title: [Histologic assessment of tissue healing of hyaline cartilage by use of semiquantitative evaluation scale].
abstract: BACKGROUND
  Articular cartilage is an avascular and aneural tissue lacking lymph drainage, hence its inability of spontaneous repair following injury. Thus, it offers an interesting model for scientific research. A number of methods have been suggested to enhance cartilage repair, but none has yet produced significant success. The possible application of the aforementioned methods has brought about the necessity to evaluate their results. The objective of this study was to analyze results of a study of the effects of the use of TGF-beta gene transduced bone marrow clot on articular cartilage defects using ICRS visual histological assessment scale.
  
  
  METHODS
  The research was conducted on 28 skeletally mature sheep that were randomly assigned to four groups and surgically inflicted femoral chondral defects. The articular surfaces were then treated with TGF-beta1 gene transduced bone marrow clot (TGF group), GFP transduced bone marrow clot (GFP group), untransduced bone marrow clot (BM group) or left untreated (NC group). The analysis was performed by visual examination of cartilage samples and results were obtained using ICRS visual histological assessment scale. The results were subsequently subjected to statistical assessment using Kruskal-Wallis and Mann-Whitney tests.
  
  
  RESULTS
  Kruskal-Wallis test yielded statistically significant difference with respect to cell distribution. Mann-Whitney test showed statistically significant difference between TGF and NC groups (P = 0.002), as well as between BM and NC groups (P = 0.002 with Bonferroni correction).
  
  
  CONCLUSIONS
  Twenty-six of the twenty-eight samples were subjected to histologic and subsequent statistical analysis; two were discarded due to faulty histology technique. Our results indicated a level of certainty as to the positive effect of TGF-beta1 gene transduced bone marrow clot in restoration of articular cartilage defects. However, additional research is necessary in the field. One of the significant drawbacks on histologic assessment of cartilage samples were the errors in histologic preparation, for which some samples had to be discarded and significantly impaired the analytical quality of the others. Defects of structures surrounding the articular cartilage, e.g., subchondral bone or connective tissue, might also impair the quality of histologic analysis. Additional analyses, i.e. polarizing microscopy should be performed to determine the degree of integration of the newly formed tissue with the surrounding cartilage. The semiquantitative ICRS scale, although of great practical value, has limitations as to the objectivity of the assessment, taking into account the analytical ability of the evaluator, as well as the accuracy of semiquantitative analysis in comparison to the methods of quantitative analysis.
  
  
  CONCLUSIONS
  Overall results of histologic analysis indicated that the application of TGF-beta1 gene transduced bone marrow clot could have measurable clinical effects on articular cartilage repair. The ICRS visual histological assessment scale is a valuable analytical method for cartilage repair evaluation. In this respect, further analyses of the method value would be of great importance.
authors.affiliation: Zavod za histologiju i embriologiju Medicinskog fakulteta Sveucilista u Zagrebu, Zagreb, Hrvatska. andreja_vukasovic@yahoo.com
authors.forename: Andreja
authors.initials: A
authors.lastname: Vukasović
authors.forename: Alan
authors.initials: A
authors.lastname: Ivković
authors.forename: Davor
authors.initials: D
authors.lastname: Jezek
authors.forename: Ivan
authors.initials: I
authors.lastname: Cerovecki
authors.forename: Drazen
authors.initials: D
authors.lastname: Vnuk
authors.forename: Mario
authors.initials: M
authors.lastname: Kreszinger
authors.forename: Damir
authors.initials: D
authors.lastname: Hudetz
authors.forename: Marko
authors.initials: M
authors.lastname: Pećina
mesh_terms: D000818:Animals; D016026:Bone Marrow Transplantation; D002358:Cartilage, Articular / Q000473:pathology* / Q000503:physiopathology; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D012756:Sheep; D014161:Transduction, Genetic; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D014182:Transplantation, Autologous; D014945:Wound Healing / Q000187:drug effects
chemical_list: D053773:Transforming Growth Factor beta1; D049452:Green Fluorescent Proteins