IADR Abstract Archives
Cell Derived Matrices as Models to Study Gingival Reintegration
Objectives: 1. Developing a simple cell-culture technique to recapitulate the ECM, which mimics aspects of the native tissue microenvironment.
2. Generation of cell-derived matrices on various substrates including hydroxyapatite, gelatin, and polymer brushes.
3. Characterisation and quantification of resulting cell-derived matrices by immunofluorescence, confocal microscopy and scanning electron microscopy.
Methods: 1. Human gingival fibroblasts were cultured for 10 days in defined supplemented media, at the surface of the various substrates studied.
2. The cells were denuded and DNA was digested to ensure complete cell lysis by biochemical methods.
3. The resultant matrix was then quantified via immunofluorescence and confocal and scanning electron microscopy and the impact of substrate chemistry correlated.
Results: 1. The data was quantified by calculating and comparing fibre density, thickness and fluorescence intensity of the ECM deposited on various substrates using ImageJ.
2. Gelatin, hydroxyapatite, polysulfopropyl methacrylate and polymethyl and ethyl acrylate brush-coated substrates show thick mature ECM deposition compared to polylysine-graft-poly ethylene glycol (PLL-PEG)-coated and, poly(oligo(ethylene glycol) methyl ether methacrylate (POEGMA)-coated substrates.
3. ECM deposition is enhanced by the use of defined supplements in the cell-culture medium during CDM generation.
Conclusions: CDMs offer the opportunity to produce microenvironments with customizable biological and biophysical properties in a controlled setting. As a result, CDMs can improve cellular functions and can be utilized as biomaterials to engineer tissues de-novo or facilitate endogenous healing and regeneration. This research provides a brief overview of the methodologies used to facilitate CDM manufacturing and explores the versatile uses of CDMs in fundamental research and particularly its therapeutic approach following gingival injury.
In conclusion, CDM-based research advancements in methodologies, as well as innovative interdisciplinary approaches, are needed to pave the way for an exciting next generation of CDMs for basic research and therapeutic approaches.
2. Generation of cell-derived matrices on various substrates including hydroxyapatite, gelatin, and polymer brushes.
3. Characterisation and quantification of resulting cell-derived matrices by immunofluorescence, confocal microscopy and scanning electron microscopy.
Methods: 1. Human gingival fibroblasts were cultured for 10 days in defined supplemented media, at the surface of the various substrates studied.
2. The cells were denuded and DNA was digested to ensure complete cell lysis by biochemical methods.
3. The resultant matrix was then quantified via immunofluorescence and confocal and scanning electron microscopy and the impact of substrate chemistry correlated.
Results: 1. The data was quantified by calculating and comparing fibre density, thickness and fluorescence intensity of the ECM deposited on various substrates using ImageJ.
2. Gelatin, hydroxyapatite, polysulfopropyl methacrylate and polymethyl and ethyl acrylate brush-coated substrates show thick mature ECM deposition compared to polylysine-graft-poly ethylene glycol (PLL-PEG)-coated and, poly(oligo(ethylene glycol) methyl ether methacrylate (POEGMA)-coated substrates.
3. ECM deposition is enhanced by the use of defined supplements in the cell-culture medium during CDM generation.
Conclusions: CDMs offer the opportunity to produce microenvironments with customizable biological and biophysical properties in a controlled setting. As a result, CDMs can improve cellular functions and can be utilized as biomaterials to engineer tissues de-novo or facilitate endogenous healing and regeneration. This research provides a brief overview of the methodologies used to facilitate CDM manufacturing and explores the versatile uses of CDMs in fundamental research and particularly its therapeutic approach following gingival injury.
In conclusion, CDM-based research advancements in methodologies, as well as innovative interdisciplinary approaches, are needed to pave the way for an exciting next generation of CDMs for basic research and therapeutic approaches.