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

Dayane Maria Braz Nogueira,Cláudio Maldonado Pastori,Paulo Sérgio Da Silva Santos,Geraldo Marco Rosa Júnior,Patrícia Lopes Alcantara,Eliana De Souza Bastos Mazuqueli Pereira,Samira Salmeron,Karina Torres Pomini,Miguel Ângelo De Marchi,Carlos Henrique Bertoni Reis,Marcelie Priscila De Oliveira Rosso,Beatriz Flavia De Moraes Trazzi,Rogerio Leone Buchaim,Daniela Vieira Buchaim,Iris Jasmin Santos German,Mariana Schutzer Ragghianti Zangrando,André Luiz De Faria Figadoli,Jéssica De Oliveira Rossi

doi:10.3390/polym14030584

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.

Highlights

Alcohol consumption is often accepted and socially encouraged, so its consumption has become excessive, causing serious damage and harm to the health of its consumers [1,2,3,4,5]
The biomaterial particles transacted the cortical bone lesion, surrounded by loose connective tissue with the presence of vascular sprouts, being more evident in non-alcoholic animals treated with Bio-Oss® xenograft (Figure 2(a1’))
This study aimed to evaluate the bone defects repair process using a Brazilian-made mixed bovine bone matrix (OrthoGen®) in relation to a biomaterial of worldwide use (BioOss®), in rats submitted to the ingestion of diet by water or 25% v/v ethanol

Summary

Introduction

Alcohol consumption is often accepted and socially encouraged, so its consumption has become excessive, causing serious damage and harm to the health of its consumers [1,2,3,4,5]. Chronic consumption can cause disturbances and damage such as organic and psychological disorders, central nervous system, muscle system, bone tissue, liver, cardiovascular diseases and (especially) socioeconomic damage [8]. One of the tissues harmed by chronic alcohol consumption is the bone tissue [9]. Alcohol acts on bone change, changing the process of bone resorption and formation, causing a serious reduction in trabecular bone volume and thickness, causing defects in mineralization and causing reduced activity and proliferation of osteoblasts [10,11]. With the reduction of osteoblastic differentiation of bone marrow cells, adipogenesis occurs. Alcohol decreases the synthesis of an ossifiable matrix, possibly due to the inhibition of cell proliferation and poor differentiation of mesenchymal cells in the repair tissue, generating deficient healing [10,12]

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