Determination of enzime activities in the peri - implant - sulcus - fluid

Steffi Schatz

The determination of enzyme activities or concentrations of gingival crevicular fluid (GCF) is applied successfully. The aim of such investigations is identifying predictors of destructiv periodontal diseases. There are some chair-side test for periodontists using enzyme activitis predicting further attachment loss and estimating the effect of therapeutic intervention.

The disease of periimplantitis is known clinically and pathohistologically. There are only few studies disputing the examination of components of the periimplant-sulcus-fluid. In our study we determined enzyme activities of acid and alkaline phosphatase, beta- glucuronidase and arylsulphatase to examine the following:

  1. Which of the elevated parameters in implants provide the same diagnostic strength like those in teeth?

  2. Is the method of determination of enzyme activities in crevicular fluid used successfully in natural teeth suitable in implants?

  3. Which correlation exist between clinical results and (Enzyme Activity) EA in (Peri-Implant-Sulcus-Fluid) PISF?



We evaluated 25 patients, 13 were male, 12 were femal. The average age of the patients was 39,7 years.

The 25 patients with 40 IMZ-implants, having been implanted and supplied with prosthetic construction from 1989 - 1992 in the School of Dentistry, Charite Berlin, were included in this study.

Natural teeth:

The natural teeth examined were localized mesially to the implant- construction. We evaluated 29 teeth.
Prosthetic construction: 12 implants served as single crowns and 28 served implants carried fixed bridgework. Course of examination: After determination of the anamnesis and the estimation of the mouth hygiene we collected GCF and PISF, for that we removed the prothetic construction. We used filter paper strips inserted into the periimplant sulcus or gingival crevice until felt a mild resistance by the method of BRILL. To determine flow volume we took the Periotron 6000, which was calibrated with a dry strip before each measurement.

Storage of filter paper strip:

To store the strips we used 1% bovine serum albumin at 0,9% of Natriumchlorid and froze it at -20 degrees Celcius.

Method of measurement of clinical parameters:

Before measurement of PD we removed the IMC. At implants implant- shoulder was the fix point basing on the method by RICHTER and CO. They evaluated IMZ-implants as well. We used the probe PCP UNC 15 (Hu- Friedy), which is calibrated in mm. 30s after measuring of PD was registrated wether or not a bleeding on probing took place. It was recorded as "0" for no bleeding and as "1" for bleeding.

Determination of enzyme activities:

Before determination the probes were thawed and eluted. The determination was taken by spectrophotometrical procedures. The per min. released activity of enzymes in GCF or PISF was pronounced as per min released nmol 4-Nitrophenol (acid and alkaline phosphatase, beta- glucorunidase or 4 Nitrocatechl (AS).


  1. Results of all parameters of measurement in teeth and implants: For the statistical comparison of natural teeth and implants one
  2. implant was compared with one tooth in the same patient by test of Wilcoxon. Also calculation of means showed accordance with parameters of flow volume of GCF and PISF and enzyme activities. Bleeding on probing could be observed at natural teeth more often than at implants, but the differences was statistically not provable. The PD's were in implants significantly lower than in teeth (1,8mm against 2,8mm at teeth).
  3. Correlations between clinical parameters and enzyme activities in PISF: The relation of clinical parameters and enzyme activities were tested with analysis of variance and with linear regression to calculate a correlation coefficient. Our findings of the relations of enzyme activities and clinical parameters in natural teeth confirm to those of earlier studies by LAMSTER and BINDER and also in our laboratory.
BOP - Enzyme - Activities:

In implant sites there were statistically higher enzyme activities of all elavated enzymes if there was bleeding on probing.

PD - EA:

For evaluation of the relation of PD and EA we devided the PD into 4 groups. On the basis of the groups the means of EA were calculated. In implant sites we detected no relation between PD and EA for all elevated enzymes. The evaluation of linear regression showed no relevant coefficients. At natural teeth sites the activities of alkaline phosphatase and Arylsulphatase were significant higher with highher PD.

Flow volume - EA:

Also for evaluation of the relation of flow volume of GCF or PISF an EA we devided the flow volume into 4 groups: In implant sites the means of EA at groups flow volumes increased significantly for all elevated EA. The calculation of coefficients of linear regression showed the following:

With the miltiple regression we detected a coefficient of r = 0,68. For natural teeth sites we found a continous rise of EA for all elevated enzymes. This relation was not statistically significant.


  1. Implant and natural teeth react similarlyregarding to the parameters of BOP, flow volume and all elevated EA. They differ significantly in PD.
  2. If there was BOP, EA of AP, SP, beta-G and AS were significantly higher than those without BOP.
  3. Flow volume of PISF correlated positiv with all elevated EA.
  4. PD at implants sites showed no correlation with any elevated EA.

The analyses of communities and differences serve applying the known conditions in etiology and pathogenesis of adult periodontitis to the situation of osseointegrated implants. The aim of such evaluations is examining which criteria could be used for diagnosing of periimplant disease.

Estimation of parameters in implants:

The significanz of BOP in implants is critically discussed.ERICSSON and co. (1992) beleave, that relating to missing resistance of the mucosa against the power of probe the top of the probe penetrate into the submucous periimplant connective tissue. There it course a bleeding also in sound tissue.

The probing depth was the only parameter differed significantly at implants and teeth. This result is similar to those of other investigators. Most of them evaluated higher PD at implant sites than at teeth sites. With this study we found higher PD in teeth than in implants. The reason for this condition is probable searching in the used method. We choosed the implant shoulder as fix point. The mean of patients age was 39,7 years and a average PD of 2,8 mm is a physiological dimension.

Already Schareyka (1978) used measurement of flow volume to estimate the situation of the periimplant mucosa. The result of this study show that the flow volume of GCF is similar to that of PISF. This condition was also find by other authors. With these findings we conclude that the parameter of flow volume is suited to estimate the clinical situation.


Only few authors use the analyse of components of PISF estimating implant prognose. There are no studies which explain the evaluation of the enzymes of AP, SP, beta-G and AS in PISF. With this study we found good relations between BOP and all elevated enzyme activities. Also between flow volume of PISF and enzyme activities exist positive functional connection. No correlation can found between PD and enzyme activities.

Because no differences were found at implants and teeth sites relating to BOP, flow volume of PISF and enzyme activities and the correlations at implants and teeth were in a same direction, we conclude the following:

  1. If the clinical parameter of flow volume was high or there was a bleeding on probing also the enzyme activities were significantly higher.
  2. The parameters BOP and flow volume of PISF are siutable estimating like at natural teeth the clinical situation.
  3. PD at implants and teeth should not be compared.
  4. With this cross sectional evaluation we can make no statement, of the EA in PISF are better suitable for implant failure or attachment loss than the usual used clinical parameter.
  5. Because of this results the suppositions seems likely that defence mechanism at implants and natural teeth are similiar evaluating. This
  6. fact contradict the histological findings of LINDHE and ERICSSON. Also DONATH (1991) estimate the incorporation of implants as a reaction to a foreign body. The scar tissue is in his opinion not capable to achieve the same defence reaction like gingival tissues. The ground substance degrading enzymes at natural teeth underly a control mechanism, i.e. through proteases, which course first of all a stabilization of situation after a gingivitis. If this mechanism can find in implants is uncertain in order to histological findings. Our biochemical findings indicate the opposite.

Please all comments to Steffi Schatz: Phone: +49 30 28023908

Fax: +49 30 28021404

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