Evaluation of biocompatibility and osteoconductivity of a hybrid cell-tissue graft for bone regenerative medicine

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Abstract

Aim – to evaluate in vitro the biocompatibility and osteoconductivity of a hybrid graft based on a bioorganic matrix, human bone marrow mesenchymal stromal cells (BM-MSC) and osteogenic growth factors.

Material and methods. Bioorganic matrices were studied for biocompatibility with human BM-MSC culture used in traumatology and orthopedics. For promoted osteogenic differentiation of BM-MSCs, allogeneic plasma enriched with soluble platelet factors was used. The osteogenic potential of BM-MSCs by the synthesis of mRNAs of early (transcription factor 2 (Run X2), alkaline phosphatase (ALP)) and late genes (osteopontin (OSP)) of osteogenesis was analyzed. The properties of cell adhesion and proliferation of MSCs in the conditions of a three-dimensional hybrid graft by the MTT test and fluorescence microscopy were assessed.

Results. The biocompatibility of the studied bioorganic matrices with human BM-MSCs was established. The collagen matrix promoted rapid cell adhesion and proliferation between the scaffold fibrils. It has also been established that allogeneic platelet-rich plasma affects the osteogenic differentiation of human BM-MSCs in vitro, increasing the expression of marker genes RunX2, ALP, OSP. When modeling a hybrid graft in vitro, the formation of a tight contact between the alloimplant and collagen biopolymer using MSCs was shown.

Conclusion. The biological properties of the developed hybrid cell-tissue graft characterize its biocompatibility and osteoconductivity of its constituent components, which makes it promising for use in regenerative medicine, especially in reconstructive surgery of bone defects.

About the authors

Nataliya N. Danilkovich

Republican Scientific and Practical Center of Transfusiology and Medical Biotechnology

Author for correspondence.
Email: nndanilkovich@gmail.com
ORCID iD: 0000-0002-1245-0426

Scientific officer of the laboratory of Stem Cell Biology and Genetics

Belarus, Minsk

Svetlana M. Kosmacheva

Republican Scientific and Practical Center of Transfusiology and Medical Biotechnology

Email: 4kosmacheva@mail.ru
ORCID iD: 0000-0002-1617-8845

PhD, Associate professor, Head of the laboratory of Stem Cells Biology and Genetics

Belarus, Minsk

Aleksandra G. Ionova

Republican Scientific and Practical Center of Transfusiology and Medical Biotechnology

Email: al_ionova96@mail.ru
ORCID iD: 0009-0000-3884-9112

Junior researcher of the laboratory of Stem Cells Biology and Genetics

Belarus, Minsk

Kirill A. Krivorot

Republican Scientific and Practical Center of Traumatology and Orthopedics

Email: kirill.doc@mail.ru
ORCID iD: 0000-0003-0456-2839

PhD, Associate professor, Deputy Director for Organizational and Methodological work, neurosurgeon of the highest qualification category

Belarus, Minsk

Andrei N. Mazurenko

Republican Scientific and Practical Center of Traumatology and Orthopedics

Email: mazurenko@mail.ru
ORCID iD: 0000-0001-7092-2615

Head of Neurosurgical Department №2, PhD, Associate professor

Belarus, Minsk

Denis G. Alekseev

Samara State Medical University

Email: D.G.Alekseev@samsmu.ru
ORCID iD: 0000-0003-4185-0709

PhD, Associate professor, Leading researcher at the BioTech Research Institute

Russian Federation, Samara

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Supplementary files

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2. Figure 1. Nuclei labeling of Hoechst 33342 BM-MSCs after one day of cultivation with different carriers: (a) CCM (control sample); (b) Osteomatrix; (c) LBM; (d) Kollapan; (e) Lyostypt. Image taken at 100x magnification.

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3. Figure 2. Proliferative activity of different concentrations of BM-MSCs on the ‘‘Lyostypt” bioorganic carrier over a period of 7-d cultivation in vitro.

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4. Figure 3. mRNA expression of osteogenic genes: (a, d) RunX, (b, e) ALP, (c, f) OSP after 4 (top row) and 7 (bottom row) days of cultivation BM-MSCs. Data are expressed as M ± SEM.

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5. Figure 4. Von Kossa staining of BM-MSCs: (a) cells cultivated in CCM; (b) cells cultivated in OM; (c) cells cultivated in OM, supplemented with 5% alPRP. Images are taken at 100x magnification.

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6. Figure 5. Co-cultivation of graft components in vitro: (a) one week, (b) three weeks. Image taken at 50x magnification.

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7. Figure 6. Live labeling of Hoechst 33342 of osteogenically predifferentiated BM-MSCs in vitro graft modeling: (a) one week, (b) two weeks, (c) tree weeks. Images are taken at 100x magnification.

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Copyright (c) 2024 Danilkovich N.N., Kosmacheva S.M., Ionova A.G., Krivorot K.A., Mazurenko A.N., Alekseev D.G.

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This work is licensed under a Creative Commons Attribution 4.0 International License.

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