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

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.

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Enhanced regeneration of bone defects using sintered porous Ti6Al4V scaffolds incorporated with mesenchymal stem cells and platelet-rich plasma

RSC Advances ◽

10.1039/d0ra10215f ◽

2021 ◽

Vol 11 (9) ◽

pp. 5128-5138

Author(s):

Ji Li ◽

Ketao Wang ◽

Xiaowei Bai ◽

Qi Wang ◽

Ningyu Lv ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Regeneration ◽

Platelet Rich Plasma ◽

Bone Defects ◽

Regeneration Of Bone

Porous Ti6AI4V scaffolds incorporated with MSC and PRP are more effective in enhancing the bone regeneration.

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The Effect of Early Application of a Combined Therapy of Bone Marrow Mesenchymal Stem Cells and Platelet-Rich Plasma on Blood and Bone Parameters in Ovariectomized Rats

Journal of Evolutionary Biochemistry and Physiology ◽

10.1134/s0022093021040219 ◽

2021 ◽

Vol 57 (4) ◽

pp. 972-990

Author(s):

G. M. Hamed ◽

N. A. Nassef ◽

R. S. A. E. Mansour ◽

M. K. E. Shawky ◽

A. A. Abo Zeid ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Platelet Rich Plasma ◽

Combined Therapy ◽

Ovariectomized Rats ◽

Early Application ◽

Bone Parameters ◽

Marrow Mesenchymal Stem Cells

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Comparative study between the effect of mesenchymal stem cells and/or platelet-rich plasma on bone parameters in ovariectomized mature rats

QJM ◽

10.1093/qjmed/hcab117 ◽

2021 ◽

Vol 114 (Supplement_1) ◽

Author(s):

Gehane M Hamed ◽

Noha A Nassef ◽

Rania S.A Eissa ◽

Mona K.E Shawky ◽

Asmaa A Abo Zeid ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Loss ◽

Platelet Rich Plasma ◽

Serum Levels ◽

Untreated Group ◽

Ovariectomized Rats ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Microscopic Studies

Abstract Background Osteoporosis is a common bone disease and is associated with considerable morbidity and mortality. Osteoporosis occurs most frequently in postmenopausal women. The inadequacy of current treatments and their side effects have driven a search for improved methods of dealing with osteoporosis. Design Experimental study. Aim of Work This study was conducted to throw light on the ability of BM-MSCs and/or PRP in improving bone formation and slow down bone loss in ovariectomized rats with osteoporosis. Materials & Methods This study was carried out on adult female Wistar rats allocated randomly into five groups: Sham-operated, OVX untreated, OVX-MSCs treated (MSCs administered once intravenously), OVX/PRP treated (PRP administered once subcutaneously), OVX-MSCs/PRP treated groups. Rats were subjected to assessment of serum ALK, CTX-1, MDA and TNF-α. In addition, specimens of tibia were taken and processed for light microscopic studies and morphometric analysis. Results OVX untreated group showed significant increases in serum levels of ALK and CTX-1. Significant elevations of serum MDA and TNF-α levels were also noticed in the OVX untreated group. Administration of BM-MSCs and PRP significantly lowered serum levels of ALK, CTX-1, MDA and TNF-α. These results were confirmed by the histopathological findings. Conclusion BM-MSCs together with PRP can partially reverse OVX-induced bone loss and halt osteoporosis progression. Abbreviations BM-MSCs: bone marrow derived mesenchymal stem cells, PRP: platelet-rich plasma, OVX: ovariectomized, ALK: alkaline phosphatase, CTX-1: cross linked c telopeptide of type 1 collagen, MDA: malondialdehyde, TNF-α: tumour necrosis factor-alpha.

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Slowly Delivered Icariin/Allogeneic Bone Marrow-Derived Mesenchymal Stem Cells to Promote the Healing of Calvarial Critical-Size Bone Defects

Stem Cells International ◽

10.1155/2016/1416047 ◽

2016 ◽

Vol 2016 ◽

pp. 1-13 ◽

Cited By ~ 4

Author(s):

