Allogeneic Bone Marrow Mesenchymal Stem Research Articles

Influence of allogeneic mesenchymal stem cells of red bone marrow culture on the development of Lewis lung epidermoid carcinoma in vivo

The relevance of this study is conditioned by the widespread use of stem cells in veterinary medicine, a wide range of studies and ambiguous data on the oncoprotective properties of stem cells of different origins. In this regard, the purpose of this study was to investigate the course of the tumour process in Lewis lung carcinoma and the specific features of the effect of allogeneic mesenchymal stem cells of red bone marrow culture on it. The leading approach to investigating this problem was the method of modelling Lewis lung carcinoma in C57BL6 mice and the use of stem cells. The use of allogeneic mesenchymal stem cells from the bone marrow culture of C57BL6 mice with transplanted epidermoid metastatic carcinoma of the Lewis lung contributed to the activation of the tumour process. Under the influence of allogeneic mesenchymal stem cells of red bone marrow culture from Day 14 to Day 24 of the study, the body weight of mice decreased by 7.0-12.1% (P < 0.05) compared to the control, the diameter of the primary tumour increased by 1.43-1.51 times (P < 0.05), which is conditioned by the activation of primary tumour growth. The number of lymphocytes as producers of vascular growth factor in primary tumour tissue under the influence of allogeneic mesenchymal stem cells of red bone marrow culture significantly increased by 1.47 and 1.52 times on Day 18 of the experiment compared to animals of the control group and placebo (P < 0.05), respectively. This promoted angiogenesis in the primary tumour node and metastasis through the circulatory system. After administration allogeneic mesenchymal cells of red bone marrow culture to mice, a larger volume of lung metastases was recorded, which was 41.52±7.9 mm3 compared to the values in the control and placebo groups, respectively, 17.94±6.59 and 16.43±5.32 mm3 . The morphological picture of the histological sections of the primary tumour of Lewis lung carcinoma confirms all the signs of qualitative and quantitative indicators of its progression. The findings obtained are of both theoretical and practical value for clinical veterinary medicine on the use of allogeneic bone marrow mesenchymal stem cells in tumour processes

Allogeneic bone marrow mesenchymal stem cell-derived exosomes alleviate human hypoxic AKI-on-a-Chip within a tight treatment window

BackgroundAcute hypoxic proximal tubule (PT) injury and subsequent maladaptive repair present high mortality and increased risk of acute kidney injury (AKI) - chronic kidney disease (CKD) transition. Human bone marrow mesenchymal stem cell-derived exosomes (hBMMSC-Exos) as potential cell therapeutics can be translated into clinics if drawbacks on safety and efficacy are clarified. Here, we determined the real-time effective dose and treatment window of allogeneic hBMMSC-Exos, evaluated their performance on the structural and functional integrity of 3D microfluidic acute hypoxic PT injury platform.MethodshBMMSC-Exos were isolated and characterized. Real-time impedance-based cell proliferation analysis (RTCA) determined the effective dose and treatment window for acute hypoxic PT injury. A 2-lane 3D gravity-driven microfluidic platform was set to mimic PT in vitro. ZO-1, acetylated α-tubulin immunolabelling, and permeability index assessed structural; cell proliferation by WST-1 measured functional integrity of PT.ResultshBMMSC-Exos induced PT proliferation with ED50 of 172,582 µg/ml at the 26th hour. Hypoxia significantly decreased ZO-1, increased permeability index, and decreased cell proliferation rate on 24–48 h in the microfluidic platform. hBMMSC-Exos reinforced polarity by a 1.72-fold increase in ZO-1, restored permeability by 20/45-fold against 20/155 kDa dextran and increased epithelial proliferation 3-fold compared to control.ConclusionsThe real-time potency assay and 3D gravity-driven microfluidic acute hypoxic PT injury platform precisely demonstrated the therapeutic performance window of allogeneic hBMMSC-Exos on ischemic AKI based on structural and functional cellular data. The novel standardized, non-invasive two-step system validates the cell-based personalized theragnostic tool in a real-time physiological microenvironment prior to safe and efficient clinical usage in nephrology.

