Surface Enginnering on Medical Devices

Surface Engineering

on

Medical Devices.

Caxias do Sul

17/04/2014

©. Property of TECNALIA.

Surface Engineering on Medical

Devices.

Problems and challenges addressed

with Surface Engineering techniques

from the research, development,

evaluation and validation points of view.


Surface Engineering on Medical Devices. Abstract

Medical Devices play an increasingly important role in clinical treatments. From surgical

tools to permanent implants, they are both the key of a successful treatment and often the

source of adverse reactions. Success or failure most of the times will depend on events

occurring on the medical device – human tissue interface, which typically are heavily

influenced by the biomaterial surface properties. Therefore, surface treatment and coating

technologies constitute powerful tools in the development of new devices and improvement

of the performance of existing ones.

We will review a number of surface technologies that are applied on medical devices, as

well as a number of case studies where issues such as infection resistance or tissue

integration of medical devices are address through the development of new surfaces.

Surface Engineering on Medical Devices.


Index

-. Tecnalia.

-. Surface engineering vs. biomedical RTD.

Problems & challenges.

Regulatory aspects.

Economic aspects.

Intellectual property rights.

Case studies.

i) Antimicrobial surfaces.

ii) Tissue integration/regeneration.



TECNALIA is the first applied

research centre in Spain and one

of the most important in Europe

with around 1.400 staff, 110

million Euro turnover and over

4.000 clients.

A unique commitment, an opportunity, a challenge.


4 APPROACHES TO THE WAY WE WORK WITH COMPANIES

TECHNOLOGICAL

SERVICES //

VENTURES //

TECHNOLOGY

AND INNOVATION

STRATEGY //

R&D PROJECTS //

YOUR

PROJECT


European main RTO’s Funding Sources

2011 data

With a strong corporate

involvement

In TECNALIA's

representation,

administration and

management.

In the strategic

market orientation of

our divisions.

In strategic and

investment planning.

In the development

and financing of

collaborative

activities.

In business creation.


Organised in 7 Business Divisions: we work from the

experience and the expertise we have acquired in the markets in

which we operate, with an efficient and proactive attitude.


International

Presence


Index

-. Tecnalia.


Problems & challenges. Regulatory aspects.

Economic aspects.


Case studies.






Medical Devices: “…any instrument, apparatus, appliance, software, material or other

article, … to be used specifically for diagnostic and/or therapeutic purposes…”

Implant: “… Any device which is intended

— to be totally introduced into the human body or,

— to replace an epithelial surface or the surface of the eye,

by surgical intervention which is intended to remain in place after the procedure.

Transient: < 60 minutes.

Short term: < 30 days.

Long term: > 30 days.

Properties: mechanical, biocompatible,…

Challenges: tissue interaction & integration,

infections, …

how to make them more safe,

durable and efficient

Solution strategies: novel surfaces & biomaterials.



With modern protocols, the incidence of infections associated to placement of implants

has become very low: e.g. estimated in 0.5–5% for total joint replacements.

Nevertheless, infections still cause a huge impact in terms of morbidity, mortality, and

medical costs. e.g. in orthopedics, the treatment of each single episode of infected

arthroplasty costs >$50,000.

Aprox. half of the nosocomial infections are related to the use of medical devices.

The pathogenesis of peri-implant infections. -> the critical dose of contaminating

microorganisms to produce infection is much lower with a foreign material at the surgical

site. Key: cell anchorage and fixation and formation of an adherent biofilms.

As in other medical fields, prevention represents a main goal, which relies on a series of

strategies on different ground levels:

-. in the past decades, control of environmental and personnel contamination has been

a principal target to cut down the rate of nosocomial and post-surgical infections.

-. the establishment of effective protocols of peri-operative antibiotic prophylaxis.

Infections vs. antimicrobial surfaces.

Problems & challenges. Surface Engineering on Medical Devices.


-. The interface biomaterial surface-surrounding tissue is where accidental

contamination can first develop into colonization and, subsequently, into the establishment

of a clinically relevant infection. -> the most convenient way to interfere with the early

phases of microbial adhesion is a modification of the chemistry or the micro/nanotopology of

the out-layer of the device.

