scholarly journals Evaluation of in vitro biocompatibility of scaffolds for the repair of bone defects

2021 ◽  
Vol 9 (2) ◽  
Author(s):  
N. Bezdieniezhnykh ◽  
◽  
Ye. Holiuk ◽  
S. Gerasymenko ◽  
K. Saulenko ◽  
...  
Keyword(s):  
Proliferative Activity ◽  
Platelet Rich Plasma ◽  
Human Fibroblasts ◽  
Bone Matrix ◽  
Bone Defects ◽  
Allogeneic Bone ◽  
Local Bone ◽  
Heat Treated ◽  
Bank Technology

The use of bone scaffolds in traumatology and orthopedics is an extremely important issue. The growing number of cases of significant bone defects, in particular after revision arthroplasty, combat trauma and due to the introduction of new methods of reconstructive surgery of bones and joints, requires more detailed studies of the using different osteoplastic materials. Materials and methods. As scaffolds used 4 types of materials that are most often used in the clinic for the correction of bone defects - ceramic hydroxylapatite, beta-tricalcium phosphate, allogeneic bone matrix treated with gamma irradiation, allogeneic bone matrix scaffold. The effect of matrices on the viability of normal human fibroblasts (M19 cell line) in cell culture in vitro was studied. The viability of cells after their co-cultivation with scaffolds was determined by colorimetric method by staining with crystal violet. To obtain an osteoinductive effect used platelet-rich plasma (PRP), standardized by the method of Araki with some modifications. The proliferative activity of fibroblasts was assessed by the level of expression of the proliferation marker Ki-67 by immunocytochemical analysis. Results. It was found that the least pronounced antiproliferative effect is shown by allogeneic bone matrix treated with gamma irradiation. Data on the complex effect of co-cultivation of fibroblasts with scaffolds in the presence of PRP on cell viability and proliferative activity were obtained. It was found that PRP improves the survival of fibroblasts by 15-30 % and increases their proliferative activity by 35-75 %. Delipidization of scaffold from allogeneic bone matrix, heat-treated by local bone bank technology, increased its biocompatibility with human fibroblasts. Conclusions. According to the results of a comparative analysis of the impact of different scaffolds on the viability of normal human fibroblasts, it was found that scaffolds from allogeneic bone matrix have the least pronounced antiproliferative effect. Platelet-rich plasma has been shown to improve fibroblast survival and increase their proliferative activity. Treatment with 70 % ethyl alcohol scaffold from allogeneic bone matrix, heat-treated by local bone bank technology, increased its biocompatibility with human fibroblasts.

Matrix Properties, Biocompatibility and Osteoplastic Potentialities of Composite Materials Based on Polylactoglycolide and Natural Coral Skeleton Granules of Various Dispersity

2013 ◽  
Vol 20 (4) ◽  
pp. 17-23
Author(s):  
N. S Sergeeva ◽  
I. K Sviridova ◽  
G. A Frank ◽  
V. A Kirsanova ◽  
S. A Akhmedova ◽  
...  
Keyword(s):  
Composite Materials ◽  
Connective Tissue ◽  
Human Fibroblasts ◽  
Bone Defects ◽  
Coral Skeleton ◽  
Matrix Properties ◽  
Natural Coral ◽  
Tibia Defect

Results of in vitro and in vivo medico5biological study of mineral-polymer composites (MPC) based on high molecular polylactoglycolide and natural A. cervicornis coral skeleton with vari5 ous dispersity (600 µm) as materials for bone defects substitution are presented. On the model of human fibroblasts in vitro it was shown that MPC were not toxic and possessed satisfactory matrix (for cells) properties. The optimum for composite size of natural coral granules made up 200-600 µm. MPC biocompatibility was shown in subcutaneous test in mice. However comparatively slow subcutaneous substitution of both polylactoglycolide and MPC on its basis by connective tissue. Study of MPC and its components’ osteoplastic potential showed that in the zone of fenestral tibia defect in rats polylactoglycolide was substituted by connective tissue. Periosteal osteogenesis that in MPC was supplemented by enchondral osteogenesis was observed around the particles of natural coral skeleton.

