5 research outputs found
Comparison of mechanical, enzymatic and mechanical-enzymatic dissagregation tecniques in extraction of lymphocytary populations and macrophagues in tissues with chronic advanced periodontitis
ANTECEDENTES: En este estudio se compararon tres t茅cnicas de disgregaci贸n en la recuperaci贸n de poblaciones linfocitarias y macr贸fagos en tejidos con periodontitis cr贸nica avanzada. OBJETIVO: comparar, por citometr铆a de flujo, el recuento de poblaciones linfocitarias y macr贸fagos de biopsias de tejidos periodontalmente afectados, con tres procesos previos de disgregaci贸n celular (mec谩nica, enzim谩tica con colagenasa y mecano-enzim谩tica) para observar cu谩l de 茅stos es m谩s efectivo en la recuperaci贸n celular y produce menos alteraciones en los marcadores de superficie. M脡TODOS: Se realiz贸 un estudio descriptivo comparativo donde se eligi贸 una muestra de 11 biopsias de tejidos con periodontitis cr贸nica avanzada. El procedimiento consisti贸 en tomar las biopsias por medio de biseles internos que se colocaban en un Eppendorf con el medio de transporte. En el grupo 1 se realiz贸 la t茅cnica de disgregaci贸n mec谩nica Medimachine庐, en el grupo 2 t茅cnica enzim谩tica (colagenasa 2%), y el en grupo 3 t茅cnica mecano-enzim谩tica (Medimachine庐 y colagenasa 2%). Posteriormente las muestras fueron analizadas por un cit贸metro de flujo FACS- Aria (Becton Dickinson San Jos茅 California). RESULTADOS: No se encontraron diferencias estad铆sticamente significativas entre las tres t茅cnicas de disgregaci贸n en la recuperaci贸n de poblaciones linfocitarias; sin embargo, hubo una mayor recuperaci贸n de macr贸fagos con la t茅cnica mecano-enzim谩tica comparada con la mec谩nica, siendo estas diferencias estad铆sticamente significativas. CONCLUSIONES: Las tres t茅cnicas estudiadas presentaron resultados comparables en cuanto a recuperaci贸n y da帽o a los marcadores de superficie, por lo cual, por tiempo y costo, se aconseja realizar una t茅cnica mec谩nica con Medimachine庐.BACKGROUND: Three disaggregation techniques for the recovery of lymphocyte and macrophage populations from tissues with advanced chronic periodontitis were analyzed. OBJECTIVE: Compare by flow cytometry the recovery of lymphocyte and macrophage populations from biopsies of periodontal tissues with periodontitis by using three processes of cellular disaggregration (mechanical, enzymatic with colagenase and mechanical-enzymatic) to observe which of these is more effective and produces fewer alterations in the surface markers. METHODS: A comparative descriptive study was carried out with a sample of 11 biopsies of tissues taken from 23sites with advanced chronic periodontitis. The procedure consisted of taking the biopsies by means of internal bevels and placing them in an Eppendorf for transportation. In group 1, the mechanical technique of disaggregation (Medimachine庐) was carried out. In group 2, the enzymatic technique (colagenase 2%) was used. In group 3, the mechanical-enzymatic technique (Medimachine庐 and colagenase 2%) was perfomred. Later on, the samples were analyzed with a FACS-Aria cytometer (Becton Dickinson San Jos茅 California). RESULTS: There were not statistically significant differences among the three disaggregation techniques for the recovery of lymphocyte populations. However, there was a bigger recovery of macrophages with the mechanical-enzymatic technique when compared to the other two techniques, being statistically significant. CONCLUSIONS: The three techniques studied produce similar results in the recovery and damage to the surface markers; hence, considering time and cost constrains, it is advisable to carry out the mechanical technique with Medimachine庐
Comparaci贸n de t茅cnicas de disgregaci贸n mec谩nica, enzim谩tica y mecano-enzim谩tica en la extracci贸n de poblaciones linfocitarias y macr贸fagos en tejidos con periodontitis cr贸nica avanzada
BACKGROUND: Three disaggregation
techniques for the recovery of lymphocyte
and macrophage populations from tissues
with advanced chronic periodontitis were
analyzed. OBJECTIVE: Compare by flow
cytometry the recovery of lymphocyte
and macrophage populations from
biopsies of periodontal tissues with
periodontitis by using three processes of
cellular disaggregration (mechanical,
enzymatic with colagenase and
mechanical-enzymatic) to observe which
of these is more effective and produces
fewer alterations in the surface markers.
METHODS: A comparative descriptive
study was carried out with a sample of 11 biopsies of tissues taken from sites
with advanced chronic periodontitis. The
procedure consisted of taking the
biopsies by means of internal bevels
and placing them in an Eppendorf for
transportation. In group 1, the
mechanical technique of disaggregation
(Medimachine庐) was carried out. In
group 2, the enzymatic technique
(colagenase 2%) was used. In group 3,
the mechanical-enzymatic technique
(Medimachine庐 and colagenase 2%)
was perfomred. Later on, the samples
were analyzed with a FACS-Aria
cytometer (Becton Dickinson San Jos茅
California). RESULTS: There were not
statistically significant differences
among the three disaggregation
techniques for the recovery of
lymphocyte populations. However, there
was a bigger recovery of macrophages
with the mechanical-enzymatic
technique when compared to the other
two techniques, being statistically
significant. CONCLU SIONS: The three
techniques studied produce similar
results in the recovery and damage to
the surface markers; hence, considering
time and cost constrains, it is advisable
to carry out the mechanical technique
with Medimachine庐.ANTECEDENTES: En este estudio se
compararon tres t茅cnicas de disgregaci贸n
en la recuperaci贸n de poblaciones
linfocitarias y macr贸fagos en tejidos con
periodontitis cr贸nica avanzada. OBJETIVO:
comparar, por citometr铆a de flujo,
el recuento de poblaciones linfocitarias
y macr贸fagos de biopsias de tejidos
periodontalmente afectados, con tres
procesos previos de disgregaci贸n celular
(mec谩nica, enzim谩tica con
colagenasa y mecano-enzim谩tica) para
observar cu谩l de 茅stos es m谩s efectivo
en la recuperaci贸n celular y produce
menos alteraciones en los marcadores
de superficie. M脡TODOS: Se realiz贸 un
estudio descriptivo comparativo donde
se eligi贸 una muestra de 11 biopsias de tejidos con periodontitis cr贸nica avanzada.
