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The aim of our study was to to evaluate the survival of apical stem cells and Success of regenerative endodontic procedure of open apex non vital anterior teeth with apical periodontitis, When three root canal medicaments are used Calcium hydroxide, Nano particle calcium hydroxide and Nano particle Calcium hydroxide and Augmentin.
Regarding part I, Measurement of cell viability and proliferation comprise the underlying basis for numerous in vitro assays directed towards the quantitation of a cell population’s response to external factors. MTT Reagent supplied ready for use were obtained from (TACSTM TREVIGEN® 8405 Hegerman Ct. Gaithersburg). It is a colorimetric assay for assessing cell metabolic activity. Using the enzymes under defined conditions which reflected the number of viable cells present. All three intracanal medicaments were tested at different concentrations.
Part II, 45 human cases selection was limited to anterior teeth of age ranging from 8-14 years. These criteria were chosen for the most predilection of immature necrotic teeth due to previous trauma. Cases were excluded due to radiographic evidence of root resorption and evidence of tooth mobility. The cases were divided randomly into three groups, group (I); where Calcium hydroxide intra canal medication was used, group (II); where Nano particle calcium hydroxide intra canal medication was used and group (III); where Nano particle calcium hydroxide and Augmentin intra canal medication was used. Cases were followed on regular intervals up to 18 months clinically and radiographically.
Local anesthesia was administrated without vasoconstrictor. During the first visit, the disinfection protocol was performed, irrigation with 20 mL 1.5% NaOCl that was followed by 10 mL 17% EDTA,adel Following the disinfection protocol Sample a was collected for further investigation for STRO-1 Gene expression analysis by RT-PCR.
Intra canal medication was injected inside the canal, While sample b was collected during the second visit just before the regenerative procedure is carried out, sample was sent for further investigation for STRO-1 Gene expression analysis by RT-PCR. Comparison between the two samples was done and between the three groups.
Results for the three groups revealed that, A statistically significant difference was found between sample a and sample b showing more positive cells to STRO 1 marker with sample b. While comparing the three groups together, STRO-1 Gene expression for sample b, showed that there was a statistically significant difference found between (NANO CaOH + Augmentin) and each of (CaOH) and (NANO CaOH), expressing more positive cells to STRO 1 marker with group III.
Radiographic evaluation has been done using CBCT imaging. CBCT imaging provides 3-dimensional images of roots, canals, and the apical area with high resolution as well as accurate measurement on the root canal and periapical area. In the present study, we applied limited field of view CBCT imaging to measure the changes in the root. In this study, CBCT imaging was used as a tool to provide better assessment of the outcomes, and the use of CBCT imaging was approved by the ethics committee for this study. Radiographic evaluation included measuring increase in length, thickness, occurrence of apical closure and decrease in volume of periapical lesion. Data were collected and tabulated for statistical analysis.
Results: Stro-1 gene expression was increased in sample (b) in second visit showing a statistically significant difference when compared to sample (a) first visit in all three groups.
Stro-1 gene expression was increased in sample (b) in group III (NANO CaOH + Augmentin) showing a statistically significant difference when compared group II (NANO CaOH) and group I (CaOH).
Radiographic evaluation showed regarding root length and thickness, there was a significant difference in all of the three groups regarding increase in root length and root thickness over the 18 months follow up period of time, with no significant difference between the three groups.
Regarding decrease in apical diameter, there was a significant difference in all of the three groups regarding decrease in apical diameter over the 18 months follow up period of time, with no significant difference between the three groups.
While evaluating decrease in volume of periapical lesion, our results showed a significant difference in all of the three groups regarding decrease in apical diameter over the 18 months follow up period of time.
group III (NANO CAOH + Augmentin) revealed superior findings when compared with the other two groups.
Under the circumstances of this study, it can be concluded that:
1. Stem cells of the apical papilla play an important role in success of regenerative endodontic treatment and the use of intra canal medications affects survival and differentiation of SCAPs.
2. Calcium Hydroxide and Nano Calcium hydroxide promoted greater survival and differentiation of SCAPs.
3. Augmentin was potent in disinfection of immature permenant teeth and was successfully used in regenerative endodontic treatment
1. Longer follow up period is needed for monitoring of the repair process
2. Further research is recommended regarding the use of Augmentin and Nano calcium hydroxide for tissue regeneration.
3. Further studies are needed for identification of the stem cell source of the regenerated tissues.
4. The addition of radio-opacifiers during preparation of Nano calcium hydroxide intracanal medication.