Purpose To identify adult human buccal epithelial stem cells (SCs) on the basis of two parameters (high p63 expression and greater nucleus/cytoplasmic (N/C) ratio) and to evaluate clinical efficacy of ex-vivo expanded autologous epithelium in bilateral limbal SC-deficient (LSCD) patients. Methods The epithelial cells were isolated from buccal biopsy and cultured on human amnion in culture inserts with 3T3 feeder layer. The SCs were identified on the basis of two-parameter analysis using confocal microscopy, surface markers, and colonyforming efficiency (CFE). The cultured epithelium was transplanted in 10 LSCD patients followed by penetrating keratoplasty in 4 patients. The clinical outcome was followed up to 3 years. Results A distinct population (3.0±1.7%) of small cells expressing high levels of p63 with greater N/C ratio was observed in buccal epithelium. The N/C ratio was found to be more appropriate than cell diameter for two-parameter analysis. These cells located in the basal layer were negative for connexin-43 and positive for melanoma-associated chondroitin sulfate proteoglycan, containing holoclones with 0.2% CFE, thus representing the SC population. After transplantation of cultured epithelium with increased (sixfold) SC content, anatomical and visual improvement was observed at 13-34 months in 3/10 LSCD patients. Conclusions The two-parameter SC marker is useful to identify and quantify buccal epithelial SCs. The transplantation of bioengineered SC-rich buccal epithelium is a strategy for corneal surface reconstruction in bilateral LSCD. However, further studies are required to optimize the culture conditions and to look for other sources of adult SCs for better visual outcome.

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Continuous development of technology in dental medicine and the high esthetic demands of the patients determined an increased awareness regarding the success of treatment among the clinical practice of dental students. The longevity of prosthesis depends on the type of luting cement, isolation techniques and the protocols used with tooth preparation. So it is important to educate students regarding this to achieve successful bonding of FPD. The aim of this study was to evaluate the knowledge and awareness of cementation protocols for Fixed Partial Dentures (FPDs) among undergraduate and postgraduate dental students in achieving success of Fixed dental prosthesis. It is a university setting study. Total of 150 students both undergraduate and postgraduates who are practicing in dental clinics in saveetha dental college were included in this study. A questionnaire containing 15 questions focused on the perceptions and experiences on protocols followed by dental students for cementation of prosthesis were given to them.The data obtained was entered in excel and imported to SPSS. Chi square association was done to compare the responses between undergraduates and postgraduates and were represented in the form of bar graphs. Out of 150 students, 70% were undergraduate and 30 % were postgraduate students. 19.33% of the postgraduates were aware about all of the protocols followed during cementation, whereas majority of the undergraduates chose isolation as the protocols to be followed 13.33% of the postgraduates and 49.33% of the undergraduates, both were equally aware that inadequate isolation majorly as led to the failure of FPD post cementation, which was statistically significant (p <0.05).20% of the postgraduates were aware about all the steps to be followed during re-cementation of FPD, whereas majority of the undergraduates chose better isolation to be followed, which shows that post graduates were more aware about re-cementation protocols than undergraduates and was statistically significant (p<0.05). This survey shows that both undergraduate and postgraduate dental students have fairly good knowledge about the cementation protocols for FPD.Through increased awareness of dental students expectations and experiences, restorations can be planned, made and placed in harmony with the hard and soft oral tissues, and in harmony with the patient's expectation as well