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Blood protein coated model biomaterials: preparation, and cell and tissue response
Linköping University, Department of Physics, Measurement Technology, Biology and Chemistry. Linköping University, The Institute of Technology.
2003 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Solid biomaterials are widely used in bone and in soft tissue applications. Problems may then arise due to a prolonged inflammation in the proximity of the implant, resulting in the formation of a fibrous capsule and a low vascularisation near the interface.

The formation of a blood plasma clot is the starting point of a normal wound healing process. One hypothesis in this thesis work was that a thin immobilised blood plasma clot may improve the integration during the early wound healing period. Model surfaces were made from titanium and silicon, and nm-μm thick blood plasma clots or protein multilayers immobilised onto the surfaces. Another hypothesis was that a submicron surface porosity further improves the integration process, and to study this some of the titanium surfaces were etched in sodium hydroxide, a treatment that resulted in 200 nm wide pores. Such porous titanium surfaces adsorbed 2 to 11 times more albumin and lgG in vitro than the corresponding smooth surfaces at varied pH and protein concentrations.

The blood plasma clot coated submicron porous titanium samples were implanted subcutaneously in the back of the rat or in rabbit bone. The soft tissue response was investigated after 3 or 24 hours and the fibrous encapsulation and vessel formation after 7 or 28 days of implantation. The bone ingrowth and the implant stability in the rabbit bone were investigated after 4 weeks of implantation.

The monocyte response on multilayer plasma protein coated surfaces was investigated through the analysis of tumor necrosis factor α (TNF-α) and interleukin-10 (IL-10) concentrations in the culture medium, the proportions of Annexin V and propidium iodide (PI) positive cells, and the amounts of nucleated cells. In parallel, the stability of the protein layers and the activation of the complement and coagulation cascades were investigated in vitro by ellipsometry.

The results from the monocyte culture and animal experiments show that the early soft tissue inflammatory response and vascularisation can be modulated through the introduction of a surface porosity and by the immobilised protein and plasma clot coatings. However, no significant differences were observed between the different surface modifications in rabbit bone with respect to bone-to-metal contact or percentage of bone area inside the threads.

Place, publisher, year, edition, pages
Linköping: Linköping University , 2003. , p. 44
Series
Linköping Studies in Science and Technology. Dissertations, ISSN 0345-7524 ; 798
National Category
Medical Biotechnology
Identifiers
URN: urn:nbn:se:liu:diva-179309Libris ID: 8861364ISBN: 9173735930 (print)OAI: oai:DiVA.org:liu-179309DiVA, id: diva2:1595199
Public defence
2003-03-14, hörsal Planck, Fysikhuset, Linköpings universitet, Linköping, 09:00
Opponent
Available from: 2021-09-24 Created: 2021-09-17 Last updated: 2023-02-28Bibliographically approved
List of papers
1. In vitro preparation and ellipsometric characterization of thin blood plasma clot films on silicon
Open this publication in new window or tab >>In vitro preparation and ellipsometric characterization of thin blood plasma clot films on silicon
2001 (English)In: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 22, no 13, p. 1803-1808Article in journal (Refereed) Published
Abstract [en]

The wound-healing process around implants differs from that of a normal healing without the inserted material. In this work, the composition of a natural wound surface was mimicked through clotting of a thin human blood plasma film with approximate ellipsometric thickness of 100nm onto differently pretreated silicon surfaces. Their stability was investigated by incubations in sodium dodecyl sulphate (SDS) solutions. The enzymatic clot degradation was induced through addition of human tissue plasminogen activator (t-PA) to the plasma and the surface protein remnants after the degradation were analyzed with polyclonal antibodies. The results show that the plasma films were not SDS resistant on hydrophilic silicon. However, stability was obtained after preparation on hydrophobic silicon or when albumin or fibrinogen was immobilized to silicon before the plasma incubations. Different surfaces bound different polyclonal antibodies after the clot film degradation. The methods indicate a simple means to improve or reestablish a normal tissue inflammatory response around biomaterials. Copyright © 2001 Elsevier Science Ltd.

