IADR Abstract Archives
An insight into the quantitative morphological characterisation of regenerated oral mucosa
Objectives: To automate computerised methods of assessing architectural information demonstrated by organotypic cultures of oral mucosa and compare the culture features with those of normal tissues.
Methods: In this study, we characterise the architectural features of the tissue based on position of cell nuclei in histological sections. These positions can subsequently be used to seed a partitioning of the whole epithelial compartment profile to produce theoretical cell extents. Oral keratinocytes were used to produce organotypic cultures on de-epidermalised dermis grown for 14 days. Five micrometre histological sections of the cultures stained with haematoxylin and eosin were colour-unmixed using colour deconvolution. We investigated 16 methods of histogram thresholding to find out which is the best method for isolating the nuclear locations on the haematoxylin channel. For this we used true-ground binary images segmented by hand and compared them those obtained by the various computerised models.
Results: Histological sections of organotypic cultures of oral keratinocytes on 3 different scaffolds DED, collagen and PET and their statistical analysis based on SIOX revealed that air-liquid interface cultures generated stratified organotypic epithelial structures after 14-days were significantly (P<0.001) different, however, regenerated tissues were characterised very much similar to the normal oral mucosa.
Conclusions: The architectural characterisation of organotypic cultures of oral keratinocytes by mean of computerised quantitative microscopy provides a better understanding of cellular feature which can aid in comparing morphological data of engineered oral mucosa with normal oral epithelium by nuclear isolation.
Methods: In this study, we characterise the architectural features of the tissue based on position of cell nuclei in histological sections. These positions can subsequently be used to seed a partitioning of the whole epithelial compartment profile to produce theoretical cell extents. Oral keratinocytes were used to produce organotypic cultures on de-epidermalised dermis grown for 14 days. Five micrometre histological sections of the cultures stained with haematoxylin and eosin were colour-unmixed using colour deconvolution. We investigated 16 methods of histogram thresholding to find out which is the best method for isolating the nuclear locations on the haematoxylin channel. For this we used true-ground binary images segmented by hand and compared them those obtained by the various computerised models.
Results: Histological sections of organotypic cultures of oral keratinocytes on 3 different scaffolds DED, collagen and PET and their statistical analysis based on SIOX revealed that air-liquid interface cultures generated stratified organotypic epithelial structures after 14-days were significantly (P<0.001) different, however, regenerated tissues were characterised very much similar to the normal oral mucosa.
Conclusions: The architectural characterisation of organotypic cultures of oral keratinocytes by mean of computerised quantitative microscopy provides a better understanding of cellular feature which can aid in comparing morphological data of engineered oral mucosa with normal oral epithelium by nuclear isolation.