Locally applied antibiotics under temporally controlled release present many advantages

over systemic clinical treatments, e.g. efficiency and side effects. Biofilm formation at the

implant tissue interface, a barrier that diminishes or disables the penetration of systemic

antibiotics. This can be achieved by a coating on top of the medical device.

Bacteriostatic and antimicrobial* surfaces: ion implantation of Ag / Si.

Drug release: ciprofloxacin.

Drug attachment: vancomicyn.

*: bacteriostatic: inhibiting growth or multiplication of bacteria.

antimicrobial: killing microorganisms or suppressing their multiplication or growth.

Antimicrobial surfaces.

Problems & challenges. Surface Engineering on Medical Devices.

Infections vs. antimicrobial surfaces.


Bone ?

-. Bio-nanocomposite made up of an organic fiber matrix (collagen,…) stiffened by ceramic

nano-crystals

(30-50nm length; 15-30nm width, 2-10nm thickness… Mj.J.Olszta et al.)

-. Non homogeneous structure.

-. Dynamic material,

that can remodel and

adapt to the

bio-mechanical

environment.

http://training.seer.cancer.gov

Surface Engineering on Medical Devices. Problems & challenges.

Tissue integration / regeneration vs. osseoinductive surfaces


Human bone AFM

. .



Human bone AFM

. .



Human bone AFM

. .



Human bone AFM

. .

Bone particle sizes:

10-30nm



Implants & prostheses:

-. Often Ti & alloys: relatively light & good mechanical prop.

-. Natural oxide layer (1.5 – 10 nm thick)

-. Tissue regeneration around titanium, i.e. osseo-integration,

“…a direct connection between living bone and a load-carrying endosseous implant at light

microscopic level…” .

Osseo integration depends on

-. Implant material

-. Implant design

-. Status of bone

-. Surgical technique

-. Implant loading conditions

-. Surface quality




.

Osseo integration depends on: Surface quality

-. Surface chemistry.

HAp, bioactive ceramics,…

Ti, Ta, ... oxide layer

Ion release

Corrosion resistance

bioactive molecules

-. Surface physical properties:

visco-elasticity,

surface energy,…

-. Surface topography.

Milimiter roughness

Micrometer “

Nanometer “





Index

-. Tecnalia.



Regulatory aspects. Economic aspects.


Case studies.





Regulatory aspects

… rules relating to the safety and performance of medical devices were harmonized

in the EU in the 1990s. The core legal framework consists of 3 directives:

• Directive 90/385/EEC regarding active implantable medical devices,

(devices that require external power sources in order to function properly)

• Directive 98/79/EEC regarding in vitro diagnostic medical devices.

(devices used for the examination of specimens taken from the human body)

• Directive 93/42/EEC regarding medical devices

and,

… requirements and procedures for the marketing authorization for medicinal products for

human use, as well as the rules for the constant supervision of products after they have

been authorized, primarily laid down in:

• Directive 2001/83/EC relating to medicinal products for human use.

Regulatory aspects.



Regulatory aspects

Some key issues:

• Identify Directives and Regulations: Is it really a medical device according to Directive

93/42/EEC?

“…any instrument, apparatus, appliance, software, material or other article, … to be used

specifically for diagnostic and/or therapeutic purposes…”

• Classify according to MDD Annex IX:

• How to “build” the Technical File demonstrating compliance?

• Enough to “convince” the Notified Body?

• How long and cost of achieving the CE marking?

Regulatory aspects.



Regulatory aspects

Some key issues:


1. Description of product family and justification for why your device falls into that family

2. Device intended use

3. Description of device components, specifications, packaging and literature

4. Device manufacturing Process

5. List of accessories to your device

6. Location of design responsibility and manufacturing facilities

7. Classification along with rationale for classification

8. Chosen compliance route according to applicable Directive(s)

9. Declaration of Conformity stating manufacturer’s compliance with applicable Directive(s)

10. Shelf life and environmental limitations of device

11. Retention of quality assurance, Competent Authority and Notified Body records

12. Vigilance reporting and Medical Device Reporting procedures

13. How and when to contact Competent Authorities

14. Name of and contract with your Authorized Representative

15. Subcontractor names and addresses if applicable

16. Essential Requirements

17. Design input specifications

18. Application and references to Standards and Guidelines

19. Testing results and clinical evaluations

20. Risk analysis

21. Instructions for Use and Labeling

Regulatory aspects.



