生理性血凝块对成骨细胞响应基于壳聚糖的3D支架的影响——一项初步研究

The Influence of Physiological Blood Clot on Osteoblastic Cell Response to a Chitosan-Based 3D Scaffold-A Pilot Investigation.

作者信息

da Costa Natacha Malu Miranda, Caetano Hilary Ignes Palma, Aguiar Larissa Miranda, Parisi Ludovica, Ghezzi Benedetta, Elviri Lisa, Zuardi Leonardo Raphael, de Oliveira Paulo Tambasco, Palioto Daniela Bazan

机构信息

Department of Oral and Maxillofacial Surgery and Periodontology, School of Dentistry of Ribeirão Preto, University of São Paulo, Avenida Do Café-Subsetor Oeste-11 (N-11), Ribeirão Preto 14040-904, SP, Brazil.

Laboratory for Oral Molecular Biology, Department of Orthodontics and Dentofacial Orthopedics, University of Bern, Freiburgstrasse 3, 3010 Bern, Switzerland.

出版信息

Biomimetics (Basel). 2024 Dec 21;9(12):782. doi: 10.3390/biomimetics9120782.

DOI:10.3390/biomimetics9120782

PMID:39727786

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11727562/

Abstract

BACKGROUND

The use of ex vivo assays associated with biomaterials may allow the short-term visualization of a specific cell type response inserted in a local microenvironment. Blood is the first component to come into contact with biomaterials, providing blood clot formation, being substantial in new tissue formation. Thus, this research investigated the physiological blood clot (PhC) patterns formed in 3D scaffolds (SCAs), based on chitosan and 20% beta-tricalcium phosphate and its effect on osteogenesis. Initially, SCA were inserted for 16 h in rats calvaria defects, and, after that, osteoblasts cells (OSB; UMR-106 lineage) were seeded on the substrate formed. The groups tested were SCA + OSB and SCA + PhC + OSB. Cell viability was checked by MTT and mineralized matrix formation in OSB using alizarin red (ARS). The alkaline phosphatase (ALP) and bone sialoprotein (BSP) expression in OSB was investigated by indirect immunofluorescence (IF). The OSB and PhC morphology was verified by scanning electron microscopy (SEM).

RESULTS

The SCA + PhC + OSB group showed greater cell viability ( = 0.0169). After 10 days, there was more mineralized matrix deposition ( = 0.0365) and high ALP immunostaining ( = 0.0021) in the SCA + OSB group. In contrast, BSP was more expressed in OSB seeded on SCA with PhC ( = 0.0033).

CONCLUSIONS

These findings show the feasibility of using PhC in ex vivo assays. Additionally, its inclusion in the experiments resulted in a change in OSB behavior when compared to in vitro assays. This "closer to nature" environment can completely change the scenario of a study.

摘要

背景

与生物材料相关的体外试验的应用可能有助于短期观察插入局部微环境中的特定细胞类型反应。血液是最先与生物材料接触的成分，它能促进血栓形成，在新组织形成过程中起着重要作用。因此，本研究调查了基于壳聚糖和20%β-磷酸三钙的三维支架（SCAs）中形成的生理性血栓（PhC）模式及其对成骨的影响。最初，将SCAs植入大鼠颅骨缺损处16小时，之后，将成骨细胞（OSB；UMR-106谱系）接种在形成的基质上。所测试的组为SCA + OSB和SCA + PhC + OSB。通过MTT检测细胞活力，并使用茜素红（ARS）检测OSB中的矿化基质形成。通过间接免疫荧光（IF）研究OSB中碱性磷酸酶（ALP）和骨唾液蛋白（BSP）的表达。通过扫描电子显微镜（SEM）观察OSB和PhC的形态。