scholarly journals Balancing Purification and Ultrastructure of Naturally Derived Bone Blocks for Bone Regeneration: Report of the Purification Effort of Two Bone Blocks

Materials ◽  
2019 ◽  
Vol 12 (19) ◽  
pp. 3234 ◽  
Author(s):  
Mike Barbeck ◽  
Ole Jung ◽  
Xin Xiong ◽  
Rumen Krastev ◽  
Tadas Korzinskas ◽  
...  
Keyword(s):  
Tissue Regeneration ◽  
Inflammatory Responses ◽  
Bone Matrix ◽  
Collagen Matrix ◽  
Bone Tissue Regeneration ◽  
Bone Block ◽  
Allogeneic Bone ◽  
Purification Process ◽  
Natural Bone

The present publication reports the purification effort of two natural bone blocks, that is, an allogeneic bone block (maxgraft®, botiss biomaterials GmbH, Zossen, Germany) and a xenogeneic block (SMARTBONE®, IBI S.A., Mezzovico-Vira, Switzerland) in addition to previously published results based on histology. Furthermore, specialized scanning electron microscopy (SEM) and in vitro analyses (XTT, BrdU, LDH) for testing of the cytocompatibility based on ISO 10993-5/-12 have been conducted. The microscopic analyses showed that both bone blocks possess a trabecular structure with a lamellar subarrangement. In the case of the xenogeneic bone block, only minor remnants of collagenous structures were found, while in contrast high amounts of collagen were found associated with the allogeneic bone matrix. Furthermore, only island-like remnants of the polymer coating in case of the xenogeneic bone substitute seemed to be detectable. Finally, no remaining cells or cellular remnants were found in both bone blocks. The in vitro analyses showed that both bone blocks are biocompatible. Altogether, the purification level of both bone blocks seems to be favorable for bone tissue regeneration without the risk for inflammatory responses or graft rejection. Moreover, the analysis of the maxgraft® bone block showed that the underlying purification process allows for preserving not only the calcified bone matrix but also high amounts of the intertrabecular collagen matrix.

scholarly journals Effect of Mesenchymal Stem Cells and Platelet-Rich Plasma on the Bone Healing of Ovariectomized Rats

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Bo Wei ◽  
Chengshuo Huang ◽  
Mingyan Zhao ◽  
Peng Li ◽  
Xiang Gao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Platelet Rich Plasma ◽  
Bone Microarchitecture ◽  
Bone Defects ◽  
Ovariectomized Rats ◽  
Marker Genes ◽  
Osteoporotic Bone ◽  
Specific Marker ◽  
Allogeneic Bone

We evaluated the efficacy of platelet-rich plasma (PRP) in combination with allogeneic bone marrow mesenchymal stem cells (BMSCs) for the treatment of osteoporotic bone defects in an ovariectomized rat model. By day 42 after injury, in vivo microcomputed tomography (micro-CT) imaging revealed that bone defects of control rats and ovariectomized rats treated with PRP and BMSCs were completely repaired, whereas those of ovariectomized rats treated with PRP or BMSCs alone exhibited slower healing. Histological data were consistent with these results. We also assessed changes to bone trabeculae in the proximal tibial growth plate. In ovariectomized rats treated with PRP or with a combination of PRP and BMSCs, the trabecular connectivity densities (Conn.D), bone volume ratios (BV/TV), and numbers (Tb.N) in the defect areas increased significantly from day 7 to day 42. These results indicate that PRP treatment enhances bone microarchitecture in osteoporosis. Moreover, expression levels of osteogenesis-specific marker genes including RUNX2, OSX, and OPN were significantly upregulated in rats treated with PRP and BMSCs compared to those of other groups. Thus, we conclude that treatment with PRP combined with BMSCs significantly promotes healing of osteoporotic bone defects. This study provides an alternative strategy for the treatment of osteoporotic bone loss.

scholarly journals Evaluation of Mesenchymal Stem Cell Sheets Overexpressing BMP-7 in Canine Critical-Sized Bone Defects

