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Skeletal stem cell and bone implant interactions are enhanced by LASER titanium modification

Skeletal stem cell and bone implant interactions are enhanced by LASER titanium modification
Skeletal stem cell and bone implant interactions are enhanced by LASER titanium modification
Purpose

To evaluate the osteo-regenerative potential of Titanium (Ti) modified by Light Amplification by Stimulated Emission of Radiation (LASER) beam (Yb-YAG) upon culture with human Skeletal Stem Cells (hSSCs1).

Methods

Human skeletal cell populations were isolated from the bone marrow of haematologically normal patients undergoing primary total hip replacement following appropriate consent. STRO-1+ hSSC1 function was examined for 10 days across four groups using Ti discs: i) machined Ti surface group in basal media (Mb2), ii) machined Ti surface group in osteogenic media (Mo3), iii) LASER-modified Ti group in basal media (Lb4) and, iv) LASER-modified Ti group in osteogenic media (Lo5). Molecular analysis and qRT-PCR as well as functional analysis including biochemistry (DNA, Alkaline Phosphatase (ALP6) specific activity), live/dead immunostaining (Cell Tracker Green (CTG7)/Ethidium Homodimer-1 (EH-18)), and fluorescence staining (for vinculin and phalloidin) were undertaken. Inverted, confocal and Scanning Electron Microscopy (SEM) approaches were used to characterise cell adherence, proliferation, and phenotype.

Results

Enhanced cell spreading and morphological rearrangement, including focal adhesions were observed following culture of hSSCs1 on LASER surfaces in both basal and osteogenic conditions. Biochemical analysis demonstrated enhanced ALP6 specific activity on the hSSCs1-seeded on LASER-modified surface in basal culture media. Molecular analysis demonstrated enhanced ALP6 and osteopontin expression on titanium LASER treated surfaces in basal conditions. SEM, inverted microscopy and confocal laser scanning microscopy confirmed extensive proliferation and migration of human bone marrow stromal cells on all surfaces evaluated.

Conclusions

LASER-modified Ti surfaces modify the behaviour of hSSCs.1 In particular, SSC1 adhesion, osteogenic gene expression, cell morphology and cytoskeleton structure were affected. The current studies show Ti LASER modification can enhance the osseointegration between Ti and skeletal cells, with important implications for orthopaedic application.
titanium surface, skeletal stem cell, tissue regeneration, bone formation, LASER
0006-291X
1-7
Sisti, Karin E.
642aac8c-cbd5-4d3b-bcb6-ad856b199a8b
de Andrés, María C.
54e87e8a-1aa2-4907-a8a0-25d0c15e5e40
Johnston, David
b41163c9-b9d2-425c-af99-2a357204014e
Almeida-Filho, Edson
d0adfd07-31e3-4232-a9af-c31d3fc16a1f
Guastaldi, Antonio C.
741c68fd-9a46-4295-8293-539f1a9ce113
Oreffo, Richard O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Sisti, Karin E.
642aac8c-cbd5-4d3b-bcb6-ad856b199a8b
de Andrés, María C.
54e87e8a-1aa2-4907-a8a0-25d0c15e5e40
Johnston, David
b41163c9-b9d2-425c-af99-2a357204014e
Almeida-Filho, Edson
d0adfd07-31e3-4232-a9af-c31d3fc16a1f
Guastaldi, Antonio C.
741c68fd-9a46-4295-8293-539f1a9ce113
Oreffo, Richard O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778

Sisti, Karin E., de Andrés, María C., Johnston, David, Almeida-Filho, Edson, Guastaldi, Antonio C. and Oreffo, Richard O.C. (2015) Skeletal stem cell and bone implant interactions are enhanced by LASER titanium modification. Biochemical and Biophysical Research Communications, 1-7. (doi:10.1016/j.bbrc.2015.10.013). (PMID:26456647)

Record type: Article

Abstract

Purpose

To evaluate the osteo-regenerative potential of Titanium (Ti) modified by Light Amplification by Stimulated Emission of Radiation (LASER) beam (Yb-YAG) upon culture with human Skeletal Stem Cells (hSSCs1).

Methods

Human skeletal cell populations were isolated from the bone marrow of haematologically normal patients undergoing primary total hip replacement following appropriate consent. STRO-1+ hSSC1 function was examined for 10 days across four groups using Ti discs: i) machined Ti surface group in basal media (Mb2), ii) machined Ti surface group in osteogenic media (Mo3), iii) LASER-modified Ti group in basal media (Lb4) and, iv) LASER-modified Ti group in osteogenic media (Lo5). Molecular analysis and qRT-PCR as well as functional analysis including biochemistry (DNA, Alkaline Phosphatase (ALP6) specific activity), live/dead immunostaining (Cell Tracker Green (CTG7)/Ethidium Homodimer-1 (EH-18)), and fluorescence staining (for vinculin and phalloidin) were undertaken. Inverted, confocal and Scanning Electron Microscopy (SEM) approaches were used to characterise cell adherence, proliferation, and phenotype.

Results

Enhanced cell spreading and morphological rearrangement, including focal adhesions were observed following culture of hSSCs1 on LASER surfaces in both basal and osteogenic conditions. Biochemical analysis demonstrated enhanced ALP6 specific activity on the hSSCs1-seeded on LASER-modified surface in basal culture media. Molecular analysis demonstrated enhanced ALP6 and osteopontin expression on titanium LASER treated surfaces in basal conditions. SEM, inverted microscopy and confocal laser scanning microscopy confirmed extensive proliferation and migration of human bone marrow stromal cells on all surfaces evaluated.

Conclusions

LASER-modified Ti surfaces modify the behaviour of hSSCs.1 In particular, SSC1 adhesion, osteogenic gene expression, cell morphology and cytoskeleton structure were affected. The current studies show Ti LASER modification can enhance the osseointegration between Ti and skeletal cells, with important implications for orthopaedic application.

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More information

Accepted/In Press date: 2 October 2015
e-pub ahead of print date: 9 October 2015
Keywords: titanium surface, skeletal stem cell, tissue regeneration, bone formation, LASER
Organisations: Human Development & Health, Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 383672
URI: https://eprints.soton.ac.uk/id/eprint/383672
ISSN: 0006-291X
PURE UUID: a3db2c61-35ee-4e80-b96d-83f1486f70bb
ORCID for David Johnston: ORCID iD orcid.org/0000-0001-6703-6014
ORCID for Richard O.C. Oreffo: ORCID iD orcid.org/0000-0001-5995-6726

Catalogue record

Date deposited: 09 Nov 2015 17:04
Last modified: 06 Jun 2018 12:53

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