Tianlin Liu ◽

Xin Zhang ◽

Yuan Luo ◽

Yuanliang Huang ◽

Gang Wu

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Growth Factors ◽

Large Volume ◽

Critical Size ◽

Bone Defects ◽

Allogeneic Bone Marrow ◽

Allogeneic Bone ◽

Trabecular Thickness

Bone tissue engineering technique is a promising strategy to repair large-volume bone defects. In this study, we developed a 3-dimensional construct by combining icariin (a small-molecule Chinese medicine), allogeneic bone marrow-derived mesenchymal stem cells (BMSCs), and a siliceous mesostructured cellular foams-poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (SMC-PHBHHx) composite scaffold. We hypothesized that the slowly released icariin could significantly promote the efficacy of SMC-PHBHHx/allogeneic BMSCs for repairing critical-size bone defects in rats. In in vitro cellular experiments, icariin at optimal concentration (10−6 mol/L) could significantly upregulate the osteogenesis- and angiogenesis-related genes and proteins, such as Runx2, ALP, osteocalcin, vascular endothelial growth factors, and fibroblast growth factors, as well as the mineralization of BMSCs. Icariin that was adsorbed onto the SMC-PHBHHx scaffold showed a slow release profile within a 2-week monitoring span. Eight weeks after implantation in calvarial critical-size bone defects, the constructs with icariin were associated with significantly higher bone volume density, trabecular thickness, trabecular number, and significantly lower trabecular separation than the constructs without icariin. Histomorphometric analysis showed that icariin was also associated with a significantly higher density of newly formed blood vessels. These data suggested a promising application potential of the icariin/SMC-PHBHHx/allogeneic BMSCs constructs for repairing large-volume bone defects in clinic.

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Mesenchymal MACF1 Facilitates SMAD7 Nuclear Translocation to Drive Bone Formation

Cells ◽

10.3390/cells9030616 ◽

2020 ◽

Vol 9 (3) ◽

pp. 616 ◽

Cited By ~ 2

Author(s):

Fan Zhao ◽

Xiaoli Ma ◽

Wuxia Qiu ◽

Pai Wang ◽

Ru Zhang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Formation ◽

Nuclear Translocation ◽

Early Stage ◽

Bone Microarchitecture ◽

Bone Diseases ◽

Conditional Knockout ◽

Osteoporotic Bone ◽

Cell Integrity

Microtubule actin crosslinking factor 1 (MACF1) is a large crosslinker that contributes to cell integrity and cell differentiation. Recent studies show that MACF1 is involved in multiple cellular functions such as neuron development and epidermal migration, and is the molecular basis for many degenerative diseases. MACF1 is highly abundant in bones, especially in mesenchymal stem cells; however, its regulatory role is still less understood in bone formation and degenerative bone diseases. In this study, we found MACF1 expression in mesenchymal stem cells (MSCs) of osteoporotic bone specimens was significantly lower. By conditional gene targeting to delete the mesenchymal Macf1 gene in mice, we observed in MSCs decreased osteogenic differentiation capability. During early stage bone development, the MACF1 conditional knockout (cKO) mice exhibit significant ossification retardation in skull and hindlimb, and by adulthood, mesenchymal loss of MACF1 attenuated bone mass, bone microarchitecture, and bone formation capability significantly. Further, we showed that MACF1 interacts directly with SMAD family member 7 (SMAD7) and facilitates SMAD7 nuclear translocation to initiate downstream osteogenic pathways. Hopefully these findings will expand the biological scope of the MACF1 gene, and provide an experimental basis for targeting MACF1 in degenerative bone diseases such as osteoporosis.

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Effect of a Decellularized Omentum Scaffold with Combination of Mesenchymal Stem Cells and Platelet-Rich Plasma on Healing of Critical-Sized Bone Defect: A Rat Model

Applied Sciences ◽

10.3390/app112210900 ◽

2021 ◽

Vol 11 (22) ◽

pp. 10900

Author(s):

Abdulsamet Emet ◽

Erdi Ozdemir ◽

Duygu Uckan Cetinkaya ◽

Emine Kilic ◽

Ramin Hashemihesar ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Platelet Rich Plasma ◽

