ion modified titanium implant surface, unicCa, employing state-of-the-art proteomics techniques ... ...

Abstract	This study delves into the osteogenic potential of a calcium-ion modified titanium implant surface, unicCa, employing state-of-the-art proteomics techniques both
MeSH term(s)	Animals ; Humans ; Rabbits ; Osseointegration/physiology ; Dental Implants ; Proteomics ; Phosphatidylinositol 3-Kinases ; Surface Properties ; Ions
Chemical Substances	Dental Implants ; Phosphatidylinositol 3-Kinases (EC 2.7.1.-) ; Ions
Language	English
Publishing date	2024-03-13
Publishing country	England
Document type	Journal Article
ZDB-ID	2702241-9
ISSN	2050-7518 ; 2050-750X
ISSN (online)	2050-7518
ISSN	2050-750X
DOI	10.1039/d3tb02694a
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Article ; Online: Proteomic evaluation of human osteoblast responses to titanium implants over time.

Romero-Gavilán, Francisco / Cerqueira, Andreia / García-Arnáez, Iñaki / Azkargorta, Mikel / Elortza, Félix / Gurruchaga, Mariló / Goñi, Isabel / Suay, Julio

Journal of biomedical materials research. Part A

2022 Volume 111, Issue 1, Page(s) 45–59

Abstract: Titanium is widely used in bone prostheses due to its excellent biocompatibility and osseointegration capacity. To understand the effect of sandblasted acid-etched (SAE) Ti implants on the biological responses of human osteoblast (HOb), their proteomic ... ...

Abstract	Titanium is widely used in bone prostheses due to its excellent biocompatibility and osseointegration capacity. To understand the effect of sandblasted acid-etched (SAE) Ti implants on the biological responses of human osteoblast (HOb), their proteomic profiles were analyzed using nLC-MS/MS. The cells were cultured with the implant materials, and 2544 distinct proteins were detected in samples taken after 1, 3, and 7 days. Comparative analyses of proteomic data were performed using Perseus software. The expression of proteins related to EIF2, mTOR, insulin-secretion and IGF pathways showed marked differences in cells grown with SAE-Ti in comparison with cells cultured without Ti. Moreover, the proteomic profiles obtained with SAE-Ti were compared over time. The affected proteins were related to adhesion, immunity, oxidative stress, coagulation, angiogenesis, osteogenesis, and extracellular matrix formation functions. The proliferation, mineralization and osteogenic gene expression in HObs cultured with SAE-Ti were characterized in vitro. The results showed that the osteoblasts exposed to this material increase their mineralization rate and expression of COLI, RUNX2, SP7, CTNNB1, CAD13, IGF2, MAPK2, and mTOR. Overall, the observed proteomic profiles can explain the SAE-Ti osteogenic properties, widening our knowledge of key signaling pathways taking part in the early stages of the osseointegration process in this type of implantations.
MeSH term(s)	Humans ; Titanium/pharmacology ; Titanium/metabolism ; Proteomics ; Tandem Mass Spectrometry ; Surface Properties ; Osteoblasts ; Osseointegration ; Osteogenesis ; Prostheses and Implants ; TOR Serine-Threonine Kinases
Chemical Substances	Titanium (D1JT611TNE) ; TOR Serine-Threonine Kinases (EC 2.7.11.1)
Language	English
Publishing date	2022-08-30
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2099989-6
ISSN	1552-4965 ; 1549-3296 ; 0021-9304
ISSN (online)	1552-4965
ISSN	1549-3296 ; 0021-9304
DOI	10.1002/jbm.a.37444
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Article: The effect of calcium–magnesium mixtures in sol–gel coatings on bone tissue regeneration

Cerqueira, Andreia / García-Arnáez, Iñaki / Muriach, María / Azkargorta, Mikel / Elortza, Félix / Izquierdo, Raúl / Romero-Gavilán, Francisco / Gurruchaga, Mariló / Suay, Julio / Goñi, Isabel

Biomaterials science. 2022 Sept. 27, v. 10, no. 19

2022

Abstract: Calcium and magnesium are two elements essential for bone structure and metabolism. However, their synergistic or competitive effects on bone regeneration are often overlooked during biomaterial development. We examined the interactions between Ca and Mg ...