Segurança e eficácia das células-tronco mesenquimais alogênicas da medula óssea para o tratamento da leucopenia canina induzida pela infecção por parvovírus canino

ABSTRACT: This study aims to establish a therapy strategy for canine leukopenia induced by canine parvovirus (CPV) infection through intravenous infusion of allogeneic bone marrow mesenchymal stem cells (BMMSCs) and to evaluate the therapeutic effect of BMMSCs on canine parvovirus. Forty healthy 2-month-old dogs were randomly divided into four groups including the BMMSC treatment group (A), conventional treatment group (B), CPV infection group (C), and a normal control group (D). Then the A, B, and C groups were orally infected with CPV (103.25 TCID50/mL) at 1mL/kg, and the D group received the same dose of saline. After the onset of infection, Group A received mesenchymal stem cells (MSCs) and rehydration as the treatment; Group B was treated with anti-inflammatory therapeutics and rehydration; and Group C and D were injected with the same dose of physiological saline. The level of leukocytes rebounded significantly after the treatment with BMMSCs and returned to reference numbers on Day 3 after treatment, which was significantly higher than that in the conventional treatment group. The concentrations of IL-2 and IFN-α were gradually increased during the treatment, and the BMMSC treatment group exhibited significantly higher IL-2 and IFN-α concentrations than the conventional treatment group on Days 3 and 4. The expression of the virus in the blood gradually decreased during the treatment, and the BMMSC treatment group displayed a faster decrease than the conventional treatment group. These results showed the advantages of BMMSC treatment over conventional treatment. This study provides a new BMMSC treatment strategy for canine leukopenia induced by CPV infection and reveals the mechanism by which BMMSC increases leukocytes after CPV infection.

Comparative Regenerative Effects of Allogeneic Bone Marrow and Patellar Ligament Fat Pad Mesenchymal Stem Cells on Experimental Superficial Digital Flexor Tendonitis in New Zealand Rabbits

Background: Cell therapy is applied in tendonitis to speed the healing process of tendon tissue and restore its functional properties. Almost all types of stem cells can differentiate from the recipient cells after transplantation. Objectives: The main goal of this study is to compare the effects of two sources of mesenchymal stem cells on tendon regeneration. Methods: This study randomly divided 32 New Zealand rabbits into 4 groups. The bacterial collagenase was induced at the superficial digital flexor tendon (SDFT) of all rabbits, and the treatment was performed 48 hours after collagenase induction. Group 1 was treated with allogeneic bone marrow mesenchymal stem cells (BMMSCs). Group 2 was treated with adipose-derived stem cells (ADSCs) from the patellar ligament fat pad. Group 3 (sham group) was treated with 0.9% normal saline, and group 4 (control group) was left with no treatment. All rabbits were euthanized 2 and 4 weeks after surgery, and tendon samples were harvested. The histopathology was assessed by hematoxylin-eosin, Masson’s trichrome, and Vangieson’s dye, and tendon structure, fiber arrangement, cell nuclei, tissue inflammation, vascularity (angiogenesis), and density were surveyed. Results: The tendon healing process in the BMMSC and ADSC groups revealed better regeneration than the control and sham groups (P≤0.05). Significant changes (P≤0.05) in some microscopic parameters were seen by comparing the BMMSC and ADSC groups. Conclusion: According to the present study, the injection of mesenchymal stem cells (BMMSCs or ADSCs) showed beneficial results in tendon tissue healing. Furthermore, ADSCs showed better regeneration of the injured tendon tissue than BMMSCs.