Regulatory aspects

Some key issues:


Testing program?

= RTD testing?

Harmonized standards?

Quality System in compliance with ISO 13485?

Regulatory aspects.



Index

-. Tecnalia.



Regulatory aspects.

Economic aspects.

Intellectual property rights. Case studies.





Economic aspects

Some key issues:

• Medical Devices where the technology could be implemented?

• Public or private health services?

• Performance improving technology?

• Cost saving technology?

• Regulatory process time and cost?

• Scaling up of technology vs. ISO 13485?

Economic aspects.



Some key issues:

• Freedom to operate?

• Effective protection?

• Time to market vs. patent life?


Index

-. Tecnalia.



Regulatory aspects.

Economic aspects.


Case studies.


Ion implantation

Antibiotic release

Plasma+click attachment ii) Tissue integration/regeneration.




M&M: Substrate: AISI 316 LVM

Si+ ion implantation.

Ref: Si-I: 5×1016 ions/cm2, 50keV, angles of ion incidence of 90º.

Ref: Si-II: 2.5×1016 ions/cm2, 50keV, angles of ion incidence of 45º.

Properties/results:

Bacteriostatic surfaces. ion implantation of Si.




Bacteriostatic surfaces:

ion implantation of Si.

Biocompatibility:

Bacteria adhesion rates in a parallel plate flow chamber microorganisms were let to adhere

under dynamic (flowing the bacterial suspension) and static conditions (stopping the flow).

for the dynamic (jo) and static (n) (↓↑:statistical significance, p <0.05). [S. aureus ATCC29213, S. epidermidis ATCC35984 (S. epidermidis4) and S. epidermidis HAM892 (S. epidermidis2)].

Bacteriostatic surfaces.


[Applied Surface Science, In Press, Accepted Manuscript, Available online 4 April 2014]


Index

-. Tecnalia.



Regulatory aspects.

Economic aspects.


Case studies.


Ion implantation

Antibiotic release

Plasma+click attachment ii) Tissue integration/regeneration.




Drug released: ciprofloxacin

M&M process parameters

Precursor: N,O-bis-tert-butyldimethylsilylated ciprofloxacin (silylciprofloxacin),

+ the low-energy plasma activation of the silylated groups

Ar or N2 RF plasma to surfaces coated with chemically modified ciprofloxacin.

+ hydrolytic profile of silyl carboxylates in aqueous media/ physiological media.

+ residual tert-butyldimethylsilanol by product arising from the hydrolysis is nontoxic.

Antimicrobial surfaces. Plasma Polymerized Silylated Ciprofloxacin




Tests / results.





Tests / results.





Tests / results.

An antibiotic, ciprofloxacin, has been disilylated and successfully incorporated into a

coating by a plasma polymerization process, keeping its antibiotic activity once released

by hydrolysis in physiological conditions.

The release dynamics are significantly influenced by the plasma processing parameters.

The plasma polymerization technique, in combination with suitably silylated prodrugs,

offers the possibility of tailoring the coating properties, e.g., thickness and degree of

polymerization, and thus the release dynamics of the antibiotic, to a wide range of

medical devices and clinical contexts.

Plasma Polymerized Silylated Ciprofloxacin as an Antibiotic Coating.

Plasma Proc. and Polymers 8 (7) (2011) 599–606.




Index

-. Tecnalia.



Regulatory aspects.

Economic aspects.


Case studies.


Ion implantation

Antibiotic release

Plasma + click attachment ii) Tissue integration/regeneration.




Prof of concept:

A) Plasma polymerization.

B) Vancomycin modification with azide linker.

C) “Click” chemistry.

D) Evaluation of antibiotic activity, against Staphylococcus epidermidis

Antimicrobial surfaces. A “Plasma-Click” Dual Procedure





M&M:

A) Plasma surface modification.

Substrate material: glass slides / KBr.

Technique: Ion Gun Inverse Magnetron (IGIM).

Optimization of plasma polymerization process.

