Multicolor Histochemical Staining for Identification of Mineralized and Non-Mineralized Musculoskeletal Tissue: Immunohistochemical and Radiological Validation in Decalcified Bone Samples
Histochemical staining of paraffin-embedded decalcified bone samples is commonly used in preclinical research of musculoskeletal diseases, enabling the visualization of multiple tissue components by the application of chromogens. The purpose of this study was to introduce a novel multicolor staining...
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MDPI AG
2022-09-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/9/10/488 |
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| author | Yu Sun Heike Helmholz Regine Willumeit-Römer |
| author_facet | Yu Sun Heike Helmholz Regine Willumeit-Römer |
| author_sort | Yu Sun |
| collection | DOAJ |
| description | Histochemical staining of paraffin-embedded decalcified bone samples is commonly used in preclinical research of musculoskeletal diseases, enabling the visualization of multiple tissue components by the application of chromogens. The purpose of this study was to introduce a novel multicolor staining protocol involving optimized chemical reagents and procedure, allowing the identification of high-mineralized bone, low-mineralized fracture callus, cartilage and skeletal muscle fibers simultaneously. Fractured femur and healthy tail vertebra samples from adult male Sprague–Dawley rats were decalcified with EDTA and formic acid, respectively, followed by paraffin embedding, tissue sectioning and multicolor staining. Conventional Movat’s pentachrome and safranin O / fast green staining were conducted in parallel for comparison. Immunohistochemical staining of collagen type-X and micro-CT analysis were included to further validate the efficacy of the staining method. The multicolor staining allowed visualization of major musculoskeletal tissue components in both types of decalcified samples, providing quality outcomes with fewer chemical reagents and simplified procedures. Immunohistochemical staining demonstrated its capacity for identification of the endochondral ossification process during fracture healing. Micro-CT imaging validated the staining outcome for high-mineralized skeletal tissue. The application of the multicolor staining may facilitate future preclinical research involving decalcified paraffin-embedded samples. |
| first_indexed | 2024-03-09T20:41:40Z |
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| id | doaj.art-3a0efcbe6de047ed83c89b0c48d26646 |
| institution | Directory Open Access Journal |
| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-09T20:41:40Z |
| publishDate | 2022-09-01 |
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| spelling | doaj.art-3a0efcbe6de047ed83c89b0c48d266462023-11-23T22:56:46ZengMDPI AGBioengineering2306-53542022-09-0191048810.3390/bioengineering9100488Multicolor Histochemical Staining for Identification of Mineralized and Non-Mineralized Musculoskeletal Tissue: Immunohistochemical and Radiological Validation in Decalcified Bone SamplesYu Sun0Heike Helmholz1Regine Willumeit-Römer2Institute of Metallic Biomaterials, Helmholtz-Zentrum Hereon, 21502 Geesthacht, GermanyInstitute of Metallic Biomaterials, Helmholtz-Zentrum Hereon, 21502 Geesthacht, GermanyInstitute of Metallic Biomaterials, Helmholtz-Zentrum Hereon, 21502 Geesthacht, GermanyHistochemical staining of paraffin-embedded decalcified bone samples is commonly used in preclinical research of musculoskeletal diseases, enabling the visualization of multiple tissue components by the application of chromogens. The purpose of this study was to introduce a novel multicolor staining protocol involving optimized chemical reagents and procedure, allowing the identification of high-mineralized bone, low-mineralized fracture callus, cartilage and skeletal muscle fibers simultaneously. Fractured femur and healthy tail vertebra samples from adult male Sprague–Dawley rats were decalcified with EDTA and formic acid, respectively, followed by paraffin embedding, tissue sectioning and multicolor staining. Conventional Movat’s pentachrome and safranin O / fast green staining were conducted in parallel for comparison. Immunohistochemical staining of collagen type-X and micro-CT analysis were included to further validate the efficacy of the staining method. The multicolor staining allowed visualization of major musculoskeletal tissue components in both types of decalcified samples, providing quality outcomes with fewer chemical reagents and simplified procedures. Immunohistochemical staining demonstrated its capacity for identification of the endochondral ossification process during fracture healing. Micro-CT imaging validated the staining outcome for high-mineralized skeletal tissue. The application of the multicolor staining may facilitate future preclinical research involving decalcified paraffin-embedded samples.https://www.mdpi.com/2306-5354/9/10/488musculoskeletal tissuefracture calluscartilagemusclehistochemistryimmunohistochemistry |
| spellingShingle | Yu Sun Heike Helmholz Regine Willumeit-Römer Multicolor Histochemical Staining for Identification of Mineralized and Non-Mineralized Musculoskeletal Tissue: Immunohistochemical and Radiological Validation in Decalcified Bone Samples Bioengineering musculoskeletal tissue fracture callus cartilage muscle histochemistry immunohistochemistry |
| title | Multicolor Histochemical Staining for Identification of Mineralized and Non-Mineralized Musculoskeletal Tissue: Immunohistochemical and Radiological Validation in Decalcified Bone Samples |
| title_full | Multicolor Histochemical Staining for Identification of Mineralized and Non-Mineralized Musculoskeletal Tissue: Immunohistochemical and Radiological Validation in Decalcified Bone Samples |
| title_fullStr | Multicolor Histochemical Staining for Identification of Mineralized and Non-Mineralized Musculoskeletal Tissue: Immunohistochemical and Radiological Validation in Decalcified Bone Samples |
| title_full_unstemmed | Multicolor Histochemical Staining for Identification of Mineralized and Non-Mineralized Musculoskeletal Tissue: Immunohistochemical and Radiological Validation in Decalcified Bone Samples |
| title_short | Multicolor Histochemical Staining for Identification of Mineralized and Non-Mineralized Musculoskeletal Tissue: Immunohistochemical and Radiological Validation in Decalcified Bone Samples |
| title_sort | multicolor histochemical staining for identification of mineralized and non mineralized musculoskeletal tissue immunohistochemical and radiological validation in decalcified bone samples |
| topic | musculoskeletal tissue fracture callus cartilage muscle histochemistry immunohistochemistry |
| url | https://www.mdpi.com/2306-5354/9/10/488 |
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