Biological Behavior of Xenogenic Scaffolds in Alcohol-Induced Rats: Histomorphometric and Picrosirius Red Staining Analysis

Abstract

In this experimental protocol, the objective was to evaluate the biological behavior of two xenogenic scaffolds in alcohol-induced rats through histomorphometric and Picrosirius Red staining analysis of non-critical defects in the tibia of rats submitted or not to alcohol ingestion at 25% v/v. Eighty male rats were randomly divided into four groups (n = 20 each): CG/B (water diet + Bio-Oss® graft, Geistlich Pharma AG, Wolhusen, Switzerland), CG/O (water diet + OrthoGen® graft, Baumer, Mogi Mirim, Brazil), AG/B (25% v/v alcohol diet + Bio-Oss® graft), and AG/O (25% v/v alcohol diet + OrthoGen® graft). After 90 days of liquid diet, the rats were surgically obtained, with a defect in the tibia proximal epiphysis; filled in according to their respective groups; and euthanized at 10, 20, 40 and 60 days. In two initial periods (10 and 20 days), all groups presented biomaterial particles surrounded by disorganized collagen fibrils. Alcoholic animals (AG/B and AG/O) presented, in the cortical and medullary regions, a reactive tissue with inflammatory infiltrate. In 60 days, in the superficial area of the surgical cavities, particles of biomaterials were observed in all groups, with new compact bone tissue around them, without complete closure of the lesion, except in non-alcoholic animals treated with Bio-Oss® xenograft (CG/B), where the new cortical interconnected the edges of the defect. Birefringence transition was observed in the histochemical analysis of collagen fibers by Picrosirius Red, in which all groups in periods of 10 and 20 days showed red-orange birefringence, and from 40 days onwards greenish-yellow birefringence, which demonstrates the characteristic transition from the formation of thin and disorganized collagen fibers initially to more organized and thicker later. In histomorphometric analysis, at 60 days, CG/B had the highest volume density of new bone (32.9 ± 1.15) and AG/O the lowest volume density of new bone (15.32 ± 1.71). It can be concluded that the bone neoformation occurred in the defects that received the two biomaterials, in all periods, but the Bio-Oss® was superior in the results, with its groups CG/B and AG/B displaying greater bone formation (32.9 ± 1.15 and 22.74 ± 1.15, respectively) compared to the OrthoGen® CG/O and AG/O groups (20.66 ± 2.12 and 15.32 ± 1.71, respectively), and that the alcoholic diet interfered negatively in the repair process and in the percentage of new bone formed.

Effects of a Biocomplex Formed by Two Scaffold Biomaterials, Hydroxyapatite/Tricalcium Phosphate Ceramic and Fibrin Biopolymer, with Photobiomodulation, on Bone Repair

“…The qualitative analysis and the arrangement of collagen fibers in the connective tissue formed in the defect was evaluated by Picrosirius-red staining, which allows the detection of different types of collagen [ 59 ]. At 14 days, all groups presented fine and disorganized collagen fibers (type III), but the G4/BFB + PBM group presented a more advanced stage of maturation with some zones of mineralization.…”

Photobiomodulation Therapy Improves Repair of Bone Defects Filled by Inorganic Bone Matrix and Fibrin Heterologous Biopolymer

“…Fundamentally, the quality of the newly formed organic matrix during the bone repair process is assessed. Depending on the nature of the fibers, they are birefringent to green when more mature, yellow and red when less mature [80]. At 42 days, a more evident cell maturation was observed, with more organized collagen fibers in the G3 group (biomaterial + fibrin biopolymer + photobiomodulation), showing a more mature regeneration in relation to the other groups.…”

Use of Plant Extracts in Polymeric Scaffolds in the Regeneration of Mandibular Injuries

“…Once the animals had died, the mandibles were removed and photodocumented to analyze the clinical conditions of the operated area, followed by image analysis using digital radiography (AJEX-240 Diagnostic X-ray System ® : 0.4–100 mA, 40–120 kVp, D-125 Inserted X-ray Tube and Pixx1717 apartment panel detector, Pixxogen). The samples were then immersed in formaldehyde solution and subjected to routine histological methods to make sections along the bone defect, and then stained with Masson’s trichrome to characterize bone neoformation and with Picrosirius red (saturated aqueous solution of picric acid plus 0.1 g of Sirius red F3B-Bayer ® , St. Louis, MO, USA) to analyze the fibrillar components by polarized light microscopy [ 20 , 53 , 54 ].…”