Precursor Acrylic Acid.

Objective: COOH functional groups at the surface.

Process parameters:

Pressure Ar Flow

Acrylic

Acid Flow

C

O

2 Time Distance Current Voltage

[mbar] [µL•min-1

] [g/h]

[

µ [min] [mm] [A] [V]

CO2H-0 E-3 70 3 - 30 117 0,07-0,08 340

CO2H-1 E-3 70 1 - 30 117 0,07-0,08 340

CO2H-2 E-3 70 1 - 30 117 0,25 340

Ref.

Process Conditions





~1706 cm-1 C=O

~ 2900–3300 cm-1

OH

C-O 1280 cm-1

~1706 cm-1 C=O C-H 1450 cm-1

1640 cm-1 C=C

M&M:


Characterization: FTIR.

Ref.

CO2H-1

CO2H-2

CO2H-0



Ion implantation and plasma based processes on Medical Devices Antimicrobial surfaces.


M&M:


Optimization processes with constant current of 0.07-0.08 A and 30 minutes.

Characterization: density of COOH groups by colorimetry with toluidine blue.

Results

Pressure Ar Flow

Acrylic

Acid Flow CO2 Flow Time Distance Current Voltage Colorimetry

[mbar] [µL•min-1

] [g/h] [µL•min-1

] [min] [mm] [A] [V] [nmol•cm-2

]

CO2H-3 E-3 70 3 - 30 117 0,07-0,08 340 0,46 (±0,01)

CO2H-6 E-3 - 3 132 30 117 0,07-0,08 337 0.99 (±0.09)

CO2H-7 E-3 - 3 70 30 117 0,07-0,08 337 0.35 (±0.16)

CO2H-9 E-3 - 3 265 30 117 0,07-0,08 337 1.44 (±0.38)

CO2H-11 E-3 - 1,5 132 30 117 0,07-0,08 337 1.87 (±0.22)

Ref.

Process Conditions

Selected process parameters:

Acrylic Acid Flow 1.5 µL min-1; CO2 gas flow 132 µL min-1; t= 30 minutes;

current 0.07 to 0.08A, voltage 337V.

Density of COOH groups: 1.87±0.22 nmol/cm2.




M&M:


Vancomycin hydrochloride in DMSO

+ DMF and 4-methylazido-benzylamine,

… mixture cooled to 0ºC

…+ HBTU in DMF and DMSO

…+ DIPEA.

… stirred overnight at room temperature.

… quenched by adding it dropwise to acetone,

precipitated, filtered, and washed.

… modified vancomycin purified by RP-HPLC.

.




M&M:


Purification by RP-HPLC.

Characterization: FTIR, MS Spectrometry.

FTIR: Vancomycin and vancomycin azide in KBr, N3 at ~2100 cm-1

Reversed phase HPLC chromatograms:

Eluent MeCN/H2O/TFA; retention time [min.]

Vancomycin, top-right

and vancomycin azide, below-right.





M&M:

C) “Click” chemistry.

Method: surface amidation with propargylamine of plasma modified surface

“click” chemistry with vancomycin-azide.6

Van- azide in PBS + alkyne glass slide + … addition of CuSO4 solution and sodium

acorbate solution, under nitrogen at room temperature for 24 h, washed with PBS.

Characterization: XPS after (ultrasound in distilled water).

The N signal corresponds to the N atom in the propargylamine, C3H5N

The Cl signal corresponds to the Cl atom in the original vancomycin molecule = C66H75Cl2N9O24

Ref. Peak BE At.% Ref. Peak BE At.%

Control C1s 285.06 70.7% "click"ed C1s 288.79 75.4%

O1s 536.53 26.7% vancomycin O1s 536.33 18.8%

N1s 404.48 2.6% N1s 404.1 5.4%

Cl2p3 0 0.0% Cl2p3 204.72 1.0%





M&M:

D) Evaluation of antibiotic activity: immersion test.

Against Staphylococcus epidermidis (CECT 231) bacteria

Surface modification prepared on inert glass; control: untreated glass.

i) immersion glass samples, at different bacterial concentrations:

5.9E+01, 1.2E+02 and 1.2E+03 c.f.u./mL, in nutrient broth, cultured at 37ºC

and counting bacterial viability after 18 hours. n= 2 or 3 per concentration.

