Departamento de Bioquímica e Biologia Tecidual, Instituto de Biologia, UNICAM, Campinas, SP Brazil.
Centro de Ciências Naturais e Humanas, Universidade Federal do ABC, São Bernardo do Campo, SP, Brazil.
Organogenesis. 2021 Oct 2;17(3-4):136-149. doi: 10.1080/15476278.2021.2003134. Epub 2021 Nov 30.
Cells with osteogenic potential are believed to be an ideal source for bone tissue bioengineering. Large bone defects require temporary substitution of the damaged parts. In this respect, the transplantation of bone cells cultured on osteogenic substrates has been investigated. To use the natural bone matrix, one approach is the so-called demineralized bone matrix (DBM). In this study, we evaluated the interaction of human fetal osteoblasts (hFOB 1.19 cells, a human fetal osteoblastic cell line) with DBM fragments. No additional bone differentiation inducer was used other than the DBM itself. The samples were processed, had adhesion pattern evaluated and analyzed by light microscopy (cytochemical and immunocytochemical analysis) and electron microscopy (scanning and transmission). The adhesion pattern of hFOB cells on DBM was similar to what was observed on the cell culture plate. Morphological analysis showed that the hFOB cells had emitted filopodia and cellular projections on both controls and DBM. On DBM, the adhered cells emitted prolongations and migrated into the matrix. The monolayer growth pattern was observed as well as the accumulation of filamentous and reticulate extracellular materials when hFOB cells were cultured on the DBM surface. EDS analysis revealed the deposition of calcium on DBM. Immunocytochemical data showed that the hFOB cells were able to secrete extracellular matrix molecules such as fibronectin and laminin on DBM. Our data indicate that DBM successfully stimulates the osteoblastic phenotype of osteoblast-like cells and corroborate with the fact that DBM is a considerable natural matrix that promotes fractured-bone healing.
具有成骨潜力的细胞被认为是骨组织工程学的理想来源。大的骨缺损需要暂时替代受损部位。在这方面，已经研究了在成骨基质上培养的骨细胞的移植。为了利用天然骨基质，一种方法是所谓的脱矿质骨基质（DBM）。在这项研究中，我们评估了人胎成骨细胞（hFOB 1.19 细胞，一种人胎成骨细胞系）与 DBM 碎片的相互作用。除了 DBM 本身外，没有使用其他额外的骨分化诱导剂。对样品进行处理，通过光镜（细胞化学和免疫细胞化学分析）和电子显微镜（扫描和透射）评估粘附模式并进行分析。hFOB 细胞在 DBM 上的粘附模式与在细胞培养板上观察到的相似。形态学分析表明，hFOB 细胞在对照和 DBM 上都发出了丝状伪足和细胞突起。在 DBM 上，粘附的细胞发出突起并迁移到基质中。当 hFOB 细胞在 DBM 表面培养时，观察到单层生长模式以及丝状和网状细胞外基质物质的积累。EDS 分析表明 DBM 上有钙沉积。免疫细胞化学数据表明，hFOB 细胞能够在 DBM 上分泌细胞外基质分子，如纤连蛋白和层粘连蛋白。我们的数据表明 DBM 成功地刺激了成骨样细胞的成骨表型，并证实了 DBM 是一种促进骨折愈合的重要天然基质。