Abstract	Calcium and magnesium are two elements essential for bone structure and metabolism. However, their synergistic or competitive effects on bone regeneration are often overlooked during biomaterial development. We examined the interactions between Ca and Mg in sol–gel coatings doped with mixtures of CaCl₂ (0.5%) and MgCl₂ (0.5, 1, and 1.5%). After physicochemical characterisation, the materials were incubated in vitro with MC3T3-E1 osteoblastic cells and RAW264.7 macrophages, and the protein adsorption was analysed using nLC-MS/MS. The incorporation of the ions did not lead to the formation of crystalline structures and did not affect the sol–gel network cross-linking. The release of the ions did not cause cytotoxic effects at any tested concentration. The proteomic analysis showed that adding the Ca and Mg ions elevated the adsorption of proteins associated with inflammatory response regulation (e.g., ALBU, CLUS, HPT, HPTR, A1AG1 and A1AG2) but decreased the adsorption of immunoglobulins. The CaMg coatings had reduced affinity to proteins associated with coagulation (e.g., FA9, FA10, FA11, FA12) but increased the adsorption of proteins involved in cell adhesion (DSG1, DESP, FBLN1, ZA2G). In vitro assays revealed that the cellular response was affected by changing the concentration of Mg. Moreover, our results show that these differences reflect the changes in the concentrations of both ions in the mix but are not a simple additive effect.
Keywords	additive effect ; adsorption ; biocompatible materials ; bones ; calcium ; cell adhesion ; coagulation ; crosslinking ; cytotoxicity ; immunoglobulins ; inflammation ; macrophages ; magnesium ; metabolism ; proteomics ; tissue repair
Language	English
Dates of publication	2022-0927
Size	p. 5634-5647.
Publishing place	The Royal Society of Chemistry
Document type	Article
ZDB-ID	2693928-9
ISSN	2047-4849 ; 2047-4830
ISSN (online)	2047-4849
ISSN	2047-4830
DOI	10.1039/d2bm00742h
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Insight into the antibacterial mechanism of Cu-enriched sol-gel coatings employing proteomics.

Romero-Gavilán, Francisco / García-Arnáez, Iñaki / Cerqueira, Andreia / Scalschi, Loredana / Vicedo, Begonya / Villagrasa, Alejandro / Izquierdo, Raúl / Azkargorta, Mikel / Elortza, Félix / Gurruchaga, Mariló / Goñi, Isabel / Suay, Julio

Biomaterials science

2023 Volume 11, Issue 3, Page(s) 1042–1055

Abstract: Advanced antibacterial biomaterials can help reduce the severe consequences of infections. Using copper compounds is an excellent option to achieve this goal; they offer a combination of regenerative and antimicrobial functions. In this study, new ... ...

Abstract	Advanced antibacterial biomaterials can help reduce the severe consequences of infections. Using copper compounds is an excellent option to achieve this goal; they offer a combination of regenerative and antimicrobial functions. In this study, new CuCl
MeSH term(s)	Humans ; Staphylococcus aureus ; Coated Materials, Biocompatible ; Copper/chemistry ; Escherichia coli ; Proteomics ; Proteins ; Anti-Bacterial Agents/pharmacology ; Anti-Bacterial Agents/chemistry
Chemical Substances	Coated Materials, Biocompatible ; Copper (789U1901C5) ; Proteins ; Anti-Bacterial Agents
Language	English
Publishing date	2023-01-31
Publishing country	England
Document type	Journal Article
ZDB-ID	2693928-9
ISSN	2047-4849 ; 2047-4830
ISSN (online)	2047-4849
ISSN	2047-4830
DOI	10.1039/d2bm01421a
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Article ; Online: Using osteogenic medium in the in vitro evaluation of bone biomaterials: Artefacts due to a synergistic effect.

Romero-Gavilán, Francisco / García-Arnáez, Iñaki / Cerqueira, Andreia / Arias-Mainer, Carlos / Azkargorta, Mikel / Elortza, Félix / Izquierdo, Raúl / Gurruchaga, Mariló / Goñi, Isabel / Suay, Julio

Biochimie

2023 Volume 216, Page(s) 24–33

Abstract: In vitro tests using bone cells to evaluate the osteogenic potential of biomaterials usually employ the osteogenic medium (OM). The lack of correlation frequently reported between in vitro and in vivo studies in bone biomaterials, makes necessary the ... ...

Abstract	In vitro tests using bone cells to evaluate the osteogenic potential of biomaterials usually employ the osteogenic medium (OM). The lack of correlation frequently reported between in vitro and in vivo studies in bone biomaterials, makes necessary the evaluation of the impact of osteogenic supplements on these results. This study analysed the proteomic profiles of human osteoblasts (HOb) cultured in the media with and without osteogenic agents (ascorbic acid and β-glycerol phosphate). The cells were incubated for 1 and 7 days, on their own or in contact with Ti. The comparative Perseus analysis identified 2544 proteins whose expression was affected by osteogenic agents. We observed that the OM strongly alters protein expression profiles with a complex impact on multiple pathways associated with adhesion, immunity, oxidative stress, coagulation, angiogenesis and osteogenesis. OM-triggered changes in the HOb intracellular energy production mechanisms, with key roles in osteoblast maturation. HOb cultured with and without Ti showed enrichment in the skeletal system development function due to the OM. However, differentially expressed proteins with key regenerative functions were associated with a synergistic effect of OM and Ti. This synergy, caused by the Ti-OM interaction, could complicate the interpretation of in vitro results, highlighting the need to analyse this phenomenon in biomaterial testing.
MeSH term(s)	Humans ; Osteogenesis ; Artifacts ; Proteomics ; Bone and Bones ; Cell Differentiation ; Osteoblasts ; Biocompatible Materials/pharmacology ; Biocompatible Materials/metabolism
Chemical Substances	Biocompatible Materials
Language	English
Publishing date	2023-09-15
Publishing country	France
Document type	Journal Article
ZDB-ID	120345-9
ISSN	1638-6183 ; 0300-9084
ISSN (online)	1638-6183
ISSN	0300-9084
DOI	10.1016/j.biochi.2023.09.015
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Article ; Online: The effect of calcium-magnesium mixtures in sol-gel coatings on bone tissue regeneration.