The effect of mesenchymal stem cells on platelet function in rats with experimental lung injury

Interstitial lung tissue diseases (pulmonary fibrosis) are accompanied by a decrease in thrombopoiesis. Stem cells are capable of differentiating into other cell types, making them a valuable material for veterinary cellular regenerative therapy. The purpose of the research – to explore changes in platelet counts in laboratory rats with experimental pulmonary fibrosis under the influence of stem cells. The research was performed on female Wistar rats, in which pulmonary fibrosis was modelled using a single transthoracic injection of bleomycin hydrochloride solution. Allogeneic mesenchymal stem cells were used to stimulate recovery processes in pathologically altered lung tissue, which were administered by different routes, and, for comparison, the conventional method of treatment. The presented results of the effect of transplanted allogeneic bone marrow mesenchymal stem cells indicate a significant change in the number and size of platelets in rats with experimental pulmonary fibrosis and an increase in the activity of regenerative processes in damaged tissues. In experimental animals, a significant increase in the number of platelets and their size was found after using allogeneic mesenchymal stem cells compared to similar data in control group animals. In addition, in the blood of animals of the experimental group, which were transplanted with mesenchymal stem cells transthoracically (directly into the lung tissue), there was a higher platelet activity than in animals with intravenous injection of mesenchymal stem cells. Platelet activation indicates an improvement in the regenerative capacity of damaged lung tissue under the influence of mesenchymal stem cells. Thus, transplanted mesenchymal stem cells stimulate platelet activity and regenerative processes in pathologically altered lung tissue in experimental fibrosis, which can be used as one of the effective methods of treating animals with this type of pathology

Cryopreserved allogeneic bone marrow mesenchymal stem cells show better osteochondral defect repair potential than adipose tissue mesenchymal stem cells

Mesenchymal stem cells (MSCs) due to their characteristic properties have a potential to treat osteoarthritis, one of the major growing joint problems. MSCs show differential ex vivo chondrogenic potential on the basis of source that remains to be validated under in vivo environment. This study compared chondrogenic potential of MSCs derived from two common sources, adipose tissue (AD) and bone marrow (BM) under ex vivo and in vivo environments. The randomized placebo controlled osteochondral defect (OCD) study divided n=72 rabbits equally into Control, AD-MSCs and BM-MSCs groups. Ex vivo chondrogenic induction resulted in an increased aggrecan fold expression in BM-MSCs and AD-MSCs. The former cell type had significantly (p<0.05) higher fold expression as compared to the latter. The cell treated OCDs had significantly reduced gene expression for inflammatory markers (IL-6, IL-8 and TNF-α) as compared to the control. In OCD study, radiography, MRI, gross observation, histopathology and SEM revealed that the cell treated defects were early filled by the tissue that had better surface architecture and matrices as compared to the control. BM-MSCs treated defects had better scores especially for gross and histopathology than the AD-MSCs. Gene expression for osteochondral regulation and cartilaginous matrices was higher in BM-MSCs group while only for matrices including the Col I in AD-MSCs as compared to the control. It was concluded that OCD in the cell treated groups are filled early with mostly a fibrocartilaginous to hyaline tissue. BM-MSCs may have an edge over AD-MSCs in OCD repair.

Heme Oxygenase-1 targeting exosomes for temozolomide resistant glioblastoma synergistic therapy.

Glioblastoma (GBM) is a highly fatal and recurrent brain cancer without a complete prevailing remedy. Although the synthetic nanotechnology-based approaches exhibit excellent therapeutic potential, the associated cytotoxic effects and organ clearance failure rest major obstacles from bench to clinics. Here, we explored allogeneic bone marrow mesenchymal stem cells isolated exosomes (BMSCExo) decorated with heme oxygenase-1 (HMOX1) specific short peptide (HSSP) as temozolomide (TMZ) and small interfering RNA (siRNA) nanocarrier for TMZ resistant glioblastoma therapy. The BMSCExo had excellent TMZ and siRNA loading ability and could traverse the blood-brain barrier (BBB) by leveraging its intrinsic brain accumulation property. Notably, with HSSP decoration, the TMZ or siRNA encapsulated BMSCExo exhibited excellent TMZ resistant GBM targeting ability both in vitro and in vivo due to the overexpression of HMOX1 in TMZ resistant GBM cells. Further, the HSSP decorated BMSCExo delivered the STAT3 targeted siRNA to the TMZ resistant glioma and restore the TMZ sensitivity, consequently achieved the synergistically drug resistant GBM treatment with TMZ. Our results showed this biomimetic nanoplatform can serve as a flexible, robust and inert system for GBM treatment, especially emphasizing the drug resistant challenge.