Keywords
Degradation, Ellipsometry, Plasma clot, Remnant, Stability, Surface coating
National Category
Engineering and Technology
Identifiers
urn:nbn:se:liu:diva-47319 (URN)10.1016/S0142-9612(00)00359-8 (DOI)
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2021-09-17
2. Adsorption of albumin and IgG to porous and smooth titanium
Open this publication in new window or tab >>Adsorption of albumin and IgG to porous and smooth titanium
2004 (English)In: Colloids and Surfaces B: Biointerfaces, ISSN 0927-7765, E-ISSN 1873-4367, Vol. 35, no 1, p. 45-51Article in journal (Refereed) Published
Abstract [en]

The possibility to load submicrometer porous titanium surfaces with relatively small proteins, albumin and immunoglobulin G (IgG) was investigated. The loading ability is of interest due to the possibility of slow release of molecules from biomaterial surfaces, and may be important for the manipulation of wound healing around prostheses. Iodine-125 (125I) labeled albumin and IgG were adsorbed onto smooth and to porous titanium with a pore diameter of 200-300 nm. The smooth and porous surfaces were divided into three groups: hydrophilic, hydrophobic, or to amine-terminated silane (3- aminopropyltriethoxysilane) that bound proteins via glutaraldehyde. The protein solution pH and protein concentrations were varied, and the adsorption experiments made without or in the presence of calcium and magnesium ions. The adsorbed amounts were quantified with a gamma counter. Two to eleven times more proteins adsorbed onto porous than smooth surfaces and the adsorbed amounts increased with increasing protein concentration (0.1-10 mg/ml) during a constant incubation time. The elutability by sodium dodecyl sulphate (SDS) was incomplete on porous surfaces. © 2004 Elsevier B.V. All rights reserved.

National Category
Natural Sciences
Identifiers
urn:nbn:se:liu:diva-41185 (URN)10.1016/j.colsurfb.2004.02.007 (DOI)55311 (Local ID)55311 (Archive number)55311 (OAI)
Available from: 2009-10-10 Created: 2009-10-10 Last updated: 2021-09-17
3. Ex vivo PMA-induced respiratory burst and TNF-a secretion elicited from inflammatory cells on machined and porous blood plasma clot-coated titanium
Open this publication in new window or tab >>Ex vivo PMA-induced respiratory burst and TNF-a secretion elicited from inflammatory cells on machined and porous blood plasma clot-coated titanium
2002 (English)In: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 23, no 13, p. 2803-2815Article in journal (Refereed) Published
Abstract [en]

The release of inflammatory mediators around implants and normal wounds may differ due to the presence of the solid surface. In this study, machined and sub-micron porous titanium implants with and without a 100nm thick blood plasma clot were inserted subcutaneously in rat for 3 or 24h. The cell recruitment to the interfaces, in vivo secretion of TNF-a and the ex vivo PMA-induced production of reactive oxygen species were subsequently investigated. The thin plasma clot coating gave rise to an increased ex vivo PMA-stimulated oxygen radical production by implant-associated cells at both implantation times, and an increased cell recruitment at 24h. The total TNF-a secretion was highest at sham sites and plasma clot-coated porous titanium at 24h. After 24h, the cell-type pattern in the exudate around the porous plasma-coated implant was more similar to that found at sham sites than that adjacent to the non-coated implants. No differences were observed between the machined Ti and the machined sub-micron porous Ti. © 2002 Elsevier Science Ltd. All rights reserved.

Keywords
Coagulation, Fibrinogen, Inflammation, Plasma, Porous surfaces
National Category
Engineering and Technology
Identifiers
urn:nbn:se:liu:diva-47012 (URN)10.1016/S0142-9612(02)00021-2 (DOI)
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2021-09-17
4. On the formation of fibrous capsule and fluid space around machined and porous blood plasma clot coated titanium
Open this publication in new window or tab >>On the formation of fibrous capsule and fluid space around machined and porous blood plasma clot coated titanium
Show others...
2001 (English)In: Journal of materials science. Materials in medicine, ISSN 0957-4530, E-ISSN 1573-4838, Vol. 12, no 10-12, p. 1019-1024Article in journal (Refereed) Published
Abstract [en]

Machined and machined submicron porous titanium, with and without a thin blood plasma coating (100 nm), were implanted for 7 or 28 days in subcutaneous pockets on the back of the rat. After explantation the specimens were analyzed by light microscopy with respect to thickness of the fibrous capsule, the fluid space width between implants and fibrous capsule, and formation of blood vessels. The results at 7 days indicate a thinnest fluid space for the plasma clot coated porous titanium surface, and the spaces vanished at the light microscopic level after 28 days outside all the analyzed surfaces. The thickness of the fibrous capsule increased outside the different surfaces at 7-28 days, and in this respect no significant differences were observed between the different surfaces at any time. Analysis of neovascularization showed that the number of vessels and proportion of vessels in the fibrous capsule increased with time at all surfaces, except machined Ti where the number instead decreased from 7 to 28 days. The average distance between the blood vessels and the fluid space increased with time for all types of surfaces. The results in the present study indicate that the healing process around titanium can be modulated by porosity and thin pre-prepared plasma coatings. (C) Kluwer Academic Publishers.