Biomaterials science

2022 Volume 10, Issue 19, Page(s) 5634–5647

Abstract	Calcium and magnesium are two elements essential for bone structure and metabolism. However, their synergistic or competitive effects on bone regeneration are often overlooked during biomaterial development. We examined the interactions between Ca and Mg in sol-gel coatings doped with mixtures of CaCl
MeSH term(s)	Biocompatible Materials/pharmacology ; Bone Regeneration ; Calcium/chemistry ; Calcium Chloride ; Ions/pharmacology ; Magnesium/chemistry ; Proteomics ; Tandem Mass Spectrometry
Chemical Substances	Biocompatible Materials ; Ions ; Magnesium (I38ZP9992A) ; Calcium Chloride (M4I0D6VV5M) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2022-09-27
Publishing country	England
Document type	Journal Article
ZDB-ID	2693928-9
ISSN	2047-4849 ; 2047-4830
ISSN (online)	2047-4849
ISSN	2047-4830
DOI	10.1039/d2bm00742h
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Article ; Online: Complex effects of Mg-biomaterials on the osteoblast cell machinery: A proteomic study.

Cerqueira, Andreia / García-Arnáez, Iñaki / Romero-Gavilán, Francisco / Azkargorta, Mikel / Elortza, Félix / Martín de Llanos, José Javier / Carda, Carmen / Gurruchaga, Mariló / Goñi, Isabel / Suay, Julio

Biomaterials advances

2022 Volume 137, Page(s) 212826

Abstract: The cell-biomaterial interface is highly complex; thousands of molecules and many processes participate in its formation. Growing demand for improved biomaterials has highlighted the need to understand the structure and functions of this interface. ... ...

Abstract	The cell-biomaterial interface is highly complex; thousands of molecules and many processes participate in its formation. Growing demand for improved biomaterials has highlighted the need to understand the structure and functions of this interface. Proteomic methods offer a viable alternative to the traditional in vitro techniques for analyzing such systems. Magnesium is a promoter of cell adhesion and osteogenesis. Here, we used the LC-MS/MS to compare the protein expression profiles of human osteoblasts (HOb) exposed to sol-gel coatings without (MT) and with Mg (MT1.5Mg) for 1, 3, and 7 days. PANTHER, DAVID, and IPA databases were employed for protein identification and data analysis. Confocal microscopy and gene expression analysis were used for further characterization. Exposure to MT1.5Mg increased the HOb cell area and the expression of SP7, RUNX2, IBP3, COL3A1, MXRA8, and FBN1 genes. Proteomic analysis showed that MT1.5Mg affected the early osteoblast maturation (PI3/AKT, mTOR, ERK/MAPK), insulin metabolism, cell adhesion (integrin, FAK, actin cytoskeleton regulation) and oxidative stress pathways. Thus, the effects of Mg on cell adhesion and osteogenesis are rather complex, affecting several pathways rather than single processes. Our analysis also confirms the potential of proteomics in biomaterial characterization, showing a good correlation with in vitro results.
MeSH term(s)	Biocompatible Materials/pharmacology ; Chromatography, Liquid ; Humans ; Integrins/metabolism ; Osteoblasts ; Proteomics ; Tandem Mass Spectrometry
Chemical Substances	Biocompatible Materials ; Integrins
Language	English
Publishing date	2022-04-25
Publishing country	Netherlands
Document type	Journal Article
ISSN	2772-9508
ISSN (online)	2772-9508
DOI	10.1016/j.bioadv.2022.212826
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Article ; Online: Influence of calcium ion-modified implant surfaces in protein adsorption and implant integration.

Anitua, Eduardo / Cerqueira, Andreia / Romero-Gavilán, Francisco / García-Arnáez, Iñaki / Martinez-Ramos, Cristina / Ozturan, Seda / Azkargorta, Mikel / Elortza, Félix / Gurruchaga, Mariló / Goñi, Isabel / Suay, Julio / Tejero, Ricardo

International journal of implant dentistry

2021 Volume 7, Issue 1, Page(s) 32

Abstract: 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 ... ...

Abstract	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.
MeSH term(s)	Adsorption ; Animals ; Chromatography, Liquid ; Ions ; Osseointegration ; Rabbits ; Surface Properties ; Tandem Mass Spectrometry
Chemical Substances	Ions
Language	English
Publishing date	2021-04-21
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2198-4034
ISSN (online)	2198-4034
DOI	10.1186/s40729-021-00314-1
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