Allogeneic Bone-Marrow Mesenchymal Stem Cell with Moldable Cryogel for Craniofacial Bone Regeneration.

Allogeneic bone-marrow mesenchymal stem cells (BMSCs) can promote bone regeneration and substitute for autologous BMSCs if autologous sources are unavailable, but the efficacy of bone regeneration by allogeneic BMSCs is still inconsistent. A Lewis rat cranium defect model was used to investigate the efficacy of bone regeneration between autologous and allogeneic BMSCs in gelatin-nanohydroxyapatite cryogel scaffolds. BMSCs from Wistar rats served as the allogeneic cell lineage. The full-thickness cranium defects were treated by either blank control, cryogel only, allogeneic BMSC-seeded cryogel, or autologous BMSC-seeded cryogel (n = 5). Bone regeneration was monitored by micro-computed tomography and examined histologically at week 12. In addition, we assessed the immune responses in vitro by mixed lymphocyte reaction (MLR) assay and CD4+ immunochemistry staining ex vivo. The MLR showed that allogeneic BSMCs elicited a weak immune response on day 14 that progressively attenuated by day 28. In vivo, the bone regeneration in allogeneic BMSCs was inferior at week 4, but progressively matched the autologous BMSCs by week 12. Our results suggest that allogeneic BMSCs can serve as an alternative source for bone regeneration.

Comparative analysis of mouse bone marrow and adipose tissue mesenchymal stem cells for critical limb ischemia cell therapy

IntroductionCritical limb ischemia (CLI) is the most advanced form of peripheral arterial disease (PAD) characterized by ischemic rest pain and non-healing ulcers. Currently, the standard therapy for CLI is the surgical reconstruction and endovascular therapy or limb amputation for patients with no treatment options. Neovasculogenesis induced by mesenchymal stem cells (MSCs) therapy is a promising approach to improve CLI. Owing to their angiogenic and immunomodulatory potential, MSCs are perfect candidates for the treatment of CLI. The purpose of this study was to determine and compare the in vitro and in vivo effects of allogeneic bone marrow mesenchymal stem cells (BM-MSCs) and adipose tissue mesenchymal stem cells (AT-MSCs) on CLI treatment.MethodsFor the first step, BM-MSCs and AT-MSCs were isolated and characterized for the characteristic MSC phenotypes. Then, femoral artery ligation and total excision of the femoral artery were performed on C57BL/6 mice to create a CLI model. The cells were evaluated for their in vitro and in vivo biological characteristics for CLI cell therapy. In order to determine these characteristics, the following tests were performed: morphology, flow cytometry, differentiation to osteocyte and adipocyte, wound healing assay, and behavioral tests including Tarlov, Ischemia, Modified ischemia, Function and the grade of limb necrosis scores, donor cell survival assay, and histological analysis.ResultsOur cellular and functional tests indicated that during 28 days after cell transplantation, BM-MSCs had a great effect on endothelial cell migration, muscle restructure, functional improvements, and neovascularization in ischemic tissues compared with AT-MSCs and control groups.ConclusionsAllogeneic BM-MSC transplantation resulted in a more effective recovery from critical limb ischemia compared to AT-MSCs transplantation. In fact, BM-MSC transplantation could be considered as a promising therapy for diseases with insufficient angiogenesis including hindlimb ischemia.

Intrathecal Transplantation of Autologous and Allogeneic Bone Marrow-Derived Mesenchymal Stem Cells in Dogs

The route used in the transplantation of mesenchymal stem cells (MSCs) can directly affect the treatment success. The transplantation of MSCs via the intrathecal (IT) route can be an important therapeutic strategy for neurological disorders. The objective of this study was to evaluate the safety and feasibility of the IT transplantation of autologous (Auto-MSCs) and allogeneic (Allo-MSCs) bone marrow mesenchymal stem cells (BM-MSCs) in healthy dogs. Based on neurodisability score, cerebrospinal fluid (CSF) and magnetic resonance imaging (MRI), no significant differences from the control group were observed on day 1 or day 5 after IT Auto- or Allo-MSCs transplantation (P > 0.05). In addition, analysis of matrix metalloproteinase (MMP)-2 and MMP-9 expression in the CSF revealed no significant differences (P > 0.05) at 5 days after IT transplantation in the Auto- or Allo-MSCs group when compared to the control. Intrathecal transplantation of BM-MSCs in dogs provides a safe, easy and minimally invasive route for the use of cell-based therapeutics in central nervous system diseases.

