In Vitro Evaluation of the Allogeneic Bone Matrix Effect on the Adipose Mesenchymal Stromal Cells Characteristics in Combined Tissue Engineering

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Abstract

The aim of the study was to evaluate in vitro the effect of native and deproteinized compact and spongy allogenic bone matrices on the characteristics of adipose mesenchymal stromal cells (ASC) in combined tissue engineering.

Material and Methods. 24 samples of native and deproteinized compact and spongy bone were examined, which were exposed to mechanical treatment, modeling, followed by sterilization of the samples by ionizing radiation and bacteriological control of sterilization. Some of the samples underwent deproteinization. The characterized cultures of human ASC were used as test cultures to assess the interaction with the bone samples. The Cytation-5 fluorescent imager and Hoechst 3334 fluorochromes (BD Pharmingen™) and calcein (Calcein AM, BD Pharmingen™) were used to characterize the degree of adhesion, migration, and viability of ASC on bone matrix samples. Matrix cytotoxicity was evaluated by MTT assay on days 1 and 7 of extraction.

Results. The bone matrix samples are characterized by the absence of cytotoxicity (rank 1). ASC demonstrated good adhesion and migration on any surface of the bone matrix and preservation of cell viability during 7 days of observation. Nuclei sizes of the cells adhered to the deproteinized bone matrix of the spongy structure increased by 25–30% compared to other samples. The cells on deproteinized bone matrix had greater size (the size of the cells from nuclei 8.8 to 11.5 μm, the average size of cells nuclei from an 86.3 μm to 129,0 μm, the average perimeter of the cells nuclei from 30.7 μm to 40.7 μm) than in the native bone matrix samples.

Conclusion. The results of the study of various allogeneic bone matrices demonstrate that deep purification of the bone matrix determines the absence of cytotoxicity and the most favorable conditions for the adhesion, migration, proliferation and viability of ASC. Also makes it possible to use tissue engineering based on bone matrices of different structures. Deproteinized spongy bone matrices are best suited for this purpose.

About the authors

L. A. Cherdantseva

Tsivyan Novosibirsk Research Institute of Traumatology and Orthopedics

Email: cherdanceff@yandex.ru
ORCID iD: 0000-0002-4729-3694

Liliya A. Cherdantseva — Cand. Sci. (Med.), Head of the Laboratory for Procurement and Preservation of Tissues

Novosibirsk

Russian Federation

E. A. Anastasieva

Tsivyan Novosibirsk Research Institute of Traumatology and Orthopedics

Email: evgeniya.anastasieva@gmail.com
ORCID iD: 0000-0002-9329-8373

Evgeniya A. Anastasieva — PhD Student

Novosibirsk

Russian Federation

D. Ya. Aleynik

Privolzhsky Research Medical University

Email: daleynik@yandex.ru
ORCID iD: 0000-0003-1482-4281

Diane Ya. Aleynik — Cand. Sci. (Med.), Senior Researcher, Laboratory of Regenerative Medicine, Institute of Experimental Oncology and Biomedical Technologies

Nizhny Novgorod

M. N. Egorikhina

Privolzhsky Research Medical University

Email: egorihina.marfa@yandex.ru
ORCID iD: 0000-0002-8815-9651

Marfa N. Egorikhina — Cand. Sci. (Biol.), Leading Researcher, Laboratory of Regenerative Medicine, Institute of Experimental Oncology and Biomedical Technologies

Nizhny Novgorod

I. A. Kirilova

Tsivyan Novosibirsk Research Institute of Traumatology and Orthopedics

Author for correspondence.
Email: IrinaKirilova71@mail.ru
ORCID iD: 0000-0003-1911-9741

Irina A. Kirilova — Dr. Sci. (Med.), Deputy Director

Novosibirsk

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