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Isolation of Human Mesenchymal Stem Cells and their Cultivation on the Porous Bone Matrix
Published on: February 9, 2015
In vivo osteoinductive effect and in vitro isolation and cultivation bone marrow mesenchymal stem cells
Amira Redzić1, Amer Smajilagić, Mufida Aljicević
1Institute for Biology and Human Genetics, Medicine Faculty, University of Sarajevo, Sarajevo, Bosnia and Herzegovina. amira.redzic@yahoo.com
Abstract:
Bone marrow contains cell type termed mesenchymal stem cells (MSC), first recognized in bone marrow by a German pathologist, Julius Cohnheim in 1867. That MSCs have potential to differentiate in vitro in to the various cells lines as osteoblast, chondroblast, myoblast and adipoblast cells lines. Aims of our study were to show in vivo capacity of bone marrow MSC to produce bone in surgically created non critical size mandible defects New Zeland Rabbits, and then in second part of study to isolate in vitro MSC from bone marrow, as potential cell transplantation model in bone regeneration. In vivo study showed new bone detected on 3D CT reconstruction day 30, on all 3 animals non critical size defects, treated with bone marrow MSC exposed to the human Bone Morphogenetic Protein 7 (rhBMP-7). Average values of bone mineral density (BMD), was 530 mg/cm3, on MSC treated animals, and 553 mg/cm3 on control group of 3 animals where non critical size defects were treated with iliac crest autologue bone graft. Activity of the Alkaline Phosphatase enzyme were measurement on 0.5, 14, 21, 30 day and increased activity were detected day 14 on animals treated with bone marrow MSCs compared with day 30 on iliac crest treated animals. That results indicates strong osteoinduction activity of the experimental bone marrow MSCs models exposed to the rhBMP-7 factor Comparing ALP activity, that model showed superiorly results than control group. That result initiates us in opinion that MSCs alone should be alternative for the autolologue bone transplantation and in vitro study we isolated singles MSCs from the bone marrow of rat's tibia and femora and cultivated according to the method of Maniatopoulos et all. The small initial colonies of fibroblast like cells were photo-documented after 2 days of primary culture. Such isolated and cultivated MSCs in future studies will be exposed to the growth factors to differentiate in osteoblast and indicate their clinically potential as alternative for conventional medicine and autologue bone transplantation. That new horizons have potential to minimize surgery and patient donor morbidity, with more success treatment in bone regenerative and metabolism diseases.
Insights
Mesenchymal stem cells (MSCs) from bone marrow show potential for bone regeneration in rabbits, outperforming traditional bone grafts. These MSCs could offer a less invasive alternative for bone repair and treating bone diseases.
Area of Science:
- Regenerative Medicine
- Biomaterials Science
- Orthopedic Surgery
Background:
- Mesenchymal stem cells (MSCs) are recognized for their differentiation potential into various cell types, including bone-forming cells.
- Surgically created bone defects present a challenge in achieving effective bone regeneration.
- Autologous bone grafts are a common but invasive method for bone repair.
Purpose of the Study:
- To evaluate the in vivo bone-producing capacity of bone marrow MSCs in rabbit mandible defects.
- To isolate and cultivate MSCs in vitro for potential cell transplantation in bone regeneration.
- To compare the efficacy of MSCs with traditional iliac crest bone grafts.
Main Methods:
- Surgically created non-critical size mandible defects in New Zealand rabbits were treated with bone marrow MSCs exposed to human Bone Morphogenetic Protein 7 (rhBMP-7).
- Control defects were treated with iliac crest autologous bone grafts.
- New bone formation was assessed using 3D CT reconstruction and bone mineral density (BMD) measurements.
- Alkaline Phosphatase (ALP) enzyme activity was measured at various time points.
Main Results:
- New bone formation was detected by day 30 in MSC-treated defects.
- MSC-treated defects showed comparable bone mineral density to the control group.
- Alkaline Phosphatase activity increased significantly by day 14 in MSC-treated animals, indicating potent osteoinduction.
- MSCs exposed to rhBMP-7 demonstrated superior osteoinductive activity compared to the control group.
Conclusions:
- Bone marrow MSCs, when exposed to rhBMP-7, exhibit significant osteoinductive capacity and potential for bone regeneration in vivo.
- MSCs represent a promising alternative to autologous bone grafting, potentially minimizing surgical morbidity.
- In vitro isolation and cultivation of MSCs provide a foundation for future cell transplantation therapies in bone regeneration and metabolic bone diseases.