Effects of local injection and intravenous injection of allogeneic bone marrow mesenchymal stem cells on the structure and function of damaged anal sphincter in rats.

Anal sphincter injury leads to damage to the anal structure and functions and has been identified as a major risk factor for fecal incontinence. Bone marrow mesenchymal stem cells (BMSCs) with capacities of multidifferentiation, paracrine, and low immunogenicity have been widely used in tissue repair and regeneration. The primary objective of this research was to compare the effects of different injection therapies of BMSCs on the injured anal sphincters. Ninety-six Sprague-Dawley female rats were randomly divided into four groups (n = 24 each): intravenous injection, local injection, sham operation, and normal control. For the first three groups, 25% removal of the anal sphincter complex was performed and 0.3-ml phosphate-buffered saline (PBS) (containing 107 green fluorescent protein-labeled allogeneic BMSCs) was given accordingly to the treatment group 24 h after operation for 7 consecutive days. The sham operation group was injected with 0.3-ml PBS only. All cases had undergone evaluation in the 1st, 7th, 14th, and 28th postoperative days. The rats were sacrificed on the 28th postoperative day, and the anal sphincters were dissected to be analyzed by morphological examination. At 14 days postoperatively, local injection of BMSC significantly improved the peak contraction pressure, electromyography amplitude, and frequency of the injured anal sphincter compared with tail vein, but there was no significant difference in resting pressure until 28 days after sphincterectomy. Masson staining results confirmed that the local injection group had significantly more new muscles on the wound. BMSC could remarkably improve peak contraction pressure, electromyography amplitude, and muscle fibers on the wound, and local injection is superior to intravenous injection.

Allogeneic Bone Marrow Mesenchymal Stem Cell Transplantation in Tooth Extractions Sites Ameliorates the Incidence of Osteonecrotic Jaw-Like Lesions in Zoledronic Acid-Treated Rats.

Medication-related osteonecrosis of the jaw (MRONJ) is defined as the exposed necrotic bone involving the maxillofacial structures in bisphosphonate treated patients, and the pathophysiology of this disease remains unclear. The aim of this study was to assess the effects of the allogeneic transplantation of bone marrow-derived mesenchymal stem cells (BM-MSCs) in a model of Wistar mice with induced MRONJ disease. BM-MSCs from five male Wistar rats were characterized and cultured on β-tricalcium phosphate (β-TCP) granules. Thirty female Wistar rats were injected intraperitoneally with zoledronic acid and afterwards upper jaw molars were extracted. The animals were randomized to receive: Group 1: 1 × 106 BM-MSCs/β-TCP construct in the alveolar socket; and Group 2: Saline solution/β-TCP construct. A clinical and histological analysis was performed. Nested polymerase chain reaction (PCR) was assessed to verify the presence of transplanted male rat cells in the female recipient jaws. Clinical and histological findings evidenced that none of the animals in Group 1 exhibited uncovered sockets or bone exposure associated to MRONJ, whereas we detected 33% of MRONJ cases in Group 2. In addition, male rat cells were detected in the maxillae site four weeks after transplantation in the BM-MSCs-group. Allogeneic BM-MSCs in extractions sites ameliorates MRONJ incidence in zoledronic acid-treated rats compared to non-MSC treatments.

A biphasic, demineralized, and Decellularized allograft bone-hydrogel scaffold with a cell-based BMP-7 delivery system for osteochondral defect regeneration.