2018 ◽  
Vol 19 (7) ◽  
pp. 2073 ◽  
Author(s):  
Yongsun Kim ◽  
Byung-Jae Kang ◽  
Wan Kim ◽  
Hui-suk Yun ◽  
Oh-kyeong Kweon
Keyword(s):  
Bone Defect ◽  
Therapeutic Potential ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Bone Defects ◽  
Trophic Factors ◽  
Gene Expressions ◽  
Cell Sheets ◽  
Demineralized Bone

The aim of this study was to investigate the in vitro osteogenic capacity of bone morphogenetic protein 7 (BMP-7) overexpressing adipose-derived (Ad-) mesenchymal stem cells (MSCs) sheets (BMP-7-CS). In addition, BMP-7-CS were transplanted into critical-sized bone defects and osteogenesis was assessed. BMP-7 gene expressing lentivirus particles were transduced into Ad-MSCs. BMP-7, at the mRNA and protein level, was up-regulated in BMP-7-MSCs compared to expression in Ad-MSCs. Osteogenic and vascular-related gene expressions were up-regulated in BMP-7-CS compared to Ad-MSCs and Ad-MSC sheets. In a segmental bone-defect model, newly formed bone and neovascularization were enhanced with BMP-7-CS, or with a combination of BMP-7-CS and demineralized bone matrix (DBM), compared to those in control groups. These results demonstrate that lentiviral-mediated gene transfer of BMP-7 into Ad-MSCs allows for stable BMP-7 production. BMP-7-CS displayed higher osteogenic capacity than Ad-MSCs and Ad-MSC sheets. In addition, BMP-7-CS combined with demineralized bone matrix (DBM) stimulated new bone and blood vessel formation in a canine critical-sized bone defect. The BMP-7-CS not only provides BMP-7 producing MSCs but also produce osteogenic and vascular trophic factors. Thus, BMP-7-CS and DBM have therapeutic potential for the treatment of critical-sized bone defects and could be used to further enhance clinical outcomes during bone-defect treatment.

scholarly journals Effects of platelet-rich plasma on in vitro proliferation and migration of fibroblasts from human chronic refractory wound granulation tissue

2021 ◽  
Vol 8 (1) ◽  
pp. 24
Author(s):  
Tie-ning Zhang ◽  
Quan Li ◽  
Te Ba ◽  
Tian-xi Shao ◽  
Fang Li ◽  
...  
Keyword(s):  
Granulation Tissue ◽  
Platelet Rich Plasma ◽  
Human Fibroblasts ◽  
In Vitro Proliferation ◽  
Scratch Assay ◽  
Fetal Bovine ◽  
And Migration ◽  
Wound Scratch Assay ◽  
Proliferation And Migration

Objective: To observe the effects of platelet-rich plasma (PRP) on in vitro proliferation and migration of fibroblasts from human chronic refractory wound granulation tissue.Methods: Fibroblasts were separated from human chronic refractory wound granulation tissue and then were identified. The obtained fibroblasts were divided into fetal bovine serum (FBS) group, hydrogel group and PRP group, and the three groups were cultured with culture mediums containing FBS, hydrogel and PRP respectively, in order to observe the growth of fibroblasts. The wound scratch assay was used to observe the migration of fibroblasts.Results: PRP group had more fibroblasts than FBS group and hydrogel group since Day 5 of culture, and exhibited greater fibroblast scratch migration area than FBS group on 48 h and 72 h of wound scratch assay (all p < .05).Conclusions: Compared with FBS, human fibroblasts cultured by PRP can more effectively promote the proliferation and migration of fibroblasts.

scholarly journals A Preliminary Study on the Potential of Manuka Honey and Platelet-Rich Plasma in Wound Healing

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
Scott A. Sell ◽  
Patricia S. Wolfe ◽  
Andrew J. Spence ◽  
Isaac A. Rodriguez ◽  
Jennifer M. McCool ◽  
...  
Keyword(s):  
Wound Healing ◽  
Culture Media ◽  
Platelet Rich Plasma ◽  
Human Fibroblasts ◽  
Healing Process ◽  
Collagen Matrix ◽  
Wound Healing Process ◽  
Manuka Honey ◽  
Matrix Production

