In vitro effects of platelet derived growth factor on human keratocyte response to excimer laser ablation

Ribeiro, João Eduardo C.; Oréfice, Fernando; Beuerman, Roger

doi:10.5935/0004-2749.19990035

SUMMARY

Purpose:

To evaluate the in vitro effects of platelet derived growth factor (PDGF) on the response ofhuman keratocytes to excimer laser ablation.

Methods:

Keratocytes were cultured to confluence in 6-well tissue culture plates and exposed to an excimer laser beam. After the laser they were mantained in one of the following media: Group I -Dulbecco's Modified Eagle Medium supplemented with heat-inactivated fetal bovine serum (FBS); Group II - DMEM + fetal bovine serum (FBS); Group III - DMEM + heat-inactivated FBS plus PDGF at a concentration of 1 ng/ml and Group IV - DMEM + heat-inactivated FBS plus PDGF at a concentration of 10 ng/ml. To quantitate the rate of wound closure micrographs were taken at O, 24, 48, 72 and 96 hours.

Results:

Ninety-six hours after the excimer laser ablation only keratocytes cultured in FBS (Group II) resurface completely. At evaluations after 48, 72 and 96 hours, the residual area in Groups I, III and IV was bigger than in Group II (p < 0.05). There was no significant difference between groups treated with different concentrations of PDGF.

Conclusion:

Cultured human keratocytes proliferate and migrate to the wounded area after excimer laser ablation and PDGF actually does not stimulate these cells in the presence ofheat-inactivated serum.

Keywords:
Platelet derived growth factor; Keratocytes; Excimer laser

RESUMO

Objetivo:

Avaliar in vitro os efeitos do fator de crescimento derivado de plaquetas (PDGF) na resposta de ceratócitos humanos à ablação com excimer laser.

Método:

Excimer laser foi aplicado na área central de culturas confluentes de ceratócitos humanos e a seguir estas foram mantidas em um dos seguintes meios de cultura: Grupo I - Dulbecco's Modified Eagle Medium (DMEM) suplementado com soro bovino fetal desnaturado (SBF d.); Grupo li - DMEM + soro bovino fetal (SBF); Grupo HI -DMEM + SBF d. enriquecido com PDGF a 1 ng/ml e Grupo IV - DMEM + SBF d. enriquecido com PDGF a 10 ng/ml. A resposta dos ceratócitos foi documentada por meio de microfotografias O, 24, 48, 72 e 96 horas após ablação.

Resultados:

Avaliação 96 horas após ablação, mostrou que apenas os ceratócitos em cultura com SBF (Grupo II) cobriram toda área ablada. Nas avaliações 48, 72 e 96 horas após aplicação do laser, a área nua residual nos Grupos 1, III e IV era maior do que no Grupo li (p < 0,05). Não se observou diferença na resposta dos ceratócitos com relação à concentração de PDGF.

Conclusão:

Conclui-se que após ablação com excimer laser, ceratócitos humanos em cultura proliferam e migram em direção à área ablada e PDGF na ausência de componentes do soro normal efetivamente não tem efeito estimulante sobre ceratócitos humanos.

Palavras-chave:
Fator de crescimento derivado de plaquetas; Ceratócitos; Excimer laser

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Part of the PhD thesis presented to the Postgraduation Course in Ophthalmology of the UFMG which conferred the title of Doctor in Medicine to the author. Perforrned at the Department of Ophthalmology of the UFMG and the Lions Eye Research Laboratories, Laboratory for the Molecular Biology of the Ocular Surface, LSU Eye Center, Louisiana State University Medical Center School of Medicine, New Orleans, Louisiana - USA.
The authors have no proprietary interest in the development or marketing of the instruments or medications referred to in the text.

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Publication Dates

Publication in this collection
Apr 1999

This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

[1] Part of the PhD thesis presented to the Postgraduation Course in Ophthalmology of the UFMG which conferred the title of Doctor in Medicine to the author. Perforrned at the Department of Ophthalmology of the UFMG and the Lions Eye Research Laboratories, Laboratory for the Molecular Biology of the Ocular Surface, LSU Eye Center, Louisiana State University Medical Center School of Medicine, New Orleans, Louisiana - USA.

[2] The authors have no proprietary interest in the development or marketing of the instruments or medications referred to in the text.

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