National Category
Engineering and Technology
Identifiers
urn:nbn:se:liu:diva-49063 (URN)
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2021-09-17
5. Bone formation after 4 weeks around blood-plasma-modified titanium implants with varying surface topographies: An in vivo study
Open this publication in new window or tab >>Bone formation after 4 weeks around blood-plasma-modified titanium implants with varying surface topographies: An in vivo study
2003 (English)In: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 24, no 2, p. 197-205Article in journal (Refereed) Published
Abstract [en]

The aim of the present study was to investigate and compare the stability and bone ingrowth capacity to screw-shaped titanium implants with five different surface treatments. The implants were: (1) standard turned with a thin blood plasma coat (TP), (2) NaOH-etched dito with pore size 0.2-0.3µm (E), (3) NaOH-etched with pore size 0.2-0.3µm and a thin blood plasma coat (EP), (4) electrochemically oxidised with pore size 1-2µm (O), (5) electrochemically oxidised with pore size 1-2µm and a thin blood plasma coat (OP). A total of 66 implants were divided into the above-described five groups and inserted for 4 weeks into tibia and femur of 11 rabbits. The implants were evaluated by resonance frequency (RF) measurements at the time of insertion and removal, and analysed histomorphometrically at removal. The RF measurements showed that the implant stability was lower in soft bone compared to dense and increased with time. No significant differences were observed between the different surface modifications. The histomorphometric analysis revealed no statistically significant differences between the implants regarding bone-to-metal contact (BMC) and bone area inside the threads (BA). The above results indicate that thin blood plasma-coated and non-coated screw-shaped titanium implants with turned, NaOH-etched and electrochemically etched surface profiles integrate similarly to bone at 1 month of implantation. © 2002 Elsevier Science Ltd. All rights reserved.

Keywords
Blood-plasma-coated, Pores, Surface characterisation, Surface modification, Titanium implants, Titanium oxide
National Category
Engineering and Technology
Identifiers
urn:nbn:se:liu:diva-46757 (URN)10.1016/S0142-9612(02)00277-6 (DOI)
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2021-09-17
6. Preparation of multilayer plasma protein films on silicon by EDC/NHS coupling chemistry
Open this publication in new window or tab >>Preparation of multilayer plasma protein films on silicon by EDC/NHS coupling chemistry
Show others...
2003 (English)In: Colloids and Surfaces B: Biointerfaces, ISSN 0927-7765, E-ISSN 1873-4367, Vol. 28, no 4, p. 261-272Article in journal (Refereed) Published
Abstract [en]

Crosslinked multilayer protein films were prepared from fibrinogen, albumin, IgG, a combination of fibrinogen and catalase, and blood plasma on silicon by ethyl-dimethyl-aminopropylcarbodiimide and N-hydroxy-succinimide coupling chemistry. The 4-70 nm thick films were placed in blood plasma and the additional protein deposition measured by null ellipsometry after 5 or 60 min of incubation. The activation of the complement system and intrinsic pathway of coagulation were indicated through the subsequent binding of anti-C3c, anti-C3d, anti-properdin and anti-HMWK on top of the surface bound blood plasma. The proportion of Annexin V, Propidium Iodide and 4,6-diamidino-2-phenylindole positive cells, and the secretion of tumor necrosis factor a (TNF-a) and interleukin-10 (IL-10) were analysed in a monocyte culture. The results show that well known protein coupling techniques can be used for the preparation of protein layers with well controlled thickness. The layers possess low contact activation of blood plasma and induce different release of TNF-a and IL-10 in monocyte cultures. © 2002 Elsevier Science B.V. All rights reserved.

Keywords
Adsorption, Cytokine, Monocyte, Multilayer, Protein
National Category
Engineering and Technology
Identifiers
urn:nbn:se:liu:diva-46560 (URN)10.1016/S0927-7765(02)00151-0 (DOI)
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2021-09-17

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