Aim. The purpose of this study was to determine thein vitroresponse of cells critical to the wound healing process in culture media supplemented with a lyophilized preparation rich in growth factors (PRGF) and Manuka honey.Materials and Methods. This study utilized cell culture media supplemented with PRGF, as well as whole Manuka honey and the medical-grade Medihoney (MH), a Manuka honey product. The response of human fibroblasts (hDF), macrophages, and endothelial cells (hPMEC) was evaluated, with respect to cell proliferation, chemotaxis, collagen matrix production, and angiogenic potential, when subjected to culture with media containing PRGF, MH, Manuka honey, and a combination of PRGF and MH.Results. All three cell types demonstrated increases in cellular activity in the presence of PRGF, with further increases in activity seen in the presence of PRGF+MH. hDFs proved to be the most positively responsive cells, as they experienced enhanced proliferation, collagen matrix production, and migration into anin vitrowound healing model with the PRGF+MH-supplemented media.Conclusion. This preliminaryin vitrostudy is the first to evaluate the combination of PRGF and Manuka honey, two products with the potential to increase regeneration individually, as a combined product to enhance dermal regeneration.

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

2021 ◽  
Vol 27 (1) ◽  
pp. 53-65
Author(s):  
L. A. Cherdantseva ◽  
E. A. Anastasieva ◽  
D. Ya. Aleynik ◽  
M. N. Egorikhina ◽  
I. A. Kirilova
Keyword(s):  
Tissue Engineering ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Bone Matrix ◽  
Allogeneic Bone ◽  
Spongy Bone ◽  
Allogenic Bone ◽  
Native Bone ◽  
Deproteinized Bone

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.

scholarly journals Dynamics of reparative histogenesis of bone tissue in presence of some osteoplastic materials in vitro

2020 ◽  
Vol 4 (34) ◽  
pp. 46-50
Author(s):  
S. Yu. Ivanov ◽  
A. V. Volkov ◽  
D. A. De
Keyword(s):  
Bone Regeneration ◽  
Bone Tissue ◽  
Bone Matrix ◽  
Three Dimensional ◽  
Bone Defects ◽  
Control Group ◽  
Bovine Bone ◽  
Reparative Osteogenesis ◽  
The Impact

Currently, to solve the bone deficiency problem in the maxillofacial region, osteoplastic materials based on allogeneic and xenogenic collagen bone matrix are used, both in pure and in activated forms, by adding growth factors. It is impossible to determine the effectiveness and mechanisms of the osteoplastic materials effect on bone regeneration without a comprehensive study, including not only histological, but also morphometric studies of the structural components and cellular reactions in the impact area. Such studies provide reliable and objective information on the main processes taking place in bone regeneration.Purpose. To determine the spatial distribution of reparative osteogenesis in the presence of some osteoplastic materials in vitro.Materials and methods. Svetlogorsk breed pigs were used as a biomodel. Depending on the osteoplastic preparations used, the animals were divided into four groups of the two in each: 1st — a preparation based on a natural bovine bone graft was injected into bone defects. 2nd — a preparation based on collagenized porcine transplant was injected into bone defects. 3rd — a preparation consisting of 60 % hydroxyapatite (HA) and 40 % beta-tri-calcium phosphate; 4th — control group — the bone defect healed under a blood clot. Animals were removed from the experiment on the 45th day. We examined sections with a thickness of 20 μm using the method of light and fluorescence microscopy.Results. The results indicate different dynamics of the reparative osteogenesis in the presence of osteoplastic materials of different classes. In group 1, the filling of the defect with newly formed bone tissue is not uniform; in group 2, the filling of the defect with newly formed bone tissue is uniform; in group 3 the filling of the defect with non-formed bone tissue is uneven due to the pronounced hyperostosis; in the control group, the filling of the defect with newly formed bone tissue is not happening.Conclusion. Stimulation, the dynamics of reparative osteogenesis and the three-dimensional organization of bone regenerate depend on the osteoplastic material class, which requires further study of the dynamics and three-dimensional organization of bone regenerate to select the optimal bone-replacing agent.

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