The reconstruction of bone and cartilage defects remains a challenge in orthopedics and tissue engineering. In this study, a mimetic natural scaffold based on a demineralized and decellularized bone and collagen type I (Col I) allograft was developed. The resulting hydrogel has the capability of loading allogeneic bone marrow mesenchymal stem cells (BMSCs) resulting in the sustained release of bone morphogenetic protein-7 (BMP-7). BMSCs transfected with lentivirus loaded with BMP-7 gene were used as the BMP-7 delivery system, and then seeded on a demineralized and decellularized allograft bone-collagen biphasic scaffold to enhance bone and cartilage regeneration. The results indicated that, after transfection, BMSCs had a higher expression of the BMP-7 gene and the sustained release of BMP-7 lasted more than 28 days. The preliminary biphasic scaffold promoted cell adhesion and proliferation. After implanting the transfected cells into the knee joints of beagle dogs, enhanced osteochondral defect regeneration was identified at 12 weeks postimplantation. Our results revealed that the new cell-loaded scaffold can avoid the side effects and the short half-life of BMP-7, and promote the reconstruction of bone and cartilage defects. Such a composite system, therefore, shows potential in bone and cartilage tissue engineering applications.

The Effect of Allogenic Bone Marrow Mesenchymal Stem Cell - Platelet Rich Plasma (Bmscs - Prp) Intra-Articular Injection Effect on the Regeneration of Full-Thickness Joint Cartilage Defect on Rabbit

Introduction: Full thickness cartilage defects are still a problem until present since the handling has not give any satisfactory results. Current handling is performed through cartilage enginering using mesenchymal stem cells alone and or in combination with growth factor. The purpose of this study was to investigate the effect of injection of intra-articular injection of Allogenic bone marrow mesenchymal stem cell - Platelet rich plasma (BMSCs-PRP) on regeneration of full thickness cartilage defect in rabbits. Method: The design of this study is a post-test only control group design using 36 New Zealand white rabbits divided into three groups treated with PRP, BMSCs and BMSCs-PRP. Results are evaluated after 10 weeks. Results: In the evaluation, macroscopic images showed the best healing in the BMSCs-PRP group. Histopathologic examination showed that in the MSCs-PRP group there was a significant increase in the amount of chondrocytes (p = 0.000), cartilage area (p = 0.000), as well as the number of Agecoprogenitor expec- tion cells (p = 0.000) and type 2 collagen (p = 0.000). Conclusions: Intra-articular injections Allogenic bone marrow mesenchymal stem cell (BMSCs-PRP) is able to regenerate and cure full-thickness joint cartilage defects through differentiation of MSCs into condroblasts.

Enhancing the Efficacy of Stem Cell Therapy with Glycosaminoglycans.

SummaryHuman mesenchymal stem cell (hMSC) therapy offers significant potential for osteochondral regeneration. Such applications require their ex vivo expansion in media frequently supplemented with fibroblast growth factor 2 (FGF2). Particular heparan sulfate (HS) fractions stabilize FGF2-FGF receptor complexes. We show that an FGF2-binding HS variant (HS8) accelerates the expansion of freshly isolated bone marrow hMSCs without compromising their naivety. Importantly, the repair of osteochondral defects in both rats and pigs is improved after treatment with HS8-supplemented hMSCs (MSCHS8), when assessed histologically, biomechanically, or by MRI. Thus, supplementing hMSC culture media with an HS variant that targets endogenously produced FGF2 allows the elimination of exogenous growth factors that may adversely affect their therapeutic potency.

Testosterone propionate can promote effects of acellular nerve allograft‐seeded bone marrow mesenchymal stem cells on repairing canine sciatic nerve