Initial bacterial Inhibition

concentration at 18h

c.f.u. / ml % S.D.

5.9 x10 99.85% (±1.4)

1.2 x 102

96.33% (±2.4)

1.2 x 103

95.80% (±4.9)

http://visualsunlimited.photoshelter.com/image?&_bqG=13&_bqH=eJxzKklNDshNcs52MTUqLzHIznMrN02rDHA2LTSxMrS0MjQwAGEg6RnvEuxsm1qQmZJalJuZklmsBhaJd_RzsS0Bsv2D3OM9XWz9QYq9TTwLEi0sTNJNs9XiHZ1DbItTE4uSMwAXACEY&GI_ID=




M&M:

D) Evaluation of antibiotic activity: direct contact.

Against Staphylococcus epidermidis (CECT 231) bacteria

Surface modification prepared on inert glass; control: untreated glass.

ii) following indications of the JISZ 2801 standard; culture at 37ºC and 100% humidity;

extraction and counting after 10, 24 and 48 hours. n= 3 per test time.

Initial bacterial Inhibition Inhibition Inhibition

concentration at 10 h at 24 h at 48h

c.f.u. / ml % % %

4 x 105 96.4%

(±2.4)

87.4% (±8.1)

87.8% (±7.1)

http://visualsunlimited.photoshelter.com/image?&_bqG=13&_bqH=eJxzKklNDshNcs52MTUqLzHIznMrN02rDHA2LTSxMrS0MjQwAGEg6RnvEuxsm1qQmZJalJuZklmsBhaJd_RzsS0Bsv2D3OM9XWz9QYq9TTwLEi0sTNJNs9XiHZ1DbItTE4uSMwAXACEY&GI_ID=




Conclusions: A “plasma-click” based coating has shown to be an effective technique for

producing surfaces with antibiotic activity.

The “click”ed antibiotic, i.e. vancomycin, showed antibiotic activity against

Staphylococcus epidermidis after all the coating processes (e.g. azidation of the

vancomycin and click chemistry).

The plasma polymerization technique allows controlling the concentration of CO2H on

the surface of the coated material, that would affect the final max. concentration of

vancomycin, and consequently the antibiotic activity.

The strategy described is feasible and could be used for several antibiotics.

Furthermore, such antibiotic coatings can be deposited on any medical device that can

withstand the plasma process.

Further work is necessary to determine the optimum vancomycin concentration at

the surface and confirm the antibiotic activity against other bacteria, e.g.

Escherichia coli, Staphylococcus aureus, Pseudomona aeruginosa,…

“Plasma-Click” Based Strategy for Obtaining Antibacterial Surfaces on Implants.

Plasma Proc. Polymers 10(4)(2013)328–335



Index

-. Tecnalia.



Regulatory aspects.

Economic aspects.


Case studies.



Ion Implantation



Tissue integration

Tissue integration.

CO ion implanted implantable musculoeskeletal implants. ..



AFM

surface

topography

Ref. E

Ref. A Ref.B

Ref. C Ref. D

Ref. E

Ref. A Ref.B

Ref. C Ref. D

Tissue integration Surface Engineering on Medical Devices.


In vitro cell culture tests:

-. Cell attachment.

-. Cell proliferation [Surf. Coat Tech vol. 196 (2005) p. 321-326]

-. Cell morphology.

-. Cell apoptosis. [Surf. Coat Tech vol. 201 (2007) p. 8091-8098]

-. ALP activity.



In vitro culture tests

Cell attachment.

16 samples of each i.i. & ctrol Ti discs,

Sterilized by UV light

placed individually into 6-well plate,

Inoculated with 1.5 x 105 human bone-cells

Incubated for 4h at 35 ºC, 5% CO2.

After rinsing and fixing

Image Analysis System used to quantify

nº attached cells.

Cell Attachment

0

400

800

1200

1600

2000

2400

2800

C A B

Sample ref.

Nu

mb

er

of

cells / c

m2)

Cell Attachment

0

400

800

1200

1600

2000

2400

2800

E DSample ref.

