EVA-enhanced embedding medium for histological analysis of 3D porous scaffold material.
When sectioning a 3D porous scaffold made of a soft elastomeric material embedded in paraffin medium, it is not easy to obtain a section because of the different mechanical properties of the paraffin and tissue/scaffold. We describe a new embedding material for histological analysis of various biomaterials that is composed of paraffin and ethylene vinyl acetate (EVA) resin (0, 3, 7, and 13 wt.%). 3D porous poly(L-lactide-epsilon-caprolactone) (PLCL) and chitosan scaffolds were fabricated to test the sectioning efficiency of the paraffin/EVA embedding material. The new embedding material was characterized by rheological analysis and solvent solubility testing in xylene and n-hexane. The hydrophilicity of the new material was assessed by contact angle measurement and its surface roughness was measured using AFM analysis. The staining efficiency of sections embedded in a paraffin/EVA mixture was determined by eosin staining of the chitosan scaffold and chitosan/collagen hybrid scaffold using a fluorescently labeled collagen. Section roughness decreased with increasing EVA content. The softening temperature of the paraffin/EVA mixture was similar to that of paraffin (50-60 degrees C by rheometer). The paraffin/EVA mixture dissolved completely in xylene after 30min at 50 degrees C, and after 30min in n-hexane at 60 degrees C. Therefore, the new embedding medium can be used for histological analysis of various biomaterials and natural tissues.
Lim
Jin Ik
JI
Department of Dental Biomaterials Science, Dental Research Institute and BK21 Program, School of Dentistry, Seoul National University, Seoul, Republic of Korea.
Lee
Yong-Keun
YK
D048271:Chitosan; D008422:Materials Testing; D010232:Paraffin / Q000737:chemistry; D011091:Polyesters; D011145:Polyvinyls / Q000737:chemistry*; D016062:Porosity; D013194:Staining and Labeling; D013499:Surface Properties; D013696:Temperature; D016610:Tissue Embedding / Q000379:methods*; D023822:Tissue Engineering / Q000295:instrumentation; D054457:Tissue Scaffolds
D011091:Polyesters; D011145:Polyvinyls; C016438:ethylenevinylacetate copolymer; C062968:lactide-caprolactone copolymer; D010232:Paraffin; D048271:Chitosan