Peripheral human nerves fail to regenerate across long tube implants (>2cm), and tissue-engineered nerve grafts represent a promising treatment alternative. The present study aims to investigate the testosterone propionate (TP) repair effect of acellular nerve allograft (ANA) seeded with allogeneic bone marrow mesenchymal stem cells (BMSCs) on 3-cm canine sciatic nerve defect. ANA cellularized with allogeneic BMSCs was implanted to the defect, and TP was injected into the lateral crus of the defected leg. The normal group, the autograft group, the ANA+BMSCs group, the ANA group, and the nongrafted group were used as control. Five months postoperatively, dogs in the TP+ANA+BMSCs group were capable of load bearing, normal walking, and skipping, the autograft group and the ANA+BMSCs group demonstrated nearly the same despite a slight limp. The compound muscle action potentials (CMAPs) on the injured side to the uninjured site in the TP+ANA+BMSCs group were significantly higher than that in the ANA+BMSCs group [CMAPs ratio at A: F(3, 20)=191.40; 0.02, CMAPs ratio at B: F(3, 20)=43.27; 0.01]. Masson trichrome staining revealed that in the TP+ANA+BMSCs group, both the diameter ratio of the myelinated nerve and the thickness ratio of regenerated myelin sheath were significantly larger than that in the other groups [the diameter of myelinated nerve fibers: F(3, 56)=13.45; P<.01, the thickness ratio of regenerated myelin sheath: F(3, 56)=51.25; P<.01]. In conclusion, TP could significantly increase the repairing effects of the ANA+BMSCs group, and their combination was able to repair 3-cm canine sciatic nerve defect. It therefore represents a promising therapeutic approach.

In vitro experimental study on co-culture of allogenic costal chondrocytes and bone marrow mesenchymal stem cells

Objective To investigate the feasibility of bone marrow mesenchymal stem cells differentiating into chondrocyte in vitro under microenvironment of costal chondrocyte, co-cultured with allogenic costal chondrocyte. Methods Firstly bone marrow mesenchymal stem cells and costal chondrocytes were isolated and cultured. Then the second generation of bone marrow mesenchymal stem cells and costal chondrocytes were co-cultured at 2∶1 for the experimental group with the final concentration of 3×108/L. The costal chondrocytes with the same concentration were taken as the control group, and the chondrocytes with the concentration of 1×108/L were taken as the low concentration group. The cells were cultured in vitro for 1, 2 and 3 months to observe the morphological changes. Two weeks later, the co-cultured cells were taken for histological identification. And after 1, 2 and 3 months, the specimens were respectively taken for gross observation and histological staining to evaluate the new cartilage. Results The two kinds of cells had good biological activity and the bone marrow mesenchymal stem cells had transformed into chondrocytes 2 weeks later in the experimental group. And histological examinationshowed that hyaline cartilages were formed in all three groups in vitro one month later. The average density of chondrocyte in the experimental group was higher than that in the other two groups in the first and second months (72.667±1.862, 98.333±1.506), and that in the control group (61.000±1.414, 84.500±1.049) was higher than that in the low concentration group (49.833±1.941, 75.000±0.894, P 0.05). Conclusion BMSCs can be induced to differentiate into chondrocyte under the condition of co-culture with allogenic costal chondrocyte in vitro. The co-cultured cells can form a thin layer of hyaline cartilage in vitro. Key words: Costal chondrocyte; Bone marrow mesenchymal stem cells; Mixed culture in vitro; Tissue engineering

THE EFFECT OF INJECTION OF INTRA ARTICULAR ALLOGENIC BONE MARROW MESENCHYMAL STEM CELL-PLATELET CELL RICH PLASMA (BMSCs-PRP) ON FULL-THICKNESS ARTICULAR CARTILAGE DEFFECT REGENERATION IN RABBIT

The joint cartilage defectfullthickness is still a problem today because its currenttreatment still has not delivered maximum results. Current treatment uses cartilage enginering using mesenchymal stem cells alone and or combining growth factor. The aim of this study was to investigate the effect of intra-articular injection of Allogenic bone marrow mesenchymal stem cell - Platelet rich plasma (BMSCs-PRP) on regeneration of cartilage defect fullthickness in rabbits. The design of this study was a post-test only control group design using 36 New Zealand white rabbits divided into three groups. Each group were treated with PRP, BMSCs and BMSCs-PRP. Results were evaluated after 10 weeks. In the evaluation, macroscopic images showed the best healing in the BMSCs-PRP group. Histopathologic examination showed that in the MSCs-PRP group, there was a significant increase in the number of chondrocytes (p = 0,000), cartilage area (p = 0,000), as well as the number of Agecoprogenitorexpec- tion cells (p = 0,000) and collagen type 2 (p = 0,000). BMSCs were able to differentiate into condroblasts which then synthesize aggressive and collagen type 2. Platelet rich plasma (PRP) contains growth factor BMP, TGF, FGF and IGF which can accelerate the occurrence of MSCs differentiation. Intra-articular injections Allogenic bone marrow mesenchymal stem cell (BMSCs-PRP) is able to regenerate and cure full-thickness joint cartilage defects through differentiation of MSCs into condroblasts.Keyword: Allogenic, Bone marrow Mesenchymal stem cell, Cartilage, Platelet rich plasma, Full-thickness.