Bone Defects ◽

Control Group ◽

Albino Rats ◽

Group 5 ◽

Group 2 ◽

Wistar Albino Rats ◽

Critical Bone Defects

The high costs and extensive time needed for the treatment of critical-sized bone defects are still major clinical concerns in orthopedic surgery; therefore, researchers continue to look for more cost and time-effective methods. This study aims to investigate the effects of a decellularized omentum scaffold with a combination of platelet-rich plasma (PRP) and mesenchymal stem cells on the healing of critical-sized bone defects. Wistar albino rats (n = 30) were investigated in five groups. Critical-sized bone defects were formed on bilateral radius shafts. No scaffold, decellularized omentum, omentum with PRP and omentum + mesenchymal stem cells was used in group 1 (control group), 2, 3 and 4, respectively. In addition, omentum with a combination of mesenchymal stem cells +PRP was used in group 5. After 6 weeks, both radiological and histological healing were evaluated comparatively among the groups. After the use of a decellularized omentum scaffold, vitality of new cells was maintained, and new bone formation occurred. When compared to the control group, radiological healing was significantly better (p = 0.047) in the omentum and omentum + PRP-treated groups. Furthermore, histological healing was better in the omentum and omentum + PRP-treated groups than the control group (p = 0.001). The use of a decellularized omentum scaffold is suitable in the healing of critical bone defects.

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227 MESENCHYMAL STEM CELL ISOLATION AND CULTURE FROM ADIPOSE TISSUE OF A DEAD DOG

Reproduction Fertility and Development ◽

10.1071/rdv28n2ab227 ◽

2016 ◽

Vol 28 (2) ◽

pp. 245

Author(s):

S. Saini ◽

V. Sharma ◽

H. N. Malik ◽

S. K. Guha ◽

D. Malakar

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Bovine Serum ◽

Fetal Bovine Serum ◽

Blue Staining ◽

Marker Genes ◽

Specific Marker ◽

The Future ◽

Fetal Bovine

Isolation of cells or stem cells from clinically dead animals may serve applications such as revival of the animal through somatic cell nuclear transfer (SCNT) or cryopreservation of their cells for a long period so that cells can be used in the future. Thus, combining isolation of cells from clinically dead animals and SCNT of germplasm of elite animals could benefit research into endangered or extinct species. In the present study, we tried to isolate and culture adipose-derived mesenchymal stem cells (ADSC) from a clinically dead dog. Adipose tissues were collected surgically from the abdomen of a dead dog after 3 h and processed tissues within 10 h of death. The isolated tissues were washed in 70% ethanol for 30 s and washed 5 times in Dulbecco’s PBS supplemented with 50 µg mL–1 gentamicin. These fat tissues were minced to very small pieces and washed in DMEM by centrifugation at 800 rpm for 3 min. The tissue pellet was subjected to enzymatic digestion (collagenase 1 mg mL–1 of Dulbecco’s PBS) at 37°C in CO2 incubator for 1 h, with intermittent shaking after every 10 min. The digestive enzyme was inactivated by equal volume of DMEM/F-12 supplemented with fetal bovine serum (20%) and centrifuged at 1000 rpm for 10 min. The pellet was resuspended in DMEM/F-12 with 10% fetal bovine serum and cultured at 1 × 106 cells mL–1 in 25-cm2 tissue culture flasks. The medium was changed after every 48 h. Mesenchymal stem cells (MSC) were observed under an inverted microscope after 6 days. These cells were subcultured and a confluent monolayer was obtained. We have already standardized the protocol of MSC culture and characterisation as we are treating wounded and paralysed dogs using these MSC in a pet clinic. Characterisation of MSC was performed with specific surface marker genes of CD44, CD29, and CD166 in PCR and by immunocytochemistry of MSC-specific marker of CD44. Differentiation of these MSC into osteogenesis and chondrogenesis were observed after 3 weeks. Chondrogenic differentiation was confirmed by positive expression of chondrocyte-specific marker genes Aggrecan F-TTGGACTTTGGCAGAATACC and R-CTTCCACCAATGTCGTATCC and Collagen II F-AACCCTGGAACTGACGGAAT and R-CTCACCCGTTTGACCTTTCG primer in PCR. The MSC were cryopreserved after 80% confluency was reached. The monolayer cells were scraped out from the culture flask and pelleted down. The pellet was resuspended in DMEM containing 10% DMSO and 20% fetal bovine serum. The number of cells was determined by trypan blue staining using an automatic cell counter and 105 cells mL–1 were added to a 2-mL cryogenic vial. The cryogenic vials were kept in a cryobox at –80°C for slow cooling. Then these vials were transferred to liquid nitrogen tanks after 12 h for long-term storage. We conclude that ADSC were successfully cultured from adipose tissue of a dog within 10 h of death and further subcultured under in vitro conditions. The cells could be used for SCNT to revive the dead animal and cryopreserve these cells for use in the future.