Nu

mb

er

of

cells /

cm

2

C & E control



Cell proliferation.

Seeded at conc. of 2,5 e4 cell/well


placed individually into 6 well plates.

Incubated for 24, 48, 144 & 192 hours

at 35 ºC and 5% CO2.

Cells washed and collected by

trypsinisation with 0.25% trypsin-EDTA

solution (SIGMA).

Cells were stained with 7-Amino-

Actinomycin D (7 AAD) (BECKTON &

DICKINSON)

Cells quantified by flow cytometry

Cell Proliferation

-1

1

3

5

24 48 144 192

Incubated time (hours)

Gro

wth

Ratio

Control Ti Treated Ti

*



ESEM

AFM

Cell morphology.



Cell apoptosis.

Seeded at a conc. of 6 x 105 cell/well


placed individually into 12 well plates.

Cells incubated for 24 and 72 h. at 35ºC and 5%CO2.

Cells were washed and collected by trypsinisation with 0.25% trypsin-EDTA solution.

Cells lysed and their apoptosis state was tested using a flow cytometry Kit, which detects

the concentrations of PARP, Bcl-2 and Caspase-3 molecules.

24 hours 72 hours



Alkaline Phosphatase

0

0,5

1

1,5

2

2,5

3 days 8 days 16 days

Rela

tive A

LP

acti

vit

y

Ion implanted

Control Ti

sample

Expression of Alkaline Phosphatase

The activity of the alkaline phosphatase enzyme was determined measuring the

final fluorescence emitted as a result of an enzyme based reaction (4-MUP). The

use of a positive control confirms that the assay conditions are accurate.



In vitro cell culture tests. Conclusions:

• Results depend on the treatment parameters.

• Osteoblast growth is favoured.

• Cell morphology indicates a better cell behaviour.

• Lower apoptosis signal.

• Higher ALP activity



In vivo tests on dental implants.

What’s a dental implant?

Typically a Ti/Ti alloy screw replacing

the tooth root and supporting a new

artificial tooth.

Requirements

It must transmit the masticating forces

to the jaw bone,

i.e. a good osseointegration is

mandatory.



How does it work ?

1. Insertion of the implant into the bone.

2. Integration on the surrounding bone (3-8 months).

3. Abutment connection.

4. Fixation of the crown

From Periodontal

Associates



Treatment example.

From www.oral-implants.com



Osseointegration (%)

30%

40%

50%

60%

70%

80%

90%

on Ti6Al4V on CP Ti

Lifenova

Untreated

Osseointegration tests

on the tibial plateau of NZW rabbits.

as machined vs. ion implanted

two base materials

Results:

Significant differences on poor

bone density areas.

Ion impl.ed

Untreated



i) Test procedure & ethical committee (Univ. Barcelona)

ii) 12 implants x 6 different surfaces.

iii) 6 implants x 6 surfaces at month 3.

6 implants x 6 surfaces at month 6.

P2I P3I P4I P2D P3D P4D

B 1 A B C D E F

B 2 F A B C D E

B 3 E F A B C D

B 4 D E F A B C

B 5 C D E F A B

B 6 B C D E F A

B 7 A B C D E F

B 8 F A B C D E

B 9 E F A B C D

B 10 D E F A B C

B 11 C D E F A B

B 12 B C D E F A

Tests in jawbone.

Case study: in vivo tests


BIC %, ESEM evaluation.

0%

10%

20%

30%

40%

50%

60%

70%

80%

Ion implantation Control Commercial

average

BIC

3 months

6 months

ESEM evaluation.

Case study: in vivo tests

Control vs. Ion Implantation

* *


BIC %, histological evaluation.

0%

10%

20%

30%

40%

50%

60%

70%

80%

Ion implantation Control Commercial

average

BIC

3 months

6 months

Histological evaluation.

Control vs. Ion Implantation

[International Journal of Oral and Maxillofacial Surgery, 37(5),(2008), 441-447]

* *



ESEM-EDS.



ESEM-EDS.

