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Statistical image analysis and escort histograms in characterization of articular cartilage repair in a skeleton animal model.

ABSTRACT: Statistical image analysis of an ensemble of digital images of histological samples is performed as an auxiliary investigation a result of the recently proposed method of articular cartilage repair utilizing growth plate chondrocytes in a skeleton animal model. A fixed-shift model of maximal likelihood estimates of image histograms applied for monochromatic (grayscale) images or their RGB components confirms the statistically significant effect of the previously proposed medical treatment. The type of staining used to prepare images of histological samples is related to the visibility of the effectiveness of medical treatment. Hellinger distance of escort distributions for maximal likelihood estimates of image histograms of medically treated and control samples is investigated to identify grayscale (or RGB) intensities responsible for statistically significant difference of the estimates. A difference of Shannon entropy quantifying informational content of the histograms allows one to identify staining and image colors which are most suitable to visualize cluster formation typical for articular cartilage repair processes.

SUBMITTER: Tomaszewski R

PROVIDER: S-EPMC8213189 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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Statistical image analysis and escort histograms in characterization of articular cartilage repair in a skeleton animal model.

Tomaszewski Ryszard R Dajka Jerzy J

PloS one 20210618 6

Statistical image analysis of an ensemble of digital images of histological samples is performed as an auxiliary investigation a result of the recently proposed method of articular cartilage repair utilizing growth plate chondrocytes in a skeleton animal model. A fixed-shift model of maximal likelihood estimates of image histograms applied for monochromatic (grayscale) images or their RGB components confirms the statistically significant effect of the previously proposed medical treatment. The t ...[more]

PMID: 34143793

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Project description:Focal cartilage injuries have poor intrinsic healing potential and often progress to osteoarthritis, a costly disease affecting almost a third of adults in the United States. To treat these patients, cartilage repair therapies often use cell-seeded scaffolds, which are limited by donor site morbidity, high costs, and poor efficacy. To address these limitations, we developed an electrospun cell-free fibrous hyaluronic acid (HA) scaffold that delivers factors specifically designed to enhance cartilage repair: Stromal Cell-Derived Factor-1α (SDF-1α; SDF) to increase the recruitment and infiltration of mesenchymal stem cells (MSCs) and Transforming Growth Factor-β3 (TGF-β3; TGF) to enhance cartilage tissue formation. Scaffolds were characterized in vitro and then deployed in a large animal model of full-thickness cartilage defect repair. The bioactivity of both factors was verified in vitro, with both SDF and TGF increasing cell migration, and TGF increasing matrix formation by MSCs. In vivo, however, scaffolds releasing SDF resulted in an inferior cartilage healing response (lower mechanics, lower ICRS II histology score) compared to scaffolds releasing TGF alone. These results highlight the importance of translation into large animal models to appropriately screen scaffolds and therapies, and will guide investigators towards alternative growth factor combinations. STATEMENT OF SIGNIFICANCE: This study addresses an area of orthopaedic medicine in which treatment options are limited and new biomaterials stand to improve patient outcomes. Those suffering from articular cartilage injuries are often destined to have early onset osteoarthritis. We have created a cell-free nanofibrous hyaluronic acid (HA) scaffold that delivers factors specifically designed to enhance cartilage repair: Stromal Cell-Derived Factor-1α (SDF-1α; SDF) to increase the recruitment and infiltration of mesenchymal stem cells (MSCs) and Transforming Growth Factor-β3 (TGF-β3; TGF) to enhance cartilage tissue formation. To our knowledge, this study is the first to evaluate such a bioactive scaffold in a large animal model and demonstrates the capacity for dual growth factor release.

Project description:To analyze the genetic and biomolecular mechanisms underlying cartilage repair, an optimized mouse model of osteochondral repair is required. Although several models of articular cartilage injury in mice have recently been established, the articular surface in adult C57Bl/6 mice heals poorly. Since C57Bl/6 mice are the most popular strain of genetically manipulated mice, an articular cartilage repair model using C57Bl/6 mice would be helpful for analysis of the mechanisms of cartilage repair. The purpose of this study was to establish a cartilage repair model in C57Bl/6 mice using immature animals. To achieve this goal, full-thickness injuries were generated in 3-week-old (young), 4-week-old (juvenile), and 8-week-old (adult) C57Bl/6 mice. To investigate the reproducibility and consistency of full-thickness injuries, mice were sacrificed immediately after operation, and cartilage thickness at the patellar groove, depth of the cartilage injury, cross-sectional width, and cross-sectional area were compared among the three age groups. The depth of cartilage injury/cartilage thickness ratio (%depth) and the coefficient of variation (CV) for each parameter were also calculated. At 8 weeks postoperatively, articular cartilage repair was assessed using a histological scoring system. With respect to the reproducibility and consistency of full-thickness injuries, cartilage thickness, depth of cartilage injury, and cross-sectional area were significantly larger in young and juvenile mice than in adult mice, whereas cross-sectional width and %depth were almost equal among the three age groups. CVs of %depths were less than 10% in all groups. With respect to articular cartilage repair, young and juvenile mice showed superior results. In conclusion, we established a novel cartilage repair model in C57Bl/6 mice. This model will be valuable in achieving mechanistic insights into the healing process of the joint surface, as it will facilitate the use of genetically modified mice, which are most commonly developed on a C57Bl/6 background.

Dataset Information

Statistical image analysis and escort histograms in characterization of articular cartilage repair in a skeleton animal model.

Publications

Statistical image analysis and escort histograms in characterization of articular cartilage repair in a skeleton animal model.

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