COLLAGEN ANALYSIS OF GRAFT IN BONE TUNNEL MODEL ANTERIOR CRUCIATE LIGAMENT (ACL) RECONSTRUCTION WITH INTRATUNNEL ALLOGENIC BONE MARROW MESENCHYMAL STEM CELLS (MSCs) AND VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF)

ACL reconstruction measures aim to obtain stable knees, and reduce the risk of further injury to the meniscus and joint surfaces. Acceleration of the integration process between the tendon graft and the bone tunnel will improve the final result of ACL reconstruction. The purpose of this study was to investigate the collagen composition of the bone tunnel graft model of anterior cruciate ligament reconstruction with intravenous allogenic bone marrow mesenchymal stem cells and vascular endothelial growth factor in experimental animals. The design of this study was Post-test only Control Group Design using 20 rabbits divided into treatment group and control group. Collagen immunohistochemical evaluation was performed at weeks 3 and 6. Evaluation at week 3 obtained the area of collagen type-1 in the higher treatment group at treatment (p &lt;0.001). In the 6th week evaluation, it was found that the area of collagen type-1 in the treatment group was higher (p &lt;0.05). Type-1 collagen at week 6 did not differ significantly with week 3 (p&gt; 0.05). Provision of allogenic bone marrow mesenchymal stem cells and intratonal vascular endothelial growth factor in ACL reconstruction enhanced the formation of collagen type-1 which is the acceleration of incorporation of the graft tendon process with bone tunnel.Keywords : Anterior Cruciate Ligament, allogenic bone marrow mesenchymal stemcells, vascular endothelial growth factor, graft and collagen.

INTRATUNNEL THE EFFECT OF ADMINISTRATION OF BONE MARROW MESENCHYMAL STEM CELLS (BM-MSCs) AND VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) TENDON-BONE TO INTERFACE HISTOLOGICAL GRAFT ANTERIOR CRUCIATE LIGAMENT APPEARANCE AFTER RECONSTRUCTION IN RABBITS

The success of the Anterior Cruciate Ligament (ACL) reconstruction using a tendon graft is determined by integration in the bone tendon-graft interface on the bone tunnel. The use of stem cells and growth factors proved to accelerate the healing of the bone tendon-graft interface. The aim of this study was to inveestigate the difference of histology picture in the tendon-bone tunnel model after ACL reconstruction with intratunnel intravenous allogenic bone marrow mesenchymalstemcells (BM-MSCs) and intratunnel vascular endothelial growth factor (VEGF). This research used Post-Test Only Control Group design with 20 rabbits divided into treatment group and control group. Each group performed histologic image evaluation (thickness of collagen fiber or sharpey fiber) at week 3 and 6. Evaluation of histology overview at week 3 and week 6 showed a significantly thicker thickness of collagen fiber or sharpey fiber in treatment group compared with control group (p &lt;0.05). Intravenous administration of BM-SCs and VEGF after ACL reconstruction can speed healing of the bone tunnel significantly from week 3 and 6. The study by Faridyan et al has concluded that intravenous BM-SCs + VEGF increased ultimate tension strength in the bone-tendon interface significantly. In this study, intravenous administration of BM-SCs and VEGF gave histologic images showing acceleration of bone tunnel healing.Keywords:Anterior cruciate ligament reconstruction, allogenic bone marrow mesenchymal stem cells, vascular endothelial growth factor, graft tunnel healing, and Sharpey fiber.