Implant

Mature

bone

New

bone

[International Journal of Oral and Maxillofacial Surgery, 38(3), (2009), 274-278]



BIC %

0

10

20

30

40

50

60

70

0 Month 1 Month 3 Month 6

Ion implanted Commercial Average Control



N patients: 19

N of mini-implants: 22

Inclusion criteria:

Healthy patients from both sexes, undergoing treatment

with conventional commercial implants, with type III or IV

bone quality (as defined by Lekhölm y Zdart)

Exclusion criteria

Minors, pregnant women, local or systemic contraindications

Clinical trial 277/06/EC



Participating Clinics

Pr. Dr. C Gay Escoda. University of Barcelona, Faculty of Odontology.

Dr. M de Maeztu. Private practice. Tolosa, Spain.

Approval and authorization

Ethical committees of the University of Barcelona and Hospital Donostia.

AGEMPS (Spanish Agency for Drugs and Medical Devices).




1.8 mm diameter

5 mm long




Surgery: drilling.



Surgery: implant insertion.



Surgery: implant extraction.



Surgery: implant extraction.



Histomorphometric study.



Histomorphometric study.

Código

Paciente

Ref. micro-

implanteLocalización

%BIC Ctrl

s/ESEM

%BIC I.I.

s/ESEM

%BIC Ctrl

s/histología

%BIC I.I.

s/histología

T001 4 3er cuadrante 22% 58% 40% 41%

5 4º cuadrante 69% 49% 53% 57%

T002 6 3er cuadrante 25% 67% 65% 63%

7 2º cuadrante

T003 8 4º cuadrante 70% 64% 47% 67%

T004 10 3er cuadrante

11 2º cuadrante 38% 43% 31% 20%

12 4º cuadrante 18% 26% 49% 31%

Código

Paciente

Ref. micro-

implanteLocalización

%BIC Ctrl

s/ESEM

%BIC I.I.

s/ESEM

%BIC Ctrl

s/histología

%BIC I.I.

s/histología

B005 14 1.8

15 2.8

B006 16 3.8

17 4.8

B007 18 1.8

19 2.8

B008 20 3.8 68% 79% 64% 87%

21 4.8 45% 40% 56% 57%

B009 22 3.8

23 4.8

B010 24 3.8 78% 66% 76% 54%

25 4.8 43% 60% 49% 47%

B011 26 1.8 49% 76% 61% 50%

27 2.8 23% 17% 14% 21%

B012 28 1.8 11% 3% 29% 23%

29 2.8

B013 32 1.8

33 2.8

B014 30 3.8

31 4.8

B015 9 3.8 56% 18% 66% 87%

13 4.8 28% 56% 71% 75%

B016 36 3.8 75% 81% 54% 92%

2 4.8 70% 81% 66% 83%

B017 41 3,8 41% 83% 99% 88%

B018 40 3.8 64% 63% 87% 90%

42 4.8 17% 36% 40% 67%

B019 34 3.8 58% 71% 94% 96%

35 4.8 49% 69% 62% 89%

Nº

pacientes

Nº

implantes

Observacione

s

%BIC Ctrl

s/ESEM

%BIC I.I.

s/ESEM

%BIC Ctrl

s/histología

%BIC I.I.

s/histología

19Total 37

valorados 22 Media 46,2% 54,8% 57,9% 62,9%

Desviación std 21% 23% 21% 25%

%BIC Histology

30,0%

40,0%

50,0%

60,0%

70,0%

80,0%

90,0%

% BIC Ctrl % BIC I.I.

Conclusions:

-. No negative reaction.

-. Larger osseointegration.


[International Journal of Oral and Maxillofacial Surgery, 42(7) (2013) 891-896]


Human bone on implant surface AFM

. .



Human bone on implant surface AFM

. .



Conclusions:

-. Higher osseointegration on poor bone density areas.

-. Faster osseointegration than untreated implants.

-. Higher osseointegration than in the case of implants with

commercial surfaces, i.e. micro-roughened TiO2.

-. Ion implanted surfaces show new bone formation arising

from the implant, unlike control samples.

Higher and faster levels of osseointegration

=

Shorter patient treatment times & treatments available for

cases/patients with lower bone quality.



Tissue integration

Tissue integration.

Neutral (Ne,Ar,Xe,Kr) ion implantation vs. cells.



Tissue integration.