El procedimiento consisti贸 en tomar
las biopsias por medio de biseles internos
que se colocaban en un Eppendorf
con el medio de transporte. En el grupo 1
se realiz贸 la t茅cnica de disgregaci贸n
mec谩nica Medimachine庐, en el grupo 2
t茅cnica enzim谩tica (colagenasa 2%), y el
en grupo 3 t茅cnica mecano-enzim谩tica
(Medimachine庐 y colagenasa 2%). Posteriormente
las muestras fueron analizadas
por un cit贸metro de flujo FACSAria
(Becton Dickinson San Jos茅
California). RESULTADOS: No se encontraron
diferencias estad铆sticamente significativas
entre las tres t茅cnicas de
disgregaci贸n en la recuperaci贸n de poblaciones
linfocitarias; sin embargo, hubo
una mayor recuperaci贸n de macr贸fagos
con la t茅cnica mecano-enzim谩tica
comparada con la mec谩nica, siendo es tas diferencias estad铆sticamente significativas.
CONCLUSIONES: Las tres t茅cnicas
estudiadas presentaron resultados
comparables en cuanto a recuperaci贸n
y da帽o a los marcadores de superficie,
por lo cual, por tiempo y costo, se aconseja
realizar una t茅cnica mec谩nica con
Medimachine
Comparison of mechanical, enzymatic and mechanical-enzymatic dissagregation tecniques in extraction of lymphocytary populations and macrophagues in tissues with chronic advanced periodontitis
ANTECEDENTES: En este estudio se compararon tres t茅cnicas de disgregaci贸n en la recuperaci贸n de poblaciones linfocitarias y macr贸fagos en tejidos con periodontitis cr贸nica avanzada. OBJETIVO: comparar, por citometr铆a de flujo, el recuento de poblaciones linfocitarias y macr贸fagos de biopsias de tejidos periodontalmente afectados, con tres procesos previos de disgregaci贸n celular (mec谩nica, enzim谩tica con colagenasa y mecano-enzim谩tica) para observar cu谩l de 茅stos es m谩s efectivo en la recuperaci贸n celular y produce menos alteraciones en los marcadores de superficie. M脡TODOS: Se realiz贸 un estudio descriptivo comparativo donde se eligi贸 una muestra de 11 biopsias de tejidos con periodontitis cr贸nica avanzada. El procedimiento consisti贸 en tomar las biopsias por medio de biseles internos que se colocaban en un Eppendorf con el medio de transporte. En el grupo 1 se realiz贸 la t茅cnica de disgregaci贸n mec谩nica Medimachine庐, en el grupo 2 t茅cnica enzim谩tica (colagenasa 2%), y el en grupo 3 t茅cnica mecano-enzim谩tica (Medimachine庐 y colagenasa 2%). Posteriormente las muestras fueron analizadas por un cit贸metro de flujo FACS- Aria (Becton Dickinson San Jos茅 California). RESULTADOS: No se encontraron diferencias estad铆sticamente significativas entre las tres t茅cnicas de disgregaci贸n en la recuperaci贸n de poblaciones linfocitarias; sin embargo, hubo una mayor recuperaci贸n de macr贸fagos con la t茅cnica mecano-enzim谩tica comparada con la mec谩nica, siendo estas diferencias estad铆sticamente significativas. CONCLUSIONES: Las tres t茅cnicas estudiadas presentaron resultados comparables en cuanto a recuperaci贸n y da帽o a los marcadores de superficie, por lo cual, por tiempo y costo, se aconseja realizar una t茅cnica mec谩nica con Medimachine庐.BACKGROUND: Three disaggregation techniques for the recovery of lymphocyte and macrophage populations from tissues with advanced chronic periodontitis were analyzed. OBJECTIVE: Compare by flow cytometry the recovery of lymphocyte and macrophage populations from biopsies of periodontal tissues with periodontitis by using three processes of cellular disaggregration (mechanical, enzymatic with colagenase and mechanical-enzymatic) to observe which of these is more effective and produces fewer alterations in the surface markers. METHODS: A comparative descriptive study was carried out with a sample of 11 biopsies of tissues taken from 23sites with advanced chronic periodontitis. The procedure consisted of taking the biopsies by means of internal bevels and placing them in an Eppendorf for transportation. In group 1, the mechanical technique of disaggregation (Medimachine庐) was carried out. In group 2, the enzymatic technique (colagenase 2%) was used. In group 3, the mechanical-enzymatic technique (Medimachine庐 and colagenase 2%) was perfomred. Later on, the samples were analyzed with a FACS-Aria cytometer (Becton Dickinson San Jos茅 California). RESULTS: There were not statistically significant differences among the three disaggregation techniques for the recovery of lymphocyte populations. However, there was a bigger recovery of macrophages with the mechanical-enzymatic technique when compared to the other two techniques, being statistically significant. CONCLUSIONS: The three techniques studied produce similar results in the recovery and damage to the surface markers; hence, considering time and cost constrains, it is advisable to carry out the mechanical technique with Medimachine庐