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225 AUTOLOGOUS TRANSPLANTATION OF MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE IN ANIMAL

Reproduction Fertility and Development ◽

10.1071/rdv28n2ab225 ◽

2016 ◽

Vol 28 (2) ◽

pp. 244 ◽

Cited By ~ 1

Author(s):

H. Malik ◽

V. Sharma ◽

S. Saini ◽

S. Guha ◽

D. Malakar

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Hip Dysplasia ◽

Autologous Transplantation ◽

Marker Genes ◽

Specific Marker ◽

Safranin O ◽

New Applications

The present study was carried out for isolation and culture of adipose tissue-derived mesenchymal stem cells of goat (gADSC) and dogs (1 dog was suffering from hip dysplasia and another dog from paraplegia) and their characterisation with different markers. Adipose tissue of goat and dog were aseptically isolated and treated with collagenase for 2 h in a CO2 incubator. The enzymatic digested cells were filtered through a 41-µm filter and cells were resuspended in cell culture flask containing medium DMEM/F12, 10% fetal bovine serum, and 50 μg mL–1 gentamycin. In vitro-cultured ADSC were characterised by amplification of mesenchymal stem cell (MSC)-specific surface marker genes of CD44, CD29, and CD166 in PCR and by immunocytochemistry of MSC-specific marker of CD44. For in vitro chondrogenesis, ADSC at passage 3 were incubated in DMEM/F12 containing 100 nM dexamethasone, 1.25 μg mL–1 BSA, and 10 ng mL–1 BMP-4 ITS (insulin-transferrin-selenium) for 3 wk. Chondrogenic differentiation cells were confirmed by Safranin O staining and positive expression of chondrocyte-specific marker genes Aggrecan: primers F-TTGGACTTTGGCAGAATACC and R-CTTCCACCAATGTCGTATCC, and Collagen II: primers F-AACCCTGGAACTGACGGAAT and R-CTCACCCGTTTGACCTTTCG in PCR. Dog ADSC-derived chondrocytes were aseptically injected at 1 × 106 cells kg–1 of BW into dogs with hip dysplasia and paraplegia. Both dogs recovered well after 1 month of autologous transplantation and were able to move freely. Then, 10 dogs having massive wounds were injected with heterologous undifferentiated mesenchymal stem cells at 1 × 106 cells kg–1 of BW and all dogs were cured in an average of 20 days. Then, the paralyzed and fractured dogs were further treated with undifferentiated MSC at 1 × 106 cells kg–1 of BW and most of the dogs were cured properly. These findings may have implications for defining the physiological roles of ADSC in arthritis, some orthopaedic problems, joint regeneration, and neurological disorders and several new applications leading to novel therapeutic opportunities.

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Effect of a Decellularized Omentum Scaffold With Combination of Mesenchymal Stem Cells and Platelet Rich Plasma on Healing of Critical-sized Bone Defect

10.21203/rs.3.rs-806042/v1 ◽

2021 ◽

Author(s):

Abdulsamet Emet ◽

Erdi Ozdemir ◽

Duygu Uckan Cetinkaya ◽

Emine Kilic ◽

Ali Celalettin Sinan Yuruker ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Platelet Rich Plasma ◽

Bone Defects ◽

Control Group ◽

Albino Rats ◽

Group 5 ◽

Group 2 ◽

New Research ◽

Critical Bone Defects

Abstract Background: High-cost and the long time needed for the treatment of critical-sized bone defects is still major clinical concern in orthopedic surgery, therefore new research continues to identify more cost and time-effective methods. In this study, we aimed to investigate the effect of a decellularized omentum scaffold with a combination of platelet-rich plasma (PRP) and mesenchymal stem cells on the healing of critical- sized bone defects. Materlials and methods: Wistar albino rats (n=30) were investigated in five groups. Critical-sized bone defects were formed on bilateral radius shafts. To increase healing, no scaffold, decellularized omentum, omentum with with PRP and omentum mesenchymal stem cells was used in group 1 (control group) , 2, 3 and 4 respectively. In addition omentum with a combination of mesenchymal stem cells and PRP was used in group 5. After 6 weeks, both radiological and histological healing were evaluated comparatively among groups. Results: After use of decellularized omentum scaffold, vitality of new cells maintained, new bone formation occurred. When compared to control group, radiological healing was statistically significantly better in omentum and omentum with PRP-treated groups (p=0.047). Furthermore histological healing was better in the omentum and omentum with PRP-treated groups than both control and omentum with mesenchymal stem cell-treated groups according to Salkeld Scoring System (p=0.001).Conclusion: The use of decellularized omentum scaffold is suitable in the healing of critical bone defects.

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