TiGr4, 1Ne40

TiGr4, 1Ne80

TiGr4, 2Ne40

TiGr4, polished

M&M:

Ion implantation of Ne.

1×1017 and 2×1017 ions/cm2,

40keV and 80keV.



M&M:

Ion implantation of Ar.

1×1017 and 2×1017 ions/cm2,

40keV and 80keV.

Tissue integration.

TiGr4, 1Ar40

TiGr4, 1Ar80

TiGr4, 2Ar40

TiGr4, polished



M&M:

Ion implantation of Kr.

1×1017 and 2×1017 ions/cm2,

40keV and 80keV.

Tissue integration.

TiGr4, 1Kr40

TiGr4, 1Kr80

TiGr4, 2Kr40

TiGr4, polished



Ion implantation and plasma based processes on Medical Devices

M&M:

Ion implantation of Xe.

1×1017 and 2×1017 ions/cm2,

40keV and 80keV.

Tissue integration.

TiGr4, 1Xe40

TiGr4, 1Xe80

TiGr4, 2Xe40

TiGr4, polished

Tissue integration / regeneration.


.

Tissue integration / regeneration. TiGr4, 1Xe80

TiGr4, 1Xe40

TiGr4, 2Xe40

TiGr4, 1Ar80

TiGr4, 1Ar 40

TiGr4, 2Ar 40

TiGr4, 1Ne80

TiGr4, 1Ne40

TiGr4, 2Ne40

TiGr4, polished

TiGr4, 1Kr80

TiGr4, 1Kr40

TiGr4, 2Kr40



Tests / results:

Wettability and roughness:

Tissue integration.

75

77

79

81

83

85

87

89

91

93

95

0 1 2 3 4 5 6 7 8

Ra (nm)

Co

nta

ct

an

gle

(º)

Ne Ar Kr Xe Control

Ref. Ra

Peak to

peak

distances

Contact

angle

(nm) (nm) (º)

Control 0.295 N.A. 82.6±5.2

1Ne80 0.56 119±5 78.6±1.6

1Ne40 0.61 67±2 85.1±2.2

2Ne40 1.49 91±2 88.2±0.9*

1Ar40 2.15 82±3 81.0±0.5

2Ar40 2.21 160±4 84.0±2.4

1Ar80 2.68 122±4 83.1±1.0

1Kr80 2.6 98±3 85.5±2.4

1Kr40 1.48 89±3 87.0±0.9

2Kr40 1.55 132±2 87.2±3.2

1Xe80 2.47 162±4 86.9±2.5

1Xe40 3.57 185±3 86.9±1.0

2Xe40 7.27 217±4 88.8±2.1*




Tests / results:

Cell culture and adhesion

hFOB 1.19 (cultured in accordance with ATCC).

1.5 x 104 cells were seeded onto the 8 mm diameter Ti discs, and incubated for 4h and 24 h

(n=3). Besides, cells were also plated on untreated Ti-discs (n=5) as a control.

Cell adhesion was assessed using the 4-[3-(4-Iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-

1,3-benzene disulfonate (WST-1) assay.

Tissue integration.




Tests / results:

The contact angle measurements: most of the treated surfaces became more

hydrophobic, as compared to the control sample, although the change was small, i.e. it

was only statistically significant for the samples 2Ne40 and 2Xe40.

The contact angle varied most for titanium samples ion implanted with Ne (78.6±1.6 to

88.2±0.9) while least for Xe ion implanted samples (86.9±1.0 to 88.8±2.1).

The in vitro cell adhesion test: differences between ion implanted samples and the

control untreated samples occurred in the short time, i.e. at 4 hours rather than at 24

hours, suggesting that nano-roughness could be related to early cell attachment.

1Ne40, 2Ne40, 1Kr40, 2Kr40, 1Xe40 and 2Xe40 samples showed statistically strongly

significant differences (p<0.01) at 4 hours as compared to untreated Ti.


Tissue integration.

Neutral (Ne,Ar,Xe,Kr) ion implantation vs. cells.

[Applied Surface Science, 27 March 2014. http://dx.doi.org/10.1016/j.apsusc.2014.03.118]

.]

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TECNALIA

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Technology

Surface Enginnering on Medical Devices